Generic relationships of selected African genera of Apiaceae
by
Anthony Richard Magee
Thesis submitted in fulfilment of the requirements for the degree
PHILOSOPHIAE DOCTOR
in
BOTANY
in the
FACULTY OF SCIENCE
at the
UNIVERSITY OF JOHANNESBURG
SUPERVISOR: PROF. B.-E. VAN WYK CO-SUPERVISOR: PROF. P. M. TILNEY CO-SUPERVISOR: PROF. S. R. DOWNIE
September 2009
AFFIDAVIT: MASTER'S AND DOCTORAL STUDENTS TO WHOM IT MAY CONCERN
This serves to confirm that I Anthony Richard Magee Full Name(s) and Surname
ID Number 8109065065089 Student number 920001887 enrolled for the
Qualification PhD (Botany)
Faculty Science Herewith declare that my academic work is in line with the Plagiarism Policy of the University of Johannesburg which I am familiar.
I further declare that the work presented in the Generic relationships of selected African genera of Apiaceae (thesis) is authentic and original unless clearly indicated otherwise and in such instances full reference to the source is acknowledged and I do not pretend to receive any credit for such acknowledged quotations, and that there is no copyright infringement in my work. I declare that no unethical research practices were used or material gained through dishonesty. I understand that plagiarism is a serious offence and that should I contravene the Plagiarism Policy notwithstanding signing this affidavit, I may be found guilty of a serious criminal offence (perjury) that would amongst other consequences compel the UJ to inform all other tertiary institutions of the offence and to issue a corresponding certificate of reprehensible academic conduct to whomever request such a certificate from the institution.
Signed at Johannesburg on this the 2 S' September 2009
Signature Print name A f-) Ti-V7 H RqC-C.
STAMP COMMISSIONER OF OATHS Affidavit certified by a Commissioner of Oaths This affidavit conforms with the requirements of the JUSTICES OF THE PEACE AND COMMISSIONERS OF OATHS ACT 16 OF 1963 and the applicable Regulations published in the GG GNR 1258 of 21 July 1972; GN 903 of 10 July 1998; GN 109 of 2 February 2001 as amended. "There is grandeur in this view of life, with its several powers, having been originally breathed into a few forms or into one; and that, whilst this planet has gone cycling on according to the fixed law of gravity, from so simple a beginning endless forms most beautiful and most wonderful have been, and are being, evolved."
— CHARLES R. DARWIN (The Origin of Species) TABLE OF CONTENTS
SUMMARY i
ACKNOWLEDGEMENTS VII
CHAPTER 1: GENERAL INTRODUCTION AND OBJECTIVES 1
1 .1 GENERAL INTRODUCTION 1
1.2 OBJECTIVES 3
CHAPTER 2: GENERAL MATERIALS AND METHODS 6
2.1 MORPHOLOGY 6
2.2 FRUIT ANATOMY AND VITTAE STRUCTURE 7
2.3 DATA CAPTURING 8
2.4 PHYLOGENETIC ANALYSES 8
CHAPTER 3: SYSTEMATICS OF THE CAPNOPHYLLUM GROUP AND PUTATIVE
RELATIVES 12
3.1 INTRODUCTION 12
3.2 MATERIAL AND METHODS 14
3.3 RESULTS 18
3.4 DISCUSSION 30
3.5 KEY TO GENERA OF THE CAPNOPHYLLUM GROUP 35 CHAPTER 4: TAXONOMIC REVISION OF THE GENUS CAPNOPHYLLUM 36
4.1 INTRODUCTION 36
4.2 VEGETATIVE AND REPRODUCTIVE MORPHOLOGY 37
4.3 TAXONOMY OF THE GENUS CAPNOPHYLLUM.. 44
CHAPTER 5: TAXONOMIC REVISION OF THE GENUS DASISPERMUM 58
5.1 INTRODUCTION 58
5.2 VEGETATIVE AND REPRODUCTIVE MORPHOLOGY 59
5.3 TAXONOMY OF THE GENUS DASISPERMUM 68
CHAPTER 6: TAXONOMIC REVISION OF THE GENUS SCARABOIDES 97
6.1 INTRODUCTION 97
6.2 VEGETATIVE AND REPRODUCTIVE MORPHOLOGY 98
6.3 TAXONOMY OF THE GENUS SCARABOIDES.. 100
CHAPTER 7: SYSTEMATICS OF AFRICAN AND MALAGASY PIMPINELLA AND RELATED
GENERA 104
7.1 INTRODUCTION 104
7.2 MATERIAL AND METHODS 105
7.3 RESULTS AND DISCUSSION 106 CHAPTER 8: SYSTEMATICS OF MALAGASY PEUCEDANUM 117
8.1 INTRODUCTION 117
8.2 MATERIAL AND METHODS 118
8.3 RESULTS AND DISCUSSION 119
8.4 TAXONOMY OF THE GENUS BILLBURTTIA 125
CHAPTER 9: SYSTEMATICS OF THE GENUS EZOSCIADIUM 128
9.1 INTRODUCTION 128
9.2 MATERIAL AND METHODS 129
9.3 RESULTS AND DISCUSSION 130
9.4 TAXONOMY OF THE GENUS EZOSCIADIUM 139
CHAPTER 10: NEW TRIBAL DELIMITATIONS OF THE EARLY DIVERGING LINEAGES OF
APIACEAE SUBFAMILY APIOIDEAE 143
10.1 INTRODUCTION 143
10.2 MATERIAL AND METHODS 144
10.3 RESULTS 148
10.4 DISCUSSION 155
10.5 TAXONOMY OF THE PROTOAPIOID TRIBES 165
CHAPTER 11: GENERAL CONCLUSIONS 171
LITERATURE CITED 177 APPENDIX A: VOUCHER SPECIMENS OF FRUIT MATERIAL STUDIED 198
APPENDIX B: DNA ACCESSIONS USED FOR THE PHYLOGENETIC ANALYSES 202
APPENDIX C: PUBLICATIONS RESULTING FROM THIS STUDY 216 SUMMARY
Recent anatomical and molecular studies have highlighted the importance of the African and Malagasy Apiaceae, many of which have been found to occupy early diverging positions within the subfamilies Apioideae and Saniculoideae. Despite the recent interest in the African contingent however, there remain several anomalous and poorly known African and Malagasy taxa in which generic boundaries remain unclear and which have yet to be incorporated within the emerging tribal classification for the family. Generic circumscriptions and affinities amongst hitherto poorly known African and Malagasy genera are here explored using anatomical, cytological, morphological and molecular sequence data. Substantial rearrangements at almost all infrafamilial levels are formalized in order to incorporate the unique African and Malagasy members for the first time.
Generic circumscriptions and phylogenetic relationships of the Cape genera
Capnophyllum, Dasispermum, and Sonderina are explored through parsimony and
Bayesian inference analyses of nrDNA ITS and cpDNA rps16 intron sequences, morphology, and combined molecular and morphological data. The relationship of these genera with the North African genera Krubera and Stoibrax is also assessed.
Analyses of both molecular data sets place Capnophyllum, Dasispermum,
Sonderina, and the only southern African species of Stoibrax (S. capense) within the newly recognized Lefebvrea Glade of tribe Tordylieae. Capnophyllum is strongly supported as monophyletic and is distantly related to Krubera. The monotypic genus
Dasispermum and Stoibrax capense are embedded within a paraphyletic Sonderina.
This complex is distantly related to the North African species of Stoibrax in tribe
Apieae, in which the type species, Stoibrax dichotomum, occurs. Consequently,
i SUMMARY
Dasispermum is expanded to include both Sonderina and Stoibrax capense. A taxonomic revision of Dasispermum s.l. is presented which includes keys to the species, complete nomenclature, typifications, descriptions as well as geographical distributions. New combinations are formalized for Dasispermum capense, 0 hispidum, D. humile, and D. tenue. In addition two new species, namely D. grandicarpum and D. perrenans, are described. As a result seven species of
Dasispermum s.l. are recognised and can be distinguished from one another by their habit (life history and growth form), leaf morphology (leaf texture, leaf colour and breadth of the ultimate leaflet segments), inflorescence structure (length of the peduncle, presence or absence and division of involucre and involucel bracts), fruit morphology (relative length of the styles, fruit size, rib prominence and relative orientation) and fruit anatomy (shape of the cells external to the vittae).
The Cape endemic genus Capnophyllum is revised. As a result of valuable recent collections and extensive fieldwork, this hitherto neglected genus was found to comprise four annual species, two of which are newly described, namely
Capnophyllum lutzeyeri and C. macrocarpum. The four species are distinguished from one another by their fruit morphology (relative length of the styles, the shape and position of the stylopodium, fruit size, surface sculpturing, and the presence or absence of a sterile apical portion) and fruit anatomy (marginal wings slightly or prominently involute and secondary ribs present or absent). A comprehensive key to the species, their complete nomenclature and typification, together with complete descriptions and known geographical distributions for all the species are presented and illustrated.
The genus Scaraboides is described herein to accommodate a new species,
S. manningii, from the Tanqua Karoo in South Africa. This monotypic genus shares
ii SUMMARY the dorsally compressed fruit and involute marginal wings with Capnophyllum, but is easily distinguished by its erect branching habit, green leaves, scabrous umbels, and fruit with indistinct median and lateral ribs, additional solitary vittae in each marginal wing, and parallel, closely spaced commissural vittae. Despite the marked fruit similarities with Capnophyllum, analyses of DNA sequence data place
Scaraboides closer to Dasispermum, with which it shares the erect habit, green
(non-glaucous) leaves, and scabrous umbels.
The phylogenetic position of the African and Malagasy species of Pimpinella are assessed using nrITS sequence data. These species are found to ally with their
Eurasian counterparts within the tribe Pimpinelleae. The genus Pimpinella is rendered paraphyletic by the inclusion of African Cryptotaenia and the small African and Malagasy endemic genera Frommia and Phellolophium. Within the paraphyletic
Pimpinella three major clades are recovered, with the African species widely separated into two of the three clades. Based on the results of the molecular analyses it is clear that the current sectional classification for the genus, based largely on fruit vestiture, is largely artificial. Chromosome base number, however, was found to be consistent with the groupings recovered in the molecular trees.
Optimisation of this cytological data supports the separation of the African species into two clades. Those African and Malagasy Pimpinella species with a chromosome base count of n=11 form the most early diverging Glade together with Frommia which also has a base count of n=11. The remaining African species ally with several
Eurasian species and share a chromosome base count x= 9. Fruit anatomical data is also explored for both African and Malagasy Pimpinella as well as Cryptotaenia,
Frommia and Phellolophium.
iii SUMMARY
The genus Billburttia is described herein to include two new Madagascan endemic species, B. capensoides and B. vaginoides. Although the species appear superficially similar to those of the southern African endemic genus Notobubon, they are easily distinguished by fruit anatomical characters, such as the narrower commissure, the six commissural vittae, the position of the vascular tissue in the tip of the ribs, and sphaerocrystals distributed in and around the epidermis. The two last mentioned characters are proposed as generic apomorphies for Billburttia. The phylogenetic position of the genus is assessed using ITS and rps16 intron sequence data. Both parsimony and Bayesian analyses place Billburttia within the tribe
Apieae, and not closely related to either Peucedanum (Selineae) or the African peucedanoid genera (Lefebvrea Glade of Tordylieae).
The hitherto poorly known Cape endemic genus Ezosciadium (Apiaceae) is revised. This genus is highly distinctive and can be distinguished from other annual genera of the region by its pilose vegetative and reproductive organs, the sessile compound umbels with conspicuously unequal rays, the non-inflexed petal tips, the relatively small, highly-inflexed stamens which appear almost sessile, and the prominent carpophores which persist on the plant. The fruit are unusual in the presence of druse crystals around the carpophore and tanniniferous substances in the epidermal cells of the ribs. The phylogenetic position of the genus within the subfamily Apioideae is assessed using rbcL, trnQ-trnK and nrITS sequence data.
Ezosciadium capense is found to form part of an early diverging lineage within the subfamily, sister group to the Annesorhiza Glade and the genera Molopospermum and Astydamia, here described as the tribe Annesorhizeae. A comprehensive taxonomic revision, including typification, detailed descriptions, geographical range and illustrations of the genus Ezosciadium, is presented.
iv SUMMARY
Phylogenetic analyses of the cpDNA trnQ-trnK 5'exon region for 27 genera and 42 species of Saniculoideae and early diverging lineages of Apioideae were carried out to assess or confirm the tribal placements of the following anomalous genera: Annesorhiza, Astydamia, Chamarea, Choritaenia, Ezosciadium, ltasina,
Lichtensteinia, Marlothiella, Molopospermum and Phlyctidocarpa. To accommodate these unique early diverging members of the Apiaceae and to reflect their relationships, a new classification system has become necessary. Many of the early diverging genera (herein referred to the as protoapioids) can readily be distinguished from the euapioids (the remaining apioids) by the presence of scattered druse crystals in the mesocarp. The major discontinuity within the family, however, lies between the combined protoapioids and euapioids (representing an expanded Apioideae s.l., including the Saniculoideae) and the subfamilies
Azorelloideae and Mackinlayoideae. The broadened subfamily Apioideae s.l. is diagnostically different from the other subfamilies in the absence of rhomboidal crystals, the presence of druse crystals scattered throughout the mesocarp
(subsequently lost in the euapioids), and the non-woody endocarp. No such diagnostic characters are available to support the traditional or recently expanded concept of Saniculoideae. The broadened concept of Apioideae is also supported by the sporadic presence of true wings. This character can be variously interpreted from a phylogenetic point of view, but nevertheless has considerable diagnostic value. A new tribal classification system for the protoapioids is proposed on the basis of molecular, morphological and anatomical evidence. It makes provision for hitherto poorly known African taxa and comprises the following eight tribes, five of which are newly described: Annesorhizeae, Choritaenieae, Heteromorpheae,
v SUMMARY
Lichtensteinieae, Marlothielleae, Phlyctidocarpeae, Saniculeae and
Steganotaenieae.
vi ACKNOWLEDGEMENTS
The following people/organisations are sincerely thanked for their contribution(s) to this study:
My supervisors Prof. Ben-Erik van Wyk, Prof. Patricia M. Tilney and Prof.
Stephen R. Downie for their enthusiasm, patient supervision, valuable input
and advice, financial support, and the many exciting opportunities made
possible. It was truly an honour working with each of them.
Mrs Deborah Katz-Downie for her exceptionally kind hospitality and logistic
support during my visit to Illinois.
Dr Samuel N. Beshers and Prof. Lynn Wiley for generously hosting me in
their home in Illinois.
Mr Heiner and Mrs Eva Lutzeyer for their enthusiasm and hospitality while
performing field studies on their reserve in Stanford and for subsequent
collections of mature fruit.
Prof. Abraham E. van Wyk for useful discussion on the identity of the crystals
in Billburtia and valuable information and material of Choritaenia.
Dr John Manning for assistance with field work and alerting us to unusual
Apiaceae collections.
Mr Jean-Pierre Reduron for supplying mature fruit of Molopospermum.
Prof. Peter Goldblatt, and Mrs Edwina Marinus for there assistance in
obtaining recent herbarium material of Ezosciadium.
Dr Antoine Nicholas and Prof. Gregory Plunkett for providing DNA aliquots of
Choritaenia.
vii ACKNOWLEDGEMENTS
Dr. Jean-Noel Labat for providing a digital image of the type of Caucalis
capense.
Dr Hugh F. Glen, Dr Ian Hedge and Dr Gerrit Koorsen for translating the
diagnoses into Latin.
The curators and staff of the cited herbaria for their kind hospitality and
assistance during visits and for making specimens available on loan.
Dr Annah N. Moteetee for the management of loans to the University of
Johannesburg Herbarium.
Dr Rebecca Liu for keen interest in and assistance with fruit anatomy of
Apiaceae.
Ms Mary Ann E. Feist, Ms Jenny M. Cordes and Mr Clark Danderson for
logistical support, advice and welcome friendship during my work in the
molecular laboratory at the University of Illinois. Mr Clark Danderson is also
thanked for providing sequences of Astydamia and Bupleurum.
Dr Carolina I. Calvin° for supplying the published trnQ-trnK dataset of the
Saniculoideae with which I could build upon, as well as one of the trnQ-trnK
sequences for Phylictidocarpa.
Dr Alexei Oskolskii for hosting us during our visit to St. Peterburg.
Dr Eleena Y. Yembaturova for receiving and guiding us in Moscow.
Mr Pieter J. D. Winter for his enthusiasm and many useful discussions on
Apiaceae systematics.
Mr Andre Marais and his team from De Hoop Nature Reserve for their kind
hospitality during field studies.
viii ACKNOWLEDGEMENTS
The National Research Foundation and the South African Biosystematics
Initiative for funding and travel grants to visit Russia (Apiales IV) and the USA
(University of Illinois).
Department of Botany and Plant Biotechnology for the use of their facilities.
The Faculty of Science at the University of Johannesburg for financial support to visit Prof. Stephen Downie's laboratory at the University of Illinois.
The Lesley Hill Laboratory, the Jodrell Laboratory and Kew Herbarium for the
DNA aliquots.
The Molecular Systematics Laboratory at the University of Johannesburg for the use of their facilities.
Fellow students in the Department of Botany and Plant Biotechnology at
University of Johannesburg for their friendship.
Ms Marianne le Roux for much treasured friendship and companionship during field studies.
Dr James S. Boatwright for his unwavering support, companionship, critical proofreading and assistance with almost all facets of this study.
My family for their love, unlimited support and understanding.
0
ix CHAPTER 1
GENERAL INTRODUCTION AND OBJECTIVES
1.1 GENERAL INTRODUCTION
The Apiaceae are a large and notoriously complex family comprising ca. 455
genera and 3600-3751 species (Pimenov and Leonov 1993) or ca. 463 genera and
3,500 species (Plunkett et al. in press). Although the family has a near cosmopolitan
distribution, the highest diversity is found within the temperate regions of the
Northern Hemisphere. Because of the distinctive umbellate inflorescence and highly
specialised bicarpellate fruit, the Umbelliferae (=Apiaceae) were the first major
natural group of flowering plants to be recognised (Constance 1971). Despite being
the focus of intensive research (particularly over the last four to five decades) from
diverse fields of study (including anatomical, chemical, cytological, morphological,
and most recently molecular approaches), there remain many problems at
practically all infrafamilial levels of classification. Burtt (1991) attributed this in part to
"the very uneven state of our knowledge of the family in different parts of the word",
and highlighted the importance of the hitherto neglected southern Africa taxa in
gaining a better understanding of the family as a whole.
Currently, the family is undergoing a renewed global research effort in which
the results of molecular sequence data is allowing for a re-exploration and re-
interpretation of anatomical, morphological and cytological data with the aim to
produce a more natural classification system to replace the outdated and largely
Eurocentric treatment of Drude (1897-98). In this classical and monumental treatment, Drude (1897-98) proposed that the family be divided into three well-
1 CHAPTER 1: GENERAL INTRODUCTION AND OBJECTIVES defined subfamilies (viz. Apioideae, Hydrocotyloideae and Saniculoideae) based primarily on fruit anatomical data. The recent treatment by Plunkett (2004), based mainly on molecular sequence data, retained Apioideae and Saniculoideae as largely natural groups (although with some rearrangements required). The members of the polyphyletic Hydrocotyloideae were accommodated within two additional subfamilies, the Azorelloideae and Mackinlayoideae. The type genus Hydrocotyle, along with a few related genera were transferred to the closely related sister family
Araliaceae. At the tribal level, however, the system of Drude (1897-98) has been shown to be highly unnatural as a result of large-scale convergence (Downie and
Katz-Downie 1996; Downie et al. 1996, 1998, 2000a, b, c, 2001; Kondo et al. 1996;
Plunkett et al. 1996a, b, 1997; Valiejo-Roman et al. 1998; Katz-Downie et al. 1999;
Plunkett and Downie 1999; Spalik et al. 2004; Calvin° and Downie 2007). As a result the current tribal classification for the family is based exclusively on molecular sequence data (Downie et al. in press) and therefore many genera have not yet been allocated to tribes due to sampling limitations.
The sub-Saharan African and Malagasy contingent of Apiaceae is comparatively small with only 76 genera and 368 species (Van Wyk and Tilney
2004) and exhibits high levels of endemism (40 and 321, respectively). The high incidence of woodiness and other unusual leaf and particularly fruit anatomical characters (Van Wyk 2001; Liu 2004), as well as the many isolated and anomalous taxa, indicate that the African genera are critical to an understanding of higher order relationships within the family (Van Wyk and Tilney 2004). Recent molecular systematic studies (Downie and Katz-Downie 1996, 1999; Plunkett et al. 1996;
Chandler and Plunkett 2004; Calvin° et al. 2006; Calvin° and Downie 2007) have shown that the African and Malagasy genera are often sister to or basally divergent
2 CHAPTER 1: GENERAL INTRODUCTION AND OBJECTIVES within the major lineages of the family (discussed in more detail in Chapters 9, 10 and 11), suggesting an African or likely southern African origin for both subfamilies
Apioideae and Saniculoideae (Calvino et al. 2006; Calvin() and Downie 2007).
Despite the increased sampling of African and particularly southern African taxa in recent phylogenetic studies (Calvino et al. 2006; Calvin() and Downie 2007;
Magee et al. 2008d; Winter et al. 2008; Nicolas and Plunkett 2009), there remain several small African genera with uncertain circumscription and affinity (Burtt 1991;
Lebrun and Stork 1992; Van Wyk and Tilney 2004). While early diverging positions within the largest subfamily Apioideae and the sister subfamily Saniculoideae have been suggested for some of these genera based on recent fruit anatomical data (Liu et al. 2003a, b, 2007b, c; Liu 2004), the majority remain either poorly known or unstudied. Due to the improved molecular sampling of African and particularly the early diverging lineages of the Apioideae-Saniculoideae Glade by Calvirio (Calvino et al. 2006, 2008a, b; Calvino and Downie 2007) and the family-wide survey of fruit anatomical characters, particularly the early diverging lineages of Apioideae and
Saniculoideae, by Liu (Liu et al. 2003a, b, 2006, 2007a, b, c, 2009; Liu 2004), comparisons between these poorly known and previously unstudied African genera and other genera throughout the family are now possible.
1.2 OBJECTIVES
This project is aimed at providing more clarity on generic circumscriptions and affinities amongst hitherto poorly known African and Malagasy genera using anatomical, morphological, and molecular sequence data. The aim was also to consider the best way in which the existing tribal classification system be formally
3 CHAPTER 1: GENERAL INTRODUCTION AND OBJECTIVES modified to allow for the incorporation of anomalous genera. The specific objectives of the study are to:
Explore the putative relationships between the Cape endemic genera,
Sonderina and Capnophyllum, and the North African genera, Stoibrax and
Krubera, as well as the placement of the only southern African species of
Stoibrax, viz. S. capense.
Clarify the generic circumscriptions of the South African genera
Capnophyllum, Stenosemis, Sonderina and Dasispermum.
Revise the species delimitations, nomenclature, typifications, diagnostic
features, and geographical distributions of the taxonomically complex genera
Sonderina, Capnophyllum and the monotypic genus Dasispermum.
Determine the systematic placement of the African and Malagasy species of
Pimpinella, as well as their putative relatives Cryptotaenia, Frommia and
Phellolophium.
Determine the correct systematic placement of the Malagasy species of
Peucedanum.
Confirm the proposed systematic placement of the Malagasy genera
Andriana, Anisopoda, Cannaboides, Pseudocannaboides and Tana within the
early diverging lineages of the Apioideae and particularly the tribe
Heteromorpheae.
Examine the generic circumscription of the poorly known South African genus
Ezosciadium and assess its systematic affinities.
Determine the systematic placement of the anomalous genera Choritaenia,
Phlyctidocarpa and Marlothiella and explore the tribal delimitations of the
4 CHAPTER 1: GENERAL INTRODUCTION AND OBJECTIVES early diverging lineages of the subfamily Apioideae and their affinities to subfamily Saniculoideae.
5 CHAPTER 2
MATERIALS AND METHODS
Authorities for scientific plant names (according to Brummitt and Powell 1992)
are given in Appendices A and B or in Table 11.1. The list of literature references
includes only those cited in the text and not the taxonomic citations included in the
relevant synonymies (abbreviated as in Stafleu and Cowan 1976).
2.1 MORPHOLOGY
During the last five years extensive field work was undertaken in which many of the South African Apiaceae were studied in situ. During these excursions fresh as well as silica- (Chase and Hill 1991) and FAA-preserved materials (formaldehyde: acetic acid : alcohol : water; Sass 1958) for molecular and anatomical studies were collected. Photographs were taken to record habit and certain features of gross morphology. All specimens collected are kept at the University of Johannesburg
Herbarium (JRAU). In addition relevant collections of African and Madagascan
Apiaceae from the following herbaria were studied: BM, BOL, JRAU, K, LE, MO,
NBG, PRE, S, SAM and THUNB-UPS (herbarium acronyms as in Holmgren et al.
1990). Information concerning habitat and phenology was obtained from these sources as well as field notes. From this material, together with information from
Leistner and Morris (1976), the recorded distribution of all the species was ascertained and mapped. The distribution data was recorded using the quarter degree reference system (Edwards and Leistner 1971). Line drawings were made
6 CHAPTER 2: MATERIALS AND METHODS
with the aid of a camera lucida attachment on a Zeiss compound microscope or a
Wild M3Z stereomicroscope.
2.2 FRUIT ANATOMY AND VITTAE STRUCTURE
Preserved (FAA) and herbarium materials were used to study fruit anatomy.
Herbarium material was first re-hydrated and then placed in FAA for a minimum of
24 h. This material was subsequently treated according to a modification of the
method of Feder and O'Brien (1968) for embedding in glycol methacrylate (GMA).
This modification involves a final infiltration in GMA for five days. Transverse
sections, about 3 1.1rn thick, were cut using a Porter-Blum ultramicrotome. The
sections were examined for the presence of crystals using a light microscope, after
which they were stained according to the periodic acid Schiff/toluidine blue (PAS/TB)
method of Feder and O'Brien (1968). A list of voucher specimens for the fruit
anatomical study is given in Appendix A1. The terminology used to describe the fruit
anatomical features follows that proposed by Kljuykov et al. (2004) and is indicated
in the relevant labelled figures.
To study the three-dimensional structure of the vittae, mature fruit were
softened by soaking in boiling water for 24 h. The exocarp was then peeled off while
keeping the fruit submerged in water to prevent desiccation. A list of voucher
specimens used to study the external vittae structure of the mature fruit is given in
Appendix A2.
7 CHAPTER 2: MATERIALS AND METHODS
2.3 DATA CAPTURING
Whole fruit and anatomical sections were photographed using a JVC KY-
F1030 digital camera mounted on either a Zeiss compound microscope or a Wild
M3Z stereomicroscope. These images were then deep etched using the computer
program Adobe Photoshop CS version 8.0. Drawings were made with the aid of a
camera lucida attachment mounted on either the before-mentioned compound
microscope or the stereomicroscope.
2.4 PHYLOGENETIC ANALYSES
2.4.1 DNA extraction, amplification and sequencing— Total DNA was
extracted from herbarium or silica material using either the 2x CTAB method of
Doyle and Doyle (1987), or one of the following commercial extraction kits: DNeasy
Plant Mini Kit (Qiagen) or PureLink TM Plant Total DNA Purification Kit (Invitrogen).
The voucher specimen information and GenBank accession numbers for the
material used in the analyses are given in Appendix B.
The internal transcribed spacers (ITS) of nuclear ribosomal DNA was
amplified using the primer combinations of Sun et al. (1994). For amplification of the
chloroplast DNA rps16 intron and the flanking exon regions (tmQ-rps16 5' exon and
rps16 3' exon-trnK 5' exon) the primers of Downie and Katz-Downie (1996) and Lee
and Downie (2006) were used, respectively. Successfully amplified PCR products were purified using either a QlAquick PCR purification kit (Qiagen Inc.) according to the manufacturer's instructions, or according to the ExoSAP protocol of Werle et al.
(1994) using 5 units of Exonuclease I (New England Biolabs, Ipswich,
8 CHAPTER 2: MATERIALS AND METHODS
Massachusetts, USA) and 0.5 units of Shrimp Alkaline Phosphatase (Promega,
Madison, Wisconsin, USA). Sequencing reactions were carried out using the BigDye
Terminator version 3.1 Cycle Sequencing Kit (Applied Biosystems Inc.) and
sequenced using either an ABI (Applied Biosystems) 3130 XL or 3730 XL
sequencer.
2.4.2 Sequence alignment and phylogenetic analyses— Complementary
strands were assembled and edited using Sequencher version 3.1.2 (Gene Codes
Corporation) and manually aligned in PAUP* version 4.0b10 (Swofford 2002), with
gaps positioned so as to minimise nucleotide mismatches. For the large trnQ - trnK
matrix of Chapter 11, the sequences were initially aligned using the default pairwise
and multiple alignment parameters in the computer program Clustal X (gap opening
cost =15.00, gap extension cost = 6.66, DNA transition weight = 0.50; Jeanmougin
et al. 1998). This alignment was then checked and adjusted manually as necessary,
with gaps positioned so as to minimise nucleotide mismatches. In the latter matrix
unambiguous gaps were scored as presence/absence characters using the simple
indel coding method of Simmons and Ochoterena (2000).
Phylogenetic analyses of all data sets were conducted initially using the
parsimony (MP) algorithm of PAUP*. Character transformations were treated as
unordered and equally weighted (Fitch parsimony; Fitch 1971). As tree search
strategies differed between the datasets they are specified in the relevant Chapters.
Branch support for the MP analyses was determined using bootstrap percentage values (BP; Felsenstein 1985). Only values greater than or equal to 50% are reported and the following scale was applied for support percentages: 74%, weak;
75%-84%, moderate; and 85%-100%, strong.
9 CHAPTER 2: MATERIALS AND METHODS
After model selection with Modeltest version 3.1.2 under the corrected Akaike
information criterion (Akaike 1974; Posada and Crandall 1998), Bayesian inference
(BI; Yang and Rannala 1997) was implemented using MRBAYES version 3.1.2
(Huelsenbeck and Ronquist 2001; Ronquist and Huelsenbeck 2003). We employed
the 'standard' model (using default parameters) for both the indel data as well as the
coded gaps (Lewis 2001). The search strategies implemented for the different
datasets are specified in the relevant Chapters. The analyses were judged to have
reached stationarity when the standard deviation between the split frequencies
stabilised below 0.009. The initial one-fourth of trees where discarded as the 'burn-
in' phase. A majority rule consensus tree was produced from the remaining trees in
order to show the posterior probabilities (PP). The following scale was used to
evaluate the PPs: 0.5-0.84, weak; 0.85-0.94, moderate; 0.95-1.0, strong.
To assess congruency of relationships within the Capnophyllum group
(Chapter 3), as inferred by separate MP analyses of the ITS, rps16 intron and
morphological data sets, the bootstrap consensus trees from each analysis were
compared. These trees were considered incongruent only if they displayed 'hard'
(i.e., incongruencies with strong bootstrap values) rather than 'soft' (i.e.,
incongruencies with weak bootstrap values) incongruence (Seelanan et al. 1997;
Wiens 1998). In addition, a partition homogeneity test (incongruence length difference test, ILD; Farris et al. 1995) was performed in PAUP*. This test was implemented with 1,000 replicate analyses, using the heuristic search option with simple addition of taxa, TBR and the MULTREES option selected. To evaluate the significance of differing topologies, we used the Shimodaira-Hasegawa test (SH;
Shimodaira and Hasegawa 1999), as implemented in PAUP* (applying the RELL re- sampling method with 1,000 bootstrap replicates).
10 CHAPTER 2: MATERIALS AND METHODS
2.4.3 Evolution of morphological characters— Selected morphological characters were reconstructed onto the respective MP trees using parsimony with
Mesquite version 2.5 (Maddison and Maddison 2008).
1 1 CHAPTER 3
SYSTEMATICS OF THE CAPNOPHYLLUM GROUP AND PUTATIVE RELATIVES
3.1 INTRODUCTION
A more natural classification of the large and taxonomically complex
cosmopolitan family Apiaceae is currently emerging as a result of molecular
systematic studies, together with rigorous comparisons of morphological and
anatomical data. Several small genera of uncertain circumscription and affinity are
evident in recent checklists of African Apiaceae (Built 1991; Lebrun and Stork 1992;
Van Wyk and Tilney 2004). The majority of these genera are either poorly known or have not been studied in recent years but may be extremely important in the understanding of relationships within the family as a whole. The South African endemic genera Capnophyllum, Dasispermum and Sonderina were identified as three such taxa. Of these, only two species (Dasispermum suffruticosum and
Sonderina humilis) have previously been included in molecular systematic studies
(Calvin() et al. 2006; Winter et al. 2008). In the phylogenetic analysis by Winter et al.
(2008) using nuclear ribosomal DNA internal transcribed spacer (ITS) sequences, both of these species were shown to be closely related to a group of recently circumscribed African peucedanoid genera, here referred to as the Lefebvrea Glade
(viz. Afroligusticum, Afrosciadium, Cynorhiza, Lefebvrea A.Rich., Nanobubon and
Notobubon) within tribe Tordylieae.
The monophyly of Sonderina has of yet not been assessed, nor has its putative relationship with Stoibrax been confirmed. The genus Sonderina was described by Wolff (1927) to accommodate four of five South African species
12 CHAPTER 3: SYSTEMATICS OF THE CAPNOPHYLLUM GROUP AND PUTATIVE RELATIVES
previously included in Ptychotis W.D.J.Koch by Sonder (1862). Wolff (1927) transferred the fifth species, Ptychotis didyma Sond., to the genus Tragiopsis Pomel
(now Stoibrax), which already included four North African species. Adamson (1939) considered this geographically disjunct treatment to be unnatural and transferred the
South African species, Tragiopsis didyma (Sond.) H.Wolff, to Sonderina. Burtt
(1989), however, argued that such a Cape and North African disjunction was not uncommon and transferred Sonderina didyma (Sond.) Adamson, along with the
North African species, back to the genus Stoibrax, as Stoibrax capense. Burtt (1989, pg 145), furthermore, expressed his doubts about the generic concept of Sonderina, stating that the genus was "probably too close to Stoibrax for it to be maintained."
Burtt (1991), in his checklist of southern African Umbelliferae, treated five species within Sonderina. One of these, the Namibian endemic Sonderina streyi Merxm., has subsequently been transferred to the early diverging African genus Anginon
(Allison and Van Wyk 1997). As a result, only four closely related species are now recognised within the taxonomically difficult genus Sonderina.
A similar disjunction has also been proposed for the genus Capnophyllum, with some authors (e.g., Tutin et al. 1968; Dyer 1975) expanding the genus to include the Mediterranean Capnophyllum peregrinum (L.) Lange. Meikle (1977), however, treated the Mediterranean species as distinct under the monotypic genus
Krubera, a decision maintained by Burtt (1991). A recent taxonomic revision of
Capnophyllum (Chapter 4; Magee et al. 2009c) recognized four species, two of which were newly described and excluded Krubera peregrina on the basis of important differences in fruit anatomy.
A thorough taxonomic study of the genera Capnophyllum, Dasispermum and
Sonderina along with extensive field work has revealed one new monotypic genus
13 CHAPTER 3: SYSTEMATICS OF THE CAPNOPHYLLUM GROUP AND PUTATIVE RELATIVES
(herein described as "Scaraboides manningii") and four new species (Magee et al.
2009c; Magee et al. unpublished). The present study is aimed at resolving generic
circumscriptions and relationships of these previously neglected South African
endemic genera. As the phylogenetic relationships of African Apiaceae genera are
often hard to predict on the basis of morphological characters alone, analyses of
both morphology and anatomy in combination with molecular data (specifically, ITS
and rps16 intron sequences) are here presented and explored.
3.2 MATERIAL AND METHODS
3.2.1 Taxon sampling— In order to assess the generic delimitations and
phylogenetic relationships of the Cape endemic genera Capnophyllum (12 new accessions), Dasispermum (two new accessions), Sonderina (13 new accessions), the undescribed monotypic genus "Scaraboides"(two new accessions) and the largely North African genus Stoibrax (five new accessions). Additional accessions of the rps16 intron region for the closely related African peucedanoid genera Cynorhiza
(two new accessions), Nanobubon (2 new accessions) and Notobubon (five new accessions) were also included. The 45 new accessions for which ITS (18 accessions) and rps16 intron (27 accessions) sequences were obtained and the previously published rps16 intron accessions are listed in Appendix B1 and previously published ITS accessions are available in Winter et al. (2008). The newly obtained ITS sequences were added to the 125 taxon ITS matrix of Winter et al.
(2008). This matrix represents all tribes and major clades of the apioid superclade plus outgroups from tribes Smyrnieae and Oenantheae (Downie et al. 2001), with those species of the latter used to root the trees. The newly obtained rps16 intron
14 CHAPTER 3: SYSTEMATICS OF THE CAPNOPHYLLUM GROUP AND PUTATIVE RELATIVES
sequences were analysed with 27 additional rps16 intron sequences from GenBank
(Appendix B1), the latter also representing several relevant major clades of the
apioid superclade. The rps16 intron trees were rooted with Sium latifolium and
Berula erecta of tribe Oenantheae. To further explore relationships within the
Capnophyllum group, combined data sets (ITS/rps16 intron, ITS/morphology and
ITS/rps16 intron/morphology) for 31 taxa of the Lefebvrea Glade were analysed, with
Lefebvrea abyssinica A.Rich. used as the outgroup. A matrix of 23 morphological
and anatomical characters was prepared based on examination of herbarium
specimens and literature (Tables 3.1 and 3.2; Magee et al. 2008a, b, 2009a, c;
Winter et al. 2008).
3.2.2 Phylogenetic analyses— Phylogenetic analyses of all data sets were
conducted initially using maximum parsimony, as described in Chapter 2, with gaps treated as missing data. Tree searches were performed using a heuristic search with
500 random sequence additions, TBR branch swapping and the MULPARS option in effect, but saving no more than five of the shortest trees from each search. These equally parsimonious trees were then used as starting trees for TBR branch swapping (MULPARS and STEEPEST DESCENT in effect) with the maximum number of trees saved set at 12,000; these trees were permitted to swap to completion (Downie et al. 1998). Bootstrap percentage values for the separate ITS and rps16 intron data sets were determined from 500,000 replicate analyses using fast stepwise addition of taxa, while BP values for the morphological and combined data sets of the Lefebvrea Glade were determined from 1,000 bootstrap replicates, holding 10 trees per replicate and with TBR and MULPARS selected. All data sets
15 CHAPTER 3: SYSTEMATICS OF THE CAPNOPHYLLUM GROUP AND PUTATIVE RELATIVES
TABLE 3.1 Morphological and anatomical characters and states used in the phylogenetic analysis of the Lefebvrea Glade.
1. Life history' (monocarpic = 0; short-lived perennial = 1; perennial = 2); 2. Habit (herbs =
0; rhizomatous = 1; suffrutices = 2; shrubs or shrublets =3); 3. Growth pattern (monopodial = 0; sympodial = 1); 4. Leaf persistence (one-seasoned or deciduous = 0; permanent, evergreen = 1); 5. Leaf arrangement (radical or if somewhat cauline then borne on deciduous branches = 0; cauline, borne on permanent branches =1); 6. Leaf texture (coriaceous = 0; flimsy = 1; sclerophyllous =2); 7. Leaf colour (concolourously green or green above = 0; glaucous =1); 8. Inflorescence vestiture (glabrous = 0; scabrous = 1); 9. Ratio of functionally male flowers (equal ratio of male to female flowers in all raylets of the umbellule = 0; inner raylets of umbellules functionally male =1); 10. Involucre and involucel bracts2 (present = 0; absent or very much reduced = 1); 11. Involucre and involucel bracts type 3 (absent or all simple = 0; at least some compound, resembling the leaves = 1); 12. Petal vestiture (leathery = 0; papillose = 1); 13. Fruit length (more than 9 mm = 0; less than 9 mm = 1); 14. Fruit compression (platyspermous = 0; isodiametric = 1); 15. Fruit in lateral view (narrowly elliptic = 0; very broadly elliptic to rotund = 1); 16. Ribs (median and lateral ribs markedly less developed than the marginal ribs = 0; median and/or lateral ribs as well developed as the marginal ribs = 1); 17. Ribs (obtusely tipped = 0; almost trifid with prominent tapering tips = 1); 18. Secondary ribs (absent = 0; usually present = 1); 19. Marginal wings (absent or flat = 0; involute = 1); 20. Commissural surface (flat = 0; concave = 1); 21. Commissure (100% from rib tip to rib tip = 0; from near rib tip to near rib tip = 1; from at most rib base to rib base = 2); 22. Rib vittae (absent = 0; present at base of all ribs = 1; present in marginal wings = 2); 23. Cells external to vittae (indistinct =0; square = 1; enlarged, upright = 2).
1 Field observations of Dasispermum suffruticosum and Sonderina sp. 1 indicate that these species are not monocarpic annuals but rather short-lived perennials lasting for only a few seasons depending on rainfall, possibly an adaptation to the dune habitat in which they both occur. Perennials include all shrubs and also species with permanent fleshy roots.
2 Some species of Sonderina are distinct in that their involucre and involucel bracts are usually absent or at best strongly reduced and rudimentary.
3 Sonderina hispida, Sonderina sp. 1 and Stoibrax capense are unusual in that at least some of the involucre and involucel bracts are pinnately divided, thus resembling the leaves.
16
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TABLE 0 0 0 0 0 0 0 0 0 ri) tl) ?) -- -g%It rc:31 -0 -CO -0 0 -0.Q000000EE() 0 Z.C.° 0. Q. .0 0 ,...-- 0 Q'..s , -2 -c, ••c, ..ca -0 'C 'Q (1) CD X Z Z Z Z Z z -0 *C 'Q *Q ci) 0) 0) F-3 ° ° Q Z '. CX .4Z1 4:1 Q Q -0 -Q .Q -Q -0 -0 -Q -Q o 2 CD (13 (1) (13 g) 0 0 :.-- 0 Q. Q. R, ° '2.....54). 4:10 2 0 0 0 0 0 0 0 0 0 0 cti Cz3 12 " -1-C3z 00 0 .1:z e .2 Fox cc,g - oc u A3 CD CO CO 0 0 0 0 0 0 0 0 0 0 ("Cj 0 0 0 0 0 CtZ C.)(ii.'. 7..()C.) ■.1 Z Z Z Z Z Z Z Z Z Z Z Z V 0)0)0)0)0) co co co (except the separate morphological data set) were subsequently analysed by
Bayesian inference, as described in Chapter 2. For the combined analysis in which morphological data were included, the "datatype = standard" option of MRBAYES for the non-nucleotide data partition was used. For the separate ITS and rps16 intron data sets, the analysis was performed for three million generations of MCMC and a sampling frequency of 100, while for the morphological and combined data sets of the Lefebvrea Glade only two million generations of MCMC and a sampling frequency of 10 was used.
Morphological characters were reconstructed on the MP trees from the combined ITS/rps16 intron/morphology data set using parsimony.
3.3 RESULTS
3.11 ITS data set— The ITS matrix consisted of 633 unambiguously aligned nucleotide positions with 382 variable and 317 parsimony informative characters.
Parsimony analyses resulted in the preset maximum tree limit of 12,000 trees, each of 2,113 steps (ensemble consistency indices [CI; Kluge and Farris 1969] of 0.33 and 0.31, with and without uninformative characters, respectively; ensemble retention index [RI; Farris 1989] of 0.72). MODELTEST selected the GTR+I+G model of evolution for use in the BI analysis. The MP strict consensus tree yielded a similar topology as the BI consensus tree (with the differences between these trees summarised in Fig. 3.1). In both analyses, the same groupings as reported previously by Winter et al. (2008) were retrieved. The Lefebvrea Glade was weakly
18
CHAPTER 3: SYSTEMATICS OF THE CAPNOPHYLLUM GROUP AND PUTATIVE RELATIVES
Notobubon galbanum 30 Notobubon capense 37 Notobubon tenuifolium 60 Notobubon tenuifolium 44 Notobubon galbanum 10 1.0 Notobubon capense 43 Notobubon galbanum 2452 1.0 1.0 Notobubon gummiferum Notobubon tenuifolium 52 0,2 Notobubon galbaniopse Notobubon striatum ... Notobubon striatum 0.98 Notobubon pungens ion Notobubon pungens 0.89 Notobubon ferulaceum 59 Notobubon ferulaceum 98 Notobubon laevigatum as Notobubon laevigatum to C Nanobubon strictum 99 Nanobubon strict= Nanobubon capillaceum _____ Cynorhiza typica Cynorhiza typica 1.0 C Afroligusticum petilianum 82 Afroliguslicum °Moth Dasispermum suffruticosum 2451
o Dasispermum suffruticosum 3137 87 so Dasispermum suffruticosum 117 01 Sonderina hispida 107 0.80 o Dasispermum suffruticosum 2292 97 0
Sonderina hispida 115 BEWICI Sonderina sp. 1 dep a
96 ep 1.o C Stoibrax capense dou 82 Stolbrax capense ep 60 -Scaraboides manninge 0.53 C Sonderina tenuls gli Sonderina sp. 2 Luni 0.99 1.0 Sonderina iruMmrs 92 6 "Capnophyllum lutzeyerr Capnophyllum leiocarpon 125 1.0 dno. 100 Capnophyllum lekcamon 6978 Capnophyllum africanum 0.67 57 Capnophyllum afrkanum Capnophyllum macrocarpum" 0.75 Afrosciadium magalismontanum 1.0 Stenosemis caffra 96 Stenosemis angustifolia 1.0 Notobubon pearsonii 100 Notobubon pearsonii 191 0.99 Lefebvrea grant!! 74 Lefebvrea grant!! 8 0.99 76 0 13 Lefebvrea grantii 89 71 Lefebvrea grantii 10 Lefebvrea abyssinica ,11L91 99 C1- Lefebvrea abyssinica 0.99 022= Ducrosia anethifolia 89 Kalakia marginata 100 Cymbocarpum anethoides 0.99 0.99 Heracteum clad° 7 1 .0 C Conium maculatum lob Conium sphaerocarpum • 0.99 Sehneae 94 0.9r_= Prangos pabularia 0 99 Ferulago galbanifera Azilia eryngioides Echinophoroae 0.98 Trachyspermum aethusifolium 0 99 = St oypma !too x ihsisapjcdp, esri
0.99 Coriandrum sativum 5 94 Bifora radians o 530 Krubera peregrina Levisticum officinal° 76 Seselopsis tianschanica Sphaenolobium tianschanicum to Deverra burchellii 1 - 0 97 Deverra denudata ssp. aphylla 1.0 93 100 Deverra triradiata Sclerosciadium noditlorum 098 ..1. 10 Apium graveolens 85 1 99 Apium prostratum 0.99 , 1 96 Naufraga balearica 76 94 Anethum graveolens 1.0 Stoibrax 0 100 'drill:it:1,71= 1 0.99 Pimpinelleae 100 0.99 Pyramidoptereae 84 099 Careae 99 o • Smymieae 100 99 Afrocarum imbricatum 0.99 99 Sium repandum 99 Seaga thunbergii 74 Sium bracteatum Helosciadium repens
FIGURE 3.1 Bayesian inference (BI) tree of ITS sequence data. Posterior probability (PP) values are presented above the branches. Bootstrap percentage (BP) values are presented below the branches. BP and PP values below 50% and 0.50, respectively, are not indicated. Branches supported only in the BI are indicated by dashed lines, those branches that differ in the parsimony strict consensus tree are indicated alongside as grey lines.
19 CHAPTER 3: SYSTEMATICS OF THE CAPNOPHYLLUM GROUP AND PUTATIVE RELATIVES supported in both the BI and MP trees (PP = 0.75, BP < 50%). Lefebvrea formed the earliest diverging lineage in the MP strict consensus tree, while in the BI tree its position was unresolved.
With the exception of Notobubon pearsonii, the woody Cape genus
Notobubon was strongly supported as monophyletic in the BI tree (PP 0.98), while in the MP strict consensus tree the latter Glade was not resolved. The Cape genera
Capnophyllum, Dasispermum and Sonderina, together with the only South African species of Stoibrax (S. capense) and the as yet undescribed monotypic genus
"Scaraboides", all formed a group (hereafter referred to as the Capnophyllum group) within the Lefebvrea Glade, which was strongly supported in the BI tree (PP 0.99); however, in the MP strict consensus tree, this Glade was supported with a BP value of less than 50%. "Scaraboides manningif was placed within a weakly supported polytomy with members of the Dasispermum—Sonderina complex (PP 0.6) in the BI tree, while in the MP strict consensus tree it was weakly supported to be sister to the
Sonderina humilis—S. tenuis group (BP < 50%). Dasispermum suffruticosum and
Stoibrax capense, together with Sonderina hispida and Sonderina sp. 1 comprise a strongly supported Glade (PP 1.0, BP 96) that arises from within a paraphyletic
Sonderina. Constraining the Dasispermum—Sonderina complex to monophyly so that "S. manningewas its sister group resulted in trees that were not significantly different (p=0.261).
The North African species Krubera peregrina and Stoibrax dichotomum, sometimes considered to be closely related to Capnophyllum and Sonderina respectively, were inferred to be distantly related to each other and to the
Capnophyllum group. Both accessions of the type species of Stoibrax (S. dichotomum) were moderately to strongly placed within tribe Apieae (PP 0.99, BP
20 CHAPTER 3: SYSTEMATICS OF THE CAPNOPHYLLUM GROUP AND PUTATIVE RELATIVES
76), while Krubera peregrina was resolved in a Glade comprising Coriandrum
sativum L., Bifora radians M.Bieb. and Levisticum officinale W.D.J.Koch.
3.3.2 rps16 intron data set— The rps16 intron matrix consisted of 936
unambiguously aligned nucleotide positions with 191 variable and 92 parsimony
informative characters. MP analyses yielded the preset maximum tree limit of
12,000 trees, each of 282 steps (CI 0.77 and 0.63, with and without uninformative characters, respectively; RI 0.84). MODELTEST selected the K81uf+G model of evolution for use in the BI analysis. Overall the MP strict consensus tree yielded a similar topology to that of the majority rule consensus tree obtained from the BI analysis (differences between the results of these analyses are presented in Fig.
3.2). Although the resolution within these trees was poor, a lineage comprising
Capnophyllum, Dasispermum, "Scaraboides", Sonderina and Stoibrax capense
(Capnophyllum group) was retrieved in both analyses (PP 0.89, less than 50% BP support). This same Capnophyllum group was resolved in the ITS trees with greater taxon sampling (Fig. 3.1). Similarly, Dasispermum suffruticosum and Stoibrax capense were again strongly embedded within a subclade of Sonderina (PP 1.0, BP
87) comprising the type species S. hispida. "Scaraboides manningii" is placed within a strongly-supported polytomy with members of the Dasispermum–Sonderina complex in the BI tree, while in the MP strict consensus tree Sonderina humilis and
Sonderina sp. 2 formed the earliest branching lineages, followed by a polytomy
(<50% BP support) comprising Sonderina tenuis, "Scaraboides manningir and the
Stoibrax capense–Dasispermum suffruticosum Glade. Constraining the
Dasispermum–Sonderina complex to monophyly so that "S. manningir was its sister group resulted in trees that were not significantly different (p=0.191).
21
CHAPTER 3: SYSTEMATICS OF THE CAPNOPHYLLUM GROUP AND PUTATIVE RELATIVES
0.99 Sonderina sp. 1 0.64 62 Sonderina hispida 112 0.6 Sonderina hispida 107 51- Dasispermum suffruticosum 1.0 Stoibrax capense 128 Stoibrax capense 131 1.0 Sonderina hispida 115 87 "Scaraboides manningii " Sonderina tenuis Sonderina sp. 2 0.89 Sonderina humilis
"Capnophyllum lutzeyeri" 07 Capnophyllum leiocarpon 125 01
0.84 1.0 — Capnophyllum leiocarpon 6978 M4 96 Capnophyllum africanum 124 "Capnophyllum macrocarpum " eal 3
Capnophyllum africanum 654 81 Nanobubon capillaceum 0P 1.0 Nanobubon stricturn 86 Nanobubon strict= 1.0 Notobubon pearsonii 0.95 0,99 97 Notobubon pearsonii Cynorhiza typica 53 Cynorhiza typica 3372 Notobubon capense 43 Notobubon tenuifolium Notobubon gummiferum Notobubon capense 37 Notobubon pungens Notobubon galbanum 0.93 Notobubon ferulaceum 1.0 Heracleum maximum 1.0 69 Heracleum lanatum Heracleum sphondylium 0.88 1.0190 871.0 Pastinaca sativa 1.0 81 Malabaila sekakul 76 Zosima orientalis Echinophora tenuifolia 0.98 0.79 Aethusa cynapium 1.0 Thaspium pinnatifidum 94 Zizia aurea Stenosemis caffra 1.0 Crithmum maritimum 88 1.0 C Conium maculatum 100 Conium sphaerocarpon 1.o Naufraga balearica 0.99 1.0 0.74 86 Apium graveolens 53 99 0. 83J Stoibrax dichotomum 0.941 Deverra burchellii Ammi majus Foeniculum vulgare Smyrnium olusatrum Berula erecta Sium latifolium
FIGURE 3.2 Bayesian inference (BI) tree of rps16 intron sequence data. Posterior probability (PP) values are presented above the branches. Bootstrap percentage (BP) values are presented below the branches. BP and PP values below 50% and 0.50, respectively, are not indicated. Branches supported only in the BI are indicated by dashed lines, those branches that differ in the parsimony strict consensus tree are indicated alongside as grey lines.
22 CHAPTER 3: SYSTEMATICS OF THE CAPNOPHYLLUM GROUP AND PUTATIVE RELATIVES
Notobubon formed a weakly supported Glade in the MP strict consensus tree,
although without N. pearsonii, which in both the BI and MP trees was placed as
sister (though supported strongly only in the BI tree with a PP value of 0.99) to an
unresolved Cynorhiza typica. Stoibrax dichotomum was once again shown to be part
of tribe Apieae in both BI (PP 0.94) and MP trees (<50% BP support) and not closely
related to its South African congener, S. capense, or to its putative relative
Sonderina. The placement of Stenosemis outside of the Lefebvrea Glade was not
consistent with that found in the ITS studies and may be due to the low resolution and limited sampling within the tribe Tordylieae.
3.3.3. Morphological data set— MP analysis of 23 morphological and anatomical characters from 31 species of the Lefebvrea Glade resulted in 10 minimal length trees each of 50 steps (CI 0.60, RI 0.88). The relationships among members of the Lefebvrea Glade were generally better resolved than those inferred from the molecular analyses, although with generally lower BP values (Fig. 3.3A). Notobubon pearsonii was included within a weakly supported Glade of Notobubon (BP 54),
Nanobubon was moderately supported as monophyletic (BP 80) and Stenosemis was strongly supported (BP 93) as monophyletic. The Capnophyllum group formed a separate lineage as in the molecular analyses. Capnophyllum was strongly supported as monophyletic (BP 89), with "Scaraboides manningii"as its sister group. Dasispermum and Stoibrax were again recovered within a paraphyletic
Sonderina (BP 72).
23
CHAPTER 3: SYSTEMATICS OF THE CAPNOPHYLLUM GROUP AND PUTATIVE RELATIVES
A. morphology Notobubon capense C. ITS/morphology 0.96 Notobubon capense 37 Notobubon galbanum 0 Notobubon tenuifchum 44 54 Notobubon tent/datum 99 Notobubon galbanum 2452 1.0 16 Notobubon gurnmiferum 87 Notobubon gummiterum 65 097. — Notobubon galbaniopse — Notobubon galbaniopse — Notobubon pungens Notobubon pungens 50 Notobubon ferutaceum Notobubon striatum 51 Notobubon faevigatum 0.79 NotobubOn ferulaceurn 2500 L Notobubon pearsonii 99 Notobubon laevigatum 2510 06 Notobubon striatum Notobubon pearsond 42
Stenosemis caffra 0.73 1.0 Stenosemis caffra 9 Stenosemis angustifolia too Stenosemis angustitolia Nanobubon striatum Nanobubon striclum 58 Nanobubon capillaceum Nanobubon caplitaccum Cynorhiza typica Cynorhiza typica 53 0.68 Afmsciadlum magaltsmontanum Afroligusticum petitianum Afroligusticum petition= Afrofigusticum elltodt Atmligusticum alba Dasispennum suffruticosum 117 10" Sondedna hispida Sondedna hispida 107 1.0 I Sondedna sp. 1 7 Stoibrax capense 5 99 Sondedna humilis Stotbrax capense 126 0. 66 dep *7 Sonderina tenuis 62 F Sondedna humilis 72 T -- 095 — Sondedna (emits 3- dou Dasfspermum suffridlcosum Sondedna sp. 2 gy
Sondedna sp. 1 ni
Scaraboides manningie w Sondedna sp. 2 - "Capnophyllum lutzeyerr B "Scaraboides manningii a Capnophyllum leiocarpon 6978 "Capnophyllum lutzeyerC dno, 100 Capnophyllum africanum 124 Capnophyllum feiocarpon "Capnophyllum macrocarpum - — T Capnophyllum of icanum 09 Afroscladium magallsinontanum "Cepnophyflum macrocarpum" Lefebvrea abyssinica Lefebvrea abyssfnica — 0.01 B. ITS/rps /6 intron D. ITS' rps16 intron/morphology 0,9 Notobubon capense 37 0.9 Notobubon capense 37 1,0 Notobubon tenurial= 44 1.0 Notobubon lenuifolium 44 100 Notobubon galbanum 2452 Notobubon galbanum 2452 66 " Notobubon gummtiefUm Notobubon gummiferum Notobubon galbaniopse Notobubon galbaniopse Notobubon pungens 50 Notobubon pungens 50 Notobubon striatum 51 Notobubon striatum 51 1.0 Notobubon ferufaceum 2500 Notobubon terulaceum 2500 100 Notobubon faevIgatum 2510 Notobubon laevigatum 2510 0.5n Nanobubon striatum 58 Notobubon peasant; 41 Cynorhiza typica 53 Stenosemis caffra Afroligusticum petitianum 100 Stenosemis angustilotia Afroligusticum entail Nanobubon striatum 58 Nanobubon capillaceum Nanobubon capillaceum 1.0 Dasispermum suffmticosum 117 Cynorhiza typica 53 10 ee Sondedna hispida 107 Afroligusticum pelitionum 1.0 Sonderina sp. 1 Afroligusticum elliotii 0,74 1 100 Stolbrax capense 128 0.79 Dasispermum suffruticosum 117 1,0 "Scaraboides manningir Sondertna hispida 107 9 1 Sondertna humNs Sondedna sp. 1 Sondedna tenths Stoibrax capense 128 1.0 0.51 Sondedna sp. 2 62 Sondedna humilis Capnophyllum lutzeyeri" a Sondedna tenuis 1.0 Capnophyllum telocarpon 6978 Sondedna sp. 2 100 "Capnophyllum macrocaTum . "Scaraboides manningii" Capnophyllum africanum 124 "Capnophyllum lutzeyeri"
0.53 Afrosciadium magalismontanum Capnophyllum leiocarpon 6978 j- 1.0 Stenosemis coif's 100 Capnophyllum atricanum 124 90 Stenosemis angustitolia "Capnophyllum maancorpum - — Notobubon peasant! 42 Afrosaiadium magatismontanum Lefebvrea abyssin/ca Lefebvrea abysanIca --- 0.01 — 0.01
FIGURE 3.3 (A), Strict consensus tree of 18 equally most parsimonious trees based on the parsimony analysis of morphological data. (B), Bayesian inference (BI) phylogram of the combined ITSIrps16 intron data sets. (C), Bayesian inference phylogram of the combined ITS/morphological data sets. (D), Bayesian inference phylogram of the combined ITSIrps16 intron/morphological data sets. Posterior probability values are presented above the branches. Bootstrap values from the parsimony analysis are presented below the branches. BP and PP values below 50% and 0.50, respectively, are not indicated. Branches supported only in the BI tree are indicated by dashed lines, those branches that differ in the parsimony strict consensus tree are indicated alongside as grey lines.
24 CHAPTER 3: SYSTEMATICS OF THE CAPNOPHYLLUM GROUP AND PUTATIVE RELATIVES
3.3.4. Combined ITS/rps16 intron data set— The combined ITS and rps16
intron matrix for 31 taxa within the Lefebvrea Glade consisted of 1,593 characters, of which 228 were variable and 112 parsimony informative. Missing data represented
13% of the entire data matrix, as rps16 intron sequences were unavailable for eight taxa. Visual inspection of the two separate bootstrap consensus trees and results of the ILD test suggested that the two matrices were not significantly incongruent
(p=1.0). Parsimony analyses of combined molecular data yielded 97 trees, each of
359 steps (CI 0.74 and 0.59, with and without uninformative characters, respectively; RI 0.77). The GTR+I+G and the K81uf+G models were retained for the
ITS and rps16 intron data partitions, respectively. Trees obtained from both the MP and BI analyses yielded the same overall topologies (Fig. 3.3B). As in the analyses of partitioned molecular data, a lineage comprising the Capnophyllum group was recovered (PP 1.0, BP 62). Capnophyllum was again strongly supported as monophyletic (PP 1.0, BP 100). Dasispermum suffruticosum and Stoibrax capense were strongly embedded within a subclade of a paraphyletic Sonderina, together with Sonderina hispida and Sonderina sp. 1 (PP 1.0, BP 100). "Scaraboides manningil' was again strongly supported as part of a Glade comprising
Dasispermum, Stoibrax capense and all species of Sonderina (PP 1.0, BP 91), although its exact position within this Glade remained equivocal. In the BI trees
"Scaraboides manningil' was weakly supported as a sister group to the subclade comprising Stoibrax-Dasispermum (PP 0.74), while in the MP strict consensus tree it formed one branch of a trichotomy. As in the separate ITS and rps16 analyses, constraining the Dasispermum–Sonderina complex to monophyly so that "S. manningir was its sister group resulted in trees that were not significantly different
(p=0.142).
25 CHAPTER 3: SYSTEMATICS OF THE CAPNOPHYLLUM GROUP AND PUTATIVE RELATIVES
3.3.5. Combined ITS/morphological data set— The combined ITS and morphological matrix for 31 taxa within the Lefebvrea Glade consisted of 712 characters, of which 206 were variable and 115 parsimony informative. Visual inspection of the two separate MP bootstrap consensus trees revealed no hard incongruence. However, the ILD test suggested that the two were significantly incongruent (p=0.001). Following the suggestions of Seelenan et al. (1997) and
Wiens (1998), the two matrices were still combined for simultaneous analyses.
Parsimony analysis yielded 200 trees, each of 364 steps (CI 0.68 and 0.55, with and without uninformative characters, respectively; RI 0.76). The GTR+I+G model was retained for the ITS data. Overall, the MP strict consensus tree yielded a similar topology as those retrieved from the BI analysis (differences between the results of these analyses are presented in Fig. 3.3C). As in the morphological analysis, subclades comprising the Cape genera Notobubon (PP 0.79, BP <50), Nanobubon
(PP 1.0, BP 77) and Stenosemis (PP 1.0, BP 100) were supported as monophyletic
(PP 0.73, BP <50). The Capnophyllum group was again recovered (PP 1.0, BP 81), with Capnophyllum strongly supported as monophyletic (PP 1.0, BP 100).
Dasispermum and Stoibrax capense again arose from within a paraphyletic
Sonderina. This broadened Dasispermum—Sonderina complex was weakly to moderately supported (PP 0.83, BP 62), with "Scaraboides" as its sister group (PP
0.85), although in the MP strict consensus tree a trichotomy comprising
Capnophyllum, "Scaraboides" and the Dasispermum—Sonderina complex was retrieved.
26 CHAPTER 3: SYSTEMATICS OF THE CAPNOPHYLLUM GROUP AND PUTATIVE RELATIVES
3.3.6. Combined ITS/rps16 intron/morphology data set— The combined
ITS, rps16 intron and morphological matrix for 31 taxa within the Lefebvrea Glade consisted of 1,616 characters, of which 251 were variable and 135 parsimony informative. Missing data represented 8.6% of the entire data matrix, as rps16 intron sequences were unavailable for eight taxa. Visual inspection of the three separate
MP bootstrap trees revealed no hard incongruencies. Parsimony analyses yielded
10 trees, each of 425 steps (CI 0.69 and 0.56, with and without uninformative characters, respectively; RI 0.77). Trees obtained from both the MP and BI analyses yielded similar overall topologies (differences between the results of these analyses are presented in Fig. 3.3D). The topology resolved from the combined ITS/rps16 intron/morphology analyses was generally similar to those obtained from analyses of the combined ITS/morphology dataset. The genera Capnophyllum (PP 1.0, BP 100),
Nanobubon (PP 0.97, BP 68) and Stenosemis (PP 1.0, BP 100) were each recovered as monophyletic. The Capnophyllum group (i.e., Capnophyllum,
Dasispermum, "Scaraboides", Sonderina and Stoibrax capense) was again retrieved
(PP 1.0, BP 81). The Glade comprising "Scaraboides" and the Dasispermum-
Sonderina group was strongly supported (PP 1.0, BP 83). As in all prior analyses,
Sonderina is rendered paraphyletic by the inclusion of Dasispermum and Stoibrax capense.
3.3.7. Morphological character evolution— Parsimony-based reconstructions of eight morphological characters supporting either the
Capnophyllum group or the Dasispermum–Sonderina complex are each summarised onto one of the ten minimal length trees inferred from MP analysis of combined ITS/rps16 intron/morphology data (Fig. 3.4).
27 CHAPTER 3: SYSTEMATICS OF THE CAPNOPHYLLUM GROUP AND PUTATIVE RELATIVES
1. Life history 3. Growth pattern 12. Petal vestiture 14. Fruit compression ■ sympodial ■ ■ perennial papillose isodiametric ■ short-lived perennial ❑ monopodial leathery platyspermous monocarpic Notobubon capense 31 Notobubon tenuifolium 44 Notobubon galbanum 2452 A C D Notobubon gummiferum INotobubon galbaniopse Notobubon pungens 50 Notobubon striatum 51 r—.1Notobubon ferulaceum 2500 I Notobubon laevigatum 2510 Notobubon pearsonii 42 Stenosemis caffra Stenosemis angustifolia Nanobubon stncturn 58 L__I Nanobubon capillaceum ICynorhiza typica 53 1,.. Afroligusticum petitianum Afrohgusticum elliotii Dasispermum suffruticosum 117 ill Sonderina hispida 107 a) Sonderina sp. 1 Stoibrax capense 128 Sonderina humilis Sonderina tenuis Sonderina sp. 2 Scaraboides manningii" a "Capnophyllum lutzeyeri" Capnophyllum leiocarpon 6978 3 Capnophyllum africanum 124 "Capnophyllum macrocarpum" lAfrosciadium magalismontanum 1Lefebvrea abyssinica
15. Fruit in lateral view 16. Median and/or lateral ribs 21. Commissure 23. Cells external to vittae ■ very broadly elliptic ■ enlarged, upright as developed as marginal rib base to rib base ■ to rotund < developed than marginal ■ near rib tip to near rib tip square narrowly elliptic rib tip to rib tip indistinct Notobubon capense 31 Notobubon tenuifolium 44 Notobubon galbanum 2452 H Notobubon gummiferum Notobubon galbaniopse Notobubon pungens 50 Notobubon striatum 51 Notobubon ferulaceum 2500 Notobubon laevigatum 2510 I Notobubon pearsonii 42 _r- Stenosemis caffra t_ Stenosemis angustifolia J- Nanobubon stnctum 58 i_ Nanobubon capillaceum ICynorhiza typica 53 Afroligusticum petitianum Afrohgusticum elliotii Dasispermum suffruticosum 117' 2 0 Sonderina hispida 107 a) Sonderina sp. 1 I a Stoibrax capense 128 • o •1:3 r 1Sonderina humilis : z- L__J L I Sonderina tenuis .< Sonderina sp. 2 o .E" I "Scaraboides manningii" "Capnophyllum lutzeyen" cc Capnophyllum /eiocarpon 6978 3 c Capnophyllum africanum 124 -0 "Capnophyllum macrocarpum" _ Afrosciadium magalismontanum Lefebvrea abYssinica
FIGURE 3.4 Reconstruction of eight morphological characters (A—H) supporting either the Capnophyllum group or the Dasispermum—Sonderina complex (indicated with a dashed line), when optimized over the ten minimal length trees inferred from MP analysis of combined ITS/rps16 intron/morphology data.
28 CHAPTER 3: SYSTEMATICS OF THE CAPNOPHYLLUM GROUP AND PUTATIVE RELATIVES
Monocarpic life history (character 1, Fig. 3.4A), sympodial growth pattern
(character 3, Fig. 3.4B) and the very broadly elliptic to rotund fruit in lateral view
(character 15, Fig. 3.4E) were reconstructed as synapomorphies for the
Capnophyllum group. Petal vestiture (character 12, Fig. 3.4C) was ambiguously reconstructed at the base of the Capnophyllum group; this character can either be interpreted as a synapomorphy for the Capnophyllum group with reversals in
"Scaraboides manningiP and Dasispermum suffruticosum, or as a convergent character supporting both Capnophyllum as well as the Dasispermum—Sonderina complex, with a reversal in Dasispermum suffruticosum. Reconstruction of the short- lived perennial habit (character 1) differed slightly among the 10 minimal length trees depending on the relative position of Sonderina sp. 1. In eight of the trees, this character was reconstructed as an autapomorphy for both Sonderina sp. 1 and
Dasispermum suffruticosum (as shown in Fig. 3.4A), while in the remaining two trees where Sonderina sp. 1 and Dasispermum suffruticosum were sister, it was reconstructed as a synapomorphy. The Dasispermum—Sonderina complex was supported by an isodiametric fruit compression (character 14, Fig. 3.4D), median and/or lateral ribs as developed as the marginal ribs (character 16, Fig. 3.4F), commissure extending from at most rib base to rib base (character 21, Fig. 3.4G) and square or enlarged, upright cells external to the vittae (character 23, Fig. 3.4H).
Flimsy leaf texture (character 6) and concolourously glaucous leaves (character 7) were both reconstructed as synapomorphies for Capnophyllum (not shown).
29 CHAPTER 3: SYSTEMATICS OF THE CAPNOPHYLLUM GROUP AND PUTATIVE RELATIVES
3.4 DISCUSSION
3.4.1. Lefebvrea Glade— Winter et al. (2008) reported that the African
species previously attributed to Peucedanum and its platyspermous allies comprise
a Glade sister to a small alliance of southwest Asian species (Ducrosia anethifolia
Boiss., Kalakia marginata (Boiss.) R.Alava, Cymbocarpum anethoides DC.) within
tribe Tordylieae. Within their "African group", here referred to as the Lefebvrea Glade,
the non-platyspermous, South African endemic species Dasispermum suffruticosum,
Sonderina humilis, Stenosemis caffra and Stenosemis angustifolia E.Mey. ex. Harv.
& Sond. were also included. Our broadened analyses show that the Lefebvrea Glade
also includes all species of Sonderina and Capnophyllum, the monotypic genus
"Scaraboides"and the Cape species Stoibrax capense. Winter et al. (2008)
separated Dasispermum, Sonderina and Stenosemis from the peucedanoid genera
by their narrower commissure, not extending to the tips of each marginal rib/wing.
Capnophyllum and "Scaraboides"also have dorsally compressed fruit with
prominent marginal wings and a broad commissure. These three characters are all plesiomorphies based on the distribution of characters states (Fig. 3.4D, F, G). The fruit of Stenosemis differ from those of Dasispermum, Sonderina and Stoibrax in that the commissure, though not as broad as found in Capnophyllum, "Scaraboides"and the peucedanoid genera, is much broader and extends to beyond the base of the marginal wings, often somewhere between the tip and the centre of the wing/rib.
This feature was reconstructed as a synapomorphy for the genus Stenosemis (Fig.
3.4G).
30 CHAPTER 3: SYSTEMATICS OF THE CAPNOPHYLLUM GROUP AND PUTATIVE RELATIVES
3.4.2. Capnophyllum and Krubera— Although the genus Capnophyllum has
sometimes been expanded to include the Mediterranean species Krubera peregrina,
Magee et al. (2009c) maintained the two genera as distinct, in agreement with
Meikle (1977) and Burtt (1991). The fruit of both genera (Fig. 3.5D, G) are superficially similar; they have dorsally compressed mericarps, a broad commissure, marginal ribs extended into wings and prominent ridges on the dorsal surface.
However, on close examination of the fruit, Magee et al. (2009c) found diagnostic differences between the two genera in terms of the size and prominence of both vittae and rib oil ducts and the shape of the marginal wings. Krubera and
Capnophyllum were very widely separated in the ITS-derived trees (Fig. 3.1), with
Krubera placed in a Glade sister to Coriandrum sativum and Bifora radians and
Capnophyllum placed in the Lefebvrea Glade of tribe Tordylieae. In all trees presented herein, Capnophyllum is strongly supported as monophyletic and occupies a position within a broader lineage comprising other largely annual, sympatric, Cape endemic genera (viz. Dasispermum, Sonderina, Stoibrax capense and "Scaraboides').
3.4.3. Dasispermum, Sonderina and Stoibrax— Burtt (1989) suggested that the South African endemic genus Sonderina may be insufficiently distinct at the generic level from the largely North African genus Stoibrax. Previous molecular systematic studies have shown the type species of Stoibrax (S. dichotomum) to be placed in tribe Apieae (e.g., Downie et al. 2001). Our analyses of both ITS and rps16 intron data clearly show Sonderina to form part of the Lefebvrea Glade together with the only South African species of Stoibrax, S. capense. Such a relationship was
31 CHAPTER 3: SYSTEMATICS OF THE CAPNOPHYLLUM GROUP AND PUTATIVE RELATIVES
11■1•1111•■
=13M121311= medr Ir \
1:10:=MIZ=18,1=
FIGURE 3.5 Transverse sections through the fruit of Stoibrax hanotei (A), Stoibrax capense (B), Sonderina hispida (C), Krubera peregrina (D), Stenosemis caffra (E), Dasispermum suffruticosum (F), and Capnophyllum africanum (G). Vouchers: A. Wall s.n. (S); B. Boatwright et al. 212 (JRAU); C. Van Wyk 3539 (JRAU); D. Lippert 22959 (PRE); E. MacOwen s.n. sub South African Exchange Club 904 (GRA); F. Winter 78 (JRAU); G. Winter 110 (JRAU). c, carpophore; cv, commissural vitta; Ir, lateral rib; mr, marginal rib; medr, median rib; rod, rib oil duct; vb, vascular bundle; vv, vallecular vitta. Scale: 1 mm.
proposed by Adamson (1939, 1950) who treated Stoibrax capense (then Stoibrax didyma) as Sonderina didyma.
The monotypic genus Dasispermum is strongly supported in all analyses as embedded within Sonderina. The single species, Dasispermum suffruticosum, is a perennial dune-endemic, distinguished by its usually fleshy leaves and often
32 CHAPTER 3: SYSTEMATICS OF THE CAPNOPHYLLUM GROUP AND PUTATIVE RELATIVES prominently winged fruit (Fig. 3.5F) which can be either homo- or heteromericarpic.
The genus is highly plastic with regard to both of these characters (Tilney and Van
Wyk 1995). Leaves may be less fleshy in some individuals within a population, especially those in more shaded sites and the ribs of some fruit may not be expanded into wings. Both characters appear to be adaptations to harsh littoral conditions and wind dispersal. Dasispermum shares the sympodial growth habit, a synapomorphy for the Capnophyllum group (Fig. 3.4B), as well as the isodiametric fruit, median and/or lateral ribs as well developed as the marginal ribs, narrow commissure (Fig. 3.5A, B, C, F) and the square or upright cells external to the vittae of the fruit (Fig. 5.4) with Sonderina and Stoibrax. These character states are all synapomorphies for the Dasispermum–Sonderina complex (Fig. 3.4D, F, G, H).
Furthermore, the close relationship between Dasispermum suffruticosum and
Stoibrax capense is supported by a chromosome number of n=9, an unusual number for the subfamily (Constance et al. 1976). An expanded circumscription of the genus Dasispermum to include Sonderina and Stoibrax capense, therefore, seems to be unavoidable.
3.4.4. "Scaraboides"— While revising the genus Capnophyllum (Magee et al. 2009c), the authors were alerted to an unusual species from the arid Tanqua
Karoo region. Although this species shares numerous fruit characters with
Capnophyllum, such as dorsally compressed mericarps, broad commissures, concave commissural surfaces and involute marginal wings (Fig. 6.1E, F, G), it also has morphological characters in common with species of Sonderina, such as an erect habit (Fig. 6.1A), green ultimate leaf segments, scabrous often sessile umbels
(Fig. 6.1A, D) and the absence of involucral and involucel bracts (Fig. 6.1D). The
33 CHAPTER 3: SYSTEMATICS OF THE CAPNOPHYLLUM GROUP AND PUTATIVE RELATIVES
sympodial habit is weakly expressed in young plants of this species and is therefore
not clearly visible in Fig. 6.1A. The species is easily distinguished by the presence of additional wing vittae (not known in any other genus within the family) and parallel, closely-spaced commissural vittae in the fruit (Fig. 6.1E, F, G).
Separate and combined analyses of ITS and rps16 intron data sets place
"Scaraboides" either within the Dasispermum–Sonderina complex (but with weak support values) or are equivocal in its placement. Shimodaira-Hasegawa tests indicate that a sister group relationship of "Scaraboides" to this complex (as retrieved in the analyses of the ITS/morphology and ITS/rps16 intron/morphology data sets) cannot be rejected. In the analysis of morphological data, the genus was moderately supported (BP 75) as sister to Capnophyllum. When combined with the molecular data sets, the position of "Scaraboides"was either unresolved (MP strict consensus tree, Fig. 3.3C) or sister to the Dasispermum–Sonderina complex (Fig.
3.3C and D). The genus is clearly morphologically distinct from the Dasispermum-
Sonderina complex, which has isodiametric fruit with a narrow commissure (Fig.
3.5B, C, F). The inclusion of "Scaraboides" within an expanded circumscription of
Dasispermum would result in a group that would be impossible to delimit based on observed morphological characters. Despite the superficial similarity between the fruits of "Scaraboides manningirand Capnophyllum, neither the molecular nor the combined molecular/morphological analyses place these taxa together. It is therefore clear that "Scaraboides" represents an independent, easily recognizable lineage.
3.4.5. Capnophyllum group—All analyses presented herein indicate a broader lineage within the Lefebvrea Glade, comprising Capnophyllum,
34 CHAPTER 3: SYSTEMATICS OF THE CAPNOPHYLLUM GROUP AND PUTATIVE RELATIVES
Dasispermum, "Scaraboides", Sonderina and Stoibrax capense. These taxa share a unique combination of characters, namely the monocarpic or rarely short-lived perennial life history, the sympodial growth pattern, the papillose petals and the very broadly elliptic to rotund fruit in lateral view. The Glade is therefore defined herein as the Capnophyllum group.
3.5 KEY TO GENERA OF THE CAPNOPHYLLUM GROUP
la. Fruit isodiametric; if homomericarpic then with the median and lateral ribs as
well developed as the marginal ribs, if heteromericarpic then with either median
or lateral ribs as well developed as the marginal ribs, prominent or winged;
commissure narrow, extending to the base of each rib Dasispermum
1 b. Fruit dorsally compressed; homomericarpic with the median and lateral ribs not
as well developed as the marginal ribs, marginal ribs prominently winged,
median and lateral ribs inconspicuous or prominent but not winged; commissure
very broad, extending to the tip of each wing:
Involucral and involucel bracts absent; rays and raylets scabrous; fruit with
indistinct median and lateral ribs; additional vittae in the marginal wings;
commissural vittae close together; ultimate leaflet segments more than 1.5
mm broad (never subterete), green Scaraboides
Involucral and involucel bracts present; rays and raylets glabrous; fruit with
prominent median and lateral ribs; additional vittae in the marginal wings
absent; commissural vittae widely separate; ultimate leaflet segments less
than 1 mm broad (often subterete), glaucous Capnophyllum
35 CHAPTER 4
TAXONOMIC REVISION OF THE GENUS CAPNOPHYLLUM
4.1 INTRODUCTION
Capnophyllum is a genus of small annual herbs endemic to South Africa. The name is derived from the Greek (kapnos = smoke, but is also the Greek name for
Fumaria; phyllon = leaf) and refers to the distinctly glaucous leaves that closely resemble those of the genus Fumaria L. (Sonder 1862; Adamson 1950).
Capnophyllum is sometimes extended to include the Mediterranean Krubera peregrinum (e.g. Tutin et al. 1968; Dyer 1975; Downie et al. 1998). However, according to Meikle (1977), Krubera can be distinguished from Capnophyllum by fruit and floral differences. Burtt (1991) followed Meikle (1977) in considering
Capnophyllum to be a South African endemic genus. Strong support for the monophyly of Capnophyllum and for its placement within the Lefebvrea Glade of the tribe Tordylieae, together with the Cape genera Dasispermum, Sonderina and
Stoibrax capense (referred to as the Capnophyllum group) was shown using molecular and morphological data in Chapter 3. In contrast, Krubera was shown to be only distantly related, occupying a position somewhere between the tribes Apieae and Selineae (Downie et al. 1998; Downie et al. 2000c; Winter et al. 2008; Magee et al. 2009b).
Until relatively recently, the genus in the strictest sense was considered to be monotypic, with two varieties of C. africanum recognised on the basis of fruit surface sculpturing. Goldblatt and Manning (2000), however, found that the two varieties were morphologically and geographically distinct and therefore raised C. africanum
36 CHAPTER 4: TAXONOMIC REVISION OF THE GENUS CAPNOPHYLLUM var. leiocarpon Sond. to the rank of species, as C. leiocarpon. As a result of valuable recent collections and extensive fieldwork, this hitherto neglected genus was found to comprise four species, two of which are as yet undescribed. We present here a detailed taxonomic treatment of the genus, including a key to the species, their complete nomenclature, typification, formal descriptions, as well as the known geographical distributions.
4.2 VEGETATIVE AND REPRODUCTIVE MORPHOLOGY
4.2.1 Vegetative morphology— The four species of Capnophyllum are all annual herbs which exhibit a sympodial growth pattern similar to that found in the closely related genus Sonderina (Magee et al. 2009b). In both these genera, the stem consists of a series of sympodial segments each ending in a terminal umbel which becomes laterally displaced by continued growth from the axillary bud of the uppermost leaf, which takes over as the main growth axis (Burtt 1991). In
Capnophyllum the plants are all well-branched sprawling herbs in which the branches become prostrate or decumbent (Fig. 4.1A-C). As the generic name implies, the leaves are always characteristically glaucous. They are pinnate to bi- pinnate and mostly cauline (Fig. 4.1A–C), with the ultimate leaflet segments flat or subterete and less than 1 mm broad (Fig. 4.1D). All the species are relatively small in stature (ranging from 50-500 mm in height). Capnophyllum lutzeyeri appears to be more robust than the other species, while C. macrocarpum (Fig. 4.1A) is generally much smaller at maturity. However, as both these species are known from only a single locality, the size of the plants may eventually prove to be of limited diagnostic value.
37 CHAPTER 4: TAXONOMIC REVISION OF THE GENUS CAPNOPHYLLUM
FIGURE 4.1 General morphology of Capnophyllum. (A) C. macrocarpum in situ (photo by B.- E. van Wyk), showing the glaucous Fumaria-like leaves characteristic of the genus as well as the sparse compound umbels and small stature typical of this species; (B) C. leiocarpon in situ (photo by A.R. Magee), showing the distinctive decumbent branches; (C) typical sympodial growth form; (D) representative lower leaf pinnae; (E) typical petal in ventral and lateral view; (F) umbellule of C. africanum showing the distinct tubercules on the ovaries. Vouchers: C. Magee et al. 125, JRAU; D. Bester 6978, JRAU; E. Williamson 3825, BOL; F. Magee et al. 124, JRAU. Scale: C = 25 mm, D and E = 5 mm, F = 1 mm.
38 CHAPTER 4: TAXONOMIC REVISION OF THE GENUS CAPNOPHYLLUM
4.2.2 Reproductive morphology— The relatively small compound umbels
appear to be laterally borne along the stems but are in fact terminal and become
leaf-opposed due to continued growth from the axillary shoots. The bracts of the
involucre and involucel may be either free or connate at their bases and the rays and raylets are invariably glabrous (Fig. 4.1F). The umbels of C. macrocarpum are always very sparse with only two or three rays consistently present (Fig. 4.1A). The other species usually have at least four rays, although two or three may rarely be present in C. leiocarpon and C. lutzeyeri.
The hermaphroditic flowers are pentamerous, with indistinct sepals and white, oblong to obovate, papillose petals with acuminate inflexed tips (Fig. 4.1 E).
The ovaries of C. africanum (Fig. 4.1F) and C. macrocarpum are distinctive in that they are prominently tubercled, while in the other species they are smooth.
The shape and position of the stylopodia in mature fruit were found to be useful diagnostic characters (Fig. 4.2). The stylopodia are either conical and raised above the fruit apex as in C. africanum (Fig. 4.2A, E), C. leiocarpon (Fig. 4.2C, G) and C. lutzeyeri (Fig. 4.2B, F), or very shortly conical to flattish and diagnostically sunken below the fruit apex in C. macrocarpum (Fig. 4.2D, H). The styles are at first erect and relatively short, but may lengthen as the fruit mature. Their relative length is an important diagnostic character. In C. macrocarpum (Fig. 4.2D, H) the styles remain erect and relatively short whereas in the other species they lengthen markedly and become reflexed. The styles of C. leiocarpon become reflexed far beyond the base of the stylopodium (Fig. 4.2C, G), while in the similar C. lutzeyeri they are reflexed only up to, at most, the stylopodium base (Fig. 4.2F). In C. africanum there appears to be some variability in that the styles may become reflexed either up to or beyond the base of the stylopodium.
39 CHAPTER 4: TAXONOMIC REVISION OF THE GENUS CAPNOPHYLLUM
11■MIMINIMON
FIGURE 4.2 Variation in fruit morphology (A—D) of Capnophyllum species with drawings of the stylopodia (E—H): (A, E) C. africanum; (B, F) C. lutzeyeri; (C, G) C. leiocarpon; (D, H) C. macrocarpum. Vouchers: Al. Bachmann s.n., BOL; A2. Bolus 2793, BOL; A3. Winter 110, JRAU; A4. Adamson 3081, BOL; A5. Van der Merwe 1767, NBG; (B1) Lutzeyher 7A, JRAU; B2. Magee et al. 106, JRAU; C1. Williamson 3825, BOL; C2. Stirton & Zantovska 11432, NBG; C3. Pillans 7899, BOL; D. (three fruit) Magee et al. 133, JRAU; E. Winter 110, JRAU; F. Lutzeyer 7A, JRAU; G. Stirton and Zantovska 11430, NBG; H. Magee 133, JRAU. Scale: A—D = 3 mm, E—H = 1 mm.
40 CHAPTER 4: TAXONOMIC REVISION OF THE GENUS CAPNOPHYLLUM
FIGURE 4.3 Transverse sections through the fruit of Capnophyllum (A—D) and Krubera (E): (A) C. lutzeyeri; (B) C. macrocarpum (almost mature fruit); (C) C. africanum; (D) C. leiocarpon; (E) K. peregrinum with inlay showing enlargement of a commissural vitta. Vouchers: A. Magee et al. 106, JRAU; B. Magee et al. 133, JRAU; C. Winter 110, JRAU; D. Williamson 3825, PRE; E. Lippert 22959, PRE. cc, commissural cavity; cv, commissural vitta; Ir, lateral rib; medr, median rib; mr, marginal rib; rod, rib oil duct; sr, secondary ribs; vv, vallecular vitta. Scale: A—E = 1 mm, inlay = 0.2 mm.
The fruit of Capnophyllum are dorsally compressed, with the marginal ribs expanded into narrow wings (Fig. 4.2) and the commissure extending over the full width of the mericarp (i.e. to the very edge of both marginal ribs). The marginal wings are distinct in that they are slightly to prominently involute, so that the commissural surface is very slightly to prominently concave (Fig. 4.3). In transverse sections of immature fruit, the commissural area between the marginal wings is
41 CHAPTER 4: TAXONOMIC REVISION OF THE GENUS CAPNOPHYLLUM composed of parenchymatous cells which do not become lignified with maturation of the fruit. Instead these cells disintegrate to create a hollow between the two mericarps (Fig. 4.3A, B). The median and lateral ribs are slightly prominent to prominent and are diagnostically tubercled in C. africanum (Fig. 4.2A) and C. macrocarpum (Fig. 4.2D). Secondary ribs are often present above the vallecular and commissural vittae in C. africanum, C. leiocarpon and C. lutzeyeri (Fig. 4.3A, C, D).
The fruit of C. macrocarpum (Fig. 4.2D) are the largest in the genus, with even the smallest fruit of this species distinctly larger than the largest fruit of C. africanum
(Fig. 4.2A). In transverse section there are two commissural vittae as well as four solitary vallecular vittae (Figs. 4.3 and 4.4) in each mericarp of all the species.
In Capnophyllum macrocarpum there is a prominent sterile apical portion on the fruit that can most easily be seen by comparing the extent of the commissural vittae (Fig. 4.4A). In this species the commissural vittae terminate well below the stylopodium; in C. lutzeyeri they terminate slightly below the stylopodium leaving a much smaller sterile portion (Fig. 4.4C). In the remaining species the commissural vittae terminate at the base of the stylopodium so that there is no conspicuous sterile portion (Fig. 4.4B, D).
4.2.3 Comparison of Capnophyllum and Krubera— The fruit of
Capnophyllum and Krubera are superficially similar in having prominent ridges on the dorsal surface, a broad commissure, dorsally compressed mericarps and marginal ribs extended into wings. On close examination of the fruit anatomy, however, the two genera are clearly distinct. The fruit of Krubera (Fig. 4.3E) have extremely small, inconspicuous commissural and vallecular vittae unlike those of the species of Capnophyllum (Fig. 4.3A-D) where the vittae are relatively large and
42 CHAPTER 4: TAXONOMIC REVISION OF THE GENUS CAPNOPHYLLUM conspicuous. The fruit of Krubera also differ markedly in the presence of large rib oil ducts. The fruit anatomical data therefore supports the separation of the two genera as found in Chapter 2.
,,,,,,.CII
C / D I--- sp
I A
FIGURE 4.4 Three-dimensional structure of the commissural vittae of Capnophyllum species: (A) C. macrocarpum; (B) C. africanum; (C) C. lutzeyeri; (D) C. leiocarpon. Vouchers: A. Magee et al. 133, JRAU; B. Van der Merwe 1767, NBG; C. Magee et al. 106, JRAU; D. Williamson 3825, BOL. cv, commissural vitta; sp, sterile portion. Scale: 1 mm.
43 CHAPTER 4: TAXONOMIC REVISION OF THE GENUS CAPNOPHYLLUM
4.3 TAXONOMY OF THE GENUS CAPNOPHYLLUM
CAPNOPHYLLUM Gaertn., Fruct. 2: 32 (1792); DC., Prodr. 4: 187 (1825); Sond. in
Harv. and Sond., Fl. Cap. 2: 562 (1862); Adamson in Adamson and Salter, Fl.
Cape Penins. 625 (1950); Dyer, Gen. S. Afr. Flowering Pl. 2: 426 (1975), p.p.
maj.; B.L. Burtt in Edinb. J. Bot. 48(2): 189 (1991). — TYPE: C. africanum (L.)
Gaertn.
Abioton Raf., Good Book 56 (1840) reimp. Scad. Gen. Omb. Pl. 56 (Amer. Midl. Nat.
Repr. No. 3, 1913), nom. illegit. — TYPE: A. africanum (L.) Raf.
Actinocladus E.Mey. in Ind. Sem. Hort. Bot. Regiomont (1846) and in Ann. Sci. Nat.
Bot. Ser. 3,7: 380 (1847). — TYPE: A. cinerascens E.Mey.
Sprawling sympodial herbs, 0.05-0.5 m tall, annual. Stems well-branched at the base, branches usually prostrate or decumbent. Leaves cauline, 15-210 mm x
7-90 mm, usually becoming smaller towards the upper part of the stem, pinnate to bi-pinnate, glabrous, glaucous (and closely resembling those of Fumaria). Petioles
5-110 mm long, angular, basal sheaths prominent, 3-12 mm x 2-9 mm, margins widely membranous. Ultimate leaflets ovate to broadly ovate or obovate, 7-17 mm x
3-13 mm, pinnatisect, venation not visible or pinnate; segments linear-oblong, 1-6 mm long, less than 1 mm broad, flat or subterete, apex obtuse to acute. Umbels compound; peduncle short; involucral bracts 2 to 4 (to 6), connate or free, lanceolate to ovate, apex acuminate, margins widely membranous, glabrous; rays (2 to) 4 to 10
(to 14), 10-45 mm long at anthesis, glabrous; involucel bracteoles 4 (to 6), connate or free, lanceolate to ovate, apex acuminate, margins widely membranous, glabrous; raylets 6 to 25, 2-8 mm long at anthesis, glabrous. Flowers
44 CHAPTER 4: TAXONOMIC REVISION OF THE GENUS CAPNOPHYLLUM
hermaphroditic; pentamerous; sepals indistinct; petals white, ± 1 mm long and
broad, broadly oblong to obovate, tips inflexed, acuminate, septum present on inner
face, apex truncate to shallowly emarginate, papillose; stamens with anthers
inflexed; ovary inferior, smooth or tubercled, stylopodium shortly conical or flat,
raised above or sunken below the fruit apex; styles at first erect, short, often longer
in mature fruit, either remaining erect or becoming reflexed up to or beyond the base of the stylopodium. Fruit a schizocarp, dorsally compressed, elliptic to broadly elliptic or ovate to broadly ovate in dorsal view, 5.5-9.0 mm x 2.5-5.5 mm; narrowly elliptic and convex in lateral view; base obtuse to truncate or shallowly concave; apex obtuse to truncate; mericarps homomorphic, very slightly to markedly concave on the commissural surface; median and lateral ribs slightly prominent to prominent, straight or flexuose, smooth or tubercled; marginal ribs narrowly winged, slightly to prominently involute, leaving a small to large commissural cavity; secondary ribs often present above the vallecular and commissural vittae; commissural vittae 2, terminating either directly below or some distance from the base of the stylopodium; vallecular vittae 4; commissure very broad, 100% of mericarp width; carpophore bipartite.
Diagnostic characters— Species of Capnophyllum are annual, often sprawling, white-flowered herbs with soft, glaucous leaves resembling those of
Fumaria. They differ from other vegetatively similar annual African genera by the dorsally compressed, narrowly-winged fruit with a broad commissure (100% of mericarp width). They are distinct from African peucedanoid genera in their sympodial growth pattern resulting in leaf-opposed umbels. All other African peucedanoid genera have a monopodial growth pattern with terminal umbels.
45 CHAPTER 4: TAXONOMIC REVISION OF THE GENUS CAPNOPHYLLUM
Furthermore, the fruit differ from those of other annual African peucedanoids in the usually prominent median and lateral ribs, the narrowly-winged marginal ribs which are slightly to prominently involute, the often conspicuous secondary ribs on both the commissural and dorsal surfaces, the slightly to prominently concave commissural surface of the mericarps and the strongly elliptical outline in both dorsal and lateral views as a result of the convex outer surface.
The species often co-occur with the superficially similar and widespread
Sonderina hispida. Although the fruit of Capnophyllum are fundamentally different from those of Sonderina (in Sonderina the fruit are isodiametric, lack marginal wings and have a narrow commissure), when in flower they may easily be confused with one another. However, even when in flower Capnophyllum species can be distinguished by their prostrate habit, glaucous leaves, tubercled ovaries in C. africanum and C. macrocarpum (glabrous or more usually scabrous or pilose in
Sonderina hispida), petals appearing truncate or at most shallowly-emarginate in dorsal view (deeply-emarginate in Sonderina) and glabrous umbels (scabrous in
Sonderina hispida).
Distribution and habitat— The four species are endemic to the Cape
Floristic Region of South Africa (Figs. 4.5 and 4.6) and occur in sandy soil near to the coast. The species are generally allopatric in their distributions; however there is some overlap between the ranges of C. africanum and C. leiocarpon around
Langebaan and Saldanha.
Phenology— Seeds germinate during winter after the first autumn rains.
Flowering occurs in spring (September and October) with mature fruit forming from
46 CHAPTER 4: TAXONOMIC REVISION OF THE GENUS CAPNOPHYLLUM
late spring to early summer (November and December). All four species are annuals that wither and die during the dry mid-summer (January).
KEY TO THE SPECIES OF CAPNOPHYLLUM
1 a. Flowers and fruit with tubercles on the median and lateral ribs (most prominent
on ovaries and young fruit):
Fruit more than 8.5 mm long; stylopodium sunken below the apex of the
fruit, very shortly conical to flat; fruit with conspicuous sterile apical portion
(commissural vittae terminating some distance from the base of the
stylopodium); rays 2 or 3 per umbel; known only from De Hoop Nature
Reserve 4. C. macrocarpum
Fruit 8 mm long or less; stylopodium not sunken below the apex of the fruit,
shortly conical; fruit without a conspicuous sterile apical portion
(commissural vittae terminating directly below the base of the stylopodium);
rays more than 3 per umbel; from Gordon's Bay northwards along the coast
to Vredenburg 1. C. africanum lb. Flowers and fruit with smooth median and lateral ribs (without tubercles):
Styles of mature fruit reflexed and extending ± to the base of the
stylopodium, short (0.7-0.8 mm long); mericarp markedly concave on the
commissural surface; known only from Stanford in the Western Cape
3. C. lutzeyeri
Styles of mature fruit reflexed and extending far beyond the base of the
stylopodium, long (1.2-1.7 mm long),; mericarp only slightly concave on the
commissural surface; known from the west coast from Langebaan
northwards to Port Nolloth 2. C leiocarpon
47 CHAPTER 4: TAXONOMIC REVISION OF THE GENUS CAPNOPHYLLUM
1 . CAPNOPHYLLUM AFRICANUM (L.) Gaertn., Fruct. 2: 32, tab. 85, f. 6 (1792); DC.,
Prodr. 4: 187 (1825); Ecklon and Zeyh., Enum. PI. Afric. Austral. 353 (1837);
Sond. in Harv. and Sond., Fl. Cap. 2: 562 (1862); Adamson in Adamson and
Salter, Fl. Cape Penins. 625 (1950). Conium africanum L., Sp. Pl. 243 (1753);
Mant. Alt. 252 (1771); Jacq., Hort. Vindob. 2: tab. 194 (1772); Thunb., Fl.
Cap. 257 (1823). Caucalis africana (L.) Crantz, Cl. Umbell. Emend. 109
(1767). Cicuta africana (L.) Lam., Encycl. 2: 4 (1786), saltem quoad
basionym. Abioton africanum (L.) Raf., Good Book 56 (1840); reimp. Scad.
Gen. Omb. 56 (Amer. Midl. Nat. Repr. 3, 1913). -TYPE: "Caucalis afra, folio
minoris Rutae"in Boerhaave, Index Pl. Alt. 1:63 (1720), (lecto., designated
by Van Wyk and Tilney in Jarvis et al. 2006).
Capnophyllum simplex Lagasca, Gen. and Sp. Nov. 13 (1816), nom. illeg.
Capnophyllum jacquinii DC., Prodr. 4: 187 (1830), p.p. min.
Actinocladus cinerascens E.Mey. in Ind. Sem. Hort. Bot. Regiomont (1846) and in
Ann. Sci. Nat. Bot. Ser. 3, 7: 380 (1847). -TYPE: Hort. Region, Zeyher ex
herb. Meyer s.n. (S!, lecto., here designated). [Note: The specimen in S is
chosen here as the label is in Meyer's handwriting and bears the annotation
"mihi".]
Sprawling or rarely somewhat erect herb, 0.05-0.3 m tall. Leaves 15-210 mm
long x 8-60 mm. Petioles 5-110 mm long, basal sheaths 3-12 mm x 3-9 mm.
Ultimate leaflets ovate to broadly ovate, 7-17 mm x 4-13 mm, venation not visible or pinnate; segments linear-oblong, 1-6 mm long, less than 1 mm broad, flat or subterete. Umbels with peduncle short, 5-60 mm long; involucral bracts (2 to) 4 (to
6), connate or free; rays 4 to 7 (to 14), 10-45 mm long at anthesis; involucel
48 CHAPTER 4: TAXONOMIC REVISION OF THE GENUS CAPNOPHYLLUM bracteoles 4 (to 6), connate or free; raylets 10 to 25, 2-7 mm long at anthesis.
Flowers with ovary tubercled; stylopodium shortly conical, raised above the fruit apex; styles lengthening as fruit matures, (0.3—) 0.5-0.8 mm long, becoming reflexed up to or beyond the base of the stylopodium. Fruit elliptic to broadly elliptic or ovate to broadly ovate, 6.5-8.0 mm x 2.5-4.0 mm; base obtuse or shallowly concave; apex obtuse to truncate; mericarps very slightly concave on the commissural surface; median and lateral ribs prominent, flexuose, tubercled; marginal ribs slightly involute; secondary ribs usually present above the vallecular and commissural vittae; commissural vittae terminating directly below the base of the stylopodium.
Diagnostic characters— Capnophyllum africanum is vegetatively similar to
C. leiocarpon and C. lutzeyeri but differs in that both the ovaries and fruit are covered with tubercles on the median and lateral ribs, as in C. macrocarpum. It differs from the latter species, however, in the larger number of rays per umbel (4 or more), the smaller fruit (less than 8 mm long) without a sterile apical portion and with the shortly conical stylopodium raised above the fruit apex.
Distribution and habitat— Capnophyllum africanum is endemic to the
Western Cape Province, occurring from Gordons Bay and the Cape Peninsula northwards along the coast to Vredenburg (Fig. 4.5). This species has also been collected on Robben Island. It occurs in open Strandveld on deep sandy soils.
49 CHAPTER 4: TAXONOMIC REVISION OF THE GENUS CAPNOPHYLLUM
Additional specimens examined.
—3217 (Vredenburg): 5 km from Vredenburg to Saldanha (—DD), Stirton 10723 (K, NBG).
—3318 (Cape Town): Elandsfontein west, second ridge W of farmhouse (—AA), Thompson
3534 (NBG, PRE); Geelbek Farm, Langebaan (—AA), Bosenberg & Rutherford 138
(NBG); flats at S end of Langebaan lagoon (—AA), Goldblatt 2711 (NBG, PRE); along
R27, 14 km from Langebaan to Cape Town, near Club Mykonos turnoff (—AA),
Magee & Boatwright 114 (JRAU); along R27, resting area near Swartberg, 33° 13'
12" S 18° 12' 15" E (—AA), Forest et al. 654 (NBG); along R27, from Velddrif to Cape
Town, 33° 07' 38" S 18° 07' 53" E (—AA), Boatwright et al. 228 (JRAU); along R27,
from Velddrif to Cape Town, 33° 10' 33" S 18° 10' 21" E (—AA), Boatwright et al. 229b
(JRAU); sand near Hopefield (—AB), Bachmann s.n. sub BOL 1627 (BOL, K); 11 km
NNE of Yzerfontein (—AD), Strid & Strid 38062 (NBG); Buffelsrivier (—CB), Van der
Merwe 1767 (K, NBG, PRE); Buffelsrivier, Hardy Joubert's sample plots near
Bokbaai (—CB), Taylor 4165 (NBG, PRE); Melkbosstrand (—CB), Dahlstrand 1064
(PRE); Clifton-on-sea (—CD), Phillips s.n. sub PRE 59930 (PRE); Milnerton (—CD),
Andreae 381 (NBG); Wall 196 (S); Robben Island (—CD), Adamson s.n. (BOL);
Sunset Beach, S of Table View (—CD), Van Slageren & Newton 842 (K); Brackenfell,
Cape Flats (—DC), Du Plessis 108 (PRE); Cape Flats Nature Reserve (—DC), Low
866a (PRE); Cape Flats (—DC); Pappe s.n. (K); Wallich s.n. (BM); sandy flats near
Kuils River (—DC), Anon ex herb. Worsdell s.n. (K); high dunes W of Sarepta (—DC),
Acocks 1112, 1294 (S); Stikland (—DC), Acocks 2253 (S); Vygekraal (—DC),
MacOwan 1751 (BM, NBG, K), Wolley Dod 706 (K).
—3418 (Simonstown): Fish Hoek (—AB), Young 27349 (PRE, fragment labelled A); near
Glencairn (—AB), Hutchinson 640 (BM, BOL, K, PRE); Kalk Bay (—AB), Levyns s.n.
sub BOL 1625 (BOL); Kommetjie, Imhoff's Gift (—AB), Winter 110 (JRAU); Oaklands
(—AB), Jameson s.n. (K); Retreat Railway Station (—AB), Bolus 2793 (BOL, PRE);
Scarborough (—AB), Adamson 3081 (BOL); Varkensvlei Experimental Farm (—BA),
Joubert 26147 (NBG); Wynberg Flats (—BA), Anon ex herb Prior s.n. (K); Gordons
50 CHAPTER 4: TAXONOMIC REVISION OF THE GENUS CAPNOPHYLLUM
Bay (—BB), Parker 3860 (BOL, K, NBG); Somerset in Hottentots Holland (—BB),
Ecklon & Zeyher 2252 (S); Strand (—BB), Parker 4349 (K, NBG).
Precise locality unknown: "Prom. b. Spei", Masson s.n. (BM); " In arenosis ad likes marls
Sinu False Bay", Bolus 4632 (BM, BOL); "In solo arenosa prope Capetown", Bolus
2793 (BOL, K, NBG); Burchell 362 (K, PRE); Zeyher 742 (K, two sheets); Zeyher
635 (K); Ecklon & Zeyher 2252 (NBG).
HEIGHT ABOVE SEA LEVEL
Over 1500 m [Lj 900 - 1500 m 300 - 900 m Under 300 m
20 0 20 40 60 80 100 km 11111/.11,111
FIGURE 4. 5 Geographical distribution of Capnophyllum africanum.
51 CHAPTER 4: TAXONOMIC REVISION OF THE GENUS CAPNOPHYLLUM
2. CAPNOPHYLLUM LEIOCARPON (Sond.) J.C.Manning and Goldblatt in Goldblatt and
Manning, Cape Pl. 705 (2000). Capnophyllum africanum var. leiocarpon
Sond. in Harv. and Sond., FL Cap. 2: 562 (1862); B.L. Burtt in Edinb. J. Bot.
48(2): 189 (1991). -TYPE: SOUTH AFRICA. Near Cape Town, DrOge 6243 (S!,
lecto., here designated; BM!, G, K!, two sheets, MO, isolecto.).
Sprawling or somewhat erect herb, 0.1-0.5 m tall. Leaves 50-200 mm x 20-
90 mm. Petioles 8-90 mm long, basal sheaths 4-12 mm x 3-6 mm. Ultimate leaflets
ovate to broadly ovate or obovate, 7-16 mm x 5-10 mm, venation not visible or
pinnate; segments linear-oblong, 1-6 mm long, less than 1 mm broad, flat or
subterete. Umbels with peduncle short, (3–) 10-60 (-105) mm long; involucral
bracts 2 to 4, connate, sometimes free; rays (2 to) 4 to 8, 10-30 mm long at
anthesis; involucel bracteoles 4 (to 6), connate; raylets 10 to 25, 2-7 mm long at
anthesis. Flowers with ovary smooth; stylopodium shortly conical, raised above the fruit apex; styles lengthening as the fruit matures, 1.2-1.7 mm long, becoming
reflexed far beyond the base of the stylopodium. Fruit elliptic to broadly ovate, 5.5-
8.0 mm x 3.0-4.0 mm; base and apex obtuse to truncate; mericarps slightly to very
slightly concave on the commissural surface; median and lateral ribs prominent, flexuose, smooth; marginal ribs slightly involute; secondary ribs usually present above the vallecular and commissural vittae; commissural vittae terminating directly below the base of the stylopodium.
Diagnostic characters— Capnophyllum leiocarpon differs from C. africanum and C. macrocarpum in the absence of tubercles on the ribs of both the ovaries and fruit. It can be distinguished from C. lutzeyeri by the longer styles of the mature fruit
52 CHAPTER 4: TAXONOMIC REVISION OF THE GENUS CAPNOPHYLLUM that become reflexed far beyond the base of the stylopodium and the only very slightly concave commissural surfaces of the mericarps.
Distribution and habitat— This widespread species occurs along the coast from near Langebaan in the Western Cape Province northwards to Port Nolloth in the Northern Cape Province (Fig. 4.6). It occurs in open Strandveld on deep sandy soil.
Additional specimens examined.
–2916 (Port Nolloth): ± 15 km E of Port Nolloth (–BB), Williamson 3508 (BOL), 3825 (BOL,
NBG, PRE).
–2917 (Springbok): Farm Zonnekwa 328, 1 km N of crossroads to Graafwater and
Sonnekwa B on road from Vaalkol (–CD), Le Roux & Lloyd 517 (NBG).
3017 (Hondeklipbaai): Farm Koingaas 475, on road from Hondeklipbaai to Koinghaas on
S banks of Swartlintjies River (–AB), Le Roux & Lloyd 327 (NBG); Farm Ghaams
492, SE of Spoeg River (–AD), Boucher 5667 (NBG); sand dunes between
Wallekraal and Hondeklipbaai (–AD), Goldblatt 4222 (K, PRE).
3117 (Lepelfontein): Brand-se-Baai (–BD), Van Rooyen 2154 (PRE).
–3217 (Vredenburg): Paternoster Bay (–DD), Lavranos 11708 (PRE); 5 km from
Vredenburg to Saldanha (–DD), Stirton 10723 (NBG).
3218 (Clanwilliam): Nortier Experimental Station, Lamberts Bay (–AB), Boucher 2539 (K,
PRE), Liengme 988 (BOL), Van Breda 4296 (K, PRE), Van der Merwe 1664 (PRE);
Elands Bay (–AD), Metelerkamp 233 (BOL), Pillans 7899 (BOL); 5 km from Elands
Bay on road to Lamberts Bay (–AD), Stirton & Zantovska 11430 (NBG); 2 km from
Redelinghuys (–BC), Stirton & Zantovska 11432 (NBG, two sheets); Dwarskersbos
(–CA), Taylor 12003 (NBG, PRE); Rocher Pan Nature Reserve (–CB), Magee &
53 CHAPTER 4: TAXONOMIC REVISION OF THE GENUS CAPNOPHYLLUM
Boatwright 116 (JRAU), Van Rooyen & Ramsey 256 (NBG); Berg River Mouth (-
CC), O'Callaghan 8/11 (NBG).
—3318 (Cape Town): along R27, from Veldrif to Cape Town, 33° 10' 33" S 18° 10' 21" E (-
AA), Boatwright et al. 229a (JRAU).
HEIGHT ABOVE SEA LEVEL Over 1500 m L„ij 900 - 1500 m LD 300 - 900 m r IUnder 300 m
20 0 20 40 60 80 100 km /1111111,1111
FIGURE 4.6 Geographical distribution of Capnophyllum leiocarpon (diamonds), C. lutzeyeri (triangle), C. macrocarpum (square).
54 CHAPTER 4: TAXONOMIC REVISION OF THE GENUS CAPNOPHYLLUM
3. CAPNOPHYLLUM LUTZEYERI Magee and B.-E.van Wyk, sp. nov., Capnophyllo
africano et C. leiocarpo stylopodio brevi costis fructus dorsalibus
secondariisque prominentibus similis sed fructu non tuberculato stylis
brevioribus et superficiebus commissurialibus valde concavis differt. -TYPE:
SOUTH AFRICA. Caledon district (3419): Grootbos Nature Reserve, Steynsbos
(–CB); 15 December 2006; H. Lutzeyer 7A (NBG!, holo.; JRAU!, iso.).
Sprawling herb, 0.2-0.5 m tall. Leaves 15-60 mm x 7-25 mm. Petioles 5-15 mm long, basal sheaths 3-8 mm x 2-5 mm. Ultimate leaflets ovate to broadly ovate,
7-15 mm x 4-9 mm, venation not visible; segments linear-oblong, 1.5-3 mm long, less than 1 mm broad, subterete. Umbels with peduncle short, 15-75 mm long; involucral bracts 2 to 4, free?; rays (2 to) 4 to 15, 10-30 mm long at anthesis; involucel bracteoles 4, free?; raylets 7 to 10, 2-6 mm long at anthesis. Flowers with ovary smooth; stylopodium shortly conical, raised above the fruit apex; styles lengthening slightly as the fruit matures, 0.7-0.8 mm long, becoming reflexed up to or just above the base of the stylopodium. Fruit broadly elliptic, 6.0-6.5 mm x 3.5-
4.0 mm; base obtuse to truncate; apex obtuse; mericarps markedly concave on the commissural surface; median and lateral ribs slightly prominent, straight, smooth; marginal ribs prominently involute; secondary ribs present above the vallecular and commissural vittae; commissural vittae terminating slightly below the base of the stylopodium.
Diagnostic characters— Capnophyllum lutzeyeri differs from C. africanum and C. macrocarpum in the smooth fruit and from C. leiocarpon in the shorter styles,
55 CHAPTER 4: TAXONOMIC REVISION OF THE GENUS CAPNOPHYLLUM which become reflexed only to the base of the stylopodium and the markedly concave commissural surface.
Distribution and habitat— The species is known only from the private nature reserve Grootbos, near Stanford in the Western Cape Province (Fig. 4.6), where it was collected on acid derived sand in Fynbos vegetation that had burnt the previous year. No plants were found in subsequent searches by Mr H. Lutzeyer, suggesting that this species may be a short-lived fireweed.
Additional specimens examined.
–3419 (Caledon): Steynsbos, Grootbos Nature Reserve (–CB), Magee et al. 106 (JRAU,
NBG, PRE).
4. CAPNOPHYLLUM MACROCARPUM Magee and B.-E.van Wyk, sp. nov., Capnophyllo
africano costis dorsalibus tuberculatis ovarii fructusque similis sed fructu
maiori portione apicali sterile longo, stylopodio immerso rudimentali et
superficiebus commissurialibus valde concavis differt. —TYPE: SOUTH AFRICA.
Caledon district (3420): De Hoop Nature Reserve, near to De Mond (–AD); 4
November 2007; A.R. Magee, B.-E. Van Wyk and J.S. Boatwright 133 (NBG!,
holo.; BOL!, JRAU!, K!, iso.).
Sprawling herb, 0.04-0.08 m tall. Leaves 25-85 mm x 10-40 mm. Petioles
10-45 mm long, basal sheaths 5-8 mm x 2-3 mm. Ultimate leaflets ovate to broadly ovate, 9-12 mm x 8-10 mm, venation not visible; segments linear-oblong, 2-5 mm long, less than 1 mm broad, flat or subterete. Umbels with peduncle short, 3-8 mm long; involucral bracts 2 or 4, connate or free; rays 2 or 3, 7-35 mm long at anthesis;
56 CHAPTER 4: TAXONOMIC REVISION OF THE GENUS CAPNOPHYLLUM
involucel bracteoles 4, connate; raylets 10 to 15, 2-5 mm long at anthesis. Flowers with ovary tubercled; stylopodium very shortly conical to flat, sunken below the fruit
apex; styles not lengthening as the fruit matures, 0.3-0.5 mm long, remaining erect or rarely becoming somewhat reflexed to just above the base of the stylopodium.
Fruit broadly elliptic to broadly ovate, 8.5-9.0 mm x 4.5-5.5 mm; base obtuse or shallowly concave; apex obtuse to truncate; mericarps markedly concave on the commissural surface; median and lateral ribs prominent, flexuose, tubercled; marginal ribs prominently involute; secondary ribs absent; commissural vittae terminating some distance from the base of the stylopodium.
Diagnostic characters— As in Capnophyllum africanum, both the ovaries and fruit are covered with tubercles on the median and lateral ribs. Capnophyllum macrocarpum differs from this species in the larger fruit (more than 8.5 mm long) with a conspicuous sterile apical portion and the rudimentary stylopodium that is sunken below the apex of the fruit. Furthermore, the umbels of this species appear to have fewer rays (2 or 3).
Distribution and habitat— This species is known only from De Hoop Nature
Reserve in the Western Cape Province (Fig. 4.6). It was collected in fynbos, on deep sandy soils. A search of the surrounding area did not reveal any further populations, indicating that the species may be highly localised.
Additional specimen examined.
—3420 (Caledon): De Hoop Nature Reserve, near to De Mond (—AD), Magee et al. 120
(JRAU).
57 CHAPTER 5
TAXONOMIC REVISION OF THE GENUS DASISPERMUM
5.1 INTRODUCTION
The concept of the genus Dasispermum is here expanded to include all five species of Sonderina (two of which are newly described), as well as the only South
African species of Stoibrax, namely S. capense. As a result, this previously monotypic genus now consists of seven South African endemic species. This conclusion came about after a thorough phylogenetic analysis of these genera using molecular as well as morphological and anatomical data (Chapter 3, Magee et al.
2009b). The geographically disjunct Stoibrax was shown to be polyphyletic, with the type of the genus from North Africa placed within the tribe Apieae and the only South
African species embedded within the genus Sonderina. Such a treatment was proposed by Adamson (1939) who considered Sonderina hispida and Stoibrax capense to be closely related (as did Sonder 1862). Sonderina was further rendered paraphyletic by the inclusion of Dasispermum suffruticosum which was strongly allied to a Glade including Sonderina hispida and Sonderina sp. 1. Dasispermum s.s. is traditionally defined by the succulent or semi-succulent leaves and often winged fruit, both of which were argued by Magee et al. (2009b) to be adaptations to wind dispersal and the harsh littoral conditions in which this species occurs. Leaf succulence and rolled margins are both common halophytic adaptations (Mucina et al. 2006b). Based on the results of separate and combined molecular/morphological analyses (Chapter 3, Magee et al. 2009b) an expanded Dasispermum s.l. was strongly supported as part of the Capnophyllum group (together with the Cape
58 CHAPTER 5: TAXONOMIC REVISION OF THE GENUS DASISPERMUM endemic genera Capnophyllum and Scaraboides) within the Lefebvrea Glade of the tribe Tordylieae. The genus, in its new circumscription, can be distinguished from all other genera within the Lefebvrea Glade by a combination of characters, namely the sympodial growth habit (resulting in leaf-opposed umbels), papillose petals, isodiametric fruit with the median and/or lateral ribs as prominent as the marginal ribs, the narrow commissure extending to, at the most, the base of each rib and the presence of square or upright cells external to the vittae of the fruit.
5.2 VEGETATIVE AND REPRODUCTIVE MORPHOLOGY
5.2.1 Vegetative morphology— The species of Dasispermum, with the exception of D. grandicarpum, exhibit a sympodial growth habit as found in the other taxa of the Capnophyllum group. Dasispermum suffruticosum and D. perennans differ in being short-lived perennial suffrutices with a permanent, slightly woody stem
(Fig. 5.1A–E) – the others are all annuals. The young plants of both these species have an erect, only slightly branched habit but become well-branched and sprawling due to the resprouting of subsequent seasonal growth from the nodes of the main stem. Dasispermum hispidum and D. capense can often be distinguished from one another by their habits. In D. hispidum (as in D. humile and D. tenue) the branches are usually erect to ascending or somewhat sprawling (Fig. 5.1G), while in D. capense they are almost always decumbent or prostrate (Fig. 5.1F), although exceptions to this have been recorded. Due to the great deal of ambiguity regarding the species concepts of many of the species, the herbarium records and previous
59 CHAPTER 5: TAXONOMIC REVISION OF THE GENUS DASISPERMUM
FIGURE 5.1 The diversity in habit found amongst the species of Dasispermum. The habit varies from short-lived perennial suffrutices, as in D. suffruticosum (A, B) and D. perennans (C—E), to annuals with a prostrate (F) or erect (G) habit, as in D. capense (F) and D. hispidum (G), respectively.
60 CHAPTER 5: TAXONOMIC REVISION OF THE GENUS DASISPERMUM
treatments of the South African flora (Goldblatt and Manning 2000, Mucina et al.
2006a) have consistently misidentified small forms of Dasispermum hispidum,
particularly in the Northern Cape Province, as D. tenue. Although D. tenue is indeed a relatively small herb, all of the species of Dasispermum exhibit a tremendous amount of variation in size depending on environmental factors. The herbaceous stems are conspicuously purple blotched in D. grandicarpum, while in the other species they are immaculate (green, glaucous or reddish in colour). The littoral D. suffruticosum is easily distinguished from the other species by the usually firm and semi-succulent leaves with the ultimate leaflet segments curled inwards (Fig. 5.1B).
The ultimate leaflet segments of the other species are herbaceous or coriaceous
(Fig. 5.1C–G) and usually less than 1 mm broad, except in D. humile where they are diagnostically broader than or equal to 2 mm.
5.2.2 Reproductive morphology— The umbels of almost all the species, except D. grandicarpum, appear leaf-opposed due to the sympodial growth habit as found also in Capnophyllum and Scaraboides. In contrast, the main axis of D. grandicarpum appears to have monopodial growth which terminates in a primary umbel. Dasispermum tenue is most easily distinguished from the other species by the consistently sessile to subsessile umbels. While subsessile umbels may occur in the other species, particularly in the closely related D. humile, only in D. tenue are all of the umbels borne on a peduncle less than 4 mm long. The presence or absence of involucral and involucel bracts (a character emphasised in the last treatment of Wolff 1927), as well as their division and texture are useful diagnostic characters. Involucral bracts are almost always absent in D. humile and D. tenue, as well as the involucel bracts in the latter species. In D. capense, D. hispidum and D.
61 CHAPTER 5: TAXONOMIC REVISION OF THE GENUS DASISPERMUM
perennans at least one of the involucral bracts is pinnately divided and thus
resembles the leaves. Similarily, the involucral and involucel bracts of D.
suffruticosum share the somewhat succulent texture typical of the leaves of this
species. Unlike the species of Capnophyllum, the rays and raylets of Dasispermum
are generally scabrous and not glabrous.
The flowers are similar in all the species: white and diagnostically with a
papillose surface sculpturing as found in most of the species of the Capnophyllum
group (except Dasispermum suffruticosum and Scaraboides manningil). This
characters (character 12, Fig. 3.4C) when optimised onto the trees inferred from MP
analysis of the combined ITS/rps16 intron/morphology data (Chapter 3) was
reconstruction as either a synapomorphy for the Capnophyllum group with reversals
in Scaraboides manningii and Dasispermum suffruticosum, or as a convergent
character supporting both Capnophyllum as well as Dasispermum s.I., with a
reversal in Dasispermum suffruticosum. The species of Dasispermum generally
have a deeply emarginate petal apex which is a useful distinguishing floral character
when compared to the usually truncate apices found in the genus Capnophyllum.
Dasispermum tenue, however, differs from the other species in that the petals are
generally smaller with truncate to only slightly emarginate apices. As in
Capnophyllum, the styles of all the species are at first erect and short, but often
lengthen and become reflexed to varying lengths as the fruit mature. In most of the
species, the styles usually extend beyond the base of the stylopodium and in D.
capense may often extend well beyond the length of the fruit. Only in D. tenue do they remain short, becoming reflexed at most only to near the base of the
stylopodium.
62 CHAPTER 5: TAXONOMIC REVISION OF THE GENUS DASISPERMUM
The size and shape of the fruit (Fig. 5.2) are important diagnostic characters.
Dasispermum grandicarpum (Fig. 5.2F) and often D. suffruticosum (Fig. 5.2G) have relatively large fruit that are more than or equal to 4 mm in length, although those of
D. suffruticosum may be much smaller (Fig. 5.2G2). D. capense (Fig. 5.2D) has the smallest fruit, with the mature fruit not much larger than the ovary. In this species the fruit bulge dorsally outwards so that in lateral view they are broader than long with a prominently cordate base. Although the fruit of D. hispidum (Fig. 5.2C) may occasionally almost approach those of D. capense in shape, becoming broadly ovate, they are never broader than long. Furthermore, D. capense is easily distinguished from the other species, including D. hispidum, by the strongly concave appearance of the marginal ribs in lateral view. The two ribs are widely separated from one another, while in all the other species they are close together and parallel, or at most only very slightly concave, and separated from one another. The ribs are usually cartilaginous and only slightly prominent, except in D. suffruticosum (Figs.
5.2G and 5.3G) where they are prominently corky (at least before desiccation of the fruit) and either very prominent or more usually extending into broad wings. In D. grandicarpum (Fig. 5.2F), D. humile (Fig. 5.2B) and D. tenue (Fig. 5.2A) the ribs differ from the other species in that they have a prominent fibrous ridge on the dorsal surface. The fruit are glabrous to only very slightly scabrous in D. grandicarpum, D. humile, D. suffruticosum and D. tenue. In D. capense (Fig. 5.2D) they may be scabrous to pustulate, while in D. hispidum (Fig. 5.2C) the fruit may be slightly scabrous to densely pilose. Traditionally Sonderina caruifolium was largely distinguished from S. hipida by the glabrous as opposed to hispid or pilose fruit.
63 CHAPTER 5: TAXONOMIC REVISION OF THE GENUS DASISPERMUM
Al A3 B4
Cl C2 C3 C4 C5 C6 C7
D1 D2 D3 D4 D5
G1 G2 IMMEEE141.1.1
FIGURE 5.2 Variation in fruit morphology (A—D) of Dasispermum species: (A) D. tenue, (B) D. humile, (C) D. hispidum, (D) D. capense, (E) D. perennans, (F) D. grandicarpum, (G) D. suffruticosum. Vouchers: A1. Van Wyk et al. 3433 (JRAU); A2. Bolus 11291 (BOL); A3. Schlechter 1703 (BOL); Bl. Flanagan 1183 (BOL); B2. Van Wyk & Van Wyk 1895 (JRAU); B3. Hilliard & Burtt 148b (PRE); B4. Rudatis 1811 (PRE); C1. Magee et al. 107 (JRAU); C2. Magee et al. 109 (JRAU); C3. Hugo 1988 (PRE); C4. Magee et al. 108 (JRAU); C5. Van Wyk et al. 3518 (JRAU); C6. Rodin 1252 (BOL); C7. Stirton 11190 (PRE); D1. Thompson 2829 (NBG); D2. Stephens & Gloven 8723 (BOL); D3. Schlechter 11404 (BOL); D4. Compton 22244 (NBG); D5. Salter 7732 (BOL); E1. Magee & Boatwright 105 (JRAU); F1. Lutzeyer s.n. (JRAU); Gl. Magee et al. 117 (JRAU); G2. Boucher 3985 (PRE). Scale: 1 mm
64 CHAPTER 5: TAXONOMIC REVISION OF THE GENUS DASISPERMUM
However, after extensive field studies of these species throughout their range, it was
observed that this character varied significantly even within populations. As a result
Sonderina caruifolium is here treated as a synonym of Dasispermum hispidum.
In transverse section the fruit of Dasispermum (Fig. 5.3A—H) are isodiametric
and either homomericarpic in D. capense (Fig. 5.3F), D. grandicarpum (Fig. 5.3G),
D. hispidum (Fig. 5.3E), D. humile (Fig. 5.3D), D. perennans (Fig. 5.3C) and D.
tenue (Fig. 5.3H), or often heteromericarpic in D. suffruticosum (Fig. 5.3A, B). As
mentioned in Chapter 3, one of the diagnostic characters for the genus is that the
median and/or lateral ribs are more or less as well developed as the marginal ribs.
In all the species there are two commissural and four vallecular vittae. The cells
external to both the commissural and vallecular vittae are prominent and either
square (Fig. 5.4C—G) or enlarged and upright (Fig. 5.4A—B), forming a distinct layer.
This layer of square or upright cells around the vittae is a distinct generic apomorphy for an expanded Dasispermum s.l. (Chapter 3, Fig. 3.4H) The cells around the vittae of D. capense (Fig. 5.4F), D. grandicarpum (Fig. 5.4G), D. hispidum (Fig. 5.4E), D. perennans (Fig. 5.4C) and D. suffruticosum (Fig. 5.4D) are square, while those in D. humile (Fig. 5.4A) and D. tenue (Fig. 5.4B) are upright. Those of D. tenue are the most prominent as they are enlarged and usually composed of two to three layers of upright cells. Unfortunately, this character is not always visible in re-hydrated fruit.
The commissure is narrow and does not extend beyond the base of the marginal ribs. All other genera within the Lefebvrea Glade have a broad commissure which extends well beyond the base of the marginal ribs and often to their tips.
65 CHAPTER 5: TAXONOMIC REVISION OF THE GENUS DASISPERMUM
affillit(11@i:inMiM30
FIGURE 5.3 Transverse sections through the fruit of Dasispermum species. (A, B) D. suffruticosum, (C) D. perennans, (D) D. humile, (E) D. hispidum, (F) D. capense, (G) D. grandicarpum, (H) D. tenue. Vouchers: A. Taylor 9896 (PRE); B. Tilney 204 (JRAU); C. Magee & Boatwright 105 (JRAU); D. Van Wyk & Van Wyk 1895 (JRAU); E. Van Wyk et al. 3539 (JRAU); F. Boatwright et al. 212 (JRAU); G. Lutzeyer s.n. (JRAU); H. Van Wyk et al. 3433 (JRAU). Scale: 0.7 mm.
66 CHAPTER 5: TAXONOMIC REVISION OF THE GENUS DASISPERMUM
FIGURE 5.4 Transverse sections through portions of the fruit of Dasispermum showing the upright (A, B) or square (C—G) cells external to the vittae. Vouchers: A. D. humile (Van Wyk and Van Wyk 1895, JRAU); B. D. tenue (Van Wyk et al. 3433, JRAU); C. D. perrenans (Magee & Boatwright 105, JRAU); D. D. suffruticosum (Taylor 9896, PRE); E. D. hispidum (Van Wyk et al. 3539, JRAU); F. D. capense (Salter 7733, BOL); G. D. grandicarpum (Lutzeyer s.n., JRAU). Scale: 0.07 mm.
67 CHAPTER 5: TAXONOMIC REVISION OF THE GENUS DASISPERMUM
5.3 TAXONOMY OF THE GENUS DASISPERMUM
DASISPERMUM Raf., Good Book 56 (1840), reimp. Scad. Gen. Omb. Pl. 56 (Amer.
Midl. Nat. Repr. no. 3. 1913); Merrill, Index Raf. 179 (1949); B.L. Burtt in
Notes RBG Edinb. 45: 92 (1988); emend. Magee and B.-E.van Wyk, emend.
nov. — TYPE: D. maritimum Raf., nom illeg. (= D. suffruticosum (Berg.)
B.L.Burtt)
Heteroptilis [E.Mey. ex.] Meisn. in J.D. Hook., Lond. J. Bot. 2: 534 (1843); Leute in
Ann. Nathist. Mus. Wien 73: 89 (1969). — TYPE: H. arenaria Meisn.
Sonderina H.Wolff in Pflanzenr. Heft 90: 92. 1927, synon. nov. — TYPE: Sonderina
hispida (Thunb.) H.Wolff (lecto., designated by Burtt 1991).
Carum sect. Brachyapium Baill., Hist. Pl. 7: 118 (1879). Brachyapium (Baill.) Maire
in Bull. Soc. Hist. Nat. Afr. Nord 23: 186. 1932, synon. nov. — TYPE:
Ptychotis didyma Sond.
Erect to sprawling or prostrate sympodial or rarely monopodial herbs or suffrutices, 0.03-0.8 m tall, annuals or short-lived perennials. Stems herbaceous or permanent and woody, purple-blotched or usually immaculate, simple to well- branched; branches erect to decumbent or prostrate, sometimes resprouting at the nodes. Leaves cauline, 15-120 mm long x 6-50 mm, becoming smaller towards the upper part of the stem, glabrous or scabrous, pinnate to 2-pinnate, herbaceous to coriaceous or firm and semi-succulent. Petioles 5-60 mm long, sheathing at their base, glabrous or scabrous. Ultimate leaflet segments linear-oblong to oblong, laminate to subterete or turgid, sometimes strongly involute (curled inwards), concolorous or discolourous, green or glaucous, apex acute to obtuse, margins
68 CHAPTER 5: TAXONOMIC REVISION OF THE GENUS DASISPERMUM entire or toothed, venation visible or obscure. Umbels compound; peduncle sessile or subsessile to long, 0 to 100 mm long; involucral bracts 0 to 6, free, lanceolate to ovate or foliose, coriaceous or slightly succulent, apex acute to acuminate, margins often membranous, glabrous or scabrous; rays 3 to 26, 5-50 mm long at anthesis, glabrous or scabrous; involucel bracts 0 to 6, free, linear to lanceolate, coriaceous or slightly succulent, apex acute to acuminate, margins often membranous, glabrous or scabrous; raylets 4 to 14, 2-10 mm long at anthesis, glabrous or scabrous. Flowers hermaphroditic, pentamerous; sepals indistinct, triangular; petals white, 0.4-1.2 mm long and broad, broadly oblong to ovate or obovate, tips inflexed, acuminate, septum present on inner face, apex truncate to deeply emarginate, smooth or papillose; stamens with anthers inflexed; ovary inferior, glabrous or pustulate, scabrous to densly pilose; stylopodium shortly conical; styles at first erect, short, often lengthening as fruit matures, 0.2-1.4 mm long, becoming reflexed up to or beyond the base of the stylopodium. Fruit a schizocarp, isodiametric, 0.6-6.0 mm x
1.0-5.0 mm, elliptic or ovate to very broadly ovate or rotund, sometimes transversely oblong (broader than long) in lateral view, base obtuse to cordate, apex obtuse; mericarps homomorphic or rarely heteromorphic, glabrous or pustulate, scabrous to densly pilose; if homomericarpic then with the median and lateral ribs as well developed as the marginal ribs, if heteromericarpic then with either median or lateral ribs as well developed as the marginal ribs, slightly to very prominent or winged, cartilaginous or corky, dorsal surface sometimes fibrously ridged, glabrous to densly pilose; marginal ribs parallel or strongly concave in lateral view; commissural vittae 2; vallecular vittae 4; commissure narrow, not extending beyond the base of the marginal ribs; carpophore bipartite.
69 CHAPTER 5: TAXONOMIC REVISION OF THE GENUS DASISPERMUM
Diagnostic characters— The genus can be distinguished from all other
genera within the Lefebvrea Glade by a combination of characters, namely the
sympodial growth habit (resulting in leaf-opposed umbels), papillose petals,
isodiametric fruit with the median and/or lateral ribs as prominent as the marginal
ribs, the narrow commissure extending to, at the most, the base of each rib and the
presence of a row of square or upright cells external to the vittae of the fruit. When in flower, the annual species may be confused with species of Capnophyllum, but they can be distinguished by the combination of the usually erect to ascending branches, the glabrous coriaceous leaves, the scabrous rays and raylets and the deeply emarginate petal apices.
Distribution and habitat— The seven species are endemic to South Africa
(Figs. 5.5 to 5.9) and occur widely in the Eastern, Western and Northern Cape
Provinces with only D. humile and D. suffruticosum extending into the KwaZulu-
Natal. The species have been collected on littoral dunes or in fynbos, renosterveld, succulent karoo and grasslands.
Phenology— The annual species begin to flower in spring with mature fruit forming from late spring to early summer, although Dasispermum tenue however, appears to flower much earlier with mature fruit recorded in early spring
(September). The perennial species, D. suffruticosum and D. perennans, flower and fruit throughout much of the year depending on favourable conditions. Although D. hispidum often co-occurs with species of Capnophyllum they appear to form a succession, with the plants of Capnophyllum already starting to wither when those of
D. hispidum begin to flower and fruit.
70 CHAPTER 5: TAXONOMIC REVISION OF THE GENUS DASISPERMUM
KEY TO THE SPECIES OF DASISPERMUM la. Short-lived perennial suffrutices, resprouting from a permanent somewhat
woody base:
2a. Fruit with very prominent, thick corky ribs or wings, often heteromericarpic;
leaves succulent to semi-succulent, firm in texture; ultimate leaflet
segments turgid or rarely laminate, often curled inwards; a dune endemic,
widely distributed along the coast from Vredendal to around Port Shepstone
6. D suffruticosum
2a. Fruit with only slightly prominent ribs, never fleshy or winged, invariably
homomericarpic; leaves coriaceous; ultimate leaflet segments subterete,
remaining erect; known only from around the Karbonkelberg on the Cape
Peninsula 5. D. perennans
1 b. Annual herbs:
Fruit more than 3.9 mm long; stem with purple blotches; plants erect, 0.5-
0.8 m tall; known only from Stanford in the Western Cape
2. D grandicarpum
Fruit less than 2.5 mm long; stem immaculate; plants erect or sprawling to
procumbent, usually less than 0.4 m tall:
Ultimate leaflet segments 2-3 mm broad; restricted to the Eastern
Cape and KwaZulu-Natal Provinces 4. D. humile
Ultimate leaflet segments less than or equal to 1 mm broad; occurring
in the Northern, Western and Eastern Cape Povinces:
5a. All umbels sessile to subsessile, peduncle less than 4 mm long;
involucral and involucel bracts absent (very rarely 4 involucel
bracts); styles not exceeding the base of the stylopodium, short
71
CHAPTER 5: TAXONOMIC REVISION OF THE GENUS DASISPERMUM
(0.2-0.3 mm long); restricted to Renosterveld vegetation between
Bonnievale and Riversdale 7. D. tenue
5b. Umbels usually on a peduncle more than 10 mm long, occasionally
some sessile to subsessile; involucral and involucel bracts present,
often foliose; styles usually far exceeding the base of the
stylopodium, rarely reaching only the base, relatively long (0.3-1.3
mm long); not occurring in Renosterveld vegetation:
Ovaries and fruit with marginal ribs markedly concave and
widely separated (in lateral view); fruit broader than long;
scabrous or pustulate; plants usually sprawling and prostrate
1. D. capense
Ovaries and fruit with marginal ribs parallel or very slightly
concave; fruit not broader than long; scabrous to densely
hispid; plants usually erect 3. D. hispidum
1. DASISPERMUM CAPENSE (Lam.) Magee and B.E.van Wyk, comb. nov. Caucalis
capensis Lam., Encycl. 1: 658 (1785). Stoibrax capense (Lam.) B.L.Burtt in
Edinb. J. Bot. 48(2): 251 (1991). — TYPE: SOUTH AFRICA. Cape, Sonnerat s.n.
(P-LAM!, holo.).
Ptychotis didyma Sond. in Harv. and Sond., Fl. Cap. 2: 258 (1862). Trachyspermum
didymium (Sond.) Drude in Engl. and Prantl, Pflanzenfam. 3(8): 189 (1898).
Tragiopsis didyma (Sond.) H.Wolff in Pflanzenr. Heft 90: 96 (1927). Sonderina
didyma (Sond.) Adamson in Adamson and Salter, J. S. Afr. Bot. 5: 55 (1939).
Stoibrax didyma (Sond.) B.L.Burtt in Tan, K., (ed.) Davis & Hedge Festschrift
145 (1989). — TYPE: SOUTH AFRICA. Cape, woods near Tulbaghskloof, Zeyher
- 72 CHAPTER 5: TAXONOMIC REVISION OF THE GENUS DASISPERMUM
730 (S!, lecto., here designated; K!, two sheets, isolecto.). [Note: The
specimen in S is chosen here as it is from Sonder's herbarium and the type
locality is clearly recorded on the label.]
Sprawling or very rarely somewhat erect sympodial herbs, 0.03-0.25 m tall, annuals. Stems herbaceous, immaculate, usually very well-branched at the base; branches prostrate to decumbent or rarely somewhat ascending to erect, not resprouting from the nodes. Leaves 15-60 mm long x 6-20 mm, glabrous or scabrous, 2-pinnate, herbaceous to somewhat coriaceous. Petioles 7-30 mm long, glabrous or scabrous. Ultimate leaflet segments linear-oblong, 1-10 mm long, less than 1 mm broad, laminate to subterete, concolorous, green or glaucous, margins entire, venation visible. Umbels with peduncle short, very rarely sessile to subsessile, (0 to 4) 6 to 30 mm long; involucral bracts 1-6, lanceolate to ovate or often foliose, coriaceous; apex acuminate; margins membranous, glabrous or scabrous; rays 4 to 12, 5-15 mm long, glabrous or scabrous; involucel bracts 2 to 6, linear to lanceolate, coriaceous, apex acuminate, margins often membranous, glabrous or scabrous; raylets 4 to 12, 2-5 mm long, glabrous or scabrous. Flowers with petals 0.7-1 mm long and broad, apex deeply emarginate, papillose; ovary scabrous to pustulate; styles lengthening as fruit matures, 0.7-1.3 mm long, becoming reflexed far beyond the base of the stylopodium and often longer than the fruit. Fruit 0.6-1.8 mm x 1.0-1.8 mm, transversely oblong (broader than long), base cordate; mericarps homomorphic, scabrous to pustulate; median and lateral ribs as well developed as the marginal ribs, slightly prominent, cartilaginous, dorsal surface not fibrously ridged, scabrous to pustulate; marginal ribs strongly concave in lateral view.
73 CHAPTER 5: TAXONOMIC REVISION OF THE GENUS DASISPERMUM
Diagnostic characters— Dasispermum capense is easily distinguished by
the transversely oblong fruit (broader than long), with the marginal ribs strongly
concave and separated from one another in lateral view. This species is most often
confused with D. hispidum from which it also differs in the usually procumbent
branches and only scabrous to pustulate, but never pilose, fruit.
Distribution and habitat— The species occurs in sandy soils (at low and
medium altitudes) from Port Nolloth in the Northern Cape Province southwards to the Western Cape Province (Cape Peninsula and eastwards to Witsands) (Fig. 5.5).
HEIGHT ABOVE SEA LEVEL
V;(/ /j Over 1500 m 900 - 1500 m t= 300 - 900 m Under 300 m
20 0 20 40 60 80 100 km 1111111111111
FIGURE 5.5 Geographical distribution of Dasispermum capense.
74 CHAPTER 5: TAXONOMIC REVISION OF THE GENUS DASISPERMUM
Additional specimens examined.
—2916 (Port Nolloth): near Port Nolloth (BB), Bolus 9510 (BOL, K).
2917 (Springbok): Spektakel (—DA), Bolus 9645 (BOL, K); Concordia (—DB), Schlechter
11404 (BOL, K).
2918 (Gamoep): Koisabes Farm (—AB), Le Roux 2529 (BOL, NBG); Goegap Nature
Reserve (—CA), Broodryk 151 (PRE).
3118 (Vanrhynsdorp): Wiedou Farm, 15 km S of Vanrhynsdorp (—DB), Thompson 2829
(NBG, PRE).
3119 (Calvinia): Nieuwoudtville (—AC), Galpin 1158 (PRE); Tuinlaagte, Oorlogskloof (-
AC), Magee et al. 128 (JRAU); Lokenburg (—CA), Acocks 17221 (PRE); along road
between farms Soutpan and Vryheid (—CD), Snijman 761 (K); between Driefontein
and Heerenlogement (—DC), Pearson 6730 (BOL, K).
3218 (Clanwilliam): near Elandsbay (—AD), Lavranos 11651 (MO, PRE); Graafwater (-
BA), Thode A2002 (PRE); near to Clanwilliam (—BB), Schlechter 5072 (BM, BOL, K,
PRE); Kriedouwkrans (—BD), Pearson 5324 (BOL, K); Papkuilsfontein Farm (—CB),
Stirton & Zantovska 11413 (MO, PRE); Het Kruis (—DA), Stephens & Glover 8723
(BOL, K).
—3219 (Wuppertal): near Citrusdal (—CA), Taylor 1335 (BOL); Knolfontein, Swartruggens (-
DC), Jardine & Jardine 274 (NBG).
—3318 (Cape Town): above Camps Bay (—CD), Salter 7732 (BOL, K, PRE); W slopes of
Lions Head (—CD), Adamson 2128 (BM), Wolley Dod 3133 (BOL); Vygekraal (—DC),
Mac Owan 1751 (PRE).
—3319 (Worcester): Platfontein (—BB), Compton 22244 (NBG).
3420 (Bredasdorp): De Hoop, just before main gate (—AD), Magee et al. 131 (JRAU); De
Hoop (—AD), Van der Merwe 1160 (PRE); De Hoop, Melkkamer (—AD), Van Wyk
1957 (NBG, PRE); Witsand, near mouth of the Breede river (—BD), Ornduff 7685
(MO, PRE); Witsand (—BD), Walgate 835 (PRE).
75 CHAPTER 5: TAXONOMIC REVISION OF THE GENUS DASISPERMUM
2. DASISPERMUM GRANDICARPUM. Magee and B.-E.van Wyk, sp. nov., Ceteris
speciebus Dasispermum cellulis quadratis prominentibus circum vittas similis,
sed annuls habitu majori erecto, caulibus purpureis maculatis fructuque mu/to
majori a ceteris speciebus differt. D. humili et D. tenui costa fructus porcata
fibrosa similis sed segmentis folioli ultimis teretiusculis subtiliter divisis
praesentiaque bracteis et involucralibus et involucellis differt. — TYPE: SOUTH
AFRICA. Caledon district (3419): Grootbos Nature Reserve, (–CB), 1
December 2006, H. Lutzeyer s.n. (NBG!, holo.; JRAU!, iso.).
Erect monopodial herbs, 0.5-0.8 m tall, annuals. Stems herbaceous, purple-
blotched, not branched at the base, remaining simple or slightly dichotomously
branched higher up; branches ascending to erect, not resprouting from the nodes.
Leaves 40-120 mm long x 25-50 mm, 2-pinnate, coriaceous. Petioles 15-40 mm long, glabrous, purple blotched. Ultimate leaflet segments linear-oblong, 3-10 mm long, less than 0.5 mm broad, laminate to subterete, concolorous, green, margins entire, venation visible or obscure. Umbels with peduncle short to long, 15 to 60 mm long; involucral bracts 4 to 6, lanceolate, coriaceous, apex acuminate, margins membranous, glabrous; rays (6) 10 to 18, 15-35 mm long, scabrous; involucel bracts 4 to 6, linear to lanceolate, coriaceous, apex acuminate, margins membranous, glabrous; raylets 6 to 14, 4-8 mm long, scabrous. Flowers with petals
0.8-1.0 mm x 0.6-1.0 mm, apex deeply emarginate, papillose; ovary scabrous; styles usually lengthening as fruit matures, 0.7-1.4 mm long, becoming reflexed far beyond the base of the stylopodium. Fruit 4.0-6.0 mm x 3 mm, elliptic in lateral view, base obtuse; mericarps homomorphic; median and lateral ribs as well developed as
76 CHAPTER 5: TAXONOMIC REVISION OF THE GENUS DASISPERMUM
the marginal ribs, prominent, cartilaginous, dorsal surface fibrously ridged, glabrous;
marginal ribs parallel to very slightly concave in lateral view.
Diagnostic characters— Dasispermum grandicarpum is easily distinguished from all other species in the larger more erect habit, the maculate (purple blotched)
stems, and the much larger fruit of more than 3.9 mm long (less than 2.5 mm in the other species). It shares the fibrously ridged fruit ribs with D. humile and D. tenue
but also differs in the finely divided subterete ultimate leaflet segments, the presence of both involucral and involucel bracts and the row of square cells external to the vittae.
Distribution and habitat— The species is known only from the private nature reserve Grootbos, near Stanford in the Western Cape Province (Fig. 5.6), where it was collected in fynbos vegetation that had burnt the previous year. No plants could be located in subsequent searches by Mr H. Lutzeyer, suggesting that this species may be a short-lived fireweed.
Additional specimens examined.
–3419 (Caledon): Grootbos Nature Reserve, (–CB), 20 October 2006, H. Lutzeyer s.n.
(NBG).
77 CHAPTER 5: TAXONOMIC REVISION OF THE GENUS DASISPERMUM
HEIGHT ABOVE SEA LEVEL
FM Over 1500 m 900 - 1500 m F. 300 900 m 1Under 300 m
20 0 20 40 80 80 100 km /11111 •1111I
FIGURE 5.6 Geographical distribution of Dasispermum grandicarpum (star), D. perennans (circle), D. tenue (triangle).
3. DASISPERMUM HISPIDUM (Thunb.) Magee and B.E.van Wyk, comb. nov. Sium
hispidumThunb., Prodr. 51 (1794). Ptychotis hispida (Thunb.) Sond. in Harv.
and Sond., Fl. Cap. 2: 537 (1862). Carum hispidum (Thunb.) Koso-Poljansky
in Bull. Soc. Natur. Mosc. 30: 286 (1917). Sonderina hispida (Thunb.) H.Wolff
in Pflanzenr. Heft 90: 94 (1927). — TYPE: SOUTH AFRICA. Cape, Thunberg s.n.
sub THUNB-UPS 7046 (UPS!, lecto. here designated). [Note: This is the only
specimen in Thunberg's herbarium and closely matches the original
protologue.]
Sium aperumThunb., Prodr. 51 (1794). Seseli asperum (Thunb.) Sond. in Harv. and
Sond., Fl. Cap. 2: 550 (1862). — TYPE: SOUTH AFRICA. Cape, Thunberg s.n.
sub THUNB-UPS 7036 (UPS!, lecto., here designated). [Note: This is the only
specimen in Thunberg's herbarium and is therefore designated here.]
78 CHAPTER 5: TAXONOMIC REVISION OF THE GENUS DASISPERMUM
Anthriscus capensis Spreng. in Roem. and Schult., Syst. Veg. 6: 526 (1820), nom.
Illeg.
Sonderina caruifolia (Sond.) H.Wolff in Pflanzenr. Heft 90:94 (1927), Ptychotis
caruifolia Sond. in Harv. and Sond., FL Cap. 2: 537 (1862). Trachyspermum
caruifolium (Sond.) Drude in Engl. and Prantl, Pfanzenfam. 3(8): 189 (1898),
synon. nov. — TYPE: SOUTH AFRICA. Cape Town district (3318):
Riebeekkasteel (–BD), Zeyher 729 (S!, lecto., here designated; BM!, LE!, K!,
two sheets, NBG!). [Note: The specimen in S is chosen here as it is from
author's collection and has the type locality clearly recorded on the sheet.]
Erect or occasionally somewhat sprawling sympodial herbs, 0.04-0.6 m tall, annuals. Stems herbaceous, immaculate, remaining simple to slightly or rarely well- branched at the base; branches erect or ascending rarely somewhat decumbent, not resprouting from the nodes. Leaves 15-90 mm long x 10-35 mm, glabrous or scabrous, 2-pinnate, herbaceous to coriaceous. Petioles 7-40 mm long, glabrous or scabrous. Ultimate leaflet segments linear-oblong, 1-10 mm long, less than 1 mm broad, laminate to subterete, concolorous, green, margins entire, venation visible or obscure. Umbels with peduncle short to long, (4) 10 to 80 mm long; involucral bracts
1-6, lanceolate to ovate or often foliose, coriaceous, apex acuminate, margins usually membranous, glabrous or scabrous; rays 3 to 25, 10-50 mm long, glabrous or scabrous; involucel bracts 2 to 6, linear to lanceolate, coriaceous, apex acuminate, margins often membranous, glabrous or scabrous; raylets 4 to 12, 2-10 mm long, glabrous or scabrous. Flowers with petals 0.7-1 mm long and broad, apex deeply emarginate, papillose; ovary almost glabrous or scabrous to densely pilose; styles lengthening as fruit matures, 0.3-1.0 mm long, becoming reflexed usually
79 CHAPTER 5: TAXONOMIC REVISION OF THE GENUS DASISPERMUM
beyond the base of the stylopodium, rarely just up to the base of the stylopodium.
Fruit 1.0-2.0 mm x 1.0-1.8 mm, elliptic to broadly ovate (not broader than long) in
lateral view, base obtuse to slightly cordate; mericarps homomorphic, almost
glabrous or scabrous to densely pilose; median and lateral ribs as well developed as
the marginal ribs, slightly prominent, cartilaginous, dorsal surface not fibrously
ridged, slightly scabrous to densely pilose; marginal ribs parallel to very slightly
concave in lateral view.
Diagnostic characters— Dasispermum hispidum is most likely to be
confused with D. capense or D. perennans with which it shares the prominent
involucel bracts (one of which is often foliose) and the usually scabrous fruit with ribs that are not fibrously ridged on the dorsal surface. It differs from D. perennans in the annual habit and from D. capense in the usually erect growth form, fruit that are narrower or at most equal to their length (in lateral view) and with the marginal ribs of the ovary and young fruit remaining parallel or only slightly concave. Very small plants of D. hispidum, as often found in the Northern Cape Province, have consistently been misidentified as D. tenue which is a Renosterveld species easily distinguished from D. hispidum by the consistently sessile to subsessile umbels and the absence of both involucral and involucel bracts.
Distribution and habitat— This widespread species is largely restricted to coastal sands, from around Lekkersing in the Northern Cape Province to St.
George's Strand in the Eastern Cape Province (Fig. 5.7).
80 CHAPTER 5: TAXONOMIC REVISION OF THE GENUS DASISPERMUM
HEIGHT ABOVE SEA LEVEL k; j Over 1500 m 900 - 1500 m 71 300 - 900 m [—I Under 300 m
20 0 20 40 60 80 100 km ,.,,,,,, ■■■ •
FIGURE 5.7 Geographical distribution of Dasispermum hispidum.
Additional specimens examined.
2816 (Oranjemund): 37 km N of Lekkersing (—DB), Germishuizen 5596 (PRE);
Richtersveld, Annisvlakte (—DB), Jiirgens 23129 (PRE); Taaibos Vlei, Little
Namaqualand (—DB), Marloth 12421 (PRE); Witbank, Little Namaqualand (—DC),
Pillans 5234 (BOL).
3017 (Hondeklipbaai): between Hondeklipbaai and Swart Lintjies River (—AD), Pillans
18056 (BOL); Farm Hardekoppie, 16 km N of Kotzesrus and 2 km W of Hardevlei (-
DC), Perold 1699 (PRE).
3117 (Lepelfontein): Brand-se-baai (—BD), Van Rooyen 2210 (PRE).
3118 (Vanrhyansdorp): Donkin's Bay (—CD), Acocks 24166 (PRE); Vanrhynsdorp (—DA),
Compton 17173 (NBG); 7 miles N of Vanrhynsdorp (—DA), Acocks 19502 (PRE, K);
81 CHAPTER 5: TAXONOMIC REVISION OF THE GENUS DASISPERMUM
7 miles N.N.E. of Vanrhynsdorp (—DA), Acocks 14746 (K); Wiedou Valley on road to
Gifberg (—DB), Hugo 702 (PRE); Wiedou, on the foot of the Gifberg (—DB), Zietsman
& Zietsman 1162 (PRE); top of Gifberg (—DC), Bayliss 6148 (MO); Vleikraal, east of
Klawer (—DC), Walters 51 (PRE, MO).
3217 (Vredenburg): Cape Columbine (—DD), Lavranos 11710 (MO); Witklip Farm 1 mile S
of Vredenburg (—DD), Thompson 2658 (PRE, 2 sheets).
—3218 (Clanwilliam): Clanwilliam (—BB), Galpin 10770 (PRE), Leipoldt 15 (BOL); Oliphants
River Valley near Platte Kloof (—BB), Pillans 8741 (BOL, K, PRE); 4km N of Algeria
turnoff on N7 (—BD), Goldblatt & Manning 10404A (PRE); between Sauer and
Velddrif (—CD), Goldblatt & Manning 11146 (MO); Het Kruis (—DA), Werdermann &
Oberdieck 441 (K, PRE).
3318 (Cape Town): Oosterwal, Langebaan (—AA), Axelson 562 (NBG); Darling Flora
Reserve (—AD), Rycroft 2009 (NBG); Mud River, N of river near bridge (—AD),
O'Callaghan 1322 (PRE); Mud River Kloof (—AD), Acocks 23381 (K, PRE), Taylor
5489 (NBG, K, PRE); Bokbaai near Darling (—CB), Hilliard & Burtt 13020 (MO, K,
PRE, S); Bok Point Gate (—CB), Compton 9418 (NBG); 9 km from Melkbosstrand to
Velddrif (—CB), Stirton 11190 (PRE); Burgers Post Farm, near Pella (—DA), Boucher
& Shepherd 4869 (K, PRE).
3319 (Worcester): Saron (—AA), Schlechter 10638 (K, LE, MO, PRE), Schlechter 10652
(PRE); De Hoek Estates (—AA), Levyns 993 (BOL).
3322 (Oudtshoorn): Wilderness (—DC), Mogg 11786 (PRE).
3324 (Steytlerville): E of Kabeljousrivier (—DD), Fourcade 4017 (BOL, K).
3325 (Port Elizabeth): Perseverance (—DC), Rodin 1252 (BOL, K, PRE, S); St George's
Strand (—DC), Long 839 (K, PRE).
3418 (Simonstown): Chapmans Bay (—AB), Wolley Dod 3606 (BOL); Fish Hoek (—AB),
Young 268 (BOL); Prince's Vlei (—AB), Mar/oth 9414 (NBG, PRE); Faure Flats (—BB),
Adamson 10367 (NBG); Red Hill (—AB), Salter s.n.(BOL); Witsands Bay (—AB),
Werdermann & Oberdieck 811 (BOL, K).
82 CHAPTER 5: TAXONOMIC REVISION OF THE GENUS DASISPERMUM
3419 (Caledon):1 km from Gansbaai on road to Stanford (–CB), Germishuizen 4140
(PRE).
3420 (Bredasdorp): De Hoop, De Mond ( –AD), Magee & Boatwright 12 (JRAU), Magee et
al. 120 (JRAU), Thompson 3265 (NBG, PRE).
3421 (Riversdale): Still Bay (–AD), Muir 4554 (BOL, K, PRE); Potberg Nature Reserve,
De Hoop (–BC), Burgers 1569 (PRE).
3422 (Mossel Bay): NE of Mossel Bay (–AB), Acocks 15394 (PRE); Belvidere, Knysna (-
BB), Duthie 1181 (BOL, NBG); Ruigte Vlei, at Swart River (–BB), Fourcade 1552
(BOL, 2 sheets); camping area close to Swartvlei, Sedgefield (–BB), Hugo 1988
(NBG).
3423 (Knysna): Plettenberg Bay (–AB), Schlechter 5937 (BM, K, S).
4. DASISPERMUM HUMILE (Meisn.) Magee and B.E.van Wyk, comb. nov. Petroselinum
humile Meisn. in J.D. Hook., Lond. J. Bot. 2: 531 (1843). Apium humile
(Meisn.) Benth. and Hook. f., Gen. Pl. 1: 888 (1867). Carum meisneri Hiroe,
Umbell. World 871 (1979). Sonderina humilis (Meisn.) H.Wolff in Pflanzenr.
Heft 90: 93 (1927). — TYPE: SOUTH AFRICA. Stanger district (2931): Natal,
near Port Natal (–CC), Krauss 418 (MO!, sheet with collection label in bottom
left corner, lecto., here designated; BM!, K!, two sheets, MO!, NBG!, S!,
isolecto.). [Note: The specimen in MO most closely matches the original
protologue and is therefore selected here as lectotype.]
Somewhat erect or rarely sprawling sympodial herbs, 0.15-0.5 m tall, annuals. Stems herbaceous, immaculate, slightly to well-branched at the base; branches ascending to erect, not resprouting from the nodes. Leaves 40-120 mm long x 30-50 mm, 2-pinnate, herbaceous. Petioles 15-60 mm long, slightly
83 CHAPTER 5: TAXONOMIC REVISION OF THE GENUS DASISPERMUM scabrous, green. Ultimate leaflet segments narrowly oblong to oblong, 7-12 mm long, 2-3 mm broad, lamiante, concolourous or discolorous, adaxial green, abaxial green or glaucous, margins entire, venation visible. Umbels with peduncle short to long, sometimes sessile to subsessile, 0 to 65 mm long; involucral bracts 0 (1), lanceolate, coriaceous, apex acuminate, margins membranous, glabrous; rays 3 to
8, 5-20 mm long, scabrous; involucel bracts (1) 2 to 4, linear to lanceolate, coriaceous, apex acute, margins sometimes membranous, glabrous; raylets 4 to 12,
2-5 mm long, scabrous. Flowers with petals 0.5-0.7 mm x 0.4-0.6 mm, apex deeply emarginate, papillose; ovary scabrous; styles usually lengthening as fruit matures,
0.4-0.8 mm long, becoming reflexed up to or more commonly beyond the base of the stylopodium. Fruit 1.6-2.0 mm long and broad, broadly elliptic to rotund in lateral view, base obtuse; mericarps homomorphic; median and lateral ribs as well developed as the marginal ribs, prominent, cartilaginous, dorsal surface fibrously ridged, glabrous; marginal ribs parallel to very slightly concave in lateral view.
Diagnostic characters— D. humile shares the non-succulent leaves and only slightly prominent fruit ribs with D. capense, D. grandicarpum, D. hispidum, D. perennans and D. tenue, but is easily distinguished from these species by the broader ultimate leaflet segments which are 2 to 3 mm broad (less than 1 mm in the other species). Of the before mentioned species, only the widespread D. hipidum may extend into the geographical range of D. humile from which it can be further differentiated by the glabrous fruit, the fibrously ridged dorsal surface of the fruit ribs and the usual absence of involucral bracts.
84
CHAPTER 5: TAXONOMIC REVISION OF THE GENUS DASISPERMUM
Distribution and habitat— The species occurs from the Zuurberg in the
Eastern Cape Province to Durban in KwaZulu-Natal (Fig. 5.8).
i8verararromm_ .,,,..114711._ lino " MAI rill IL I 1111116111: 1111111111111111111 1 ligl MIMI 21111111111111MIN ismounamin i_ two mriciationg 2 11111111111111 i 112 111111111111,11 4611011111 11 riargrilart112
tar FEW TIMMINIMill I 6 8011 mg • • - 111 Illikla IN pall= Mg pail NMI 11128 mi. 1114;11 1.1 A VICIIII „ irl ii on wet MOM III p, _ ulposi 11■1 __ ail,,, rli, %II '11,32
,,,iiiiii ‘ RilieLeas ' 1 itilWrIi ,., ,2 14 16 18 120 ..2 24 26 28 30 32 34 --
FIGURE 5.8 Geographical distribution of Dasispermum humile.
Additional specimens examined.
-2930 (Pietermaritzberg): Umlaas, Natal (-DA), Krauss 418 (BM, K, 2 sheets, S).
-3225 (Somerset East): Boschberg (-DA), MacOwan 390 (BOL, K).
85 CHAPTER 5: TAXONOMIC REVISION OF THE GENUS DASISPERMUM
3227 (Stutterheim): King Willaim's Town (—CD), Sim s.n. (PRE, 2 sheets); banks of the
Kabusi River near Komga (—DB), Flanagan 1183 (BOL, 3 sheets, K, PRE, S).
3228 (Butterworth): Kobinquba Forest (—CB), Pegler 1212 (BOL).
3325 (Port Elizabeth): Zuurberg National Park (—AD), B.-E. & M. Van Wyk 1759 (JRAU, 2
sheets); Enon (—BC), Thode A2670 (K, PRE); Ouboskloof (—BC), B.-E. & M. van
Wyk 1883 (JRAU, 2 sheets), B.-E. & M. van Wyk 1895 (JRAU, 2 sheets).
—3326 (Grahamstown): Bucklands (—BA), Acocks 23793 (PRE); Hopewell (—DA), Acocks
23899 (K, PRE).
3327 (Peddie): road through Fort Grey forest (—BB), Hilliard & Burtt 14861 (K, PRE, S).
Precise locality unknown: Drege 9545 (S); Thode A2670 (K); Umpampanyani-Dumiso,
Rudatis 2166 (PRE).
5. DASISPERMUM PERENNANS Magee and B.-E.van Wyk, sp. nov., A Dasispermo
hispido habitu perenni repullanti et caule ligneo permanenti differt et a D.
suffruticoso foliis teretiusculis coriaceis ceraceis viribus (in D. suffruticoso
foliis succulentis, plerumque glaucis), petalis papillosis et fructu costis modo
leviter prominentibus (in D. suffruticoso manifeste costato vel alato). — TYPE:
SOUTH AFRICA. Simonstown district (3418): sandy flats between Noordhoek
and Kommetjie (–AB), Esterhuysen 34312 (BOL!, holo.; BOL!, 2 sheets, K!,
S!, iso.).
Sprawling (erect when young) sympodial suffrutices, 0.15-0.5 m tall, resprouting, short-lived perennials. Stems permanent, woody, immaculate, at first simple becoming slightly to well-branched; branches ascending to somewhat decumbent, resprouting at the nodes. Leaves 20-90 mm long x 10-40 mm, glabrous, 2-pinnate, coriaceous. Petioles 5-40 mm long, glabrous. Ultimate leaflet
86 CHAPTER 5: TAXONOMIC REVISION OF THE GENUS DASISPERMUM
segments linear-oblong, 1-3 mm long, less than 1 mm broad, subterete,
concolorous, bright green, margins entire, venation visible or obscure. Umbels with
peduncle short to long, 20 to 100 mm long; involucral bracts 1-6, lanceolate to ovate
or often foliose, coriaceous, apex acuminate, margins usually membranous, glabrous; rays (4) 10 to 20, 15-30 mm long, scabrous; involucel bracts 2 to 6, linear to lanceolate, coriaceous, apex acuminate, margins often membranous, glabrous;
raylets 8 to 10, 3-5 mm long, glabrous or scabrous. Flowers with petals 0.8-1.2 mm
long and broad, apex deeply emarginate, papillose; ovary glabrous; styles lengthening as fruit matures, ± 1.0 mm long, becoming reflexed beyond the base of the stylopodium. Fruit 2.0-2.5 mm x 1.5-2.5 mm, broadly ovate in lateral view, base obtuse to very slightly cordate; mericarps homomorphic, glabrous; median and lateral ribs as well developed as the marginal ribs, slightly prominent, cartilaginous, dorsal surface not fibrously ridged, glabrous; marginal ribs parallel to very slightly concave in lateral view.
Diagnostic characters— Dasispermum perennans is most similar to D. hispidum and D. suffruticosum but differs from the former in the resprouting perennial habit and permanent woody stem and from the latter in the subterete coriaceous, waxy green leaves (succulent, glaucous leaves in D. suffruticosum), papillose petals and fruit with only slightly prominent ribs (very prominently ribbed or winged in D. suffruticosum).
Distribution and habitat— The species is known only from the Cape
Peninsula between Noordhoek and Karbonkelberg (Fig. 5.6) where it occurs on very deep sandy soils often amongst Thamnochortus erectus.
87 CHAPTER 5: TAXONOMIC REVISION OF THE GENUS DASISPERMUM
Additional specimens examined.
–3418 (Simonstown): Hout Bay (–AB), Bond 434 (NBG); Karbonkelberg, on S.E. slopes (-
AB), Esterhuysen 28979 (BOL, K); Sandy Bay, Table Mountain National Park (–AB),
Magee & Boatwright 105 (JRAU, NBG); Winter 3850 (JRAU!, PRE!).
6. DASISPERMUM SUFFRUTICOSUM (Berg.) B.L.Burtt in Notes RBG Edinb. 45: 93
(1988). Conium suffruticosum Berg., Pl. Cap. 77 (1767, September). Cicuta
suffruticosum (Berg.) Poir., Encycl. Suppl. 2: 261 (1811). Cnidium
suffruticosum (Berg.) Cham. and Schlechtd. in Linnaea 1: 387 (1826).
Selinum suffruticosum (Berg.) Benth. and Hook. f., Gen. Pl. 1: 914 (1867).
Heteroptilis suffruticosa (Berg.) Leute in Ann. Nathist. Mus. Wien 73: 89
(1969). — TYPE: SOUTH AFRICA. Cape of Good Hope, Grubb s.n. (STB!,
holo.).
Conium rigens L., Mant. 56 (1767, October); Mantissa altera 352, 512 (1771). Cicuta
rigidens (L.) Lam., Encycl. Meth. 2: 4 (1786). Bunium rigens (L.) Spreng. in
Roem. and Schult., Syst. Veg. 6: 500 (1820). Trachyspermum rigens (L.)
G.Don in Loudon, Hort. Brit. 103 (1830). Trachydium rigens (L.) Hiroe,
Umbell. World 702 (1979). — TYPE: SOUTH AFRICA. "Habitat ad Cap. b. spei."
Herb. Linn. No. 343.2 (LINN!, lecto., designated by Burtt 1988, isolecto. E).
Sium patulum Thunb., Prodr. 51 (1794). — TYPE: SOUTH AFRICA. Cape, in littore
Houteniquas, Thunberg s.n. sub THUNB-UPS 7055 (UPS!, lecto., here
designated). [Note: This is the only specimen in Thunberg's herbarium, it is
annotated on the reverse by him and so is here designated]
88 CHAPTER 5: TAXONOMIC REVISION OF THE GENUS DASISPERMUM
Capnophyllum jacquinii DC., Prodr. 4: 187 (1830), p.p. max. Conium jacquinii (DC)
D.Dietr., Syn. Pl. 2: 988 (1840). — TYPE: Cult. Hort. Trianon (G-DC, lecto.,
designated by Burtt 1991).
Dasispermum maritimum Raf., Good Book 56 (1840) and Scad. Omb. Pl. 56 (Amer.
Midl. Nat. Repr. No. 3, 1913), nom illegit.
Heteroptilis arenaria [E.Mey. ex] Meisn. in J.D. Hook., Lond. J. Bot. 2: 534 (1843). -
TYPE: SOUTH AFRICA. Oudtshoorn district (3322): Cape, George, Zwartevallei
(–DC), Krauss 1196 (specimen not located).
Sprawling to prostrate (erect when young) sympodial suffrutices, 0.10-0.4 m tall, resprouting, short-lived perennials. Stems permanent, woody, immaculate, at first simple becoming slightly to well-branched; branches decumbent to prostrate, resprouting at the nodes. Leaves 15-100 mm long x 10-40 mm, glabrous, pinnate to 2-pinnate, firm and semi-succulent. Petioles 5-40 mm long, glabrous. Ultimate leaflet segments oblong, 1-4 mm long, 1-3 mm broad, turgid or rarely laminate, usually strongly curled inwards, concolorous, glaucous or sometimes green, margins entire or toothed, venation usually obscure. Umbels with peduncle short to long, 10 to 80 mm long; involucral bracts 4 to 6, lanceolate to ovate, slightly succulent, apex acute to acuminate, margins usually membranous, glabrous; rays 6 to 26, 8-32 mm long, glabrous or scabrous; involucel bracts 4 to 6, lanceolate, slightly succulent, apex acute to acuminate, margins often membranous, glabrous; raylets 6 to 14, 2-5 mm long, glabrous. Flowers with petals 0.8-1 mm long and broad, apex slightly to deeply emarginate, smooth; ovary glabrous; styles lengthening as fruit matures,
0.8-1.0 mm long, becoming reflexed up to or beyond the base of the stylopodium.
Fruit 2.5-6.0 mm x 3.0-5.0 mm, ovate to very broadly ovate in lateral view, base
89 CHAPTER 5: TAXONOMIC REVISION OF THE GENUS DASISPERMUM obtuse; mericarps homomorphic or heteromorphic, glabrous; if homomericarpic then with the median and lateral ribs as well developed as the marginal ribs, if heteromericarpic then with either median or lateral ribs as well developed as the marginal ribs, very prominent to winged, corky, dorsal surface not fibrously ridged, glabrous; marginal ribs parallel in lateral view.
Diagnostic characters— This highly specialised littoral species can readily be distinguished from the other species of Dasispermum by the perennial suffrutescent habit (often with slender stems adapted to emerge and form new plants when covered by moving sand), the succulent leaves with the ultimate leaflet segments often curled inwards and the prominently ribbed or often winged fruit, with the ribs or wings corky.
Distribution and habitat— Dasispermum suffruticosum is a halophytic species with a very wide distribution along the South African coastline, occurring on littoral dunes from Vredendal in the Western Cape Province to Port Shepstone in
KwaZulu-Natal (Fig. 5.9).
90 CHAPTER 5: TAXONOMIC REVISION OF THE GENUS DASISPERMUM
12 14 16 18 20 22 24 26 28 30 32 34 --Willia 1111111 [SNOT 11113. ir 18 18M111 ■ III■irumagrol a is IT 1 20 MN ■ ■ to • MI 11 22 al 1 i 111111211111MI a 2° ii ELPlimr11111112 240 Imo ■ gm non gimmi 4 11.I 1111 NM ITELIIN 11-511111/1,81111I 26 ■ 1: 111r■ Aa Notmnteaa minim l I so111115(611 115 REM IN 10128 281.s.i li■s 2 on■ El -n 111 -12,21meA • s 111 11.---/ . Lori 32,. in 1 I -.11111111 PIE ■1--- I mi.. o i.orsrow pumili_r... um 34 ... hiLPill Ili III MINN, eu 10 12 14 16 18 20 22 24 26 28 30 32 34 36
FIGURE 5.9 Geographical distribution of Dasispermum suffruticosum.
Additional specimens examined.
3030 (Port Shepstone): lsipingo beach (—BB), Forbes & Obermeyer 45 (PRE); Port
Shepstone (—CB), Bayliss 2199 (NBG), Mogg s.n. (PRE); between Ramsgate and
Margate (—CD), Whellan 1065 (PRE).
3118 (Vanrhynsdorp): district Papendorp, Farm Geelwalkaroo 262 (—CA), Sachse 472
(PRE); Vredendal, dunes opposite Rob Island, N of Oliphants River Mouth (—CA), Le
Roux & Ramsey 77 (PRE).
91 CHAPTER 5: TAXONOMIC REVISION OF THE GENUS DASISPERMUM
3129 (Port St Johns): Umngazi River Mouth (—CB), Taylor 9000 (PRE, S); Bluff (—DA),
Wood s.n. (PRE).
3217 (Vredenburg): Shell Bay near Veldrif (—DB), Marsh 1283 (NBG).
3218 (Clanwilliam): Papkuilsfontein, 5 1/2 miles NW of L' Agulhas (—CB), Rycroft 1833
(NBG); Rocher Pan Nature Reserve (—CB), Brandt 8 (NBG); beach N of Berg River
Mouth (—CC), O'Callaghan 1184 (NBG).
3227 (Stutterheim): near King Williams Town (—CD), Sim s.n (PRE).
3228 (Butterworth): The Haven (—BB), Wisura 2681 (NBG); Mazeppa Bay (—BC), Hilnes
508 (PRE); Kei Mouth (—CB), Flanagan 257 (PRE, S); Strey 11239 (PRE); Kentani (-
CB) Pegler 892 (PRE).
3317 (Saldanha): Hoedjies Point (—BB), Van Rensburg 126 (PRE).
3318 (Cape Town): Saldanha Bay between Churchhaven and South Point (—AA), Brenan
14020A (NBG), Hall 855 (NBG); Sandveld Forestry Area (—AC), Low 815 (NBG);
Ysterfontein (—AC), Glen 1090 (PRE), Reyneke 32 (PRE); Koeberg beach (—CB),
Barnes 2441 (PRE); Bloubergstrand (—CD), Compton 8920 (NBG), Manning &
Reeves 2819 (NBG), Pole Evans 4389 (PRE), Raitt s.n. (NBG); Milnerton (—CD),
Andreae 381 (NBG).
3323 (Willowmore): Nature's Valley (—DC), Stirton 9628 (PRE).
3325 (Port Elizabeth): Van Stadens River Mouth (—CC), Klein 118 (PRE); Maitland River
Mouth (—CD), Boucher 3347 (NBG); Fishwater Flats, Swartkopsrivier (—DC), Olivier
4881 (PRE); Humewood (—DC), Compton 12144 (NBG); Markman Industrial Area,
sand flats off Swartkopsrivier (—DC), Dahlstrand 3032 (NBG).
3326 (Grahamstown): Kenton-on-sea, mouth of Kariega river (—DA), Hilliard & Burtt
10875 (NBG, S); Kenton-on-sea, Joan Muirhead Nature Reserve (—DA), Mucina
LM6161/2 (PRE); near Kenton-on-sea, Kasuka River Mouth (—DA), Phillipson 355
(PRE); Kowie (—DB), Britten 527 & 1894 (PRE); Port Alfred (—DB), Hutton 2736
(PRE).
92 CHAPTER 5: TAXONOMIC REVISION OF THE GENUS DASISPERMUM
—3327 (Peddie): Fish River Mouth (—AC), Phillipson 178 (PRE); East London (—BB),
Commins 1034 (PRE), Galpin 3301 (PRE).
3418 (Simonstown): Cape Peninsula, on sand dunes, Millers Point (—AB), Bond 287
(NBG); Hout Bay (—AB), Thode 6243 (NBG); Scarborough (—AB), Goldblatt 6753
(PRE); Schuster's River Mouth (—AB), Whellan 1741 (PRE); Simon's Bay (—AB),
Prior s.n. (PRE); Cape of Good Hope Nature Reserve, Platboom Bay (—AD), Taylor
6533 (NBG, PRE); Cape Point (—AD), Marloth 3246 (PRE), Phillips s.n. (PRE), Wall
166 (S); Skilpadvlei flats (—AD), Van der Merwe 6069 (NBG); near Slangkop (—AD),
Marloth 11868 (PRE); Macassar beach (—BA), Downing 402 (NBG); Macassar dunes
(—BA), De Bruyn s.n. (NBG); Seekoei River Mouth (—BA), Parsons 26 (NBG, PRE);
Strandfontein (—BA), Henderson 1164(NBG), Rycroft 2363 (NBG), Taylor 3180
(PRE); Strand (—BB), Parker 4395 (NBG, PRE), Strand 4350 (NBG); Hol Bay
Peninsula, E of Cape Hangklip (—BD), Rourke 696 (NBG).
—3419 (Caledon): Kleinmond (—AC), Van Wyk 6069 (NBG); Hermanus, Grotto Beach (-
AD), Taylor 9876 (NBG, PRE); Hermanus, Mossel River (—AD), Guthrie s.n. (PRE);
Dawidskraal (—BD), Walsh s.n. (PRE); Pearly beach (—CB), Maguire 24 (NBG),
Meyer 2388 (PRE), Taylor 4045 (NBG, PRE).
3420 (Bredasdorp): De Hoop, De Mond (—AD), Jordaan s.n. (NBG); Stilbaai (—BD),
Beyleveld s.n. (NBG), Bohnen 412.7 (NBG), Rycroft 3079 (NBG, S); Witsand river
mouth (—BD), Hugo 1908 (NBG, PRE); Arniston (—CA), Galpin 11348 (PRE);
Waenhuiskrans (—CA), Greuter 21965 (PRE), Westhuizen 125 (PRE); Skipskop (-
CB), Scott 478 (NBG); Agulhas (—CC), Fellingham 381 (NBG, PRE), Taylor 1588
(PRE).
3421 (Riversdale): Still Bay (—AD), Bakker 5880 (PRE), Bohnen 412.7 (PRE), Muir 111 &
4533 (PRE).
3422 (Mossel Bay): Blinde River Estuary (—AA), O'Callaghan, FeHingham & Van Wyk 263
(NBG, PRE); Hartenbos river mouth (—AA), Parsons 321 (NBG, PRE); Kleinbrakrivier
(—AA), Du Plessis 14 (PRE), Meyer 2288 (PRE); Mossel Bay (—AA), Rogers 4243
93 CHAPTER 5: TAXONOMIC REVISION OF THE GENUS DASISPERMUM
(NBG, PRE), Glentana beach (AB), Goldblatt & Manning 9522A (NBG), Wilderness
(—BA), Compton 10727 (NBG), Marloth s.n. (PRE), Mogg 11884 (PRE); Goukamma
Nature Reserve (—BB), Anon. 187 (PRE); Kingfisher Creek, Sedgefield (—BB),
Huddlenost 2048 (NBG).
—3423 (Knysna): Brenton beach (—AA), Bos 969 (NBG, PRE); Keurboomstrand (—AB),
Codd 3570 (PRE), Morris 432 (NBG), Taylor 4332 (NBG); Plettenberg Bay (—AB),
Kapp 120 (PRE).
—3424 (Humansdorp): Jeffreys Bay (—BB), Stirton 9601 (PRE), Taylor 5146 (NBG); St
Francis Bay (—BB), Brand 197 (PRE).
3425 (Skoenmakerskop): Sea View (—AB), Long 253 (PRE).
7. DASISPERMUM TENUE (Sond.) Magee and B.E.van Wyk, comb. nov. Ptychotis
tenuis Sond. in Harv. and Sond., Fl. Cap. 2: 537 (1862). Trachyspermum
tenue (Sond.) Engler, Pflanzenwelt Afrikas 3(2): 809 (1921). Sonderina tenuis
(Sond.) H.Wolff in Pflanzenr. Heft 90: 93 (1927). — TYPE: SOUTH AFRICA.
Bredasdorp district (3420): Cape, Buffeljagdrivier to Rietkuil (–BA), Zeyher
2672 (S!, lecto. here designated; LE!, isolecto.). [Note: The specimen in S is
chosen here as it is from the author's collection, has an authentic indication
of the type locality and closely matches the original protologue.]
Sprawling or somewhat erect sympodial herbs, 0.04-0.3 m tall, annuals.
Stems herbaceous, immaculate, usually well-branched at the base; branches decumbent or ascending, not resprouting from the nodes. Leaves 30-95 mm long x
10-35 mm, 2-pinnate, herbaceous to very slightly coriaceous. Petioles 15-50 mm long, slightly scabrous, green. Ultimate leaflet segments linear-oblong, 2-7 mm long, less than or equal to 1 mm broad, laminate, concolorous, green, margins
94 CHAPTER 5: TAXONOMIC REVISION OF THE GENUS DASISPERMUM entire, venation visible. Umbels with peduncle sessile to subsessile, less than 4 mm long; involucral bracts 0; rays 3 to 6, 5-20 mm long, scabrous; involucel bracts 0 (to
4), linear to narrowly ovate, coriaceous, apex acute, margins not membranous, glabrous; raylets 4 to 9, 2-5 mm long, scabrous. Flowers with petals 0.4-0.6 mm long and broad, apex truncate to shallowly emarginate, papillose; ovary scabrous; styles not markedly elongated in mature fruit, 0.2-0.3 mm long, remaining erect or becoming reflexed almost up to the base of the stylopodium. Fruit 1.7-2.2 mm x
1.7-2.0 mm, broadly elliptic to rotund and convex in lateral view, base obtuse; mericarps homomorphic; median and lateral ribs as well developed as the marginal ribs, prominent, cartilaginous, dorsal surface slightly fibrously ridged, scabrous; marginal ribs parallel to very slightly concave in lateral view.
Diagnostic characters— Dasispermum tenue is similar to D. humile and D. grandicarpum in that the fruit ribs are usually fibrously ridged on the dorsal surface but differs from the former in the narrower ultimate leaflet segments and from the latter in the much smaller fruit and the absence of involucel bracts. It is further distinguished from both of the before mentioned species by the invariably sessile to subsessile umbels and the short styles which do not extend beyond the base of the stylopodium.
Distribution and habitat— This easily overlooked Renosterveld endemic species is restricted to shale soils in the Western Cape Province from Worcester in the west to Riversdale in the east (Fig. 5.6).
95 CHAPTER 5: TAXONOMIC REVISION OF THE GENUS DASISPERMUM
Additional specimens examined.
3319 (Worcester): Karoo Garden Veld (—CB), Perry 497 (NBG).
3320 (Montagu): hills between Buffeljagsrivier and Rietkuil (—BA), Zeyher 2672 (BM photo,
S); hills near Bonnievale (—CC), Esterhuysen 23797 (BOL); Touwsberg, farm
Wolwefontein, kloof above house (—DB), Van Wyk, Winter & Tilney 3433 (JRAU,
PRE).
—3420 (Bredasdorp): Windhoek, shale slopes at Apoisfontein (—AD), Burgers 2559 (CPA).
—3421 (Riversdale): Riversdale (—AB), Bolus 11291 (BOL, K); Schlechter 1703 (BOL).
Precise locality unknown: Linde, Zeyher s.n. (S).
96 CHAPTER 6
TAXONOMIC REVISION OF THE GENUS SCARABOIDES
6.1 INTRODUCTION
Scaraboides is here described to accommodate an unusual new species from the Succulent Karoo in the Western Cape Province of South Africa. Scaraboides manningii was first brought to our attention during a revision of the genus
Capnophyllum with which it shared the dorsally compressed fruit with a broad commissure, concave commissural surface and involute marginal wings. This species was however, distinctly different from those of Capnophyllum due, amongst other characters, to the additional pair of vittae within the marginal wings and the parallel, closely-spaced commissural vittae in the fruit. In a recent phylogenetic analysis of the Lefebvrea Glade (tribe Tordylieae) using separate and combined morphological, anatomical and molecular sequence data (Chapter 3), the genus was placed within the Capnophyllum group (Magee et al. 2009b) together with the annual southern African endemic genera Capnophyllum and Dasispermum s.l.
Despite the numerous similarities between the fruit of Capnophyllum and
Scaraboides, the separate molecular and combined molecular/morphological analyses placed Scaraboides most closely allied to Dasispermum. The genus is morphologically distinct from the other members within the Capnophyllum group and clearly represents an isolated, easily recognisable lineage.
97 CHAPTER 6: TAXONOMIC REVISION OF THE GENUS SCARABOIDES
6.2 VEGETATIVE AND REPRODUCTIVE MORPHOLOGY
6.2.1 Vegetative morphology— Scaraboides manningii is an erect and only
slightly dichotomously branched annual herb. The species shares the sympodial
growth habit with other members of the Capnophyllum group, albeit only weakly
expressed in young plants. The pinnately divided leaves of S. manningii differ from
species of Capnophyllum in the dark green colour, with the ultimate leaflet segments flat and relatively broad, appearing much like those of Dasispermum humile.
6.2.2 Reproductive morphology— In Scaraboides the umbels are often
sessile or rarely born on a short peduncle. The species lacks both involucre and
involucel bracts and has scabrous rays and raylets similar to those found in some
species of Dasispermum. The petals are notably smaller in Scaraboides manningii
(± 0.5 mm long and broad — Fig. 6.1H) than those of both Capnophyllum and
Dasispermum (± 1 mm long and broad — Fig. 6.1D) and are obtusely tiped without a septum on the inner face (Fig. 6.1 H). The stylopodium is prominently flat and may
be either level or slightly sunken below the fruit apex. The styles are usually erect and remain relatively short in mature fruit. The broadly elliptic fruit are dorsally compressed, with the median and lateral ribs indistinct and the marginal ribs expanded into narrow wings. The marginal wings, as found also in Capnophyllum, are prominently bent inwards so that the commissural surface is very strongly concave. The commissure is very broad and extends to the edge of both marginal ribs. In addition to the two commissural and four vallecular vittae, the fruit of
Scaraboides manningii have additional solitary vittae in each of the marginal wings.
This is an unusual feature, to our knowledge, found in no other species. The position
98 CHAPTER 6: TAXONOMIC REVISION OF THE GENUS SCARABOIDES of the commissural vittae in S. manningii differs furthermore in that they remain parallel to one another, running vertically down along either side of the carpophore.
FIGURE 6.1 Scaraboides manningii. (A), habit. (B), lower leaf pinnae. (C), petal in ventral and lateral view. (D), umbellule. (E), fruit, dorsal surface. (F), fruit, commissural surface. (G), transverse section through the mature fruit. Vouchers: A—D. Manning 3010 (NBG); E—G. Manning 3061 (NBG). cv, commissural vitta; vv, vallecular vitta; wv, additional wing vitta. Scale: A = 10 mm, B = 5 mm, C, E—G = 1 mm, D = 2.5 mm.
99 CHAPTER 6: TAXONOMIC REVISION OF THE GENUS SCARABOIDES
6.3 TAXONOMY OF THE GENUS SCARABOIDES
SCARABOIDES Magee and B.-E.van Wyk, gen. nov. Capnophyllo Gaertn. habitu
annuali, commissura lata fructus, mericarpiis dorsaliter compressis,
superficiebus commissurialibus concavis, alis marginalibus involitis similis
sed ramis erectis (non prostratis nec decumbentibus), segmentis ultimis
foliorum latis viridis (non angustis glaucis), umbellis scabris (non glabris)
saepe sessilibus, bracteis involucralibus involucellaribusque deficientibus,
costis dorsalibus fructus indistinctis, vittis additis solitariis in quoque alo
marginali fructus et vittis paralelis crebris commissurialibus differt. — TYPE: S.
manningii Magee & B.-E.van Wyk.
Etymology— The generic name is derived from the distinctive dark colour and the strongly convex, smooth outline of the fruits (making them distinctly "beetle- like" in appearance), hence scaraboides [from the Greek, scarabeus (beetle) and – oides (like)].
1. SCARABOIDES MANNINGII Magee and B.-E.van Wyk, sp. nov., Species unica,
characteribus generis. — TYPE: SOUTH AFRICA. Sutherland district: Tanqua
Karoo National Park, NE slopes of Elandsberg; 06 August 2006; B. Sachse
116 (PRE!, holo.; BOL!, JRAU!, K!, KMG!, KSAN!, NBG!, iso.).
Erect herb, 0.2-0.4 m tall. Stem single, rarely slightly branched at the base, erect. Leaves 50-120 mm x 20-60 mm, pinnate, glabrous, green. Petioles 20-60 mm long, basal sheaths 7-12 mm x 3-5 mm. Ultimate leaflets broadly ovate, 12-30
100 CHAPTER 6: TAXONOMIC REVISION OF THE GENUS SCARABOIDES mm x 10-28 mm, venation pinnate; segments narrowly oblong, 2-9 mm x 1.5-3 mm, flat. Umbels compound; peduncle sessile or rarely short, 0(-30) mm long; involucre absent; rays 4 to 6, 10-30 mm long at anthesis, slightly scabrous; involucel absent; raylets 6 to 9, 5-8 mm long at anthesis, scabrous. Flowers pentamerous; petals ± 0.5 mm long and broad, papillose, inflexed tips obtuse, septum absent on inner face, apex truncate; ovary glabrous; stylopodium flat, level with or slightly sunken below the fruit apex; styles not markedly elongated in mature fruit, 0.2-0.4 mm long, remaining erect or very rarely becoming somewhat reflexed up to the base of the stylopodium. Fruit broadly elliptic, 5.5-6.0 mm x 3.0-3.5 mm; base obtuse or shallowly concave; apex obtuse; mericarps strongly concave on the commissural surface; median and lateral ribs indistinct; marginal ribs distinctly involute; additional solitary vittae present in the marginal wings.
Diagnostic characters —The fruit is most similar to those of Capnophyllum in the broad commissure, the dorsally compressed mericarps with concave commissural surfaces and the involute marginal wings, but differs in the indistinct median and lateral ribs, the additional solitary vittae in each marginal wing and the parallel, closely spaced commissural vittae. When in flower, this species may be confused with Dasispermum humile but it is geographically isolated and can easily be distinguished by the flattened stylopodium and the petals which are not keeled on the adaxial face and which have only a short, obtuse tip (not slender and attenuate — the typical lobulum inflexum as is found in most other Apioideae).
101 CHAPTER 6: TAXONOMIC REVISION OF THE GENUS SCARABOIDES
HEIGHT ABOVE SEA LEVEL
I-1 Over 1500 m 1:=3 SOO - 1500 m ri 300 - 900 m 1-1 Under 300 m
20 0 20 40 60 80 100 km I f l/111 li
FIGURE 6.2 The known geographical distribution of Scaraboides manningii.
Distribution and habitat— Scaraboides manningii is restricted to the
Succulent Karoo of the Western Cape Province, where it is known from only two localities, the Tanqua Karoo National Park and Mauwerskop near Vanrhynsdorp
(Fig. 6.2). The species grows in seasonally damp, dolerite or clay soils.
Etymology— This species is named in honour of Dr. John Manning (NBG) who brought it to our attention and who also provided the first complete collection.
102 CHAPTER 6: TAXONOMIC REVISION OF THE GENUS SCARABOIDES
Additional specimens examined.
—3118 (Calvinia): Mauwerskop, NW of Matsikammaberg (—DB), Snijman 1056 (PRE, MO).
3219 (Wuppertal): Top of Boulderkoppie, E of Leeuberg (—BB), Sachse 71 (PRE), Tanqua
Karoo National Park.
3220 (Sutherland): NE slopes of Remhoogte, ca. 1 km S from Maansedam, Tanqua
Karoo National Park (—AA), Bester 7171 (PRE); E foot of Elandsberg, Tanqua Karoo
National Park (—AA), Manning 3010 (NBG); E foot of Elandsberg, Tanqua Karoo
National Park (—AA), Manning 3061 (NBG); Elandsberg, Wilderness chalets (—AA),
Sachse 628 (PRE).
103 CHAPTER 7
SYSTEMATICS OF AFRICAN AND MALAGASY PIMPINELLA AND RELATED
GENERA
7.1 INTRODUCTION
Pimpinella is one of the largest genera within the family Apiaceae, comprising about 150 species (Pimenov and Leonov 1993) distributed throughout much of the
Old World. Due to its size and geographical range, coupled with the high levels of infraspecific variation, the generic boundary of the genus and its infra-generic classification remains unclear (Abebe 1992; Spalik and Downie 2007). The genus was last revised in its entirety by Wolff (1927), who subdivided it into three sections based on petal colour and the vestiture of the fruit. In this system, the African and
Malagasy species were accommodated into two subsections within sections
Tragium and Tragioselinum, depending on whether the fruit are papillose to hairy or glabrous, respectively. More recent taxonomic studies of the African representatives of Pimpinella have revealed several new tropical African species (Abebe 1989,
1992; Townsend 1985), and as a result ca. 49 sub-Saharan (Van Wyk and Tilney
2004) and five Malagasy (Sales and Hedge 2004) species are generally recognised.
In a recent analysis of nrITS sequence data, Spalik and Downie (2007) found that the African species of Cryptotaenia comprised a Glade within the tribe
Pimpinelleae, together with the Malagasy endemic Phellolophium, Malagasy
Pimpinella and the African endemic genus Frommia. As this African Glade was sister to the Glade comprising the Eurasian members of Pimpinella, the latter authors highlighted the need for the inclusion of African Pimpinella in a molecular
104 CHAPTER 7: SYSTEMATICS OF AFRICAN AND MALAGASY PIMPINELLA AND RELATED GENERA phylogenetic study to ascertain whether they were more closely related to the
Eurasian or to the Malagasy members. The present study is aimed at assessing the phylogenetic relationships of the African and Malagasy species of Pimpinella and related genera through comparisons of fruit anatomical, cytological and nrITS sequence data.
7.2 MATERIAL AND METHODS
7.2.1 Taxon sampling—The 18 nrITS sequences (17 newly obtained) of
African and Malagasy Pimpinella were analysed together with 29 accessions of
Eurasian Pimpinella and a comprehensive sampling of the tribe Pimpinelleae (from
Spalik and Downie 2007). Two accessions from the closely related tribe Apieae and four accessions from the tribe Pyramidoptereae were included as outgroups, with the latter used to root the trees. Sources of material used in the study, together with their corresponding GenBank accession numbers, are listed in Appendix B2.
7.2.2 Phylogenetic analyses— Phylogenetic analyses of all data sets were conducted initially using maximum parsimony, as described in Chapter 2, with gaps treated as missing data. Tree searches were performed using a heuristic search with
1,000 random sequence additions, TBR branch swapping and the MULPARS option in effect. A limit of 10 trees per replicate was set to reduce the time spent on swapping in each replicate. Internal support was assessed with 1,000 bootstrap replicates using TBR swapping and holding 10 trees per replicate. Bayesian inference was performed, as outlined in Chapter 2, for 3 million generations of
105 CHAPTER 7: SYSTEMATICS OF AFRICAN AND MALAGASY PIMPINELLA AND RELATED GENERA
Monte Carlo Markov Chains and a sampling frequency of 100, with the SYM+G model implemented.
7.3 RESULTS AND DISCUSSION
7.3.1 Phylogenetic relationships— The matrix consisted of 623 unambiguously aligned nucleotide positions with 317 variable and 231 parsimony informative characters. The MP analysis (Fig. 7.1) resulted in 36 equally most parsimonious trees with a TL of 788 steps (CI of 0.59 and 0.53, with and without uninformative characters, respectively; RI of 0.81). The respective topologies of the
MP strict consensus tree and the BI consensus tree were congruent (with the differences between the trees summarised in Fig. 7.1) and retrieved similar groups to those shown by Spalik and Downie (2007). The tribe Pimpinelleae was strongly recovered to be monophyletic (PP 1.0, BP 98). Within the tribe, the largest genus,
Pimpinella was rendered paraphyletic by the inclusion of African Cryptotaenia and the small African and Malagasy endemic genera Frommia and Phellolophium.
Within the paraphyletic Pimpinella lineage (PP 1.0, BP 94), three major clades were recovered. The African species of Pimpinella were widely separated into two of the three major clades. The majority of the Eurasian species of Pimpinella sampled were recovered together with the type species (P. saxifraga) within a strongly supported (PP 1.0, BP 96), but poorly resolved Eurasian Glade. Successively sister was a strongly supported Glade (PP 1.0, BP 98) comprising both African and
Eurasian species of Pimpinella. Within this Glade the African species formed a strongly supported group (PP 1.0, BP 99 — hereafter the P. hirtella group) sister to the Eurasian species. A third Glade (PP 1.0, BP 80), which was the most early diverging, comprised only African and Malagasy species. Within this Glade African
106 CHAPTER 7: SYSTEMATICS OF AFRICAN AND MALAGASY PIMPINELLA AND RELATED GENERA
Pimpinella section: 0.83 Pimpinella niitakayamensis ❑ Reutera 0.98 0.57 Pimpinella peregrina 78 53 Pimpinella eriocatpa OTragium Pimpinella diversifolia iiiTragoselinum 1.0 1.0 1--- Pimpinella huillensis 98 100 I-- Pimpinella huillensis 1.0 0.82 Pimpinella Melia 74r 55 Pimpinella oreophila Pimpinella krookii P. hirtella group 0.98 1.0 Pimpinella caffra 84 100 Pimpinella transvaalensis Pimpinella ledermannii Pimpinella kyimbilaensis Pimpinella lutea Pimpinella kotschyana -Pimpinella oliverioides - Pimpinella cappadocica 0.90 1.0 Pimpinella aurea 63 -Pimpinella anisum 99 0.96 53 -Pimpinella anisetum kotschyana corymbosa aromatica 0.76L.....= PPPPpiiiimmipPPPiiiiinnnneeeeeillillillaaa: tragium .0.._ 1r-- 19-01 tragium 73 1 Pimpinella tragium 0.94 Pimpinella affinis 0.99 1.0I- Pimpinella saxifraga 78 99 I-- Pimpinella rhodantha Pimpinella peucedanifolia 0.99 Pimpinella nudicaulis 1.0 Pimpinella isaurica 0.78 Pimpinelleae 96 Pimpinella flabellifolia Pimpinella puberula 1.0 Pimpinella paucidentata 94 Pimpinella sintenisii 1.0 1-Pimpinella cretica 100 1- Pimpinella cretica 0.91 Pimpinella trifurcata
1 . Pimpinella rigidistyla 750 1.0 70 Pimpinella alismatifolia 0.85 100 1.0 Pimpinella buchananii 96 Pimpinella buchananii P buchananii group 1.0 1.0 Pimpinella favifolia 98 100 1.0 Pimpinella favifolia 1.0 1- Pimpinelle sp. B 1.0 79 '- Frommia ceratophylloides 78 1.0 Pimpinella perrieri 100 Pimpinella betsileensis 0.96 0.84 Malagasy Pimpinella 60 1 0 Phellolophium madagascariense 53 1.0 88 Phellolophium madagascariense and Phellolophium 1.0 95 Phellolophium madagascariense 1.0 80 Cryptotaenia calycina 98 0.96 Cryptotaenia africana African Cryptotaenia 61 Cryptotaeniary africana 1.0 f-Araftte aromatica 100 1-Arefoe aromatica 1.0 1.0 Psammogeton canescens 100 1.0 99 Psammogeton bitematum 1.0 99 Aphanopleura capillifolia 100 1.0 i-Nothosmymium japonicum 84 Aph a nopleun3 trachysperma 1.0 Deverra burchellii 97 = Stoibrax dichotomum ] Apieae 1.0 Pyramidoptera cabulica 0.80 100 Oedibasis platycarpa 60 Bunium bulbocastanum ] Pyramidoptereae rrithmtim marilimum
FIGURE 7.1 Bayesian inference tree of ITS sequence data for Pimpinella species and some related taxa (tribe Pimpinelleae). Posterior probability (PP) values are presented above the branches. Bootstrap percentage (BP) values are presented below the branches. BP values below 50% are not indicated. Branches supported only in the BI are indicated by dashed lines. The current
sectional classification for Pimpinella is indicated with coloured text. Those species of Pimpinella not included by Wolff (1927) or subsequently by Abebe (1992) are indicated in black text.
107 CHAPTER 7: SYSTEMATICS OF AFRICAN AND MALAGASY PIMPINELLA AND RELATED GENERA
Cryptotaenia was, as shown by Spalik and Downie (2007), paraphyletic and the
Malagasy species of Pimpinella (PP 1.0, BP 100) were weakly supported as sister group to the Malagasy genus Phellolophium (PP 0.84, BP 60). In addition the monotypic, African endemic genus Frommia was placed within a group of tropical
African Pimpinella species (hereafter the P. buchananii group) in which it was moderately to strongly supported as sister to Pimpinella sp. B. (PP 1.0, BP 79).
7.3.2 African and Malagasy Pimpinella— Based on the results of the molecular analyses, three major clades could be identified within Pimpinella. The three sections proposed by Wolff (1927), do not correspond to any of the clades recovered and appear to be largely artificial (Fig. 7.1). In the latter classification, the
African and Malagasy species were, with the exception of P hirtella, separated from the Eurasian species based on geographical distribution. Although the African and
Malagasy species were recovered within two of the three major clades, they were consistently found to ally together. Within the African species, the separation based on the presence or absence of hairs on the fruit appears to be only slightly useful. In general, the P. buchananii group and the Malagasy species have glabrous fruit
(although variable in P. rigidistyla), while the P. hirtella group have hairy fruit (except
P. kyimbilaensis and P. oreophila, variable in the P. caffra complex). While the presence or absence of hairs is largely a conservative species-specific character for the tropical African species, in the South African members the character is well known to be of limited value (Townsend 1985) due to the large amount of infra- specific variation. However, the placement of these species (viz. P. caffra, P. krookii and P. transvaalensis) within the P. hirtella group suggests that the sporadic loss of hairs is likely to be a secondary adaptation.
108 CHAPTER 7: SYSTEMATICS OF AFRICAN AND MALAGASY PIMPINELLA AND RELATED GENERA
FIGURE 7.2 Transverse sections through the fruit of African (A—J) and Malagasy (K—N) species of Pimpinella, Phellolophium (0), Frommia (P) and Cryptotaenia (Q, R). A. P acutidentata (Fanshawe s.n., K); B. P hirtella (Ash 2677, MO); C. P oreophila (Townsend 2428, K); D. P transvaalensis (Miller 5910, K); E. P caffra (Hilliard & Burtt 9777, K); F. P caffra subsp. conopodioides (Pawek 9268, K); G. P huillensis (Milne-Redhead 10894, K); H. P buchananii (Milne-Redhead 9045A, K); I. P trifurcata (Jean Pawek 13923, MO); J. P lindblomii (Gilbert 6531, K); K. P tenuicaulis (Baron 3241, MO); L. P betsileensis (Humbert 28087, P); M. P perrieri (Bosser 8622, P); N. P ebracteata (Baron 929, K); 0. Ph. madagascariensis (Pettersen & Nilsson 359, K); P. F ceratophylloides (Hooper et al 1844, K); Q. C. africana (Verdcourt 2481, PRE); C. canadensis (PE0866713). Scale: 0.4 mm. 109 CHAPTER 7: SYSTEMATICS OF AFRICAN AND MALAGASY PIMPINELLA AND RELATED GENERA
There are some useful fruit anatomical differences (Fig. 7.2) between the
species of Pimpinella, and the data seems to be more useful at the species level, as
previously suggested by Abebe (1992), than for the recognition of infra-generic groups. While most of the species have multiple vallecular vittae between the ribs, in
P. transvaalensis (Fig. 7.2D) and P oreophila (Fig. 7.2C) only a single vallecular vitta
between each rib was observed. This may prove particularly useful to distinguish between the closely related P caffra and P transvaalensis. However, fruit from many more populations of both these species and those studied by Abebe (1992) need to be included before any definite conclusions can be made.
Although the morphological and anatomical characters investigated were only poorly correlated with the results of the molecular phylogeny, the optimisation of cytological data (Table 7.1) onto the MP trees revealed very interesting results. This is in contrast to the findings of Abebe (1992) who could not find any correlation between the cytological variation and the subgeneric divisions based on fruit and leaf morphology within Pimpinella. In the Apioideae the predominant chromosome base number is x=11 (Moore 1971). Within Pimpinella however, x=8, 9, 10 and 11 have been recorded, with x=9 being the most prevalent. In all the species (for which counts are available) within the early diverging Glade comprising the P buchananii group, including Cryptotaenia africana, a count of x=11 has been recorded, while those species in the P hirtella group consistently have a count of x=9, as found also in the Eurasian species sister to this Glade. Optimisation of the cytological data (Fig.
7.3) suggests x=10 to be the plesiomorphic state for the genus. The chromosome count of x=11 found in the early diverging lineage comprising the P buchananii group is recovered as an apomorphy for this lineage, while x=9 appears to be
110
CHAPTER 7: SYSTEMATICS OF AFRICAN AND MALAGASY PIMPINELLA AND RELATED GENERA
Pimpinella niitakayamensis Chromosome counts [1:1 Pimpinella peregrina 0 Pimpinella eriocarpa Elx=11 Pimpinella diversifolia Pimpinella huillensis Elx= .10 Pimpinella huillensis Pimpinella hirtella II x=9 Pimpinella oreophila Pimpinella krookii P hirtella group 'x=8 Pimpinella caffra Pimpinella transvaalensis Pimpinella ledermannii ■ Pimpinella kyimbilaensis 1 ❑ Pimpinella lutea Pimpinella kotschyana 1 weed Pimpinella oliverioides ipI Pimpinella cappadocica Pimpinella aurea 01 Pimpinella anisum Pimpinella anisetum Pimpinella kotschyana I. Pimpinella corymbosa Pimpinella aromatica GI Pimpinella tragium 0 Pimpinella tragium T ■III Pimpinella tragium Pimpinella affinis 1: II Pimpinella saxifraga II Pimpinella rhodantha Pimpinella peucedanifolia Pimpinella nudicaulis Pimpinella isaurica Pimpinella flabellifolia ■ Pimpinella puberula Pimpinella paucidentata Pimpinella sintenisii ❑ Pimpinella cretica ❑ Pimpinella cretica t ❑ Pimpinella trifurcata Pimpinella rigidistyla Pimpinella alismatifolia ❑ Pimpinella buchananii ❑ Pimpinella buchananii ❑ Pimpinella favifolia P buchananii group t I ❑ Pimpinella favifolia ❑ Pimpinella sp. B r 1 ❑ Frommia ceratophylloides Pimpinella perrieri Pimpinella betsileensis I7-1 Phellolophium madagascariense Malagasy Pimpinella I Phellolophium madagascariense Phellolophium madagascariense and Phellolophium I Cryptotaenia calycina 1 ❑ Cryptotaenia africana African Cryptotaenia 1 ❑ Cryptotaenia africana I Ara foe aromatica Ara foe aromatica 1 ❑ Psammogeton canescens I 1=011• Psammogeton canescens 1 ❑ Aphanopleura capillifolia ❑ Nothosmymium japonicum I I Aphanopleura trachysperma I ❑ Deverra burchellii I I ❑ Stoibrax dichotomum I Pyramidoptera cabulica I Oedibasis platycarpa ❑ Bunium bulbocastanum 1 ❑ Crithmum maritimum
FIGURE 7.3 Parsimony-based reconstructions of chromosome base numbers (Table 7.1) in Pimpinella and related taxa when optimized over the 36 minimal length trees inferred from MP analysis of ITS data.
111 CHAPTER 7: SYSTEMATICS OF AFRICAN AND MALAGASY PIMPINELLA AND RELATED GENERA apomorphic for the African-Eurasian Glade comprising the P hirtella group, with a subsequent decrease to x=8 in P eriocarpa. The ancestor to the entirely Eurasian
Glade, including the type species (P saxifraga), appears to have a basic chromosome number of x=10, although subsequent decreases to x=9 have occurred in many of the species (Fig. 7.3). The possibility that x=11 (the predominant number in the Apioideae) is the plesiomorphic state within the genus, with subsequent aneuploidy decrease to x=10, 9, 8 in the more derived lineages seems to be a plausible hypothesis. Chromosome data should be explored across a broader sample of species, including the outgroup taxa.
7.3.3 Frommia— The genus Frommia is an anomalous African endemic genus with a limited distribution in Malawi. The most striking feature of the genus is reflected in the epithet of its only species, F ceratophylloides. The leaves resemble those of Ceratophyllum due to the sessile pinnae and finely divided and linear ultimate leaf segments (Cannon 1973). Although Cannon (1978) suggested a possible affinity to the genus Carum, the generic placement has remained unclear.
In the analysis of Spalik and Downie (2007), Frommia was placed within the tribe
Pimpinelleae where it was sister to the Glade comprising the Malagasy endemics
Phellolophium madagascariense and Pimpinella betsileensis. In the current analysis, with the broader sampling of African Pimpinella, the genus was moderately to strongly placed sister to the undescribed Pimpinella sp. B., known only from a few collections in Zambia (Cannon 1973), within the P buchananii group. Interestingly, the latter species also has finely divided, linear ultimate leaf segments as found in
Frommia, although without sessile pinnae. Pimpinella lineariloba Cannon from
Angola may form part of this group as it also has linear ultimate leaf segments and
112 CHAPTER 7: SYSTEMATICS OF AFRICAN AND MALAGASY PIMPINELLA AND RELATED GENERA
according to Cannon (1973) a possible affinity to Pimpinella sp. B. In transverse
section the fruit of Frommia (Fig 7.2P) appear similar to those of the P buchananii
group (Fig. 7.2H—J), with the multiple vallecular (three in each furrow) and
commissural (four, sometimes appearing confluent in mature fruit) vittae, as well as
the prominently concave commissural surface of the seed. Furthermore, a
chromosome count of x=11, as found in Pimpinella sp. B and the other species from
the P buchananii group (Fig. 7.3H—J), has also been recorded for Frommia
ceratophylloides (Constance et al. 1971, 1976).
7.3.4 Phellolophium— The Malagasy endemic genus Phellolophium was
described by Baker (1884), who proposed an affinity to Seseli L. and Foeniculum
Mill. Recently a second as yet undescribed species (Sales et al. 2004) was
discovered and an affinity to Pimpinella speculated (Sales and Hedge in press). In
the analysis of Spalik and Downie (2007), the genus was shown to be closely
related to the African members of the tribe Pimpinelleae, together with P
betsileensis, the only Malagasy species sampled. In the current analysis, the sister
group relationship of Phellolophium and Malagasy Pimpinella was confirmed, with
this group successively sister to the P buchananii group. The genus shares the
broad, pinnately divided leaves with some of the Malagasy species of Pimpinella but
differs in the presence of prominent involucre and involucel bracts, fruit with thick
corky ribs and four vallecular vittae as well as another five slightly smaller vittae
usually situated more or less below the ribs (Fig. 7.20).
7.3.5 Cryptotaenia— The geographically disjunct genus Cryptotaenia was shown by Spalik and Downie (2007) to be polyphyletic, with the species divided
113 CHAPTER 7: SYSTEMATICS OF AFRICAN AND MALAGASY PIMPINELLA AND RELATED GENERA between the tribes Oenantheae (Cryptotaenia s.s.), Scandiceae (C. elegans Webb ex Bolle) and Pimpinelleae (C. africana, C. calycina C.C.Towns.). As in the analyses of Spalik and Downie (2007), both the MP and BI analyses presented herein, place the African species of Cryptotaenia (C. africana, C. calycina) successively sister to the lineage comprising Phellolophium, the Malagasy species of Pimpinella and the
P. buchananii group. The fruit of C. africana (Fig. 7.2Q) appear only superficially similar to those of Cryptotaenia s.s. In Cryptotaenia canadensis (Fig. 7.2R) the fruit are diagnostically different with only one vallecular vittae between each rib and two additional vascular bundles located near to the commissure. The fruit of C. africana
(Fig. 7.2Q), in contrast, have multiple vittae between the ribs and only five vascular bundles, as found also in most species of Pimpinella.
7.3.6 Taxonomic implications— The preliminary analyses of Spalik and
Downie (2007) suggested that the African and Malagasy members of the
Pimpinelleae occupied an isolated position separate from the Eurasian species.
These authors also suggested that the African taxa were derived from a common ancestor of Middle Eastern origin which extended into Africa through a Middle-East-
East African tract and subsequently into Madagascar. With the inclusion of more
African species of Pimpinella in the phylogenetic analysis, it is now apparent that they are not as isolated as previously suggested. Although it is likely that the species from Madagascar are derived from a single ancestor of African origin, it would appear that more than one dispersal event into Africa from Eurasia must have taken place. While it is tempting to simply include the anomalous African genera
Cryptotaenia, Frommia and Phellolophium into an expanded and already rather poorly defined Pimpinella, there remains a need for a broader sampling of
114 CHAPTER 7: SYSTEMATICS OF AFRICAN AND MALAGASY PIMPINELLA AND RELATED GENERA particularly the eastern Asiatic species, which according to Spalik and Downie
(unpublished data), are only distantly related. Formal taxonomic and nomenclatural changes should only be implemented once the full range of diversity has been studied. Although the fruit characters traditionally used to define infra-generic groups within Pimpinella appear to be only of limited use to define natural groups, the results of this study suggest that cytological data should be explored in more detail as this is likely to be very useful in understanding evolutionary relationships within the genus.
115 CHAPTER 7: SYSTEMATICS OF AFRICAN AND MALAGASY PIMPINELLA AND RELATED GENERA
TABLE 7.1 Chromosome numbers of Pimpinella species and other taxa included in the nrITS analyses. Taxa for which chromosome counts were unavailable where coded as missing in the optimisation analyses (Fig. 7.3). Chromosome Taxa Sources numbers Aphanopleura capillifolia n=11 Vasil'eva et al. 1991, 1993 Buttler 1985; Vasil'eva et al. 1985; Verlaque and Bunium bulbocastanum 2n=20 Filosa 1992 Cryptotaenia africana 2n=22 Auquier and Renard 1975; Morton 1993 Al-Bermani et al. 1993; Vasil'eva et al. 1993; Crithmum maritimum n=10; 2n=20 Pimenov et al. 1998 Deverra burchellii n=11 Al-Eisawi 1989 (for Pituranthos triradiatus) Frommia ceratophyfioides n=11 Constance et al. 1971, 1976 Nothosmyrnium japonicum 2n=20 Pan et al. 1995 Pimpinella affinis n=9; 2n=16 Constance et al. 1976; Yurtseva 1988 Pimpinella anisum 2n=18,19 Pimenov et al. 2003 Pimpinella buchananii n=11; 2n=22 Constance and Chuang 1982; Abebe 1992 Pimpinella caffra n=9 Constance and Chuang 1982 Pimpinella corymbosa n=10; 2n=20 Al-Eisawi 1989; Pimenov et al. 1996 Pimpinella cretica n=10; 2n=10 Al-Eisawi 1989 Pimpinella diversifolia n=9; 2n=18 Cauwet-Marc 1982 Pimpinella eriocarpa n=8 Al-Eisawi 1989 Pimpinella favifolia n=22 Constance and Chuang 1982 Pimpinella hirtella 2n=18 Abebe 1992 (as P. volkensii) Pimpinella huillensis n=9 Constance and Chuang 1982 Pimpinella ledermannii n=9 Constance and Chuang 1982 (as P. engleriana) Pimpinella lutea 2n=20 Verlaque et al. 1992 Hedberg and Hedberg 1977 (as P. Pimpinella oreophila 2n=18 kilimandscharica) Pimpinella peregrina n=9; 2n=16,18,20 Yurtseva 1988; Abebe 1992; Pimenov et al. 1996 Pimpinella puberula 2n=18 Yurtseva 1988 n=9, 10; 2n=20, 36, Constance et al. 1976; Pimenov and Vassiljeva Pimpinella rhodantha 40 1983; Yurtseva 1988; Daushkevich et al. 1995 Pimpinella saxifraga 2n=18, 20, 36, 40 Gawlowska 1967 Pimpinella sp. B n=11 Constance and Chuang 1982 Yurtseva and Tikhomirov 1998; Pimenov et al. Pimpinella tragium 2n=18, 20, (22) 2003 Pimpinella trifurcata 2n=22 Abebe 1992 Psammogeton biternatum 2n=18 Ahmad and Koul 1980 Psammogeton canescens n=11 Khatoon and All 1993 Stoibrax dichotomum n=10 Silvestre 1990
116 CHAPTER 8
SYSTEMATICS OF MALAGASY PEUCEDANUM
8.1 INTRODUCTION
Although the family Apiaceae is relatively poorly represented in Madagascar
[15 genera (11 endemic) and 31 species (10 endemic)], many of the Malagasy taxa
represent important component of early diverging lineages of subfamily Apioideae
(Van Wyk et al. 1999; Calvin() et al. 2006; Calvifio et al. in prep.). These Malagasy
species display a diverse range of unusual habit and fruit anatomical characters
(Sales et al. 2004). Earlier treatments under estimated the total number of endemic
genera, largely due to inadequate material, and many of the isolated Malagasy
species had been included in African genera. Humbert (1956) placed eight of the
woody Malagasy species in the arboreal African genus Heteromorpha. However,
after extensive morphological and anatomical studies, the Malagasy species were
later excluded from Heteromorpha (Winter et al. 1993; Winter and Van Wyk 1994,
1996) and largely accommodated in four new Malagasy endemic genera (Van Wyk
et al. 1999), with one of the species transferred to the tropical African genus
Pseudocarum. Sales and Hedge (in press) recently described five new Malagasy species. Two of these species were tentatively placed within the problematic
Peucedanum (Sales et al. 2004; Peucedanum sp. A and Peucedanum sp. B). The former species, subsequently described as Billburttia capensoides (Magee et al.
2009d), has been relatively well-collected, but has consistently been misidentified as part of the "Peucedanum capense (Thunb.) Sond. complex" (Burtt 1991) due to surprising morphological similarities. Recent detailed studies on African
117 CHAPTER 8: SYSTEMATICS OF MALAGASY PEUCEDANUM
Peucedanum have shown that all the African species are unrelated to Eurasian
Peucedanum and, therefore, were accommodated in six new African endemic genera (Magee et al. 2008a,b, 2009a; Winter et al. 2008). As a result, the species in
Madagascar represented the last remaining challenge to the so-called
"Peucedanum problem" in Africa. Using fruit anatomical and molecular sequence data, we show here that the two species (described in Magee et al. 2009d, as B. capensoides and B. vaginoides) are unrelated to Peucedanum and other peucedanoid taxa but represent a distinct new genus described herein as Billburttia.
8.2 MATERIAL AND METHODS
8.2.1 Taxon sampling— To assess the phylogenetic positions of B. capensoides (3 specimens) and B. vaginoides (1 specimen), the newly obtained ITS
(three accessions of B. capensoides and one accession of B. vaginoides) and rps16 intron (two accessions of B. capensoides) sequences were incorporated into the respective ITS and rps16 intron data matrices of Magee et al. (2009b). These matrices include most of the relevant major clades of the "apioid superclade," including a representative sampling of the Lefebvrea Glade (Magee et al. 2009b) which comprises many African peucedanoid genera (Winter et al. 2008). Preliminary analyses indicated an affinity between the two Billburttia species and the tribe
Apieae, and therefore all available accessions of the latter were also included. The trees were rooted with taxa from tribes Smyrnieae and Oenantheae (Downie et al.
2001). In total, 150 accessions of ITS and 60 accessions of rps16 were considered for the phylogenetic study (Appendix B3).
118 CHAPTER 8: SYSTEMATICS OF MALAGASY PEUCEDANUM
8.2.2 Phylogenetic analyses— Phylogenetic analyses of all data sets were conducted initially using maximum parsimony, as described in Chapter 2, with gaps treated as missing data. Tree searches were performed using a heuristic search with
500 random sequence additions, TBR branch swapping, and the MULPARS option in effect, but saving no more than 5 of the shortest trees from each search. These equally parsimonious trees were then used as starting trees for TBR branch swapping (MULPARS and STEEPEST DESCENT in effect) with the maximum number of trees saved set at 12,000; these trees were permitted to swap to completion (Downie et al. 1998). Bootstrap percentage values were determined from
500,000 replicate analyses using fast stepwise addition of taxa. Bayesian inference was performed for four million generations of Monte Carlo Markov Chains with a sampling frequency of 100.
8.3 RESULTS AND DISCUSSION
8.3.1 Morphology— Billburttia capensoides and B. vaginoides are glabrous shrubs or subshrubs up to 2 m tall. Their leaves are cauline and glaucous. The species are easily separated from each other by differences in their ultimate leaf segments. In B. capensoides these are linear-elliptic to elliptic (Fig. 8.1A) and closely resemble those of Notobubon laevigatum (Aiton) Magee (Fig. 8.11), from which it can be distinguished by the lack of a prominent midrib and the presence of well-developed lateral venation. The ultimate leaf segments of B. vaginoides, in contrast, are filiform to linear with only the main vein visible (Fig. 8.1B). Each stem terminates in a much branched inflorescence borne on a long peduncle. The relatively small fruit (Fig. 8.1G) are superficially similar to those found in species of
119 CHAPTER 8: SYSTEMATICS OF MALAGASY PEUCEDANUM
Notobubon (Fig. 8.1L), but differ in the prominent dorsal ribs and several other
important details, as described below.
8.3.2 Fruit anatomical description— In transverse sections (Fig. 8.1C-E)
the fruit of B. capensoides and B. vaginoides are slightly dorsally compressed and
homomericarpic. The marginal ribs are expanded into narrow or scarcely developed wings and the dorsal ribs are very prominent. The commissure differs notably from
that of species of both Eurasian Peucedanum (Fig. 8.1K) and the African
peucedanoid taxa (e.g. Notobubon, Fig. 8.1J) in that it extends only to the base of
each marginal wing (and not to the tip of each wing, as in the latter genera). The
epidermal cells are periclinally elongated with a striate outer surface. There is some
lignification of the mesocarp in the ribs. The cells of the endocarp are
parenchymatous and periclinally elongated. The vascular tissue is located at the tip of the dorsal and marginal ribs (Fig. 8.1C-E) and as such is an unusual and diagnostic character for both species. Rib oil ducts were not observed in the mature fruit. The mericarp has 10 to 12 regular vittae. Along the commissural length there are six vittae of unequal length, while in each vallecula there may be one or two vittae present. Crystals were observed in groups associated with the epidermis and around the vittae in unstained sections of both species (Fig. 8.1F). These crystals were initially thought to be calcium oxalate druses, as reported in some taxa of the
Apioideae-Saniculoideae Glade (Rompel 1895; Drude 1897-1898; Van Wyk and
Tilney 2004; Liu et al. 2006, 2007c; Magee et al. 2008c). However, the crystals did not dissolve during staining with PAS/TB and thus it became apparent that they were not composed of calcium oxalate. On closer examination of both stained and unstained
120 CHAPTER 8: SYSTEMATICS OF MALAGASY PEUCEDANUM
A D ••118■■■■■••■••■•11
F r
FIGURE 8.1 Diagnostic morphological and anatomical characters of Billburttia (A—H) in comparison to the vegetatively similar Notobubon laevigatum (I, J, L, M) and the type species of Peucedanum (P. officinale) (K). A and B ultimate leaf segment of B. capensoides (A) and B. vaginoides (B), C—E transverse sections through the fruit of B. vaginoides (C) and B. capensoides (D, E), F marginal wing of B. capensoides showing the presence of sphaerocrystals (indicated with arrows) associated with the epidermis, G and H mature mericarp of B. capensoides in dorsal view (G) and in commissural view (H) showing six commissural vittae of unequal length, I ultimate leaf segment of N. laevigatum, J and K, transverse sections through the fruit of N. laevigatum (J) and P officinale (K), L and M mature mericarp of N. laevigatum in dorsal view (I) and in commissural view (M) showing two commissural vittae. Vouchers: A. Du Puy et al. M660 (P); B. Perrier de la Bathie 6799 (P); C. Decary 7583 (P); D. Rakotozapy 631 (P); E. and F. Humbert 3634 (P); G. and H. Humbert 1727 (P); I., L. and M. Magee 7 (JRAU); J. Botha 3547 (PRE); K. PE 328993. Scale: A, B, G, H, L = 1 mm, C—E, J, K = 0.5 mm, F = 0.2 mm. 121 CHAPTER 8: SYSTEMATICS OF MALAGASY PEUCEDANUM transverse sections they were identified as sphaerocrystals of unknown composition. This appears to be the first report of sphaerocrystals in Apiaceae and, as such, represents a potentially useful diagnostic character at the generic level.
8.3.3 ITS and rps16 intron data sets— The ITS dataset included 634 unambiguously aligned nucleotide positions of which 384 were variable and 321 parsimony informative. MP analyses resulted in the preset maximum tree limit of
12,000 trees, each of 2,207 steps [consistency indices (CI) of 0.33 and 0.30, with and without uninformative characters, respectively; retention index (RI) of 0.73].
MODELTEST selected the GTR+I+G model of evolution for use in the BI analysis.
The MP strict consensus and BI majority-rule consensus trees (Fig. 8.2A) were largely topologically congruent and retrieved the same clades as those reported by
Magee et al. (2009b). The Glade comprising the two species of Billburtia (three accessions of Billburttia capensoides and one accession of B. vaginoides) was strongly supported (PP 1.0, BP 100) and formed part of the tribe Apieae (PP 1.0, BP
84). The two species were sister group to a Glade comprising Sclerosciadium nodiflorum Coss., Ammi majus. and Petroselinum crispum (PP 0.91, BP 61).
The rps16 intron dataset included 865 unambiguously aligned nucleotide positions of which 179 were variable and 98 parsimony informative. MP analyses resulted in the preset maximum tree limit of 12,000 trees, each of 277 steps (CI of
0.75 and 0.63, with and without uninformative characters, respectively; RI of 0.86).
MODELTEST selected the K81uf+G model of evolution for use in the BI analysis.
The MP strict consensus tree and the BI majority-rule consensus trees (Fig. 8.2B) were largely topologically congruent and retrieved the same clades as those reported by Magee et al. (2009b), although the topologies recovered within the Glade
122 CHAPTER 8: SYSTEMATICS OF MALAGASY PEUCEDANUM
comprising the tribe Apieae differed slightly as discussed below. The position of the
Malagasy species, B. capensoides, within the tribe Apieae (PP 0.97, BP <50) was
confirmed (Fig. 8.2B), although rps16 intron data for the closely related B.
vaginoides was unavailable due to amplification difficulties. In both the BI and MP
consensus trees, the two accessions of B. capensoides formed a Glade which was
sister group to Stoibrax dichotomum. (PP 0.85, BP <50). However, in the BI
consensus tree B. capensoides and Stoibrax dichotomum formed part of a larger,
weakly supported Glade (PP 0.82) including Deverra, Ammi majus, Petroselinum
crispum, Apium graveolens and Naufraga balearica (BI 0.82), while in the MP strict
consensus tree B. capensoides and Stoibrax formed part of a Glade (BP <50%)
comprising Anethum graveolens and Foeniculum vulgare.
8.3.4 Phylogenetic position— The two species are unrelated to both
Eurasian Peucedanum and the African peucedanoid taxa. Analyses of both ITS and
rps16 intron data strongly place Billburttia within the tribe Apieae (Fig. 8.2). Although
B. vaginoides was not included within the analyses of the rps16 intron dataset, this
species is clearly closely related to B. capensoides based on morphological,
anatomical and ITS sequence data. It is unclear what the direct sister group of
Billburttia is, as either Stoibrax or Ammi, Petroselinum and Sclerosciadium as
retrieved as sister groups by either cpDNA or nrITS sequence data, respectively.
This may be due to the low resolution within the rps16 intron dataset or the result of
hybridization and introgression of the genome. Although the fruit of Billburttia are
superficially similar to the previously mentioned peucedanoid genera, anatomically they are markedly dissimilar. The position of the vascular bundles in the tips of the
ribs and the presence of sphaerocrystals are both unusual fruit anatomical
123
CHAPTER 8: SYSTEMATICS OF MALAGASY PEUCEDANUM
characters which lend support to both the isolated position of the two species and
their recognition as a distinct and monophyletic new genus.
A. ITS 98 Deverra burchellfi B. rps16 intron 100 1 0 Deverra denudata 1.0 Deverra triradiata 79 931 621 Sclerosciadium nodiflorum 1.0 Ammi majus 901 Naufraga balearica 1 0 0-77 61 Petroselinum crispum 1 01 Apium graveolens Bilburtia capensoides 0.91 0.54 94 Petroselinum crispum 100 Billburtia capensoides 1.0 1 Ammi majus 85 1.0 Billburtia capensoides 0.74 1.0 Deverra triradiata Billburtia vaginoides Ff." 52 99 Apium graveolens sv 0.95 78 I Deverra denudata "C3 97 CD cT 11.0 Apium prostratum 0.82 1.0 I Deverra burchellii 0) 1.0 Naufraga balearica 84 97 Billburtia capensoides CD 1.0 Anethum graveolens 1.0 Billburtia capensoides 0.97 Foeniculum vulgare 0.97 0.85 93 70 Stoibrax dichotomum 0.61 I Foeniculum vulgare 1.0 0.99 Ridolfia segetum 77 Anethum graveolens 1.0 100 r— Stoibrax dichotomum Foeniculum vulgare 1.0 Stoibrax dichotomum
FIGURE 8.2 Portions of Bayesian inference trees, showing the position of Billburttia within the tribe Apieae, based on ITS (A) and rps16 intron (B) sequence data. Posterior probability (PP) values are presented below the branches. Bootstrap percentage (BP) values from parsimony analysis are presented above the branches. BP values below 50% are not indicated. Branches supported only in the BI are indicated by dashed lines.
124 CHAPTER 8: SYSTEMATICS OF MALAGASY PEUCEDANUM
8.4 TAXONOMY OF THE GENUS BILLBURTTIA
BILLBURTTIA Magee and B.-E.van Wyk gen. nov., Forma foliorum, mericarpiorum et
habitu Peucedanum L. et praecipue Notobubon B. -E. van Wyk similis sed ab
eorum vittis vallecularibus 6, margine mericarpiorum indistincto, commissuris
angustis, sphaerocrystallis in regione epidermidis bene differt. [to be
translated into Latin] — TYPE: B. capensoides Sales & Hedge.
Shrub or subshrub, clump-forming, glabrous, 0.5-2.0 m tall. Rootstock thick
to very thick, woody, vertical or oblique. Stems erect, branched in region of
inflorescence, terete, clearly and finely ridged, solid, blue-green or reddish. Leaves
cauline, persisting, 1- to 3-pinnate. Ultimate leaf segments filiform–linear or linear-
elliptic to elliptic; margins entire; base narrow to broad and cuneate; apex acute to
mucronulate; firm to thick-textured; concolorous, glabrous to glaucous; midrib and
lateral veins equally developed or only midrib prominent, flush with or sunken below
the lamina. Petioles terete, sheathing almost along their entire length. Inflorescence
of 1 to 6, lax hermaphrodite and smaller male compound umbels; terminal umbels
hermaphrodite; peduncle long, more than two times longer than the diameter of the primary umbel; striate. Primary umbel rounded; rays 15 to 30,15-40 mm long, ± equal, glabrous; involucre present; bracts numerous, linear-oblong, unequal, apex acute to acuminate, glabrous, papery; raylets glabrous, glaucous; involucel present; bracteoles numerous, linear-oblong, unequal, apex acute to acuminate, ± fused at base, glabrous, papery, shorter than raylets; umbellule many-flowered. Flowers pentamerous, predominantly hermaphrodite, those in the lateral umbels entirely or in part functionally male; sepals minute, apex truncate to acuminate, glabrous; petals
/ 125 CHAPTER 8: SYSTEMATICS OF MALAGASY PEUCEDANUM
1.5 mm long, greenish yellow, yellow or white, elliptic to obovate, acuminate, with inflexed tips, glabrous; stylopodium broadly conical; styles 2, short; ovary bilocular.
Fruit slightly dorsally compressed, narrowly ellipsoid or oblong-ellipsoid, 3.5-4.5 mm x c. 1.5 mm, glabrous; mericarps homomorphic; median and lateral ribs prominent; marginal ribs narrowly-winged, not saccate at base; styles becoming strongly reflexed up to or beyond the base of the stylopodium; commissural vittae 6; vallecular vittae 4 to 6, solitary or in pairs; commissure broad, extending to the base of each wing; carpophore bipartite. Endosperm flat.
Etymology— The genus is named in honour of Brian Laurence ("Bill") Burtt
(1913-2008) of the Royal Botanic Garden Edinburgh (Weber and Noltie 2008) whose paper on southern African Apiaceae (Burtt 1991) has contributed greatly to our understanding of the family in that region. He was also a co-author of the account of Gesneriaceae for Flore de Madagascar.
Diagnostic characters— The species of Billburttia are superficially similar to some species of Notobubon. They are small shrubs or subshrubs with a woody rootstock and have relatively small, slightly dorsally compressed fruit. Billburttia is easily distinguished from Notobubon and members of the tribe Apieae by a combination of distinct fruit characters, namely the narrower commissure (not extending beyond the base of each wing, as in Notobubon), the six commissural vittae (vs. two commissural vittae), ribs with the vascular bundles located near the tips, and the presence of sphaerocrystals associated with the epidermis and vittae.
Distribution— The species are endemic to Madagascar.
126 CHAPTER 8: SYSTEMATICS OF MALAGASY PEUCEDANUM
KEY TO THE GENUS BILLBURTTIA AND ITS SPECIES:
1a. Mericarps isodiametric or laterally compressed in transverse section
All other Madagascan Aplaceae
1 b. Mericarps slightly dorsally compressed Billburttia:
Ultimate leaf segments broad, more than 3 mm wide, linear-elliptic to
elliptic; shrub or subshrub, 1-2 m tall 1. B. capensoides
Ultimate leaf segments narrow, less than 1 mm wide, linear-filiform;
subshrub, c. 60 cm tall 2. B. vaginoides
BILLBURTTIA CAPENSOIDES Sales and Hedge in Magee et al. in Plant Syst. Evol.
(2009d). — TYPE: MADAGASCAR. De la Bathie 6807 (P!, holotype).
BILLBURTTIA VAGINOIDES Sales and Hedge in Magee et al. in Plant Syst. Evol.
(2009d). — TYPE: MADAGASCAR. De la Bathie 13557 (P!, holotype).
127 CHAPTER 9
SYSTEMATICS OF THE GENUS EZOSCIADIUM
9.1 INTRODUCTION
Ezosciadium is a poorly known and collected, monotypic genus endemic to
the Eastern Cape Province of South Africa. As with many other small African
endemic Apiaceae its correct placement within the new emerging tribal classification
of the family remains untested by molecular data (Calvin° et al. 2006). The genus
was included in a large rbcL analysis by Forest et al. (2007) in assessing the
phylogenetic diversity of the Cape Flora. Although the genus occupied a position
sister group to Annesorhiza and ltasina, the limited sampling of the family did not
allow for any conclusions to be drawn on possible tribal affinities. Ezosciadium
capense is traditionally placed in the large tribe Apieae (Pimenov and Leonov 1993).
De Candolle (1830) and Sonder (1862) included it within the genus Helosciadium
W.D.J.Koch, a genus now placed in the tribe Oenantheae (Downie et al. 2001).
Wolff (1927) followed Ecklon and Zeyher (1837) in recognising the genus as distinct and postulated an affinity rather to Sonderina or Apium, genera traditionally placed in the tribe Apieae. Recent molecular systematic studies, however, have shown
Sonderina to form part of an African peucedanoid Glade together with Dasispermum and other African species of tribe Tordylieae previously treated in the genus
Peucedanum (Winter et al. 2008). As a result, the three genera with which
Ezosciadium is traditionally associated are scattered in various tribes throughout the subfamily Apioideae. This paper is aimed at ascertaining the phylogenetic placement of the genus within the subfamily using both DNA sequence and anatomical data as
128 CHAPTER 9: SYSTEMATICS OF THE GENUS EZOSCIADIUM well as providing the first comprehensive taxonomic revision, including nomenclature, typification, detailed description, geographical distribution and illustrations, of this anomalous and poorly known African genus.
9.2 MATERIAL AND METHODS
9.2.1 Taxon sampling— In order to assess the phylogenetic position of
Ezosciadium, DNA sequences of the chloroplast genes rps16 intron and rbcL were compiled using available sequences from GenBank. Due to the high divergence and difficulty in aligning the nuclear ribosomal DNA internal transcribed spacer (ITS) region across the subfamily, a reduced sampling was compiled representing the
Glade within which Ezosciadium was shown to be placed based on results of phylogenetic analyses of the aforementioned chloroplast genes. Sources of material used in the study are listed in the Appendix B4. In total, 52 accessions of rps16 (two new), 19 accessions of rbcL and 12 accessions of ITS (one new) were considered for phylogenetic study. For the broader analyses using plastid markers, representatives of both tribes of the subfamily Saniculoideae as well as many of the major clades and tribes of the subfamily Apioideae were included. The genus
Hermas L., a member of the subfamily Azorelloideae, was selected as outgroup for the chloroplast datasets, based on previous molecular analyses (Calvin° et al.
2006), while the ITS dataset was rooted using Lichtensteinia obscura based on the results of the chloroplast analyses.
129 CHAPTER 9: SYSTEMATICS OF THE GENUS EZOSCiAD/UM
9.2.2 Phylogenetic analyses— Maximimum parsimony was performed as described in Chapter 2 on all three datasets using a heuristic search with 1000 random sequence additions, TBR branch swapping and the MULPARS option in effect. A limit of 10 trees per replicate was set to reduce the time spent on swapping in each replicate. Internal support was assessed with 1000 bootstrap replicates using TBR swapping and holding 10 trees per replicate. Bayesian inference was performed as outlined in Chapter 2 for 1 million generations of Monte Carlo Markov
Chains and a sampling frequency of 10, with the TrN+G model implemented.
9.3 RESULTS AND DISCUSSION
9.3.1 Morphology— Ezosciadium is a markedly pilose (Fig. 9.1A), erect annual herb up to 0.35 m tall. The leaves are cauline, with the lower ones prominently tri-lobed (Fig. 9.1C) and the upper ones digitately compound (Fig.
9.1 D). The cotyledons are simple and narrowly oblanceolate (Fig. 9.1B), fitting the L type, as defined by Cerceau-Larrival (1962) as commonly found in the subfamily
Apioideae. The very sparse, compound umbels are sessile and borne in the leaf axils. They are composed of 2 to 4 markedly unequal rays (Fig. 9.1A) that are of diagnostic value amongst African annual genera. The flowers are also unusual in that the petals are not inflexed at their tips (Fig. 9.1 H) and the relatively small stamens remain highly inflexed, so that they appear almost sessile (Fig. 9.11).
130 CHAPTER 9: SYSTEMATICS OF THE GENUS EZOSC1A MUM
FIGURE 9.1 Ezosciadium capense. (A), portion of a fruiting stem. (B), cotyledon. (C), first leaf. (D), mature leaf. (E), mature fruit. (F), transverse section through the mature fruit. (G and H), petals in dorsal view (G) and in ventral view (H). I, stamens in dorsal view. Vouchers: A, E, F. Acocks 20021 (PRE); B—D, G—I Fries et al. 1156 (PRE). Scale: A—D = 5 mm, E = 1 mm, F = 0.2 mm, G—I = 0.5 mm.
131 CHAPTER 9: SYSTEMATICS OF THE GENUS EZOSCIADIUM
9.3.2 Fruit anatomy— The fruit are distinctly pilose and oblong in shape (Fig.
9.1E). They are borne on a partly bifid carpophore (Fig. 9.1E), one half of which
remains attached to the umbellate rays even after the fruit have fallen off (Fig. 9.1A).
In transverse section (Figs. 9.2A and 9.1F) the fruit are similar to many other Apioid
genera. They are isodiametric and homomericarpic with five equal, prominent ribs.
The commissure is very narrow, being confined to the carpophore region. The
epidermal cells are bottle-shaped and extend into hairs; those of the ribs contain
tanniniferous substances (Fig. 9.2B). This appears to be the first record in Apiaceae
and has previously only been reported in members of Araliaceae, viz. Hydrocotyle L.
and Trachymene Rudge (Liu 2004). The mesocarp is composed of parenchymatous
cells that are usually periclinally elongated. The endocarp is also parenchymatous, the cells being narrow and strongly periclinally elongated. The vascular tissue occupies a large portion within each of the five ribs and is distinctly circular. Rib oil ducts were not observed in the fruit. The vittae are arranged as in most members of the subfamily Apioideae – two commissural and four vallecular. The endosperm is isodiametric and flattened to slightly concave on the commissural face. The testa is prominent in transverse section and may be comprises of up to three layers of cells especially in the commissural region, with the outer periclinal cell walls of the outermost layer being conspicuously thickened.
Druse crystals, restricted to the region surrounding the carpophore in the commissural area (Fig. 9.2C), were found to be present in all fruit of Ezosciadium examined. The presence of scattered druse crystals in the mesocarp (i.e., with crystals occurring around the seed and not restricted to the carpophore region) has a high predictive value for determining ancestral lineages within Apioideae (Van Wyk and Tilney 2004). This type of crystal distribution is known to occur in many of the
132 CHAPTER 9: SYSTEMATICS OF THE GENUS EZOSCIADIUM
early diverging branches of the Apioideae-Saniculoideae Glade (Van Wyk and Tilney
2004), including most members of the Saniculoideae (Liu et al. 2007c), all genera of
the Annesorhiza Glade (i.e. Annesorhiza, Chamarea, ltasina), the tribe
Heteromorpheae, as well as Molopospermum W.D.J.Koch (Liu et al. 2006) and
Astydamia D.C. In the remaining Apioideae, crystals are usually completely absent
but, for example, are frequently present around the carpophore in some members of
the tribe Scandiceae, such as species of Anthriscus, Caucalis, Chaerophyllum L.,
Daucus, Osmorhiza Raf., Scandix and Torilis (Drude 1897-1898). The presence of
druse crystals, even though restricted to the carpophore region in Ezosciadium, may
suggest an affinity to the early diverging lineages of the subfamily as shown by
analyses of all three molecular data sets (Figs. 9.3 and 9.4).
FIGURE 9.2 Transverse sections through the fruit of Ezosciadium capense. (A), mature mericarp; (B), rib of mature fruit with epidermal cells containing tanniniferous substances (indicated with arrow); (C), immature fruit showing presence of druse crystals around the carpophore (crystals indicated with an arrow, similarly dark structures located at the base of the ribs are highly lignified vascular tissue). Vouchers: A ,B Goldblatt & Porter 12578 (NBG); C Fries et al. 1156 (PRE). Scale: A, C = 0.2 mm, B = 0.07 mm.
133 CHAPTER 9: SYSTEMATICS OF THE GENUS EZOSCIADIUM
9.3.3 rps16 intron dataset— The rps16 intron dataset included 1113 characters of which 304 were variable and 169 parsimony informative. The MP analysis (Fig. 9.3) resulted in 2208 equally most parsimonious trees with a tree length (TL) of 534 steps, a consistency index (CI) of 0.70 (including uninformative characters) and a retention index (RI) of 0.85. The overall topologies obtained from both the MP and BI analyses were similar, with the general branching order similar to that found in previous analyses (Calvino et al. 2006). Amongst the early diverging lineages of Apioideae the genus Lichtensteinia formed the earliest branching Glade
(BP 98; PP 1.0). Following this was the lineage comprised of the Annesorhiza Glade plus Ezosciadium and Molopospermum (BP 65; PP 0.91). In the BI tree,
Ezosciadium capense grouped with Molopospermum (PP 0.67), sister group to the
Annesorhiza Glade (Annesorhiza, Chamarea and ltasina). The tribe
Heteromorpheae (BP 100; PP 1.0) was a subsequent sister group, followed by the tribes Bupleureae (BP 97; PP 1.0) and Pleurospermeae (BP 100; PP 1.0). Higher up on the trees the tribes Oenantheae (BP 98, PP 1.0), Smyrnieae, Scandiceae and other members of the Apioid superclade, including the tribe Apieae (BP 91, PP 1.0), were retrieved. The placement of Chamaesciadium acaule differed in the two analyses. In the MP tree, it was found to be sister group to a Glade comprising
Apieae, Echinophoreae, Selineae and Tordylieae, while in the BI tree (not shown) it was sister group to a Glade comprising Echinophoreae, Selineae and Tordylieae (PP
1.0).
134
CHAPTER 9: SYSTEMATICS OF THE GENUS EZOSCIADIUM
Heracleum sphondylium Pastinaca sativa Notobubon galbanurn Tordylieae 84 0.69 rps 1 6 0.871- Dasispermum suffruticosum 1.0 Aethusa cynapium I Selineae Echinophora tenuifolia I Echinophoreae Deverra burchellii Foeniculum vulgare 91 57 Ammi majus 1 0 Apieae Apium graveolens 68 Stoibrax dichotomum 1.0 Chamaesciadium acaule 0.92 Torilis arvensis I Scandiceae 88 Smyrnium olusatrum I Smymieae 1.0 L Helosciadium nodifiorum 100 98 Berula erecta 1.0 1.0 Oenantheae 89 Sium latifolium 0.99 Komarovia anisosperma I Komarovia Glade 99 100r Pleurospermum uralens Pleurospermeae 1.0 1.01 Pleurospermum foetens I 97 r-Bupleurum falcatum Bupleureae 1.0 1 Bupleurum fruticosum 72 I 1001 Heteromorpha papillose 0.65 1.0 Heteromorpha involucra 88 Polemannia simplicior 100 67 1 0 Polemannia montana Heteromorpheae 1.0 0.94 Anginon verticillatum Anginon difforme 100 Pseudocarum laxiflorum 1.0 1 Pseudocarum eminii 95 72 Annesorhiza fllicaulis 1.0 _90.97L- Chamarea snijmaniae 1.0 0.63 Chamarea longipedicellata Annesorhiza latifolia Annesorhiza macrocarpa 99 Annesorhiza Glade Annesorhiza altiscapa 76 1.0 0.93 1.0 Chamarea aff. gracillima Itasina filifolia 65 Chamarea sp.1 0.91 Ezosciadium capense 100 0.67 M lopospermum peloponnesiacum 1.0 98 r- Lichtensteinia obscura Lichtensteinia Glade 1.0 Lichtensteinia lacera 60 Polemanniopsis marlothii Steganotaenieae 0.94 Steganotaenia araliacea I 69 Sanicula arctopoides ideae
0.64 lo 54 0.75 Arctopus echinatus Saniculeae icu
0.98 100 Alepidea serrata n 1.0 Alepidea amatymbica Sa Hermas quinquedentata Hermas quercifolia Hermas gigantea
FIGURE 9.3 Strict consensus tree of the 2208 equally most parsimonious trees with a length of 534 steps, based on the MP analysis of rps16 sequence data (CI= 0.70 and RI= 0.85). BP's are given above the branches and PP's are given below the branches. Branches supported only in the BI are indicated as dotted lines, while those that differed in the BI are indicated as grey lines.
135 CHAPTER 9: SYSTEMATICS OF THE GENUS EZOSCIA t'
9.3.4 rbcL dataset— The rbcL dataset included 1238 characters of which
126 were variable and 57 parsimony informative. The MP analysis (Fig. 9.4A)
resulted in 3 equally parsimonious trees with a TL of 176 steps, a CI of 0.76 and a
RI of 0.75. The overall topology of the MP trees retrieved in this analysis was found
to be similar to that shown in the rps16 intron trees. Although rbcL sequences for
Molopospermum and Astydamia were not available, the position of Ezosciadium
capense as sister group to the Annesorhiza Glade (represented in this analysis by
Annesorhiza and ltasina) is moderately supported (BP 84).
Berula erecta
A 2i Slum serra B rbcL 84 85 Apium graveolens ITS Heracleum sphondylium 66 Torilis arvensis 63 Itasina filifolia Pleurospermum camtschaticum co co Annesorhiza filicaulis 86 97 Bupleurum fruticosum 57 Annesorhiza macrocarpa Annesorhiza altiscapa 9 Heteromorpha arborescens 52 Annesorhiza Chamarea snijmaniae Glade 89 Anginon rugosum _E 100 Anriesorhiza fibrosa 96 Itasina hrtfolia Annesorhiza Chamarea sp. 63 84 Glade Annesorhiza altiscapa H_ Annesorhiza latifolia Ezosciadium capense 58 Astydamia latifolia 54 Lichtensteinia lacera Molopospermum peloponnesiacum Ezosciadium capense
AESanicula europaea Lichtensteinia obscura 72 Eryngium giganteum ideae
Arctopus echinatus lo
Steganotaenia araliacea icu
6 C Polemanniopsis marlothii San Hermas villosa
FIGURE 9.4 (A), Strict consensus tree of the 3 equally most parsimonious trees with a length of 176 steps, based on MP analysis of rbcL sequence data (CI= 0.76 and RI= 0.75). (B), Single most parsimonious tree with a length of 220 steps, based on ITS sequence data (CI= 0.78 and RI= 0.52).
136 CHAPTER 9: SYSTEMATICS OF THE GENUS EZOSCIAMUM
9.3.5 ITS dataset— The ITS dataset included 638 characters of which 136 were variable and 57 parsimony informative. The MP analysis (Fig. 9.4B) resulted in a single parsimonious tree with a TL of 220 steps, a CI of 0.78 and a RI of 0.52. In this tree, Ezosciadium capense is sister group to the Glade of Astydamia and
Molopospermum (BP 54) which, in turn, is sister group to the Annesorhiza Glade.
9.3.6 Basal African Apiaceae— Burtt (1991) argued that the southern
African taxa of Apiaceae are of importance beyond their relatively small size. Recent molecular systematic studies have indeed shown that African genera are often sister to (or basally divergent within) other major lineages and therefore essential to understanding of relationships within the family as a whole. The basally divergent position of African genera was first shown by early molecular systematic studies which suggested that Heteromorpha and Anginon formed basal-branching lineages within the subfamily Apioideae (Downie et al. 1996, 1998; Plunkett et al. 1996a, b).
Downie and Katz-Downie (1999) subsequently retrieved a broader Heteromorpha
Glade, comprising numerous woody African genera (Anginon, Dracosciadium, Glia,
Heteromorpha and Polemannia). This Glade was later recognised by Downie et al.
(2000b) as the tribe Heteromorpheae. More recently, an analysis focusing on the phylogenetic position of African Apiaceae (Calvino et al. 2006) indicated that several other African genera occupied early diverging positions within the Apioideae.
Lichtensteinia was shown to be the most early diverging lineage sister group to other genera of the subfamily Apioideae. Successively sister to the Lichtensteinia
Glade was a Glade comprising largely African herbaceous genera, the Annesorhiza
Glade (comprising Annesorhiza, Chamarea and Itasina). This Annesorhiza Glade was closely related to Molopospermum and Astydamia, however, these relationships
137 CHAPTER 9: SYSTEMATICS OF THE GENUS EZOSCIAD112,11
were not very strongly supported. Even within the subfamily Saniculoideae, its
relatively small African contingent has been shown to be basally divergent. Downie
and Katz-Downie (1999) demonstrated that two woody African genera (viz.,
Steganotaenia and Polemanniopsis) traditionally placed within the Apioideae, formed a sister group relationship with the subfamily Saniculoideae. These two
genera have subsequently been included in an expanded Saniculoideae by Calvin()
and Downie (2007) as the tribe Steganotaenieae. Furthermore, within their tribe
Saniculeae two African endemic genera (Alepidea and Arctopus) have now also
been shown to occupy early diverging positions, both successively sister to the
remaining genera (Calvin() and Downie 2007; Magee et al. 2008d). In this study, we
report on yet another early diverging branch of subfamily Apioideae from South
Africa. In the results of all molecular analyses, Ezosciadium capense shows a close affinity with the Annesorhiza Glade, the latter representing a group of deciduous, perennial herbs endemic to southern Africa. However, in all cladograms currently available for the group, the relationships among Ezosciadium, the Annesorhiza
Glade, the European genus Molopospermum and the Canary Islands endemic genus
Astydamia are not yet clear and demand further study.
It seems that the basally divergent lineages are morphologically as diverse as the more derived lineages as they include woody and herbaceous elements, with various leaf and fruit types. It is therefore not easy to ascertain the phylogenetic position of most genera without molecular systematic evidence. The lineage comprised of the Annesorhiza Glade, Astydamia, Ezosciadium and Molopospermum has no obvious morphological synapomorphies, but the combined presence of heteromericarpic fruit (in Annesorhiza and Molopospermum), scattered druse
138 CHAPTER 9: SYSTEMATICS OF THE GENUS EZOSCIADIUM
crystals in the mesocarp and the highly lignified vascular bundles (in Annesorhiza
and Ezosciadium) do, to some extent, provide support for this Glade.
9.4 TAXONOMY OF THE GENUS EZOSCIADIUM
EZOSCIADIUM B.L.Burtt in Edinb. J. Bot. 48(2):207, 268 (1991). Trachysciadium
H.Wolff in Pflanzenr. Heft 90: 108 (1927), non Trachysciadium (DC.) Eckl.
and Zeyh. (1837). — TYPE: E. capense (Eckl. and Zeyh.) B.L.Burtt. [Note:
Burtt (1991) gives a detailed argument for considering Trachysciadium as a
synonym of Pimpinella L. and hence the need for the new generic name,
Ezosciadium. De Candolle (1830) described Trachysciadium as a section of
the genus Helosciadium to accommodate two Himalayan species now
included in Pimpinella. The publication of the genus Trachysciadium by
Ecklon and Zeyher (1837) had been considered valid by some authors (e.g.
Wolff, 1927), who regarded it as being validated by the combined generico-
specific description and taxonomically independent of Helosciadium sect.
Trachysciadium (in which case the name Ezosciadium would be superfluous).
We however, prefer to follow Burtt (1991), who argued that there is no
reasonable doubt that Ecklon and Zeyher (1837) adopted De Candolle's
taxon and simply altered its rank. As such, Burtt's proposed Ezosciadium
nom. nov. is valid and necessary]
This highly distinctive genus appears to be an isolated basal lineage within
Apiaceae subfamily Apioideae sister group to the Annesorhiza Glade. DNA sequence data also support a possible close relationship with Astydamia and
139 CHAPTER 9: SYSTEMATICS OF THE GENUS EZOSCIADIUM
Molopospermum, as do some morphological and fruit anatomical characters. The
genus is endemic to the Eastern Cape Province of South Africa within the eastern
part of the Cape Floristic Region.
1. EZOSCIADIUM CAPENSE (Eckl. and Zeyh.) B.L.Burtt in Edinb. J. Bot. 48(2): 207
(1991). Trachysciadium capense Eckl. and Zeyh., Enum. Pl. Afric. Austral.
341 (1837); H.Wolff in Pflanzenr. Heft 90: 108 (1927). Helosciadium capense
(Eckl. and Zeyh.) Sond. in Harv. and Sond., Fl. Cap. 2: 536 (1862).
Trachydium capense (Eckl. and Zeyh.) Drude in Engl. and Prantl,
Pflanzenfam. 3(8): 189 (1898). — TYPE: SOUTH AFRICA. Uitenhage district
(3325): 'Coega Kopje' not far from `Zwartkopsrivier' (–DC), Ecklon and Zeyher
2196 (SAM!, lecto., here designated; MO, photo!, P, photo!, S!, isolecto.).
Erect, annual herb, 0.1-0.35 m tall, pilose. Cotyledons simple, narrowly
oblanceolate, 15 mm x 2 mm, margins entire. Stem solitary, dichotomously
branched. Leaves cauline, 10-37 x 7-26 mm, first leaves tri-lobed, upper leaves
digitately compound. Petioles 5-18 mm long, sheathing slightly at the base. Pinnae
12-24 mm x 5-13 mm, 2- or 3-lobed; lobes linear-oblong to narrowly oblong, 5-12
mm x 1-2 mm, flat, apex acute, margins and venation pilose, venation pinnate.
Umbels compound, sparse, axillary, sessile (peduncle absent, rays arising directly from each node); involucre present; bracts 2, 2-4 mm long, becoming prominently
recurved, lanceolate, apex acute, pilose; rays 2 to 4, unequal in length, with at least one remaining markedly shorter, longer rays 11-31 mm long and shorter ray 3-10
mm long at anthesis, pilose; involucel present; bracteoles 2, 1-2 mm long, becoming prominently recurved, lanceolate, apex acute, pilose; umbellate rays 2 to
140 CHAPTER 9: SYSTEMATICS OF THE GENUS EZOSCIADIU11
4, short, less than 1 mm long at anthesis, pilose. Flowers subsessile, pentamerous, hermaphroditic; sepals obsolete; petals yellow, 0.6-0.7 mm long, ovate to obovate, tips not inflexed, acute, septum absent on inner face, pilose on the dorsal surface; stamens with anthers highly inflexed, small, 0.2-0.4 mm tall; ovary densely pilose, stylopodium flat; styles erect, very short. Fruit isodiametric, oblong, 2.5-3.0 mm x
±1.5 mm broad; mericarps homomorphic, distinctly pilose; median, lateral and marginal ribs 5, equal, prominent; commissural vittae 2; vallecular vittae 4; commissure very narrow; carpophore bifid for the upper two thirds of its length, persisting partly on the plant. Figure 9.1.
Diagnostic characters— Ezosciadium capense is a highly distinctive species, easily distinguished from all other annual species by the pilose vegetative and reproductive organs, the sessile compound umbels with a few, markedly unequal rays (Fig. 9.1A), the non-inflexed petal tip (Fig. 9.1H), the relatively small, highly inflexed stamens which appear almost sessile (Fig. 9.11) and the prominent bifid carpophores, one half of which persists on the plant (Fig. 9.1A, E).
Distribution and habitat— This poorly collected species is endemic to the
Eastern Cape Province of South Africa and is known from only four recorded populations (Fig. 9.5), three of which have only come to light recently. Originally known only from Coega koppie near Uitenhage, the distribution range has been expanded to include Perdepoort (the westernmost locality), Joubertina and
Bethelsdorp.
141
CHAPTER 9: SYSTEMATICS OF THE GENUS EZOSC/ADM1/
Additional specimens examined.
—3322 (Outshoorn): Perdepoort, N of Camfer (—CD), Goldblatt & Porter 12578 (NBG).
3323 (Willowmore): 4 miles N by W of Joubertina (—DD), Acocks 20021 (PRE).
3325 (Uitenhage): Bethelsdorp, salt pan (—DC), Fries et al. 1156 (BM, PRE).
Precise locality unknown: Anon s.n. herb. S sheet 243 (S).
HEIGHT ABOVE SEA LEVEL WA Over 1500 m 900 - 1500 m 300 - 900 m Under 300 m
20 0 20 40 60 80 100 km .11111 , 11111j
FIGURE 9.5 The known geographical distribution of Ezosciadium capense.
142 CHAPTER 10
NEW TRIBAL DELIMITATIONS FOR THE EARLY DIVERGING LINEAGES OF
APIACEAE SUBFAMILY APIOIDEAE
10.1 INTRODUCTION
The Apiaceae are a large and taxonomically complex family comprising 463
genera and ca. 3,500 species (Plunkett et al. in press), with a near cosmopolitan
distribution. The classification system of Drude (1897-98) made little provision for
the comparatively small sub-Saharan African and Madagascan contingent (80
genera and 354 species), many of which were unknown or poorly studied at the
time. As suggested by Burtt (1991) and later by molecular systematic studies
(Calvin() and Downie 2007; Calvin° et al. 2006; Downie et al. 1996, 1998, 2000b;
Downie and Katz-Downie 1999; Magee et al. 2008a, b; Plunkett et al. 1996a, b) the
African genera are crucial to an understanding of the evolutionary history of the
family and its full range of morphological diversity. Many African taxa have indeed
been found to occupy early diverging positions within Apiaceae, suggesting a
southern African origin for both Apioideae (Calvin° et al. 2006) and Saniculoideae
(Calvin° and Downie 2007). In recent years tribal delimitations within the largest and
most complicated subfamily, the Apioideae, have been the focus of intensive
phylogenetic research (see Downie et al. 2001; Downie et al. in press). The most
recent treatment of the "core" apioid lineages by Downie et al. (in press), based
largely on nrITS sequence data, recognized 14 tribes and 12 major clades. In this treatment the earliest diverging lineages of the subfamily were excluded due to their highly divergent ITS sequences and consequent alignment difficulties. Although
143 CHAPTER 10: NEW TRIBAL DELIMITATIONS there has been much progress on the generic placement of several anomalous taxa within these early diverging lineages (Calvino et al. 2006; Calvino et al. in prep.;
Magee et al. 2008a) the correct placement of several critical African taxa has remained uncertain. This paper is aimed at resolving the early diverging lineages of the Apioideae, assessing their relationship to their sister subfamily Saniculoideae and proposing a new tribal classification for these lineages using anatomical, morphological and DNA sequence data.
10.2 MATERIAL AND METHODS
10.2.1 Taxon sampling— The cpDNA trnQ-trnK region for 46 accessions, representing 27 genera and 42 species of Saniculoideae and early diverging lineages of Apioideae and two accessions of the South African azorelloid genus
Hermas. were examined. Sources of material, together with their corresponding taxonomic authorities and GenBank accession numbers, are listed in Appendix 1.
Data from the trnQ-trnK region for 27 of these 46 accessions were available from previous studies (Calvino and Downie 2007; Calvin() et al. 2008; Downie et al. 2008;
Tilney et al. 2009). Sequence data from the rps16 intron region were available for a further ten accessions (Calvino et al. 2006; Downie and Katz-Downie 1999; Magee et al. 2008a). Rps16 intron data for the remaining nine accessions and data from the tmQ-rps16 and rps16-tmK intergenic spacers for 15 and 17 accessions, respectively, were obtained specifically for this study. The monotypic southern
African genera Choritaenia, Marlothiella and Phlyctidocarpa were included as previous authors had suggested their placements within the early diverging lineages of the Apioideae-Saniculoideae Glade (Calvino et al. 2006; Lui et al. 2003, 2007a, b;
144 CHAPTER 10: NEW TR1 BA L DELIMITATIONS
Nicolas and Plunkett 2009; Tilney et al. 2009). The genus Hermas (Azorelloideae) was used to root the trees, based on results from previous molecular phylogenetic analyses (Calvino et al. 2006).
10.2.2 Phylogenetic analyses— For analyses of the entire cpDNA trnQ-trnK region, the portion of the matrix representing the tmQ-rps16 5' exon region was scored as missing in three accessions (Polemannia grossulariifolia, Chamarea snijmaniae and Steganotaenia commiphoroides) and the portion representing the rps16 3' exon-trnK region was scored as missing in one accession (Polemanniopsis sp. 1) because of difficulties in amplifying these regions in these taxa. In addition, portions of the tmQ-rps16 5' exon region for one accession of Choritaenia capense
(1618 bp) and of the rps16 3' exon-trnK region for one accession of Phlyctidocarpa flava (580 bp) were also unobtainable despite our best efforts. In Molopospermum peloponnesiacum we were unable to amplify both intergenic spacer regions. Overall, missing data represented approximately 10% of the entire matrix.
Phylogenetic analyses were conducted initially using the parsimony (MP) algorithm as described in Chapter 2, with gaps treated as binary scored characters.
Tree searches were performed using a heuristic search with 1,000 random sequence additions, tree bisection-reconnection (TBR) branch swapping, and the
MULPARS option in effect. Bootstrap percentage values were determined from
1,000 bootstrap replicates, holding 10 trees per replicate, with TBR and MULPARS selected. Bayesian inference was performed, as outlined in Chapter 2, for 2,000,000 generations of Monte Carlo Markov Chains and a sampling frequency of 100, with the K81uf+G model and the 'standard' model (using default parameters) implemented for the nucleotide and indel data, respectively.
145 CHAPTER 10: NEw TRIBAL DELIMITATIONS
Fourteen morphological and anatomical characters (Table 10.1 and 10.2) considered important for defining early diverging lineages within the Apioideae-
Saniculoideae Glade were scored for the 46 taxa included in the molecular analyses.
Character data were taken from field observations of the South African taxa, herbarium specimens and literature (Table 10.1 and 10.2; Winter et al. 1993; Vessio
2001; Lui 2004; Lui et al. 2003, 2007a, 2007b, 2009; Magee et al. 2008a, 2008b;
Tilney and Van Wyk 2001; Tilney et al. 2009; Yembaturova et al. 2009, Van Wyk et al. 1999). These morphological data were then reconstructed on the MP trees.
TABLE 10.1 Morphological and anatomical characters and states examined during this investigation and optimised onto the molecular trees.
1. Habit (herbaceous = 0; woody = 1); 2. Leaf persistence (not proteranthous = 0; proteranthous = 1); 3. Umbel (compound = 0; simple = 1); 4. Flowers (pedicellate = 0; sessile or subsessile = 1); 5. Involucre bracts (not forming a pseudanthium = 0; forming a prominent pseudanthium = 1); 6. Fruit surface (glabrous = 0; surface vesicles = 1; spines/bristles = 2; stellate hairs =3); 7. Fruit wing/rib development (pseudo-wings/ribs = 0; true wings/ribs = 1); 8. Fruit ribs shape (simple = 0; bifurcate = 1); 9. Fruit symmetry (homomericarpic = 0; heteromericarpic = 2); 10. Endocarp (woody = 0; parenchymatous/lignified = 1); 11. Regular vittae (absent = 0; present = 1; oil vesicles = 2); 12. Rib oil ducts (small = 0; large = 1; forming cavities = 2); 13. Rhomboidal crystals (present = 0; absent = 1); 14. Druse crystals (absent = 0; scattered throughout mesocarp = 1; restricted to commissure = 2).
146 CHAPTER 10: NEW TRIBAL DELIMITATIONS
TABLE 10.2 Matrix of morphological and anatomical character states optimised onto the molecular trees. Dashes represent non-applicable character states that were scored as missing in the analysis. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 Hermas gigantea 0 0 0 0 0 0 0 0 0 0 0 0 1 0 Hermas quinquedentata 0 0 0 0 0 0 0 0 0 0 0 0 1 0 Sium suave 0 0 0 0 0 0 1 0 0 1 1 0 0 0 Physospermum cornubiense 0 0 0 0 0 0 0 0 1 1 0 0 0 Bupleurum angulosum 0 0 0 0 0 0 - 0 0 1 1 0 0 0 Bupleurum fruticosum 1 0 0 0 0 0 - 0 0 1 1 0 0 0 Polemannia grossulariifolia 1 0 0 0 0 0 1 0 0 1 1 0 0 1 Dracosciadium italae 0/1 0 0 0 0 0 - 0 0 1 1 0 0 1 Anginon paniculatum 1 0 0 0 0 0 - 0 0 1 1 0 0 1 Anginon difforme 1 0 0 0 0 0 1 0 0 1 1 0 0 1 Heteromorpha arborescens 1 0 0 0 0 0 1 0 1 1 1 0 0 1 Annesorhiza altiscapa 0 0 0 0 0 0 1 0 1 1 1 0 0 1 Annesorhiza macrocarpa 0 1 0 0 0 0 1 0 1 1 1 0 0 1 Chamarea snijmaniae 0 1 0 0 0 0 - 0 0 1 1 0 0 1 Itasina filifolia 0 0 0 0 0 0 0 0 1 1 0 0 1 Ezosciadium capense 0 0 0 0 0 0 - 0 0 1 1 0 0 2 Astydamia latifolia 0 0 0 0 0 0 1 0 0 1 1 0 0 1 Molopospermum peloponnesiacum 0 0 0 0 0 0 1 0 1 1 1 0 0 1 Choritaenia capensis 0 0 0 0 0 0 1 0 0 0 2 0 0 0 Choritaenia capensis 0 0 0 0 0 0 1 0 0 0 2 0 0 0 Marlothiella gummifera 1 0 0 0 0 3 0 1 1 0 1 0 1 Lichtensteinia obscura 0 1 0 0 0 0 - 0 0 1 0 1 0 1 Lichtensteinia trifida 0 1 0 0 0 0 - 0 1 1 0 1 0 1 Lichtensteinia globosa 0 1 0 0 0 0 - 0 0 1 0 1 0 1 Lichtensteinia lacera 0 1 0 0 0 0 - 0 1 1 0 1 0 1 Phlyctidocarpa flava 0 0 0 0 0 1 - 1 0 1 1 1 0 1 Phlyctidocarpa flava 0 0 0 0 0 1 - 1 0 1 1 1 0 1 Polemanniopsis sp. 1 1 1 0 0 0 0 1 0 1 1 0 2 0 1 Polemanniopsis marlothii 1 1 0 0 0 0 1 0 1 1 0 2 0 1 Steganotaenia araliacea 1 1 0 0 0 0 1 0 1 1 0 2 0 1 Steganotaenia commiphoroides 1 1 0 0 0 0 1 0 1 1 0 2 0 1 Alepidea amatymbica 0 0 1 1 1 0 - 1 0 1 1 1 0 1 Alepidea capensis 0 0 1 1 1 2 0 0 1 1 1 0 1 Alepidea peduncularis 0 0 1 1 1 2 - 0 0 1 0 1 0 1 Arctopus echinatus 0 0 1 1 1 2 - 0 0 1 0 0 0 0 Actinolema macrolema 0 0 1 1 1 2 - 0 0 1 0 1 0 1 Astrantia major 0 0 1 0 1 1 - 0 0 1 0 1 0 1 Astrantia minor 0 0 1 0 1 1 - 0 0 1 0 1 0 1 Hacquetia epipactis 0 0 1 1 1 2 - 0 0 1 0 1 0 1 Petagnaea gussonei 0 0 1 1 1 0 0 0 1 0 0 0 0 Sanicula canadensis 0 0 1 1 1 2 0 0 1 0 1 0 1 Sanicula europaea 0 0 1 1 1 2 0 0 1 0 0 0 1 Eryngium maritimum 0 0 1 1 1 2 1 0 0 1 0 1 0 1 Eryngium giganteum 0 0 1 1 1 2 1 0 0 1 0 1 0 1 Eryngium palmatum 0 0 1 1 1 2 0 0 1 0 1 0 1 Eryngium planum 0 0 1 1 1 2 0 0 1 0 1 0 1
147 CHAPTER 10: NEW TRIBAL DELINIITATIONS
10.3 RESULTS
10.3.1 Phylogenetic analyses— The matrix, representing the entire trnQ- trnK region, consisted of 5,072 unambiguously aligned positions and 587 binary scored indels, and resulted in 2,045 variable and 1,285 parsimony informative characters. MP analyses of these data yielded two minimal length trees, each of
2,113 steps [ensemble consistency indices (CI; Kluge and Farris 1969) of 0.75 and
0.66, with and without uninformative characters, respectively; ensemble retention index (RI; Farris 1989) of 0.86]. The MP strict consensus tree was identical in topology to that inferred from the BI analysis (Fig. 10.1). In both analyses, the same clades and relationships as reported previously (Calvin() et al. 2006; Ca'vino and
Downie 2007; Magee et al. 2008a) were retrieved. The subfamily Saniculoideae sensu Calvin° & Downie is monophyletic (PP 0.99, BP 74), with tribes Saniculeae
(i.e., Saniculoideae s.s.) and Steganotaenieae strongly supported (PP 1.0, 1.0, BP
100, 99, respectively). Phlyctidocarpa is sister group to Polemanniopsis and
Steganotaenia (tribe Steganotaenieae) with strong or weak support depending on the analysis (PP 0.97, BP 71). Subfamily Apioideae sensu Ca!vino & Downie is also supported as monophyletic (PP 1.0, BP 76) and is sister group to subfamily
Saniculoideae sensu Calvin° & Downie (PP 1.0, MP 100). Within the apioid Glade a strongly supported subclade comprising Choritaenia, Lichtensteinia Cham. &
Schltdl. and Marlothiella (PP 1.0, MP 100) is sister group to all remaining Apioideae lineages. The accessions of Bupleurum (tribe Bupleureae), Physospermum Cusson
(tribe Pleurospermeae) and Sium L. (tribe Oenantheae), representing the upper lineages of the subfamily, are a sister group to a strongly supported tribe
Heteromorpheae (PP 1.0, BP 100). Successively sister to this group is a strongly
148 below thebranches. presented abovethebranches. Bootstrappercentage(BP)values(>50%) arepresented subfamilial circumscriptionsindicatedalongside. Posterior probability(PP)valuesare FIGURE 10.1 100 1.0 0.99 74 100 1.0 76 1.0 Bayesian inference(BI)treeof 0.87 50 100 1.0 100 1.0 0.97 100 71 100 1.0 89 1.0 53 1.0 1.0 90 0.93 0.71 100 79 100 1.0 100 100 100 1.0 100 1.0 1.0 1.0 1 100 . 1.0 0.84 0 100 100 100 1.0 1.0 1.0 67 1.0 96 99 1.0 95 1.0 99 1.0 72 .69 100 C 1.0 r E r E 0.69 100 1.0 1.0 1.0 1.0 1.0 100 1.0 100 98 96 1.0 .82 63 71 1.0 1.0 95 70 92 I-- Hermasgigantea 1-Hermas quinquedentata = = = = 1- trnQ-trnK ,
Astrantia minor Astrantia major Actinolema macrolema Petagnaea gussonei Eryngium palmatum Eryngium giganteum Eryngium planum Arctopus echinatus Alepidea peduncularis Eryngium maritimum Alepidea capensis Alepidea amatymbica Sanicula canadensis Sanicula europaea Hacquetia epipactis Astydamia latifolia Steganotaenia commiphoroides Polemanniopsis marlothii Polemanniopsis sp.1 Phlyctidocarpa flava Phlyctidocarpa flava Annesorhiza macrocarpa Lichtensteinia trifida Lichtensteinia obscura Annesorhiza altiscapa Steganotaenia araliacea Lichtensteinia globosa Lichtensteinia lacera Marlothiella gummifera Anginon difforme Anginon paniculatum Molopospermum peloponnesiacum Ezosciadium capense Choritaenia capensis Choritaenia capensis Itasina filifolia Dracosciadium italae Heteromorpha arborescens Chamarea snijmaniae Polemannia grossulariifolia Bupleurum fruticosum Bupleurum angulosum Physospermum cornubiense Sium suave sequence datawiththeprevious CHAPTER 10: NEW TRIBALDELIMITATIONS
Azorelloideae
s•s eeeppinopes s•s eeep!o!dv
149
e e owni D & ° n vi al C u sens eae e owni d oi ul c ani S D & C u s ° ° n vi al en s eae d oi pi A CHAPTER 10: NEW TRIBAL DELIMITATIONS supported Annesorhiza Glade (PP 1.0, BP 89), comprising Annesorhiza, Astydamia,
Chamarea, Ezosciadium, ltasina and Molopospermum.
10.3.2 Reconstruction of morphological and anatomical characters-
Parsimony-based reconstructions of 14 morphological and anatomical characters
(Table 10.1) considered important for defining early diverging lineages within the
Apioideae-Saniculoideae Glade are each summarised onto one of the two minimal length trees inferred from MP analysis of trnQ-trnK nucleotide substitution and scored indel data (Fig 10.2). True wings/ribs on the fruit (character 7, Fig. 10.2G;
Fig. 10.3E, H–K), parenchymatous or lignified endocarp (character 10, Fig. 10.2J;
Fig. 10.3C, D, F–K), absence of rhomboidal crystals (character 13, Fig. 10.2M), and the presence of druse crystals scattered throughout the mesocarp (character 14,
Fig. 10.2N) were reconstructed as synapomorphies for the Apioideae-Saniculoideae
Glade. The base of the trees was reconstructed as ambiguous for each of the aforementioned characters. However, if a further outgroup of either Azorelloideae or
Mackinlayoideae had been included, this node would certainly be reconstructed for the pleiseomorphic state as in Hermas. As shown by Calvin° et al. (2008),
Saniculoideae s.s. (i.e. tribe Saniculeae sensu Calvin° & Downie) is supported by the presence of simple umbels (character 3, Fig. 10.2C), sessile or subsessile fructiferous flowers (character 4, Fig. 10.2D), large involucre bracts forming a prominent pseudanthium (character 5, Fig. 10.2E), and fruit with surface outgrowths
(character 6, Fig. 10.2F). No characters were found to support the clades comprising either Saniculoideae sensu Calvin° & Downie (Phlyctidocarpa-
Petagnaea) or Apioideae sensu Calvin() & Downie (Sium–Lichtensteinia). The presence of regular vittae (character 11, Fig. 10.2K; Fig. 10.31–K) was reconstructed
150 CHAPTER 10: NEW TRIBAL, DELIMITATIONS
as a synapomorphy for the lineages Sium—Molopospermum and two species of
Alepidea (Fig. 10.3D), and as an apomorphy for Phlyctidocarpa (Fig. 10.3G). Large
rib oil ducts (character 12, Fig. 10.2L; Fig. 10.3C, D, F—H), traditionally a character
for Saniculoideae s.s., was ambiguously reconstructed as either a synapomorphy for
the Apioideae—Saniculoideae Glade with secondary reversals in the upper lineages
(from the Annesorhiza Glade upwards), Arctopus, Sanicula europaea, and
Petagnaea or as a convergent character state in Saniculoideae and in Lichtensteinia
and Marlothiella. The woody habit (character 1, Fig. 10.2A) was reconstructed as
convergent for both Marlothiella and the tribe Steganotaenieae, and was ambiguous
for the ancestor to tribe Heteromorpheae. Although Dracosciadium is a rhizomatous
herb, character 1 (Table 10.2) was coded as polymorphic to reflect the missing
Madagascan taxa with which it is allied (Calvin° et al. in prep.). As this genus forms
a Glade with the woody Madagascan genera, it is likely that with more accessions
the woody habit may be reconstructed as a synapomorphy for the Heteromorpheae,
with the herbaceous habit of Dracosciadium reconstructed as a reversal. Bifurcate
ribs (character 8, Fig. 10.2H), surface vesicles (character 6, Fig. 10.2F), and regular
vittae (character 11, Fig. 10.2K) seem to have evolved independently in
Phlyctidocarpa (Fig. 10.3G) and other lineages. The proteranthous leaves (character
2, Fig. 10.2B) in Lichtensteinia and Steganotaenieae were reconstructed as synapomorphies for each lineage. Steganotaenieae was also supported by the presence of heteromericarpic fruit (character 9, Fig. 10.21), where only the sepaline ribs are well-developed (Fig. 10.3H). This character was also found in the genus
Heteromorpha (Fig. 10.3K).
The number of parsimony informative indels and reconstructed morphological apomorphies are indicated on a BI phylogram (Fig. 10.4) above and below
151 -
CHAPTER 10: NEW TRIBAL DELIMITATIONS
1. Habit 2. Leaf persistence 3. Umbel herbaceous not proteranthous o compound ■ woody proteranthous ® simple 11. Hermas gigantea Hennas quinquedentata , Sium suave Physospermum comubiense Bupleurum angulosum Bupleutum fruticosum Li tz1teemamnno„ipa goasrsutagfoliaooeens
Angmon putumanicla Anginon difforme •, Dracosciadium italae Annesorhiza altiscapa E•, Annesorhiza macmcarpa = Chamarea snijmaniae ., ltasina filifolia ., Astydamia latifolia ,_I, Ezosciadium capense it_u Molopospermum peloponnesiacum Choritaenia capensis I-I Choritaenia capensis Marlothiella gummifere Lichtensteinia obscura Lichtensteinia bifida H _F Lichtensteinia lacera --1-11 71 1 Lichtensteinia globosa 7_1 Phlyctidocarpa Sava Phlyctidocarpa Sava Polemanniopsis sp. 1 Polemanniopsis marlothit Steganotaenia arafracea Steganotaenia commiphoroides Alepidea amatymbica Alepidea capensis Alepidea peduncularis Arctopus echinatus Actinolema macrolema Astrantia major Astrantia minor Hacquetia epipactis Sanicula europaea Sanicula canadensis Eryngium maritimum Eryngium plenum Eryngium giganteum A B C Eryngium palmatum Petagnaea gussonei
4. Flowers 5. Involucre bracts 6. Fruit surface o pedicellate not forming a pseudanthium glabrous surface vesicles ■ sessile or subsessile forming a prominent spines/bristles pseudanthium ■ stellate hairs 7,,, Hermes gigantea Hermes quinquedentata .1 Sium suave Physospermum comubiense Bupleurum angulosum Bupleurum fruticosum Iteemm amnonrpia gross° ulmansicteelnias Li He Anginon paniculatum Anginon diffomre ., Dracosciadium italae Annesorhiza altiscapa Annesorhiza macrocarpa Chamarea snijmaniae ltasina filifolia , Astydamia latifolia ,i. Ezosciadium capense Molopospermum peloponnesiacum Choritaenia capensis Choritaenia capensis Martothiella gummifera Lichtensteinia obscure Lichtensteinia trifida Lichtensteinia lacera Lichtensteinia globosa r , Phlyctidocarpa Sava Phlyclidocarpa flare Polemanniopsis sp. 1 Polemannlopsis marlothii Steganotaenia araliacea Steganotaenia commiphoroides Alepidea amatymbica ' o Alepidea capensis Alepidea peduncularis Arctopus echinatus Actinolema macrolema Astrantia major Th Astrantia minor Hacquetia epipactis Sanicula europaea LI Sanicula canadensis Eryngium maritimum L' i Eryngium planum D F Eryngium giganteum Eryngium palmatum 1 Petagnaea gussonei
FIGURE 10.2 Reconstruction of morphological characters 1 to 14, when optimized over one of the two minimal length trees inferred from MP analysis of the trnQ-trnK data. The proposed new classification is indicated alongside Fig. 2N.
152 -
CHAPTER 10: NEW TRIBAL DELIMITATIONS
7. Fruit wing/rib development 8. Fruit ribs shape 9. Fruit symmetry pseudo-wings/ribs ❑ simple homomericarpic true wings/ribs ■ bifurcate heteromericarpic
Hernias gigantea Hermas quinquedentata Sium suave Physospermum comubiense Bupleurum angulosum Bupleurum fruticosum Polemannia grossuladifolia Heteromorpha arborescens Anginon paniculaturn Anginon difforme Dracosciadium italae Annesorhiza attiscapa Annesorhiza macrocarpa Chamarea snijmaniae ltasina WOO p- Astydamia latifolia Ezosciadium capense Molopospemium peloponnesiacum Choritaenia capensis Choritaenia capensis Marlothiella gummifera Lichtenstelnia obscura Lichtensteinia bifida Lichtensteinia lacers Lichtensteinia globosa Phlyctidocarpa 'lava Phlyctidocarpa flora Polemanniopsis sp. I Polemannsopsrs martotho Steganotaerva araliacea Steganotaenia commiphoroides Alepidea amatymbica o Alepidea capensis Alepidea peduncularis Arctopus echinatus Actinolema macrolema Astrantia major Astrantia minor Hacquetia epipactis Sanicula euroPaea Sanicula canadensis Eryngium maritimum Eryngium plenum Eryngium giganteum G H Eryngium palmatum Petegnaea gussonei
10. Endocarp 11. Regular vittae 12. Rib oil ducts woody absent small parenchymatous/lignified present large ■ oil vesicles forming cavities
1,--1.1 Hernias gigantea L_' a, Hermas quinquedentata Sium suave I-13 Physospermum comubiense Bupleurum angulosum Bupleurum fruticosum __1.. , Polemannia grossulanifolia Heteromorpha arborescens Anginon paniculatum Anginon difforme Dracosciadium italae Annesorhiza altiscapa Annesorhiza macrocarpa Chamarea snijmaniae Itasina (Molls Astydamia latifolia i__..0 Ezosciadium capense I—JJ Molopospermurn peloponnesiacum -,., Choritaenia capensis L._,,, Choritaenia capensis , Marlothiella gummifera . Lichtensteinia obscura Lichtensteinia bifida --- Lichtensteinia lacers 1 Lichtensteinia globosa , Phlyctidocarpa tiara li.., I ■ Phlyctidocorpo flora Polemanniopsis sp. 1 Polemanniopsis marlothil Steganotaenia araliacea Steganotaenia commiphoroides j----■ Alepidea amatymbica , Alepidea capensis , Alepidea pedunculatis •i Arctopus echinatus Actinolema macrolema I I , Astrantia major ---11 Astrantia minor . Hacquetia epipactis 1171±=0 Sanicula europaea .j Sanicula canadensis i , Eryngium maritimum __r L— Eryngium plenum L Eryngium giganteum J Eryngium palmatum • I 1 Petagnaea gusscnei
FIGURE 10.2 (Cont.)
153
CHAPTER 10: NEW TRIBAL DELINHTATIONS
13. Rhomboidal crystals 14. Druse crystals present absent absent scattered throughout mesocarp restricted to commissure Hermes gigantea Hermes quinquedentata Azorelloideae Sium suave J Oenantheae m — Physospermum comubiense 3 Pleurospermeae Bupleurum angulosum Bupleurum fruticosum Bupleureae a.§: Polemannia grossularlifolia — Heteromorpha arborescens Anginon paniculatum Heteromorpheae Anginon difforme Dracosciadium italae Annesorhiza altiscapa 1 Annesorhiza macrocarpa Chamarea snljmaniae Itasina filifolia Annesorhizeae Astydamia latilotia Ezosciadium capense Moloposperrnum peloponnesiacumJ Chonfaenia capensis Choritaenia capensis Choritaenieae Marlothiella gummlfera Marlothielleae Lichtensteinia obscure Lichtensteinia trifida Lichtensteinieae Lichtensteinia lacera -o Lichtensteinia globose Apioideae s.I. Phlyctidocarpa lava 0 Phlyctidocarpa !lava Phlyctidocarpeae -o Polemanniopsis sp. I 0 Polemanniopsis martothii a: Steganotaenia araliacea Steganotaenieae Steganotaonia commiphoroides MEI Alepidea amatymbica Alepidea capensis Alepidea peduncularis Arctopus echinatus Actinolema macrolema Astrantia major Astrantia minor Hacquetia epipactis Saniculeae Sanicula eumpaea Sanicula canadensis Eryngium mantimum Eryngium plenum M ■ N Eryngium giganteum I Eryngium palmatum Petegnaea gussonei
FIGURE 10.2 (Cont.)
branches, respectively. The branch leading to the Apioideae-Saniculoideae
Glade was relatively long and supported by 31 parsimony informative indels
(although until more outgroup taxa from the Azorelloideae are included, we cannot
be certain how many of these indels are synapomorphic for Hermas) and four
morphological synapomorphies. In contrast, the branches leading to Saniculoideae sensu Calvin° & Downie (Phlyctidocarpa—Petagnaea) and Apioideae sensu Calvin°
& Downie (Sium—Lichtensteinia) were both relatively short, supported by only three and two parsimony informative indels, respectively (Fig. 10.4), and without any reconstructed morphological synapomorphies.
154 CHAPTER 10: NEW TRIBAL DELINIITATIONS
10.4 DISCUSSION
10.4.1 Euapioids and Protoapioids— The early diverging lineages (a
paraphyletic assemblage herein referred to as the protoapioids) of the Apioideae-
Saniculoideae Glade are readily separated from the remaining apioid lineages (the
euapioids) by the presence of scattered druse crystals of calcium oxalate in the
mesocarp of the fruit (character 14, Fig. 10.2N). Optimization of this character by
Calvin° et al. (2008) suggested that the scattered druse crystals in the mesocarp
were either absent or ambiguously reconstructed in the ancestor of the Apioideae-
Saniculoideae Glade. This was, however, the result of the incorrect scoring of this
character as absent in some of the protoapioid taxa (viz. Anginon, Polemanniopsis,
Steganotaenia and several species of Lichtensteinia). Druse crystals are often dissolved and therefore easily overlooked in PAS/TB stained anatomical sections.
Further confounding this problem, they may be only sparsely distributed throughout the fruit mesocarp, so that many sections need to be studied. After careful examination of both stained and unstained (where necessary) fruit sections, we are now able to confirm the presence of scattered druse crystals in almost all protoapioid taxa. Druse crystals were absent only in Arctopus, Choritaenia and
Petagnaea, and restricted to the commissure in Ezosciadium. These results confirm the suggestion by Liu et al. (2006) that the presence of scattered druse crystals in the mesocarp is a synapomorphy for the Apioideae-Saniculoideae Glade.
10.4.2 Phylictidocarpa— In both the BI and MP trees the two accessions of
Phlyctidocarpa flava comprised a strongly supported monophyletic group (PP 1.0,
BP 100), weakly to strongly supported as a sister group to the tribe Steganotaenieae
155 CHAPTER 10: NEW TRIBAL DELIMITATIONS
(PP 0.97, BP 71). This monotypic Namibian endemic genus had previously been
placed within Drude's (1897-98) subfamily Apioideae, tribe Ammineae, by Theobald
and Cannon (1973), based on the presence of crystals in the commissure and at the
base of the fruit ribs, as well as the arrangement of the vittae. These authors
mentioned the similarity of the scattered druse crystals with members of the
Saniculoideae s.s., but the significance of this similarity remained unclear to them. It
has subsequently become evident that the presence of scattered druse crystals in
the fruit is not restricted to the Saniculoideae s.s. (Liu & al., 2006; character 14, Fig.
10.2N), but that this character extends also into the early diverging lineages of
Apioideae s.s. Calving, & Downie (2007) suggested the placement of Phlyctidocarpa
within Saniculoideae based on a preliminary molecular phylogenetic study but the
genus was not yet formally included in their broadened circumscription of
Saniculoideae. The isolated position of Phlyctidocarpa within the Glade comprising
both Steganotaenieae and Saniculeae, as found in this study, has subsequently also
been confirmed by Nicolas and Plunkett (personal communication) using trnT-trnD
and rp116 intron data. However, the results from these authors place Phlyctidocarpa
successively sister to Steganotaenieae and Saniculeae, as opposed to the direct sister relationship with Steganotaenieae recovered in our analyses. Phlyctidocarpa
is unusual in that, like many of the early diverging protoapioid lineages, it combines characters traditionally used to define both Apioideae s.s. and Saniculoideae s.s.
The fruit (Fig. 10.3G) have unusual surface blisters on the ribs and large rib oil ducts
(as in most Saniculoideae s.s.), together with prominent regular vittae (as in most
Apioideae s.s.). Although the surface blisters are superficially similar to those found in the saniculoid genus Astrantia L.(Fig. 10.3C), they differ in that the ribs on which they are borne are bifurcate in transverse section (Fig. 10.3G) due to a longitudinal
156 CHAPTER 10: NEW TRIBAL DELIMITATIONS
' •••■•, ‘.4..__ Iffai) ,„ 4olfil •,,. ... 444,-;,,,iiiiffliniii.",,, : - ' .4 .° Wor•,,Pailihr ''''''''''''''''''''''77:
FIGURE 10.3 A, Myodocarpus involucratus; B, Hermas villisa; C, Astrantia major; D, Alepidea cordifolia; E, Choritaenia capensis; F, Lichtensteinia trifida; G, Phlyctidocarpa flava; H, Polemanniopsis marlothii; I, Annesorhiza macrocarpa; J, Anginon difforme; K, Heteromorpha arborescens. Vouchers: A, Lowry & Oskolski 4639 (MO); B, Anon. s.n. (JRAU); C, Pauca 165b (PE); D, Van Wyk 4232 (JRAU); E, Hanekom 1834 (PRE); F, Winter & Tilney 4164 (JRAU);G, Gress et al. 6075 (PRE); H, Taylor 11269 (PRE); I, Rourke 1700 (NBG); J, Van Wyk 2944 (JRAU); K, Greenway 12558 (PRE). br, bifurcate rib; Ir, lateral rib; medr, median rib; mr, marginal rib; ov, oil vesicle; rod, rib oil duct; sv, surface vesicle; vb, vascular bundle; vv, vallecular vitta; wc, wing cavity; we, woody endocarp. Scale: A—E, I—K=0.8 mm; F=0.5 mm; G=1 mm H=0.4 mm. 157 CHAPTER 10: Ni' TRIBAL DELIMITATIONS division or groove. As a result, both the vallecular vittae and rib oil ducts appear, in transverse section, to be located in the valleculas between the ribs. Somewhat bifurcate ribs are also found in Alepidea woodii Oliv. and A. cordifolia B.-E.van Wyk
(Yembaturova et al. in press).
10.4.3 Choritaenia, Lichtensteinia and Marlothiella— Previous analyses of molecular sequence data (Calvin° et al. 2006; Calvin° and Downie 2007) placed the
South African endemic genus Lichtensteinia as the earliest diverging lineage within
Apioideae sensu Calvin° & Downie. Recently, in an assessment of the subfamily
Azorelloideae, Nicolas and Plunkett (2009) using molecular data showed that the anomalous genus Choritaenia was not related to the other genera of the
Azorelloideae but rather formed a Glade together with Lichtensteinia, that was weakly supported as sister group to Saniculoideae sensu Calvin° & Downie. In the present study, both the MP and BI trees recover a strongly supported Glade (PP 1.0,
BP 100) comprising Choritaenia, Lichtensteinia and Marlothiella, with this Glade as sister group to the remaining members of Apioideae sensu Calvin° & Downie.
Choritaenia is an annual herb endemic to the dry interior of southern Africa (Lui et al.
2007a) and clearly represents a highly adapted lineage. Traditionally placed within the Hydrocotyloideae (now largely included in Azorelloideae and Mackinlayoideae) due to the presence of a woody endocarp, Choritaenia differs from the other members of Azorelloideae in the conspicuous oil vesicles located in its wings (an autapomorphy for the genus), absence of rhomboidal druse crystals, marginal wings composed entirely of the mesocarp, and further lignification of the mesocarp (Fig.
10.3E). Amongst Apiaceae included in this investigation wings may develop in one of two ways (Calvin° et al. 2008). In the first, wings are due to the compression or
158 CHAPTER 10: NEW TRIBAL DELIMITATIONS folding of the carpel and are as such composed of both the mesocarp and endocarp with the vascular bundle located at the margin (Fig. 10.3A and B). This type
(hereafter referred to as pseudo-wings) seems to be the plesiomorphic state, present in wing-fruited members of Azorelloideae, Mackinlayoideae, and even in the closely related families Araliaceae and Myodocarpaceae. In the second wing configuration type, the wings are due to an expansion of the mesocarp and are as such composed only of mesocarp, with the vascular bundle usually located at its base (Fig. 10.3E—J). This type (hereafter referred to as true wings) is restricted to winged or prominently ribbed members of the Apioideae-Saniculoideae Glade and was reconstructed in this study as a synapomorphy for this group (character 7, Fig.
10.2G). Calvin() et al. (2008) also described a third wing type composed entirely of exocarp in some lineages of Eryngium L., but this state was not observed in any of the fruit investigated in this study. It clearly represents a secondary apomorphy for individual lineages within Eryngium.
Tilney et al. (2009) suggested that the Namibian endemic genus Marlothiella may be the closest relative of Lichtensteinia, due to the shared presence of concentric rings of cells around the large rib oil ducts and the marked differences that may be present in the size of the rib ducts (Fig. 10.3F) so that the fruit may appear heteromericarpic in transverse section. Indeed, our analyses of the trnQ-trnK region support a close relationship between these two otherwise very different genera. Marlothiella was strongly supported (PP 1.0, BP 100) as a sister group to
Choritaenia, with Lichtensteinia sister group to this Glade. Liu et al. (2004) suggested that both Lichtensteinia and Marlothiella be placed within an expanded
Saniculoideae based on the presence of large rib oil ducts and the absence of regular vittae (Fig. 10.3F). Calvin° & al. (2008a) considered the presence of rib oil
159 CHAPTER 10: NEW TRIBAL DELIMITATIONS
ducts in the fruit as a plesiomorphy in Saniculoideae due their presence in both
Azorelloideae and Mackinlayoideae. Moreover, they considered the loss (or
reduction) of rib oil ducts to be a synapomorphy for the upper lineages of Apioideae
(from the Annesorhiza Glade upwards). Large rib oil ducts (traditionally a character
for Saniculoideae s.$) when reconstructed onto the MP trees (character 12, Fig.
10.2L) could most parsimoniously be considered as either a synapomorphy for the
Apioideae—Saniculoideae Glade with a secondary reversal in the upper lineages of
Apioideae (upwards from the Annesorhiza Glade), or the result of convergence in the
Phlyctidocarpa—Petagnaea and Marlothiella—Lichtensteinia clades. Since relatively
large rib oil ducts also occur in some members of the Azorelloideae, there are
various ways of interpreting the evolution of the character (see Calvin° et al. 2008a).
10.4.4 Annesorhiza Glade— Initially the Annesorhiza Glade was recognised
by Calvirio et al. (2006) as comprising Annesorhiza, Chamarea and ltasina.
Although Astydamia and Mo/opospermum were weakly recovered as being closely
related, the authors excluded them from the Annesorhiza group until their
placements had been confirmed with further data. Magee et al. (2008a) subsequently showed that the annual, South African endemic genus Ezosciadium was also related to this Glade. In both the MP and BI analyses, all accessions of these six genera were strongly supported to comprise a monophyletic group (PP
1.0, BP 89), successively sister to the tribe Heteromorpheae and subsequent lineages of the Apioideae. The Annesorhiza Glade comprises herbaceous members largely typical of the euapioid tribes with regular vittae and compound leaves, but differs most prominently in the presence of scattered druse crystals [except in
Ezosciadium, where they are restricted to the commissural area (Magee et al.
160 CHAPTER 10: NEW TRII3A1. DELIMITATIONS
2008a)], the thick and highly lignified vascular bundles in the fruit (Fig. 10.31), and
the proteranthous or deciduous leaves.
10.4.5 Paraphyly of Apioideae— The subfamily Apioideae s.s. as
circumscribed by Drude (1897-98) is clearly not monophyletic when the African taxa
are considered (Downie & Katz-Downie, 1999; Downie & al., 2001; Liu & al., 2003a;
2006; Calvin° & al., 2006; Calvin() & Downie, 2007), and the characters traditionally
used to segregate this subfamily are reconstructed to be either plesiomorphic (e.g.,
compound umbels) or synapomorphic and restricted to the upper lineages of the
subfamily (e.g., inconspicuous rib oil ducts, presence of regular vittae, absence of
scattered druse crystals); Calvin° & al. (2008) and this study. Calvin° & Downie
(2007) based on phylogenetic studies, accommodated many of the conflicting
African taxa in Apioideae or Saniculoideae resulting in two recircumscribed
monophyletic subfamilies. However, as shown by Calvin° et al. (2008), the
subfamilies Apioideae sensu Calvin° & Downie and Saniculoideae sensu Calvirio &
Downie are each not supported by any morphological or anatomical
synapomorphies studies to date. The nodes supporting Apioideae sensu Calvirio &
Downie (Sium–Lichtensteinia)and Saniculoideae sensu Calvin() & Downie
(Phlyctidocarpa–Petagnaea) are both strongly to moderately supported with a low
rate of nucleotide substitutions per site in the BI trees (Fig. 10.4). The Saniculoideae s.s have traditionally been defined by the presence of simple spinescent or setiferous leaves, simple umbels, showy involucre bracts forming a pseudanthium, sessile or subsessile fructiferous flowers, fruit with exocarp outgrowths, distinct rib oil ducts, scattered druse crystals and the absence of regular vittae (Drude 1897-
1898). In contrast, the Apioideae s.s. have been circumscribed traditionally by their
161 CHAPTER 10: NEW TRIBAL. DELINIITATIONS usually compound leaves, prominent compound umbels, fruit with inconspicuous rib oil ducts, and regular vittae (Drude 1897-1898). Many of the protoapioid genera, however, share characters with both subfamilies. Lichtensteinia, Marlothiella,
Polemanniopsis and Steganotaenia have prominent compound umbels typical of
Apioideae s.s., but also saniculoid-like fruit with large rib oil ducts and without regular vittae (Fig. 10.3F and H). The fruit of Phlyctidocarpa (Fig. 10.3G) share the blistered exocarp and large prominent rib oil ducts with members of Saniculoideae s.s., together with a compound umbel and regular vittae as in Apioideae s.s. It is interesting to note the recent discovery of regular vittae in some species of the most early diverging saniculoid genus Alepidea (Yembaturova & al., 2009; Fig. 3D) which further narrows the taxonomic distance between the two subfamilies. Due to the presence of equally isolated and morphologically divergent lineages within the protoapioids, the rank of subfamily for Saniculoideae no longer seems to be appropriate. Drude's (1897-98) classification system reflects incomplete knowledge of several other lineages (mostly African) that are morphologically as distinct as the
Saniculoideae. The only logical way to delimit a morphologically congruent
Apioideae would be to treat the Saniculoideae s.s. as a tribe (the Saniculeae) within a more widely delimited subfamily Apioideae. The union of Apioideae and
Saniculoideae into a single subfamily was also proposed by Koso-Poljansky (1916) who included both subfamilies within his Ligusticoideae, based on the parenchymatous endocarps. Optimization of morphological and anatomical data onto one of the two minimal length trees obtained from the MP analyses of the trnQ- trnK region (Fig. 10.2) shows that such an expanded Apioideae s.l. is well-supported by four synapomorphies (Fig. 4), namely the presence of true wings/ribs (wings or prominent ribs consisting only of mesocarp and exocarp — Fig. 10.2G), the
162
CHAPTER 10: NEW TRIBAL DELIMITATIONS
Sium suave - Physospermum cornubiense 14 Bupleurum angulosum Bupleurum fruticosum A
— Polemannia grossularifolia pi
8 Anginon paniculatum oi d
Anginon difforme eae ,-1 Heteromorpha arborescens
Dracosciadium italae s 4nnesorhiza altiscapa en
1 su C 1 Annesorhiza macrocarpa t Chamarea snijmaniae — Itasina filifolia alvin Astydamia latifolia () & 1 Ezosciadium capense 1 Molopospermum peloponessiacum D
4 1 Choritaenia capensis owni 3 I Choritaenia capensis
, Marlothiella gummifera e
Lichtensteinia obscura 4 dv 2 f-- Lichtensteinia trifida oi 31 1 Lichtensteinia lacera 4 Lichtensteinia globosa —
15 iPhlyctidocarpa flava eeepl s
3 I Phlyctidocarpa flava T • Polemanniopsis sp. 1 S
7 Polemanniopsis marlothii ani 4 Steganotaenia araliacea 1 c Steganotaenia commiphoroides ul rlepidea amatymbica oi d
27 Alepidea capensis ea
Alepidea peduncularis e sen 1 Arctopus echinatus 4 Actinolema macrolema 10 E s 1 Astrantia major u C 2 Astrantia minor al
epipactis vi
3 5 SaniculaSanicula europaea europaea n ° & Sanicula canadensis 1 Eryngium maritimum D Eryngium planum ow 3 11 Eryngium giganteum ni Eryngium palmatum e Petagnaea gussonei 2 -Hermas gigantea Hermas quinquedentata 0.1
FIGURE 10.4 Bayesian inference phylogram of trnQ - trnK sequence data. Numbers above the branches indicate the distributions of parsimony informative indels and those below the branches the number of apomorphies as constructed from character optimisation. The scale indicates branch length.
163 CHAPTER 10: NEW TRIBAL DELImITATIoNs
parenchymatous or sometimes lignified endocarp (becoming secondarily woody in
Choritaenia – Fig. 10.2J), the absence of rhomboidal crystals (Fig. 102M), and the
presence of druse crystals of calcium oxalate scattered throughout the mesocarp
(subsequently lost in the euapioids – Fig. 102N).
10.4.6 Tribal delimitations— While morphological and anatomical
characters support the lineage comprising the Apioideae s.I., as well as many of the
terminal clades within the protoapioids (herein recognized as tribes), there are no
available morphological or anatomical characters to support the hierarchical
relationships among the tribes (i.e., the clades comprising the Lichtensteinieae-
Oenantheae, Lichtensteinieae–Choritaenieae, Steganotaenieae–Phlyctidocarpeae
or Saniculeae–Phlyctidocarpeae). Therefore, in order to reflect the available
morphological, anatomical and molecular data, the protoapioids are here segregated
into eight small and morphologically isolated tribes (each well-supported by several
morphological and/or anatomical characters and/or molecular sequence data),
which probably represent relicts from a once more numerous African apioid flora.
The tribal affinities of all euapioid and protoapiod genera from sub-Saharan Africa is
summarised in Table 11.1 (Chapter 11), and a taxonomic key to the subfamilies of
Apiaceae and tribes of the protoapioids is provided below.
164
CHAPTER 10: NEW TRIBAL DELIMITATIONS
10.5 TAXONOMY OF THE PROTOAPIOID TRIBES
KEY TO THE SUBFAMILIES OF APIACEAE AND TRIBES OF THE PROTOAPIOIDS (APIOIDEAE S.L.)
la. Endocarp woody, adjacent mesocarp usually parenchymatous; rhomboidal
crystals usually present, druse crystals absent; true wings absent; wing-like
structures (pseudo-wings) or prominent ribs comprising endocarp and mesocarp
with vascular tissue at the tip:
Fruits laterally compressed; mericarps not separating at maturity;
carpophore absent Mackinlayoideae
Fruits isodiametric or dorsally compressed; mericarps separating at
maturity; carpophore present Azorelloideae
1 b. Endocarp parenchymatous or sometimes becoming lignified, if somewhat
woody then with adjacent mesocarp lignified; rhomboidal crystals absent; druse
crystals present or absent; true wings often present; wings or prominent ribs
comprising only mesocarp, with vascular tissue almost always at the base:
Fruit with oil vesicles in the wings Choritaenieae
Fruit without oil vesicles in the wings:
Inflorescence a pseudanthium; regular vittae absent, if rarely present
then with larger rib oil ducts and amphi-seminal druse crystals in the
fruit Saniculeae
Inflorescence umbellate, if rarely a pseudanthium then with regular
vittae, poorly developed rib oil ducts, and without amphi-seminal druse
crystals in the fruit:
5a. Druse crystals of calcium oxalate absent or commissural only;
without tanniniferous epidermal cells euapioids
165 CHAPTER 10: NEW TRIBAL DELIMITATIONS
5b. Druse crystals of calcium oxalate amphi-seminal, if reduced to
commissure only, then with tanniniferous epidermal cells and only
partly bifid carpophore:
6a. Regular vittae absent:
Fruit winged; wing cavities present; woody shrubs or trees
Steganotaenleae
Fruit not winged; large rib oil ducts surrounded by a
concentric ring of cells; herbs or small woody shrublets:
Rhizomatous herbs; leaves large, coriaceous
Lichtensteinieae
Woody shrublets; leaves small, succulent
Marlothielleae
6b. Regular vittae present:
Fruit surface blistered; ribs bifurcate; vittae and rib oil ducts
equal in size Phlyctidocarpeae
Fruit surface smooth; ribs not bifurcate; vittae larger than
rib oil ducts:
Perennial or annual herbs; leaves proteranthous or
deciduous Annesorhizeae
Woody trees, shrubs, suffrutices or lianas; if rarely
herbaceous then leaves persistent
Heteromorpheae
166 CHAPTER 10: NEW TRIBAL DELINHTATIoNs
1 ANNESORHIZEAE Magee, C.I.Calvirio, M.Liu, S.R.Downie, P.M.Tilney and B.-E.van
Wyk trib. nov. Heteromorpheis similis umbello multiplexo, fructu crystallis
drusaceis omnino in mesocarpio dispersis et vittis regularibus, sed habitu
herbaceo, foliis proteranthis vel deciduis et fructu fasciculis vascularibus
valde lignosis differt. — TYPE: Annesorhiza Cham. & Schlechtd.
Annesorhizeae is similar to the Heteromorpheae in that the species have compound umbels, fruit with druse crystals scattered throughout the mesocarp and regular vittae. However, they differ from members of the Heteromorpheae in the herbaceous habit (rarely annual in Ezosciadium), the proteranthous or deciduous leaves and fruit with strongly lignified vascular bundles. The tribe includes
Annesorhiza, Astydamia, Chamarea, Ezosciadium, ltasina and Molopospermum.
Although most of the genera are southern African endemics, Molopospermum is a
European genus and Astydamia is restricted to North Africa and the Canary Islands.
2. CHORITAENIEAE Magee, C.I.CalVinO, M.Liu, S.R.Downie, P.M.Tilney and B.-E.van
Wyk trib. nov. Tribubus multis Apioidearum umbello multiplexo alisque
marginalibus veris similis, sed unicus vesiculis olei in alis marginalibus,
carpophoro hygroscopico, endocarpio lignoso cum lignificatione adiuncta
mesocarpii. — TYPE: Choritaenia Benth.
Choritaenieae shares the compound umbels and true marginal wings with many other tribes of the Apioideae, but is unique in that the fruit have oil vesicles in the marginal wings, a hygroscopic carpophore as well as a woody endocarp with further lignification of the mesocarp. It also differs from most other protoapioids in
167 CHAPTER 10: NEW TRIBAL DELIMITATIONS the annual and ephemeral habit and the absence of druse crystals scattered throughout the mesocarp of the fruit. The tribe is monogeneric and includes the southern African genus Choritaenia.
LICHTENSTEINIEAE Magee, C.I.Calvirio, M.Liu, S.R.Downie, P.M.Tilney and B.-
E.van Wyk trib. nov. Marlothieffieis similis umbellis muftiplexis, ductis olei
maximis costalibus annulis concentricis cellularum circumcinctis, crystaffis
drusaceis omnino in mesocarpio dispersis, sed habitu herbaceo deciduo,
foliis non succu/entis saepe proteranthis marginibus dentatis setaceis, fructu
sine pills stellatis va/de differt. — TYPE: Lichtensteinia Cham. & Schlechtd.
Lichtensteinieae is similar to Marlothielleae in the compound umbels, very large rib oil ducts surrounded by concentric ring of cells and druse crystals scattered throughout mesocarp, but differs markedly in the herbaceous deciduous habit, non- succulent leaves (often proteranthous, with dentate, setaceous margins) and fruit without stellate hairs. It is also similar to the tribe Annesorhizeae in the herbaceous and deciduous habit but differs markedly in the large ribs oil ducts and the absence of vittae in the fruit. The tribe is monogeneric and includes the South African genus
Lichtensteinia.
MARLOTHIELLEAE Magee, C.I.Calvino, M.Liu, S.R.Downie, P.M.Tilney and B.-E.van
Wyk trib. nov. Lichtensteinieis similis umbeffis multiplexis, crystaffis drusaceis
omnino in mesocarpio dispersis et ductis o/ei maximis costalibus annulis
concentricis cellularum circumcinctis, sed habitu lignoso, foliis succulentis et
fructu pills stellatis differt. — TYPE: Marlothiella H.Wolff.
168 CHAPTER 10: NEW TRIBAL DELIMITATIONS
Marlothielleae is similar to Lichtensteinieae in the presence of compound
umbels, druse crystals scattered throughout the mesocarp, and the very large rib oil
ducts surrounded by concentric rings of cells, but differs in the woody habit,
succulent fleshy leaves and fruit with stellate hairs. Although it shares the woody
habit and heteromericarpic fruit with members from both Heteromorpheae and
Steganotaenieae, it differs from the former in the large rib oil ducts and the absence
of regular vittae in the fruit and from the latter in the isodiametric fruit, ribs that do
not develop into wings and that are without wing cavities. The tribe is monogeneric
and includes the Namibian endemic genus Marlothiefia.
5. PHLYCTIDOCARPEAE Magee, C.I.Calvirio, M.Liu, S.R.Downie, P.M.Tilney and B.-
E.van Wyk trib. nov. Annesorhizeis similis habitu herbaceo, vittis regularibus
et crystaffis drusaceis omnino in mesocarpio dispersis, sed ductis olei
costalibus magnis, costis bifurcatis vesiculis superficialibus magnis et
fasciculis duobus ventralibus non evolutis differt. — TYPE: Phlyctidocarpa
Cannon & Theobald.
Phlyctidocarpeae is similar to Annesorhizeae in the herbaceous habit, regular vittae and druse crystals scattered throughout the mesocarp, but differs in the large rib oil ducts, bifurcate ribs with large surface vesicles and the two undeveloped ventral bundles. Although it shares the large rib oil ducts, undeveloped ventral bundles and surface vesicles with members of the tribe Saniculeae it differs markedly in the absence of a pseudanthium, the pedunculate compound umbels, the presence of regular vittae and the bifurcate ribs. Phlyctidocarpa has previously
169 CHAPTER 10: NEW T RI BA L DELIMITATIONS been associated with superficially similar members of the tribe Apieae and other euapioid tribes but differs in the presence of druse crystals scattered throughout the mesocarp of the fruit as well as large rib oil ducts, bifurcate ribs with large surface vesicles and undeveloped ventral bundles. The tribe is monogeneric and includes the Namibian endemic genus Phlyctidocarpa.
170 CHAPTER 11
GENERAL CONCLUSIONS
The multidisciplinary approach followed in this study has led to an improved understanding of relationships, particularly within the African and Malagasy members of the Apiaceae. As these taxa had been largely unstudied until relatively recently, little provision was made for them in the classification system of Drude
(1897-98). It is, however, clear that they represent an important component of the
Apiaceae, and substantial rearrangements at almost all infrafamilial levels are required in order to incorporate them into the new emerging familial classification.
While molecular data clearly has an important role to play in teasing out such a classification, this study has illustrated the necessity of a thorough investigation and in some cases reassessment of anatomical, cytological and morphological data if natural, readily definable groups are to be circumscribed. The new insights gained from the study of selected African and Malagasy genera has resulted in the re- circumscription of the subfamily Apioideae, the erection of five new tribes, two new genera and five new species, new combinations for four species and the reduction of one genus and one species into synonymy. It is now possible to assign all but five
African genera to their correct (or most likely) tribal positions (Table 11.1). The following general conclusions regarding the selected genera investigated can be drawn:
The South African endemic genera Capnophyllum, Sonderina and the Cape endemic Stoibrax capense are closely related but only distantly related to their putative North African relatives, Krubera and Stoibrax s.s. Fruit anatomy, morphology and molecular sequence data indicate that their similarities to the North
171 CHAPTER 11: GENERAL CONCLUSIONS
TABLE 11.1 Suprageneric affinities of the Apiaceae genera indigenous to sub-Saharan Africa and Madagascar.
Subfamily Tribe Saniculeae: Tribe Scandiceae: Mackinlayoideae: Alepidea La Roche Agrocharis Hochst. Centella L. Arctopus L. Ammodaucus Coss. & Eryngium L. Durieu Subfamily Azorelloideae: Sanicula L. Anthriscus Pers. Hermas L. Caucalis L. Tribe Steoanotaeniae: Cuminum L. Subfamily Apioideae: Polemanniopsis Daucus L. B.L.Burtt Scandix L. Protoapioids: Steganotaenia Hochst. Torilis Adans.
Tribe Annesorhizeae: Euapioids: Tribe Selineae: Annesorhiza Cham. Anethum L. &Schltdl. Tribe Apieae: Chamarea Eckl. & Ammi L. Tribe Tordvlieae: Zeyh. Apium L. Afroligusticum Ezosciadium B.L.Burtt Billburttia Magee & C.Norman ltasina Raf. B.E.van Wyk Afrosciadium Deverra DC. P.J.D.Winter Tribe Choritaenieae: Foeniculum Mill. Capnophyllum L. Choritaenia Benth. Haplosciadium Hochst. Cynorhiza Eckl. & Petroselinum Hill. Zeyh. Tribe Heteromorpheae: Dasispermum Raf. Andriana B.-E.van Tribe Bupleurieae: Heracleum L. Wyk Bupleurum L. Lefebvrea A.Rich. Anisopoda Baker Nanobubon Magee Anginon Raf. Tribe Careae: Notobubon B.-E.van Cannaboides B.-E.van Carum L. Wyk Wyk Pastinaca L. Dracosciadium Hilliard Tribe Coriandreae: Scaraboides Magee & & B.L.Burtt Coriandrum L. B.-E.van Wyk Aframmi C.Norman Stenosemis Harv. & Glia Sond. Tribe Echinophoreae: Sond. Heteromorpha Cham. Pycnocycla Lindl. &Schltdl. Conium Glade: Polemannia Eckl. & Tribe Oenantheae: Conium L. Zeyh. Berula W.D.J.Koch Pseudocannaboides Ferula L. Diplolophium Glade: B.-E.van Wyk Helosciadium Diplolophium Turcz. Pseudocarum W.D.J.Koch C.Norman Lilaeopsis Greene Nirarathamnos Glade: Tana B.-E.van Wyk Oenanthe L. Seseli L. p.p. Trachyspermum Link Tribe Lichtensteinieae: Tribe Pimpinelleae: p.p. Lichtensteinia Cham. Cryptotaenia DC. p.p. &Schltdl. Frommia H.Wolff Incertae sedis: Phellolophium Baker Afrosison H.Wolff Tribe Marlothielleae: Pimpinella L. Angoseseli Chiov. Marlothiella H.Wolff Physotrichia Hiern Tribe Pvramidoptereae: Pseudoselinum Tribe PhIvctidocarpeae: Ciclospermum Lag. C.Norman Phlyctidocarpa Cannon Oreoschimperella Spuriodaucus & Theobald Rauschert C.Norman
172 CHAPTER 11: GENERAL CONCLUSIONS
African genera are only superficial. The data supports the placement of these South
African taxa within the Capnophyllum group of the Lefebvrea Glade, together with
the genus Dasispermum. The extensive analyses of morphological, anatomical and
molecular sequence data generated for these poorly known South African annuals
or short-lived perennials allowed the unravelling of natural relationships and
revealed the need for substantial rearrangements in order to re-classify them into
more natural, monophyletic genera. The characters traditionally used to define the
monotypic Dasispermum are suggested to represent secondary adaptations to the
dry littoral habitat to which it is endemic. As a result, the five species of Sonderina
(two of which are newly described) and Stoibrax capense are included in an
expanded Dasispermum. The superficially similar genus Stenosemis was found to
be more closely related to the Cape peucedanoid genera Notobubon and
Nanobubon than to the members of the Capnophyllum group, and could readily be
distinguished from all genera within the Lefebvrea Glade by the breadth of the fruit
commissure, which extends beyond the base of the marginal wings somewhere
between the tip and the centre of the wing/rib.The genus Capnophyllum was found
to comprise four species. An unusual new species from the arid Tanqua Karroo
region was recognised at the generic level as Scaraboides. All three of these genera
are now well-defined and supported by several morphological and anatomical
characters. After extensive fieldwork and a thorough taxonomic study, it is apparent that these annual species are poorly known and collected, and consequently five new species have now been described. It is interesting that two of them appear to be fireweeds, germinating and flowering in the year following a fire. This fire-survival strategy has not yet been described for African Apiaceae. It is surprising that these new species have remained unknown for so long, as the South African flora is
173 CHAPTER 11: GENERAL CONCLUSIONS
relatively well-collected. It therefore seems likely that such, short-lived plants are
easily overlooked and that there are more undescribed taxa still to be discovered.
The African and Malagasy species of Pimpinella are clearly related to their
Eurasian counterparts, with the three anomalous African genera Cryptotaenia,
Frommia and Phellolophium embedded within. Unexpectedly, the African species do
not form a single monophyletic lineage but are rather divided among two of the three major clades within the genus, indicating at least two separate dispersal events into
Africa. Fruit morphological and anatomical data appear to be of little use to define natural groups within Pimpinella, as indicated by the paraphyly of the current infrageneric classification. However, optimisation of cytological data onto the molecular trees suggests that chromosome numbers may prove useful in combination with other characters to identify natural groups within Pimpinella.
Chromosome counting of more species could yield interesting results. Although there remains much work before the circumscription and subdivision of this notoriously unwieldy genus can be attempted, the inclusion of the African contingent within the molecular phylogeny is clearly an important step towards unravelling this long term problem.
The Malagasy species of Peucedanum represent the last remaining challenge to solving the so-called "Peucedanum problem" in Africa. The similarity of the Malagasy species to Eurasian Peucedanum and particularly to the South African peucedanoid genus Notobubon, is clearly only superficial. The two Malagasy species were found to differ from all other genera within the family by the position of the vascular tissue at the tip of the ribs and the presence of sphaerocrystals distributed in and around the epidermis and vittae. The latter character has not, to our knowledge, been found in any other members within the family. The molecular
174 CHAPTER 11: GENERAL CONCLUSIONS
data clearly place the species within the tribe Apieae, far removed from either the
Eurasian species of Peucedanum in the Selineae or the African peucedanoid genera
in the Tordylieae. Based on this evidence, the new genus Billburttia was described
to accomodate the two Malagasy endemic species, B. capensoides and B.
vaginoides.
Ezosciadium represents yet another early diverging apioid lineage close to
the Annesorhiza Glade and the genera Astydamia and Molopospermum, although
the genus does not seem to have any close relatives. A position within the newly
described tribe Annesorhizeae is proposed (Table 11.1).
The anomalous southern African genera Choritaenia, Phlyctidocarpa and
Marlothiella are clearly members of the early diverging lineages within the
Apioideae-Saniculoideae Glade. Through a re-investigation of morphological,
anatomical and molecular sequence data, alterations at both the tribal and
subfamilial level are proposed in order to reflect the diversity observed in these
genera. We propose the recognition of two informal groups, the protoapiods
(referring to the early lineages with scattered druse crystals in the fruit mesocarp)
and the euapioids (the remaining upper lineages where druse crystals are usually
absent or where occasionally present, restricted to the commissure). As the genera
within the protoapioids comprise a gradient between the traditional concepts of the
subfamilies Apioideae and Saniculoideae, it is suggested herein that the latter two groups comprise an expanded Apioideae, which can easily be distinguished from the other two subfamilies, the Azorelloideae and Mackinlayoideae, by the absence of rhomboidal crystals, the non-woody endocarp and the presence of true wings. In order to make provision for the protoapioid taxa, eight tribes are recognised (viz.
Annesorhizeae, Choritaenieae, Heteromorpheae, Lichtensteinieae, Marlothielleae,
175 CHAPTER 11: GENERAL CONCLUSIONS
Phlyctidocarpeae, Saniculeae and Steganotaenieae) of which five are newly described (Table 11.1). Such a system reflects the molecular phylogeny and also allows for the circumscription and identification of both the subfamily Apioideae and the protoapioid tribes using morphological and anatomical characters.
Within Africa the majority of the genera are from the Apioideae with only three genera from the Azorelloideae and Mackinlayoideae (Table 11.1). As a result of the work presented in this study and that of several other recent studies, the majority of the African Apioideae have now been placed within some 22 tribes or as yet undescribed major clades, with about a third of the genera within the protoapioid tribes (Table 11.1). Further studies should focus on the systematically unplaced genera Afrosison, Ammodaucus, Angoseseli, Physotrichia, Pseudoselinum and
Spuriodaucus, should material for such studies become available. This study has not only confirmed the notion that the African Apiaceae taxa are of systematic significance beyond their relatively small number but also revealed important new lineages that should be incorporated in comparative studies in the future.
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197 APPENDIX A
APPENDIXA1. Voucher specimens for fruit material studied in transverse section. In addition to those listed below, transverse sections of taxa in the protoapioid tribes from the following studies were also re-examined: Liu et al.
2003a, b, 2006a, b, c; Winter et al. 1993; Van Wyk and Tilney 1994; Van Wyk et al. 1997; Tilney and Van Wyk 2001; Vessio 2001; Liu 2004.
Astydamia DC.: A. latifolia (L.f.) Kuntze — Murrey s.n. sub Alex Prior (K).
Billburttia Magee & B.-E.van Wyk: B. capensoides Sales & Hedge—
Humbert 3634 (P); Rakotozafy 631 (P). B. vaginoides Sales & Hedge —
Decary 7583 (P). Capnophyllum Gaertn.: C. africanum (L.) Gaertn. — Van der Merwe 1767 (NBG); Winter 110 (JRAU); Anon. sub. NBG 754111. C. leiocarpon (Sond.) J.C.Manning & Goldblatt — Stirton & Zantovska 11430
(NBG); Taylor 12003 (PRE); Williamson 3825 (PRE). C. lutzeyeri Magee &
B.-E.van Wyk — Magee et al. 106 (JRAU). C. macrocarpum Magee & B.-
E.van Wyk — Magee et al. 133 (JRAU). Choritaenia Benth.: C. capense
Benth — Smith 540 (PRE); Zietsman 427 (PRE); Hanekom 18341 (PRE).
Cryptotaenia DC.: C. africana (Hook. f.) Drude — Verdecourt 2481 (PRE).
C. canadensis (L.) DC.— PRE0866713 (PRE); Howler 11047 (PRE).
Dasispermum Raf.: D. capense (Lam.) Magee & B.-E.van Wyk — PRE7732
(PRE); Salter 7733 (BOL); Boatwright et al. 212 (JRAU). D. grandicarpum
Magee & B.-E.van Wyk — Lutzeyer s.n. (JRAU). D. hispidum (Thunb.)
Magee & B.E.van Wyk — Van Wyk 3518 (JRAU); Van Wyk 3539 (JRAU). D. humile (Meisn.) Magee & B.E.van Wyk — Van Wyk 1759 (JRAU); Van Wyk
198 APPENDIX Al
1895 (JRAU). D. perrenans Magee & B.E.van Wyk — Magee & Boatwright
105 (JRAU). D. suffruticosum (Berg.) B.L.Burtt — Du Plessis 14 (PRE);
Reyneke 32 (PRE); Retief 1196 (PRE); Tilney 204 (JRAU); Arnold 1170
(PRE); Winter 78 (JRAU); Winter & Tilney 153 (JRAU); Taylor 9876 (PRE). D.
tenue (Sond.) Magee & B.E.van Wyk — Van Wyk 3433 (JRAU); Zeyher s.n.
(K).Ezosciadium B.L.Burtt: E. capense (Eckl. & Zeyh.) B.L.Burtt — Fries et
al. 1156 (PRE); Acocks 20021 (PRE); Goldblatt & Porter 12578 (NBG).
Frommia H.Wolff: F ceratophylloides H.Wolff — Hooper et al. 1844 (K).
Krubera Hoffm.: K. peregrina Lowe — Lippert 22969 (PRE); Sandwith 6215
(K). Molopospermum W.D.J.Koch: M. peloponnesiacum Koch — Lazare 12
(JRAU). Pimpinella L.: P. acutidentata Norman— Fanshawe s.n. (K). P.
betsileensis Sales & Hedge — Humbert 28087 (P). P. buchananii H.Wolff —
Milne-Redhead 9045A (K). P. caffra subsp. caffra D.Dietr. — Hilliard & Burtt
9777 (K). P. caffra subsp. conopodioides C.C.Towns. — Pawek 9268 (K). P.
ebracteata Baker — Baron 929 (K). P. hirtella A.Rich. — Ash 2677 (MO). P. huillensis Welw. ex. Engl. — Milne-Redhead 10894 (K). P. humbertii Sales
& Hedge — Lewis 967 (P). P. ledermannii H.Wolff — Jacques Felix 2858 (P);
Pawek 7458 (MO). P. lindblomii H.Wolff — Gilbert 6531 (K). P. oreophila
Hook.f. — Townsend 2428 (K). P. perrieri Sales & Hedge— Bosser 8622 (P).
P. tenuicaulis Baker — Baron 3241 (MO). P. transvaalensis H.Wolff — Miller
5910 (K). P. trifurcate H.Wolff — Jean Pawek 13923 (MO). Phellolophium
Baker: P. madagascariense Baker — Pettersson & Nillssan 359 (K).
Phlyctidocarpa Cannon & Theobald: P. flava Cannon & Theobald — Gress et al. 6075 (PRE); Manheimer 2889 (JRAU); Winter & Leistner 5739 (PRE).
Polemanniopsis B.L.Burtt: P. sp. 1 — Burke 7002 (WIND). Scaraboides
199 APPENDIX Al
Magee & B.E.van Wyk: S. manning!! Magee & B.E.van Wyk — Manning
3061 (NBG). Stenosemis [E.Mey.] ex. Sond. : S. caffra Sond. — MacOwen
904 (PRE); Phillipson 3677 (PRE); Story 3345 (PRE).Stoibrax Raf.: S. dichotomum (L.) Raf. — Font Quer s.n. (S). S. hanotei (Br.-BI. & Maire)
B.L.Burtt — Wall s.n. (S).
200 APPENDIX A2. Voucher specimens of fruit material used to study external vittae
structure.
Billburttia Magee & B.-E.van Wyk: B. capensoides Sales & Hedge—
Humbert 3634 (P); Rakotozafy 631 (P). Capnophyllum Gaertn.: C.
africanum (L.) Gaertn. — Van der Merwe 1767 (NBG); Winter 110 (JRAU). C.
leiocarpon (Sond.) J.C.Manning & Goldblatt — Williamson 3825 (PRE). C.
lutzeyeri Magee & B.-E.van Wyk — H. Lutzeyer 7A (JRAU); Magee et al. 106
(JRAU). C. macrocarpum Magee & B.-E.van Wyk — Magee et al. 133
(JRAU). Dasispermum Raf.: D. hispidum (Thunb.) Magee & B.E.van Wyk —
Magee et al. 109 (JRAU). D. suffruticosum (Berg.) B.L.Burtt — Magee et al.
117 (JRAU). Krubera Hoffm.: K. peregrina Lowe — Sandwith 6215 (K).
Phlyctidocarpa Cannon & Theobald: P. flava Cannon & Theobald —
Manheimer 2889 (JRAU).
201 APPENDIX B
APPENDIX Bl. Voucher information of ITS and rps16 intron sequences used in the analyses presented in Chapter 3, with corresponding voucher specimens
(for the new accessions) and GenBank reference numbers. For the new accessions the information is listed as follows: taxon — ITS, rps16 intron; voucher specimen. Taxa where the ITS sequence data has been published
(Winter et al. 2008) are indicated with an asterix. For the previously published rps16 intron accessions the information is listed as follows: taxon — rps16 intron.
NEW ACCESSIONS. Capnophyllum Gaertn.: C. africanum (L.) Gaertn. —
FM201528, FM201546; Magee 124 (JRAU). C. africanum (L.) Gaertn. —
FM201527, FM201548; Forest et. al. 654 (NBG). C. leiocarpon (Sond.)
J.C.Manning & Goldblatt — FM201525, FM201545; Bester 6978 (PRE). C. leiocarpon (Sond.) J.C.Manning & Goldblatt — FM201526, FM201544; Magee
& Boatwright 125 (JRAU). C. lutzeyeri Magee & B.-E.van Wyk — FM201524,
FM201543; Magee et al. 106 (JRAU). C. macrocarpum Magee & B.-E.van
Wyk — FM201529, FM201547; Magee et al. 133 (JRAU). Conium L.: C. sphaerocarpum Hilliard & Burtt— FM201530, FM201558; Magee at al. 129
(JRAU). Cynorhiza Eckl. & Zeyh.: C. typica Eckl. & Zeyh. — *, FM201556;
Magee et al. 53 (JRAU). C. typica Eckl. & Zeyh. — *, FM201557; Van Wyk
3372 (JRAU). Dasispermum Raf.: D. suffruticosum (Berg.) B.L.Burtt —
FM201514, FM201541; Magee & Boatwright 117 (JRAU). Nanobubon
Magee: N. capillaceum (Thunb.) Magee — *, FM201549; Magee & Boatwright
202 APPENDIX B1
14 (JRAU). N. strictum (Spreng.) Magee — *, FM201550; Magee et al. 58
(JRAU). Notobubon B.-E.van Wyk: N. capense (Eckl. & Zeyh.) Magee — *,
FM201551; Magee et al. 43 (JRAU). N. capense (Eckl. & Zeyh.) Magee — *,
FM201555; Magee et al. 37 (JRAU). N. gummiferum (L.) Magee — *,
FM201554; Magee et al. 61 (JRAU). N. pearsonii (Adamson) Magee — *,
FM201552; Magee et al 42 (JRAU). N. tenuifolium (Thunb.) Magee — *,
FM201553; Magee et al. 44 (JRAU). Scaraboides Magee & B.-E.van Wyk: S. manningii Magee & B.-E.van Wyk — FM201523, FM201542; Manning 3010
(NBG). Sonderina H.WoIff : S. hispida (Thunb.) H.WoIff — FM201520,
FM201537; Magee & Boatwright 115 (JRAU). S. hispida (Thunb.) H.Wolff --
, FM201536; Magee et al. 112 (JRAU). S. hispida (Thunb.) H.WoIff —
FM201521, FM201535; Magee et al. 107 (JRAU). S. humilis (Meisn.) H.WoIff
— FM201518, FM201538; Van Wyk & Van Wyk 1883 (JRAU). S. sp.1 —
FM201519, FM201534; Magee & Boatwright 105 (JRAU). S. sp. 2 —
FM201522, FM201540; Lutzeyer s.n. (JRAU). S. tenuis (Sond.) H.WoIff —
FM201517, FM201539; Van Wyk et al. 3433 (JRAU). Stoibrax Raf.: S. capense (Lam.) B.L.Burtt — FM201516, FM201532; Magee et al. 128 (JRAU).
S. capense (Lam.) B.L.Burtt — FM201515, FM201533; Magee et al. 131
(JRAU). S. dichotomum (L.) Raf. — FM201531, –; Sanchez-Meta & Molina
Abril s.n. (K).
PUBLISHED RPSI6 INTRON GENBANK ACCESSIONS. Aethusa cynapium L. —
AF110539a . Ammi majus L. —AF164814b. Apium graveolens L. —
AF110545a . Berula erecta (Huds.) Coville —AF164819 b. Conium maculatum
L. —AF110546a. Crithmum maritimum L. —AF110540a. Deverra burchelli
(DC.) Eckl. & Zeyh. —AY838418d . Echinophora tenuifolia L. —AF164812 b.
203 APPENDIX B1
Foeniculum vulgare Mill. —AF110543 a . Heracleum lanatum Michx. —
AF110537a. Heracleum maximum Bartr. — EF426691 f. Heracleum spondylium L. — AF164800 b. Malabaila sekakul Boiss. — AF164802 b.
Nanobubon strictum (Spreng.) Magee — AY838438 d. Naufraga balearica
Constance & Cannon —AF164816 b. Notobubon ferulaceum (Thunb.) Magee
—AY838434d. Notobubon galbanum (L.) Magee —AY838435 d. Notobubon pearsonii (Adamson) Magee —AY838436 d. Notobubon pungens (E.Mey. ex
Sond.) Magee —AY838437d . Pastinaca sativa L. —AF110538a . Sium latifolium L. — AF110552a. Smyrnium olusatrum L. —AF110551 a. Stenosemis caffra Sond. — AY838444 d. Stoibrax dichotomum (L.) Raf. —AM982518 a.
Thaspium pinnatifidum (Buckley) A.Gray — AY372896 c. Zizia aurea Koch —
AF110535a. Zosima orientalis Hoffm. —AF164806 b.
Downie and Katz-Downie (1999) a; Downie et al. (2000)b; Sun and Downie
(2004)`; Calvino et al. (2006) d; Magee et al. (2008c)e; McNeill and Kemper
(unpubl.)f.
204 APPENDIX B2. Voucher information of ITS sequences used in the analyses presented in Chapter 7, with corresponding voucher specimens (for the new accessions) and GenBank accession numbers.
NEW ACCESSIONS. Pimpinella L.: P. alismatifolia C.C.Towns. — FM986448;
Bidgood et al. 3564 (K). P. buchananii H.Wolff — FM986455; Winter 4179
(PRE). P buchananii H.Wolff — FM986456; Winter 4188 (PRE). P caffra
D.Dietr. — FM986447; Van Wyk et al. 4233 (JRAU). P favifolia C.Norman —
FM986458; Winter 3992 (PRE). P favifolia C.Norman — FM986453; Phillips
1612 (MO). P. huillensis Welw. ex. Engl. — FM986443; Biegel 3461 (PRE). P. huillensis Welw. ex. Engl. — FM986454; Fanshawe 1011 (P). P hirtella
A.Rich. — FM986444; De Wide 6578 (PRE). P krookii H.Wolff — FM986445;
Hilliard & Burtt 14439 (K). P kyimbilaensis H.Wolff — FM986452; Gereau et al. 3150 (MO). P ledermannii H.Wolff subsp engleriana (H.Wolff) C.C.Towns.
— FM986457; de Witte 6205 (PRE). P oreophila Hook.f. — FM986450;
Abebe 1124 (MO). P perrieri Sales & Hedge — FM986460; Bosser 16.027
(P). P rigidistyla C.C.Towns — FM986459; Gereau & Kayombo 3987 (MO). P transvaalensis H.Wolff — FM986449; Winter s.n. (PRE). P trifurcata H.Wolff
— FM986446; LaCroix 4284 (PRE). P sp. B — FM986451; Brummit et al
16990 (MO).
PUBLISHED GENBANK ACCESSIONS. Aphanopleura capillifolia (Regel &
Schmalh.) Lipsky — DQ516368 h. Aphanopleura trachysperma Lipsky —
AF008629e, AF009108e. Arafoe aromatica Pimenov & Lavrova — AF077874 b.
Arafoe aromatica Pimenov & Lavrova — U78383b, U78443b. Bunium bulbocastanum L. — DQ443722', DQ4437244 . Crithmum maritimum L. —
205 APPENDIX B2
AH003474a. Cryptotaenia africana (Hook. f.) Drude - DQ516370 h.
Cryptotaenia africana (Hook. f.) Drude - DQ516371 h . Cryptotaenia calycina
C.C.Towns. - DQ516372 h. Deverra burchellii (DC.) Eckl. & Zeyh. -
AM408887'. Frommia ceratophylloides Wolff - DQ647630h. Nothosmyrnium japonicum Miq. - DQ516367 h. Oedibasis platycarpa Koso-Pol. -
AF008632d, AF009111 d. Phellolophium madagascariense Baker -
DQ647627h. Phellolophium madagascariense Baker - DQ647628 h.
Phellolophium madagascariense Baker - DQ647629h. P affinis Ledeb. -
AY581780g. P. anisetum Boiss. & Balansa -AY581781g. P. anisum L. -
AY581782g. P. aromatica M.Bieb. -AY581784g. P aurea DC. -AY581785g.
P betsileensis Sales & Hedge - DQ647626h. P. cappadocica Boiss. &
Balansa -AY581786g. P corymbosa Boiss. -AY581787g. P cretica Poir. -
AY581789g. P cretica Poir. -AY581788g. P diversifolia DC. - DQ516369 h.
P eriocarpa Sol. -AY581790g. P flabellifolia (Boiss.) Benth. & Hook. f. -
AY581791g. P isaurica Matthews - AY581792g. P. kotschyana Boiss. -
AY5817939. P. kotschyana Boiss. - DQ516373h. P lutea Desf. -
DQ516374h. P. niitakayamensis Hayata - DQ516375 h. P nudicaulis Trautv.
-AY581794g. P. oliverioides Boiss. & Hausskn. -AY581795g. P paucidentata Matthews - AY581796g. P. peregrina L. -AY581797g. P. peucedanifolia Fischer ex Ledeb. -AY581798g. P puberula (DC.) Boiss. -
AY5817999. P rhodantha Boiss. -AY581800c. P. saxifraga L. -AY581801g.
P sintenisii H.Wolff -AY581802g. P tragium Vill. - AY581803g. P tragium
Vill. -AY5818049. P tragium Vill. -AY581805g. Psammogeton biternatum
Edgew. -AF164839f, AF164864f. Psammogeton canescens (D.C.) Vatke -
206 APPENDIX B2
AF008630e, AF009109e. Pyramidoptera cabulica Boiss. —AF008631 d.
Stoibrax dichotomum (L.) Raf. — DQ516366 h.
Downie & Katz-Downie (1996)a; Downie et al. (1998)b; Valiejo-Roman et al.
(1998)c; Katz-Downie et al. (1999)d; Lee and Downie (1999) e; Downie et al.
(2000c)f; Tabanca et al. (2005)g; Spalik and Downie (2007) h; Winter et al.
(2008)'; Degtjareva et al. (unpublished)i.
207 APPENDIX B3. Voucher information of ITS and rps16 intron sequences added to the matrices of Magee et al. (2009b), for the analyses presented in Chapter
8, with corresponding voucher specimens (for the new accessions) and
GenBank accession numbers.
NEW ACCESSIONS. Billburttia Magee & B.-E.van Wyk: B. capensoides Sales &
Hedge — FM986437, FM986441; Du Puy et al. M660 (P). B. capensoides
Sales & Hedge — FM986438, FM986442; Keraudren 119 (P). B. capensoides
Sales & Hedge — FM986439, —; Razafindrabe 193 (P). B. vaginoides Sales
& Hedge — FM986440, —; Bojer s.n. (P).
PUBLISHED GENBANK ACCESSIONS. Ammi majus L. — U78386a & U78446a, —.
Anethum graveolens L. — —, AF110542 b. Deverra denudata (Viv.) R.Pfisterer
& Podlech — —, AY838419e. Deverra triradiata Hochst. Ex Boiss. — —,
AF164815C• Diplolophium somaliense Verdc. — DQ368843 e, —. Foeniculum
vulgare Mill. —AY581806d, —. Foeniculum vulgare Mill. — EF421428 f, —.
Petroselinum crispum (Mill.) Fuss — U78387 a & U78447a, —. Ridolfia segetum Moris — U78384a & U78444a, —.
Downie et al. (1998) a; Downie and Katz-Downie (1999) b; Downie et al.
(2000)°; Tabanca et al. (2005)d, Calvino et al. (2006)e; Kersten and Knoess
(unpublished)f.
208 APPENDIX B4. List of taxa included in the analyses presented in Chapter 9 with
GenBank accession numbers.
ITS. Annesorhiza altiscapa Schltr. ex H.Wolff - DQ368830'. Annesorhiza fibrosa B.-E.van Wyk - DQ368831'. Annesorhiza filicaulis Eckl. & Zeyh. -
DQ368832'. Annesorhiza latifolia Adamson - DQ368833'. Annesorhiza macrocarpa Eckl. & Zeyh. - DQ368835'. Astydamia latifolia (L.f.) Kuntze -
DQ368836'. Chamarea snijmaniae B.L.Burtt - DQ368839'. Chamarea sp. 1
- DQ368840'. Ezosciadium capense (Eckl. & Zeyh.) B.L.Burtt - AM982517".
Itasina filifolia (Thunb.) Raf. - DQ368857'. Lichtensteinia obscura (Spreng.)
Koso-Pol. - DQ368858'. Molopospermum peloponnesiacum Koch -
AF074335e, AF074336e. rbcL. Anginon rugosum (Thunb.) Raf. - U50222c.
Annesorhiza altiscapa Schltr. ex H.Wolff -AM234812'. Apium graveolens L.
- L01885 L11165a. Arctopus echinatus L. - AY1884149. Berula erecta
(Huds.) Coville - AM234813 1. Bupleurum fruticosum L. - D44556".
Eryngium giganteum M.Bieb. - DQ133808h. Ezosciadium capense (Eckl. &
Zeyh.) B.L.Burtt - AM234818 1. Heracleum sphondylium L. - AY39554d.
Hermas villosa Thunb. - DQ133810 h. Heteromorpha arborescens Cham. &
Schltdl. - DQ133811 h . Itasina filifolia (Thunb.) Raf. -AM234821 1.
Lichtensteinia lacera Cham. & Schltdl. -AM234822'. Pleurospermum camtschaticum Hoffm. - D44583b. Polemanniopsis marlothii (H.Wolff)
B.L.Burtt - AM234824I. Sanicula europaea L. - DQ133820 h. Sium serra
(Franch. & Savat.) Kitag. - D44587 b. Steganotaenia araliacea Hochst. -
EU213518m. Torilis arvensis Link - AM234827I. rps16 intron. Aethusa cynapium L. - AF110539 d. Alepidea amatymbica Eckl. & Zeyh. -
209 APPENDIX B4
DQ832338k. Alepidea serrata Eckl. & Zeyh. - DQ832349k. Ammi majus L. -
AF164814f. Anginon difforme (L.) B.L.Burtt - AY838391 e. Anginon
verticillatum (Sond.) B.L.Burtt - AY838404 e. Annesorhiza altiscapa Schltr. ex
H.Wolff - AY838405e. Annesorhiza filicaulis Eckl. & Zeyh. - AY838407e.
Annesorhiza latifolia Adamson - AY838408e. Annesorhiza macrocarpa Eckl.
& Zeyh. - AY838410 e. Apium graveolens L. - AF110545d. Arctopus echinatus L. - DQ832351 k. Berula erecta (Huds.) Coville - AF164819f.
Bupleurum falcatum L. - AF110566d. Bupleurum fruticosum L. -
AF110569d. Chamaesciadium acaule C.A. Mey. - AY838411 e. Chamarea longipedicellata B.L.Burtt - AY83841 3e. Chamarea snijmaniae B.L.Burtt -
AY838414e. Chamarea sp.1 - AY838415e. Chamarea aff. gracillima
(H.Wolff) B.L.Burtt - AY83841 6e. Dasispermum suffruticosum (Bergius)
B.L.Burtt - AY83841 7e. Deverra burchellii (DC.) Eckl. & Zeyh. - AY838418e.
Echinophora tenuifolia L. - AF164812f. Ezosciadium capense (Eckl. & Zeyh.)
B.L.Burtt - AM982518". Foeniculum vulgare Mill. - AF110543d.
Helosciadium nodiflorum Koch - AF164820f. Heracleum sphondylium L. -
AF164800f. Hermas gigantea L.f. - AY838420e. Hermas quercifolia Eckl. &
Zeyh. - AY838421 e. Hermas quinquedentata L.f. - AY838422e.
Heteromorpha involucrata Conrath - AF110577d. Heteromorpha papillosa
C.C.Towns. - AY838425e. Itasina fififolia (Thunb.) Raf. - AY838426e.
Komarovia anisosperma Korov. - AF110555 d. Lichtensteinia lacera Cham. &
Schltdl. - AY838427e. Lichtensteinia obscura (Spreng.) Koso-Pol. -
AY838428e. Mo/opospermum peloponnesiacum Koch - AY838432e.
Notobubon ga/banum (L.) Magee - AY838435e. Pastinaca sativa L. -
AF110538d. Pleurospermum foetens Franch. - AF110559 d. Pleurospermum
210 APPENDIX B4 uralense Hoffm. — AF110560 d. Polemannia montana Schltr. & H.Wolff —
AF110570d. Polemannia simplicior Hilliard & B.L.Burtt — AF110571 d.
Polemanniopsis marlothii (H.Wolff) B.L.Burtt — DQ832374 k. Pseudocarum eminii H.Wolff —AY838441 e. Pseudocarum laxiflorum (Baker) B.-E.van Wyk
— Y838442e. Sanicula arctopoides Hook. &Arn. — DQ832375 k. Sium latifolium L. — AF110552d. Smyrnium olusatrum L. — AF110551 d.
Steganotaenia araliacea Hochst. — DQ832384k. Stoibrax dichotomum (L.)
Raf. — AM982519". Torilis arvensis Link —AF110548d.
Olmstead et al. (1992)a, Kondo et al. (1996) b, Plunkett et al. (1996b)°, Downie and Katz-Downie (1999)d, Downie et al. (2000a) e, Downie et al. (2000c) f,
Chandler and Plunkett (2004)g, Andersson et al. (2006) h, Calvin° et al. (2006)',
Silvertown et al. (2006)j, Calvin° and Downie (2007) k, Forest et al. (2007) 1 ,
Lahaye et al. (2008)`", Magee et al. (in press)".
211 APPENDIX B5. Voucher information of cpDNA trnQ-trnK sequences used in the analyses presented in Chapter 10, with corresponding voucher specimens and GenBank accession numbers.
Actinolema macrolema Boiss. — Coode & Jones 2425 (E); tmQ-rps16
DQ832386b, rps16 DQ832337b, rps16-trnK DQ832473c. Alepidea amatymbica
Eckl. & Zeyh. — Mohle 480 (MO); tmQ-rps16 DQ832387c, rps16 DQ832338c, rps16-tmK DQ832474c. Alepidea capensis (Berg.) R.A.Dyer — Hilliard & Burtt
16645 (E); tmQ-rps16 DQ832389c, rps16 DQ832340b, rps16-trnK DQ832476c.
Alepidea peduncularis Steud. ex. A. Rich. — Gereau & Kayombo 4094 (MO); tmQ-rps16 DQ832386c, rps16 DQ832337c, rps16-tmK DQ832473c. Anginon
difforme (L.) B.L.Burtt — Magee 130 (JRAU); tmQ-rps16 FM986461 h , rps16
FM986476h, rps16-trnK FM986485h. Anginon paniculatum (Thunb.) B.L.Burtt
— Downie 2458 (ILL); trnQ-rps16 DQ832400c, rps16 AY838397b, rps16-tmK
DQ832487c. Annesorhiza altiscapa H.Wolff — Magee 97 (JRAU); tmQ-rps16
FM986462h, rps16 FM986477h, rps16-tmK FM986486h. Annesorhiza macrocarpa Eckl. & Zeyh. — Downie 2454 (ILL); trnQ-rps16 DQ832400c, rps16AY838397b, rps16-tmK DQ832487b. Arctopus echinatus L. — Ornduff
7380 (UC); tmQ-rps16 DQ832402c, rps16 DQ832351 c, rps16-tmK
DQ832489c. Astrantia major L. — Schilling 2937 (E); trnQ-trnK DQ832443c.
Astrantia minor L. — Vautier, Mermoud & Bersier s.n. (ILL); trnQ-trnK
DQ832444c. Astydamia latifolia (L.f.) Kuntze — Hildenbrand s.n. (ILL); tmQ- rps16 FM986463h, rps16-tmQ FM986487h. Bupleurum angulosum L. —
Younger 2565 (E); trnQ-rps16 FM986464h, rps16AF110568 a, rps16-trnK
FM986488h. Bupleurum fruticosum L. — McBeath 2592 (E); trnQ-rps16
212 APPENDIX B5
FM986465h, rps16 AF110569a, rps16-tmK FM986489h. Chamarea snijmaniae
B.L.Burtt — MacGregor s.n. (NBG); tmQ-rps16 rps16 AY838414b, rps16-
tmK FM986490h. Choritaenia capensis Benth.— Zeitman 271 (PRE); tmQ- rps16 FM986467h, rps16 FM986479h, rps16-tmK FM986492h. Choritaenia
capensis Benth.— Van Wyk 545 (PRU); tmQ-rps16 FM986466h, rps16
FM986478h, rps16-tmK FM986491 h . Dracosciadium italae Hilliard & B.L.Burtt
— Ngwenya & Singh 1279 (PRE); EU4346579, EU4346529 . Eryngium giganteum M.Bieb. — Mc Neal 472 (UC); tmQ-rps16 EU070440d, rps16
EU070502d, rps16-tmK EU070564d. Eryngium maritimum L. — Medina et al.
MP1656 (TEX); tmQ-rps16 EU070454d, rps16 EU070516d, rps16-tmK
EU070578d. Eryngium palmatum Pan iá & Vis. — Constance C-99 (UC);
trnQ-rps16 DQ832416c, rps16 DQ832365b, rps16-tmK DQ832504c. Eryngium planum L. — Downie 191 (ILL); trnQ-trnK DQ832456b. Ezosciadium capense
(Eckl. & Zeyh.) B.L.Burtt — Goldblatt & Porter 12578 (NBG); tmQ-rps16
FM986468h, rps16 AM982518f rps16-trnK FM986493h. Hacquetia epipactis
DC. — M. F & S. G. Gardner 2590 (E); tmQ-rps16 DQ832423c, rps16
DQ832372c, rps16-tmK DQ832511 c. Hermas gigantea L. — McDonald 1769
(NBG); tmQ-rps16 DQ832425b, rps16 AY838420b, rps16-trnK DQ832513b.
Hermas quinquedentata L.f. — Burman 1080 (BOL); tmQ-rps16 DQ832426b, rps16 AY838422b, rps16-tmK DQ832514b. Heteromorpha arborescens Cham.
& Schltdl.— Van Wyk 4122 (JRAU); trnQ-rps16 FM986469h, rps16
AY838424b, rps16-tmK FM986494h. ltasina filifolia (Thunb.) Raf. — Downie
2453 (ILL); trnQ-rps16 FM986470 h , rps16 AY838426b, rps16-tmK
FM986495 h. Lichtensteinia globosa B.-E.van Wyk & Tilney — Downie 2462
(ILL); tmQ-rps16 DQ832429c, rps16 AY838429b, rps16-tmK DQ832517c.
213 APPENDIX B5
Lichtensteinia lacera Cham. & Schltdl. — Downie 2464 (ILL); tmQ-rps16
DQ832427c, rps16 AY838427b, rps16-tmK DQ832515c. Lichtensteinia obscura
(Spreng.) Koso-Pol.— Downie 2457 (ILL); trnQ-rps16 DQ832428c, rps16
AY838428b, rps16-tmK DQ832516c. Lichtensteinia trifida Cham. & Schltdl. —
Downie 2460 (ILL); tmQ-rps16 DQ832430c, rps16 AY838430b, rps16-tmK
DQ832518c. Marlothiella gummifera H.WoIff — Manheimer 2987 (JRAU); tmQ-rps16 FM986471 h, rps16 FM986480h, rps16-tmK FM986496h.
Molopospermum peloponnesiacum Koch — Argent ML2 (E); tmQ-rps16 —, rps16 AY838432 b, rps16-tmK —. Petagnaea gussonei (Spreng.) Rauschert —
Donila 2005 (PAL); trnQ-trnK DQ832466c. Phlyctidocarpa flava Cannon &
Theobald — Merxmueller & Giess 30626 (WIND); tmQ-rps16 FM986473h, rps16 FM986482h, rps16-trnK FM986498h. Phlyctidocarpa flava Cannon &
Theobald — Manheimer 2889 (JRAU); tmQ-rps16 FM986472h, rps16
FM986481 h , rps16-tmK FM986497h. Physospermum cornubiense DC. —
Pimenov et al. s.n. (MW); tmQ-rps16 FM986474h, rps16AF110556a, rps16- tmK FM986499h. Polemannia grossularifolia Eckl. & Zeyh.— De Castro 274
(JRAU); tmQ-rps16 —, rps16 AY838439b, rps16-tmK FM986500h.
Polemanniopsis marlothii (H.Wolff) B.L.Burtt — Downie 2459 (ILL); tmQ-rps16
DQ832432c, rps16 DQ832374c, rps16-trnK DQ832520c. Polemanniopsis sp. 1
— Manheimer 2769 (JRAU); tmQ-rps16 FM986475h, rps16 FM986483h, rps16-tmK —. Sanicula canadensis L. — Downie 737 (ILL); trnQ-trnK
DQ832467c. Sanicula europaea L. — Catalan 1896 (JACA); trnQ-trnK
DQ832468c. Sium suave Walter — Downie 12 (ILL); trnQ-trnK EF185274 e.
Steganotaenia araliacea Hochst.— Downie 2456 (ILL); tmQ-rps16
DQ832442c, rps16 DQ832384c, rps16-trnK DQ832530c. Steganotaenia
214 APPENDIX B5
commiphoroides Thulin — Friis et al. 4889 (K); tmQ-rps16 —, rps16
FM986484h, rps16-tmK FM986501 h .
Downie and Katz-Downie (1999) a; Calvirio et al. (2006) b; Calvirio and Downie
(2007)c; Calvin° et al. (2008b)d; Downie et al. (2008)e; Magee et al. (2008c)f,
Tilney et al. (2009)g; present study h.
215 APPENDIX C
APPENDIX C1. MAGEE, A. R., B.-E. VAN WYK, P. M. TILNEY and S. R. DOWNIE
2008. Ezosciadium (Apiaceae): a taxonomic revision of yet another early diverging South African apioid genus. Plant Systematics and Evolution 276:
167-175.
APPENDIX C2. MAGEE, A. R., B.-E. VAN WYK, P. M. TILNEY and S. R. DOWNIE
2009. A taxonomic revision of Capnophyllum (Apiaceae: Apioideae). South
African Journal of Botany 75: 283-291.
APPENDIX C3. MAGEE, A. R., B.-E. VAN WYK, P. M. TILNEY and S. R. DOWNIE
2009. Generic delimitations and relationships of the Cape genera
Capnophyllum, Dasispermum and Sonderina, the North African genera
Krubera and Stoibrax, and a new monotypic genus of the subfamily Apioideae
(Apiaceae). Systematic Botany 34(3): in press.
APPENDIX C4. MAGEE, A. R., B.-E. VAN WYK, P. M. TILNEY, F. SALES, I. HEDGE and
S. R. DOWNIE. Billburttia, a new genus of Apiaceae (tribe Apieae) endemic to
Madagascar. Plant Systematics and Evolution. In press.
APPENDIX C5. MAGEE, A. R., C. I.CALviNo, M. Liu, S. R. DOWNIE, P. M. TILNEY and B.-E. VAN WYK. New tribal delimitations for the early diverging lineages of
Apiaceae subfamily Apioideae. Taxon. Submitted.
216 APPENDIX Cl
MAGEE, A. R., B.-E. VAN WYK, P. M. TILNEY and S. R. DOWNIE 2008.
Ezosciadium (Apiaceae): a taxonomic revision of yet another early diverging
South African apioid genus. Plant Systematics and Evolution 276: 167-175. Plant Syst Evol (2008) 276:167-175
DOI 10.1007/s00606 - 008 - 0086 - z
ORIGINAL ARTICLE
Ezosciadium (Apiaceae): a taxonomic revision of yet another early diverging South African apioid genus
A. R. Magee • B.-E. van Wyk • P. M. Tilney • S. R. Downie
Received: 16 April 2008 /Accepted: 12 August 2008 /Published online: 26 September 2008 © Springer-Verlag 2008
Abstract The hitherto poorly known Cape endemic Introduction genus Ezosciadium (Apiaceae) is revised. This genus is highly distinctive and can be distinguished from other Ezosciadium B.L. Burtt is a poorly known and collected annual genera of the region by its pilose vegetative and monotypic genus endemic to the Eastern Cape Province reproductive organs, the sessile compound umbels with of South Africa. As with many other small African conspicuously unequal rays, the non-inflexed petal tips, the endemic Apiaceae, its correct placement within the new relatively small, highly-inflexed stamens which appear emerging tribal classification of the family remains almost sessile, and the prominent carpophores which per- untested by molecular data (Calvirio et al. 2006). The sist on the plant. The fruit are unusual in the presence of genus was included in a large rbcL analysis by Forest druse crystals around the carpophore and tanniniferous et al. (2007) in assessing the phylogenetic diversity of the substances in the epidermal cells of the ribs. The phylo- Cape Flora. Although the genus occupied a position sister genetic position of the genus within the subfamily group to Annesorhiza Cham. and Schltdl. and Itasina Apioideae was assessed using rbcL, rps16 intron (2 new Raf., the limited sampling of the family did not allow for accessions) and nrITS (1 new accession) sequence data. any conclusions to be drawn on possible tribal affinities. Ezosciadium capense was found to form part of an early Ezosciadium capense is traditionally placed in the large diverging lineage within the subfamily, sister group to the tribe Apieae (Pimenov and Leonov 1993). De Candolle Annesorhiza Glade and possibly also closely related to the (1830) and Sonder (1862) included it within the genus genera Molopospermum and Astydamia. A comprehensive Helosciadium W.D.J. Koch, a genus now placed in the taxonomic revision, including typification, detailed tribe Oenantheae (Downie et al. 2001). Wolff (1927) descriptions, geographical range and illustrations, is followed Ecklon and Zeyher (1837) in recognising the presented. genus as distinct and postulated an affinity rather to Sonderina H. Wolff or Apium L., genera traditionally Keywords Crystals • Ezosciadium capense • ITS • placed in the tribe Apieae. Recent molecular systematic rbcL • rps16 intron • Fruit anatomy • Morphology • studies, however, have shown Sonderina to form part of Phylogeny an African peucedanoid Glade together with Dasispermum Raf. and other African species of tribe Tordylieae previ- ously treated in the genus Peucedanum L. (Winter et al. A. R. Magee ( 121 ) • B.-E. van Wyk • P. M. Tilney 2008). As a result, the three genera with which Ezoscia- Department of Botany and Plant Biotechnology, dium is traditionally associated are scattered in various University of Johannesburg, P.O. Box 524, Auckland Park 2006, Johannesburg, South Africa tribes throughout the subfamily Apioideae. This paper is e-mail: [email protected] aimed at ascertaining the phylogenetic placement of the genus within the subfamily using both DNA sequence and S. R. Downie anatomical data as well as providing the first compre- Department of Plant Biology, University of Illinois at Urbana-Champaign, hensive taxonomic revision, including nomenclature, Urbana, IL 61801, USA typification, detailed description, geographical distribution
Springer 168 A. R. Magee et al. and illustrations, of this anomalous and poorly known immature fruit, from Fries et al. 1156 (PRE), as well as African genus. mature fruit, from Acocks 20021 (PRE) and Goldblatt & Porter 12578 (NBG). Following rehydration, the fruit were placed in FAA for a minimum of 24 h and subsequently Materials and methods treated according to a modification of the method of Feder and 0' Brien (1968) for embedding in glycol methacrylate Taxonomic study (GMA). This modification involves a final infiltration in GMA of 5 days. Transverse sections, about 3 i.tm thick, Specimens from the following herbaria were studied: BM, were cut using a Porter—Bliim ultramicrotome. The sections K, NBG, PRE and S. From this material, together with were examined for the presence of crystals using a light information from Leistner and Morris (1976), the recorded microscope, after which they were stained according to the distribution of E. capense was ascertained and mapped. All periodic acid Schiff/toluidine blue (PAS/TB) staining illustrations (Fig. 1) were made by the first author with the method of Feder and 0' Brien (1968). Crystals tend to be aid of a camera lucida attachment on a Zeiss compound dissolved during this procedure, hence the need to study microscope or a Wild M3Z stereomicroscope. unstained sections.
Fruit anatomy DNA sampling, isolation, amplification and sequencing
Fruit from three specimens of E. capense, representing In order to assess the phylogenetic position of Ezoscia- three of the four known populations, were rehydrated in dium, DNA sequences of the chloroplast genes rps16 order to study the anatomy. This material included both intron and rbcL were compiled using available sequences from GenBank. Due to the high divergence and difficulty in aligning the nuclear ribosomal DNA internal tran- scribed spacer (ITS) region across the subfamily, a reduced sampling was compiled representing the Glade within which Ezosciadium was shown to be placed based on results of phylogenetic analyses of the aforementioned chloroplast genes. Sources of material used in the study are listed in the Appendix. In total, 52 accessions of rps16, 19 accessions of rbcL, and 12 accessions of ITS were considered for phylogenetic study. For the broader analyses using plastid markers, representatives of both tribes of subfamily Saniculoideae as well as many of the major clades and tribes of subfamily Apioideae were included. The genus Hermas L., a member of subfamily Azorelloideae, was selected as outgroup for the chloro- plast datasets, based on previous molecular analyses (Calvitio et al. 2006), while the ITS dataset was rooted using Lichtensteinia obscura based on the results of the chloroplast analyses. An aliquot of total DNA from E. capense was obtained from the Leslie Hill Laboratory at Kirstenbosch Botanical Garden, South Africa. The rps16 intron was amplified using the primers of Oxelman et al. (1997), while the ITS region was amplified using the Sun et al. (1994) AB101 and AB102 primers. Amplified PCR products were purified using a QIAquick PCR purification kit (Qiagen Inc.) according to the manufacturer's instructions and directly sequenced on a 3130 x/ Genetic Analyzer (Applied Bio- Fig. 1 Ezosciadium capense. a Portion of a flowering stem, b systems Inc.) using BigDye Terminator version 3.1 cotyledon, c first leaf, d mature leaf, e mature fruit, f transverse (Applied Biosystems Inc.). The newly obtained ITS and section through the mature fruit, g and h petals in dorsal view (g) and in ventral view (h), i stamens in dorsal view. a, e and f from A cocks rps16 intron sequences of E. capense from Goldblatt & 20021 (PRE); b—d and g—i from Fries et al. 1156 (PRE) Porter 12578 (NBG) and Stoibrax dichotomum from
4) Springer Ezosciadium (Apiaceae): a taxonomic revision 169
Sanchez-Mata & Molina Abril s.n. (K) were submitted to were imported into PAUP* and a majority rule consensus Genbank. tree was produced in order to show the posterior proba- bilities (PP) of all observed bi-partitions (only PPs above Sequence alignment and phylogenetic analyses 0.5 are reported).
For each data set, complementary strands were assembled and edited using Sequencher version 3.1.2 (Gene Codes Results and discussion Corporation) and aligned manually in PAUP* version 4.0b10 (Swofford 2002), with gaps positioned so as to Morphology minimise nucleotide mismatches. Data sets are available on request from the corresponding author. Phylogenetic Ezosciadium is a markedly pilose (Fig. I a), erect annual analyses were conducted using the maximum parsimony herb up to 0.35 m tall. The leaves are cauline, with the (MP) algorithm of PAUP*. Character transformations were lower ones prominently tri-lobed (Fig. 1 c) and the upper treated as equally likely (Fitch parsimony; Fitch 1971). ones digitately compound (Fig. I d). The cotyledons are Tree searches were performed using a heuristic search with simple and narrowly oblanceolate (Fig. lb), fitting the L 1,000 random sequence additions, tree bisection-recon- type, as defined by Cerceau-Larrival (1962) as commonly nection (TBR) branch swapping and the MULPARS option found in the subfamily Apioideae. The very sparse, com- in effect. A limit of ten trees per replicate was set to reduce pound umbels are sessile and borne in the leaf axils. They the time spent on swapping in each replicate. Internal are composed of 2 to 4 markedly unequal rays (Fig. I a) support was assessed with 1,000 bootstrap replicates that are of diagnostic value amongst African annual genera. (Felsenstein 1985) using TBR swapping and holding ten The flowers are also unusual in that the petals are not trees per replicate. Only values greater than 50% are inflexed at their tips (Fig. 1h) and the relatively small reported. stamens remain highly inflexed, so that they appear almost Bayesian inference (BI) was performed on the rps16 sessile (Fig. 1 i). intron dataset using MRBAYES v. 3.1.2 (Huelsenbeck and Ronquist 2001; Ronquist and Huelsenbeck 2003). The Fruit anatomy TrN+G model, as indicated to be the best model by MODELTEST v. 3.06 (corrected AKAIKE information The fruit are distinctly pilose and oblong in shape (Fig. l e). criterion; Posada and Crandall 1998), was implemented. They are borne on a partly bifid carpophore (Fig. le), one The analysis was performed for 1 million generations of half of which remains attached to the umbellate rays even Monte Carlo Markov Chains and a sampling frequency of after the fruit have fallen off (Fig. la). In transverse section ten. The resulting trees were plotted against their likeli- (Fig. 2a, If) the fruit are similar to many other apioid hoods in order to determine where the likelihoods converge genera. They are isodiametric and homomericarpic with on a maximum value. All the trees before this convergence five equal, prominent ribs. The commissure is very narrow, were discarded as the turn-in' phase. The remaining trees being confined to the carpophore region. The epidermal
b
A 02 mm,
Fig. 2 Transverse sections through the fruit of Ezosciadium capense. (crystals indicated with an arrow, similarly dark structures located at a Mature mericarp, b rib of mature fruit with epidermal cells the base of the ribs are highly lignified vascular tissue). a and b from containing tanniniferous substances (indicated with arrow), c imma- Goldblatt & Porter 12578 (NBG), c from Fries et al. 1156 (PRE) ture fruit showing presence of druse crystals around the carpophore
Springer 170 A. R. Magee et al. cells are bottle-shaped and extend into hairs; those of the topologies obtained from both the MP and BI analyses ribs contain tanniniferous substances (Fig. 2b). This is were similar, with the general branching order similar to unusual in Apiaceae, but their presence has been recorded that found in previous analyses (Calvifio et al. 2006). in Hydrocotyle L. and Trachymene Rudge (Liu 2004). Amongst the early diverging lineages of Apioideae the The mesocarp is composed of parenchymatous cells that genus Lichtensteinia Cham. and Schltdl. formed the ear- are usually periclinally elongated. The endocarp is also liest branching Glade (BP 98; PP 1.0). Following was the parenchymatous, the cells being narrow and strongly lineage comprised of the Annesorhiza Glade plus Ezos- periclinally elongated. The vascular tissue occupies a large ciadium and Molopospermum (BP 65; PP 0.91). In the BI portion within each of the five ribs and is distinctly cir- tree, E. capense grouped with Molopospermum (PP 0.67), cular. Rib oil ducts were not observed in the fruit. The sister group to the Annesorhiza Glade (Annesorhiza, vittae are arranged as in most members of the subfamily Chamarea and Itasina). The tribe Heteromorpheae (BP Apioideae—two commissural and four vallecular. The 100; PP 1.0) was a subsequent sister group, followed by endosperm is isodiametric and flattened to slightly concave the tribes Bupleureae (BP 97; PP 1.0) and Pleurospermeae on the commissural face. The testa is prominent in trans- (BP 100; PP 1.0). Higher up on the trees the tribes Oe- verse section and may be comprised of up to three layers of nantheae (BP 98, PP 1.0), Smyrnieae, Scandiceae, and cells especially in the commissural region, with the outer other members of the Apioid superclade, including the periclinal cell walls of the outermost layer being conspic- tribe Apieae (BP 91, PP 1.0), were retrieved. The place- uously thickened. ment of Chamaesciadium acaule differed in the two Druse crystals, restricted to the region surrounding the analyses. In the MP tree, it was found to be sister group carpophore in the commissural area (Fig. 2c), were found to a Glade comprising Apieae, Echinophoreae, Selineae to be present in all fruit of Ezosciadium examined. The and Tordylieae, while in the BI tree (not shown) it was presence of scattered druse crystals in the mesocarp (i.e. sister group to a Glade comprising Echinophoreae, Selin- with crystals occurring around the seed and not restricted eae and Tordylieae (PP 1.0). to the carpophore region) has a high predictive value for determining ancestral lineages within Apioideae (Van rbcL dataset Wyk and Tilney 2004). This type of crystal distribution is known to occur in many of the early diverging branches The rbcL dataset included 1,238 characters of which 126 of the Apioideae–Saniculoideae Glade (Van Wyk and were variable and 57 parsimony informative. The MP Tilney 2004), including most members of the Saniculoi- analysis (Fig. 3b) resulted in three equally parsimonious deae (Liu et al. 2007), all genera of the Annesorhiza Glade trees with a TL of 176 steps, a CI of 0.76 and a RI of 0.75. (i.e. Annesorhiza, Chamarea Eckl. and Zeyh., Itasina), the The overall topology of the MP trees retrieved in this tribe Heteromorpheae, as well as Molopospermum W.D.J. analysis was found to be similar to that shown in the rps16 Koch (Liu et al. 2006) and Astydamia DC. (Liu pers. intron trees. Although rbcL sequences for Molopospermum comm.). In the remaining Apioideae, crystals are usually and Astydamia were not available, the position of E. ca- completely absent but, for example, are frequently present pense as sister group to the Annesorhiza Glade (represented around the carpophore in some members of the tribe in this analysis by Annesorhiza and Itasina) is moderately Scandiceae, such as species of Anthriscus Pers., Caucalis supported (BP 84). L., Chaerophyllum L., Daucus L., Osmorhiza Raf., Scandix L. and Torilis Adans. (Drude 1897-1898). The ITS dataset presence of druse crystals, even though restricted to the carpophore region in Ezosciadium, may suggest an The ITS dataset included 638 characters of which 136 were affinity to the early diverging lineages of the subfamily as variable and 57 parsimony informative. The MP analysis shown by analyses of all three molecular data sets (Fig. 3c) resulted in a single parsimonious tree with a TL (Fig. 3a–c). of 220 steps, a CI of 0.78 and a RI of 0.52. In this tree, E. capense is sister group to the Glade of Astydamia and rps16 intron dataset Molopospermum (BP 54) which, in turn, is sister group to the Annesorhiza Glade. The rps16 intron dataset included 1,113 characters of which 304 were variable and 169 parsimony informative. Basal African Apiaceae The MP analysis (Fig. 3a) resulted in 2,208 equally most parsimonious trees with a tree length (TL) of 534 steps, a Burtt (1991) argued that the southern African taxa of consistency index (Cl) of 0.70 (including uninformative Apiaceae are of importance beyond their relatively small characters) and a retention index (RI) of 0.85. The overall size. Recent molecular systematic studies have indeed
Springer
Ezosciadium (Apiaceae): a taxonomic revision 171
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Fig. 3 a Strict consensus tree of the 2,208 equally most parsimonious differed in the BI tree are indicated as grey lines. b Strict consensus trees with a length of 534 steps, based on the MP analysis of rpsl6 tree of the 3 equally most parsimonious trees with a length of 176 sequence data (CI = 0.70 and RI = 0.85). BP's are given above the steps, based on MP analysis of rbcL sequence data (CI = 0.76 and branches and PP's are given below the branches. Branches supported RI = 0.75). c Single most parsimonious tree with a length of 220 only in the BI tree are indicated as dotted lines, while those that steps, based on ITS sequence data (Cl = 0.78 and RI = 0.52)
shown that African genera are often sister to (or basally African genera occupied early diverging positions within divergent within) other major lineages and therefore the Apioideae. Lichtensteinia was shown to be the most essential to understanding of relationships within the early diverging lineage sister group to other genera of the family as a whole. The basally divergent position of subfamily Apioideae. Successively sister to the Lichten- African genera was first shown by early molecular sys- steinia Glade was a Glade comprising largely African tematic studies which suggested that Heteromorpha Cham. herbaceous genera, called the Annesorhiza Glade (com- and Schltdl. and Anginon Raf. formed basal-branching prising Annesorhiza, Chatnarea, and Itasina). This lineages within the subfamily Apioideae (Downie et al. Annesorhiza Glade was closely related to Molopospermum 1996, 1998; Plunkett et al. 1996a, b). Downie and Katz- and Astydamia, however, these relationships were not very Downie (1999) subsequently retrieved a broader Hetero- strongly supported. Even within the subfamily Saniculoi- morpha Glade, comprising numerous woody African deae, its relatively small African contingent has been genera (Anginon, Dracosciadium Hilliard and B.L. Burtt, shown to be basally divergent. Downie and Katz-Downie Glia Sond., Heteromorpha and Polemannia Eckl. and (1999) demonstrated that two woody African genera (viz., Zeyh.). This Glade was later recognised by Downie et al. Steganotaenia Hochst. and Polemanniopsis B.L.Burtt) (2000b) as the tribe Heteromorpheae. More recently, an traditionally placed within the Apioideae, formed a sister analysis focusing on the phylogenetic position of African group relationship with the subfamily Saniculoideae. Apiaceae (Calvino et al. 2006) indicated that several other These two genera have subsequently been included in an
1 1 Springer 172 A. R. Magee et al. expanded Saniculoideae by Calvin() and Downie (2007) as Helosciadium sect. Trachysciadium (in which case the name the tribe Steganotaenieae. Furthermore, within their tribe Ezosciadium would be superfluous). We however, prefer to Saniculeae two African endemic genera (Alepidea Delar. follow Burn (1991), who argued that there is no reasonable and Arctopus L.) have now also been shown to occupy doubt that Ecklon and Zeyher (1837) adopted De Candolle' s early diverging positions, both successively sister to the taxon and simply altered its rank. As such, Burtt's proposed remaining genera (Calvin() and Downie 2007; Magee et al. Ezosciadium nom. nov. is valid and necessary]. 2008). In this study, we report on yet another early This highly distinctive genus appears to be an isolated diverging branch of subfamily Apioideae from South basal lineage within Apiaceae subfamily Apioideae sister Africa. In the results of all molecular analyses, E. capense group to the Annesorhiza Glade. DNA sequence data also shows a close affinity with the Annesorhiza Glade, the support a possible close relationship with Astydamia and latter representing a group of deciduous, perennial herbs Molopospermum, as do some morphological and fruit endemic to southern Africa. Such a relationship between anatomical characters. The genus is endemic to the Eastern Ezosciadium and the Annesorhiza Glade has also been Cape Province of South Africa within the eastern part of proposed independently (C. Calvifio, unpublished data) on the Cape Floristic Region. the basis of MP analysis of cpDNA trnQ-rps16 sequences Ezosciadium capense (Eckl. and Zeyh.) B.L. Burtt in obtained from one of the same collections examined Edinb. J. Bot. 48(2): 207 (1991). Trachysciadium capense herein (Goldblatt & Porter 12578, MO). However, in all Eckl. and Zeyh., Enum. Pl. Afric. Austral. 341 (1837); cladograms currently available for the group, the rela- H. Wolff in Pflanzenr. Heft 90: 108 (1927). Helosciadium tionships among Ezosciadium, the Annesorhiza Glade, the capense (Eckl. and Zeyh.) Sond. in Harv. and Sond., Fl. European genus Molopospermum, and the Canary Islands Cap. 2: 536 (1862). Trachydium capense (Eckl. and Zeyh.) endemic genus Astydamia are not yet clear and demand Drude in Engl. & Prantl, Pflanzenfam. 3(8): 189 (1898). further study. Type: South Africa. Uitenhage district (3325): `Coega It seems that the basally divergent lineages are mor- Kopje' not far from `Zwartkopsrivier' (—DC), Ecklon and phologically as diverse as the more derived lineages as Zeyher 2196 (SAM!, lectotype here designated; MO, they include woody and herbaceous elements, with vari- photo!, P, photo!, S!, isolectotypes). ous leaf and fruit types. It is therefore not easy to Erect, annual herb, 0.1-0.35 m tall, pilose. Cotyledons ascertain the phylogenetic position of most genera without simple, narrowly oblanceolate, 15 mm x 2 mm, margins molecular systematic evidence. The lineage comprised of entire. Stem solitary, dichotomously branched. Leaves the Annesorhiza Glade, Astydamia, Ezosciadium and cauline, 10-37 x 7-26 mm, first leaves tri-lobed, upper Molopospermum has no obvious morphological synapo- leaves digitately compound. Petioles 5-18 mm long, morphies, but the combined presence of heteromericarpic sheathing slightly at the base. Pinnae 12-24 mm x 5- fruit (in Annesorhiza and Molopospermum), scattered 13 mm, 2- or 3-lobed; lobes linear-oblong to narrowly druse crystals in the mesocarp, and the highly lignified oblong, 5-12 mm x 1-2 mm, flat, apex acute, margins vascular bundles (in Annesorhiza and Ezosciadium) do, to and venation pilose, venation pinnate. Umbels compound, some extent, provide support for this Glade. sparse, axillary, sessile (peduncle absent, rays arising directly from each node); involucre present; bracts 2, 2-4 mm long, becoming prominently recurved, lanceolate, Taxonomic treatment apex acute, pilose; rays 2 to 4, unequal in length, with at least one remaining markedly shorter, longer rays Ezosciadium B.L. Burtt in Edinb. J. Bot. 48(2):207, 268 11-31 mm long and shorter ray 3-10 mm long at anthesis, (1991). Trachysciadium H. Wolff in Pflanzenr. Heft 90: 108 pilose; involucel present; bracteoles 2, 1-2 mm long, (1927), non Trachysciadium (DC.) Eckl. and Zeyh. (1837). becoming prominently recurved, lanceolate, apex acute, TYPE: E. capense (Eckl. and Zeyh.) B.L. Burtt. [Note: Burn pilose; umbellate rays 2 to 4, short, less than 1 mm long at (1991) gives a detailed argument for considering Trachys- anthesis, pilose. Flowers subsessile, pentamerous, her- ciadium as a synonym of Pimpinella L. and hence the need maphroditic; sepals obsolete; petals yellow, 0.6-0.7 mm for the new generic name, Ezosciadium. De Candolle (1830) long, ovate to obovate, tips not inflexed, acute, septum described Trachysciadium as a section of the genus Helos- absent on inner face, pilose on the dorsal surface; stamens ciadium to accommodate two Himalayan species now with anthers highly inflexed, small, 0.2-0.4 mm tall; ovary included in Pimpinella. The publication of the genus Tra- densely pilose, stylopodium flat; styles erect, very short. chysciadium by Ecklon and Zeyher (1837) had been Fruit isodiametric, oblong, 2.5-3.0 mm x ±1.5 mm considered valid by some authors (e.g. Wolff 1927), who broad; mericarps homomorphic, distinctly pilose; median, regarded it as being validated by the combined generico- lateral and marginal ribs 5, equal, prominent; commissural specific description, and taxonomically independent of vittae 2; vallecular vittae 4; commissure very narrow;
4-.1 Springer Ezosciadium (Apiaceae): a taxonomic revision 173
HEIGHT ABOVE SEA LEVEL 1-71 Over 1500 m 71 900 - 1500 m CD 300 - 900 m 1—I Under300 m
Fig. 4 The known geographical distribution of Ezosciadium capense carpophore bifid for the upper two-thirds of its length, been expanded to include Perdepoort (the westernmost persisting partly on the plant (Fig. 1). locality), Joubertina and Bethelsdorp.
Diagnostic characters Additional specimens examined
Ezosciadium capense is a highly distinctive species, easily South Africa, Eastern Cape Province. —3322 (Oudshoorn): distinguished from all other annual species by the pilose Perdepoort, N of Camfer (—CD), Goldblatt & Porter 12578 vegetative and reproductive organs, the sessile compound (NBG). —3323 (Willowmore): 4 miles N by W of Jou- umbels with a few, markedly unequal rays (Fig. I a), the bertina (—DD), Acocks 20021 (PRE). —3325 (Uitenhage): non-inflexed petal tip (Fig. lh), the relatively small, highly Bethelsdorp, salt pan (—DC), Fries et al. 1156 (BM, PRE). inflexed stamens which appear almost sessile (Fig. I i), and Precise locality unknown: Anon s.n. herb. S sheet 243 (S) the prominent bifid carpophores, one half of which persists on the plant (Fig. I a, e). Acknowledgment The authors would like to thank Prof. Peter Goldblatt, Dr John Manning and Mrs Edwina Marinus for providing recent material; the Lesley Hill Laboratory for the DNA aliquot of Distribution and habitat Ezosciadium capense, the Jodrell Laboratory and Kew Herbarium for the DNA aliquot of Stoibrax dichotomum, the curators and staff from This poorly collected species is endemic to the Eastern the cited herbaria who kindly made their specimens available for study, the molecular systematics laboratory at the University of Cape Province of South Africa and is known from only Johannesburg for the use of their facilities and Prof. Michael Pimenov four recorded populations (Fig. 4), three of which have and one anonymous reviewer for helpful comments on the manu- only come to light recently. Originally known only from script. Funding from the University of Johannesburg and the National Coega koppie near Uitenhage, the distribution range has Research Foundation of South Africa is gratefully acknowledged.
l Springer 174 A. R. Magee et al.
Appendix 1
List of taxa included in this study with GenBank accession numbers and authors
ITS Annesorhiza altiscapa Schltr. ex H.Wolff, DQ368830i; Annesorhiza fibrosa B.-E.van Wyk, DQ368831'; Annesorhiza filicaulis Eckl. and Zeyh., DQ368832'; Annesorhiza latifolia Adamson, DQ368833'; Annesorhiza macrocarpa Eckl. and Zeyh., DQ368835'; Astydamia latifolia (L.f.) Kuntze, DQ368836'; Chamarea snijmaniae B.L.Burtt, DQ368839'; Chamarea sp. I, DQ368840'; Ezosciadium capense (Eckl. and Zeyh.) B.L.Burtt, AM982517"; Itasina filifolia (Thunb.) Raf., DQ368857'; Lichtensteinia obscura (Spreng.) Koso-Pol., DQ368858'; Molopospermum peloponnesiacum Koch, AF074335c, AF074336a . rbcL Anginon rugosum (Thunb.) Raf., U50222 a; Annesorhiza altiscapa Schltr. ex H.Wolff, AM234812 Apium graveolens L. L01885 L11165"; Arctopus echinatus L. AY188414g; Berula erecta (Huds.) Coville, AM234813'; Bupleurum fruticosum L., D44556"; Eryngium giganteum M.Bieb., DQ133808 h; Ezosciadium capense (Eckl. and Zeyh.) B.L.Burtt, AM2348181; Heracleum sphondylium L., AY395540'; Hennas villosa Thunb., DQ133810h; Heteromorpha arborescens Cham. and Schltdl., DQ133811 h; Itasina filifolia (Thunb.) Raf., AM234821'; Lichtensteinia lacera Cham. and Schltdl., AM234822'; Pleurospennum camtschaticum Hoffm., D44583 b; Polemanniopsis marlothii (H.Wolff) B.L.Burtt, AM234824'; Sanicula europaea L., DQ133820h; Sium serra (Franch. and Savat.) Kitag., D44587"; Steganotaenia araliacea Hochst., EU2135I8"'; Torilis arvensis Link, AM234827'. rpsl 6 intron Aethusa cynapium L., AFI 105394; Alepidea amatymbica Eckl. and Zeyh., DQ832338 k; Alepidea serrata Eckl. and Zeyh., DQ832349k; Atnmi majus L., AFI64814f; Anginon diffonne (L.) B.L.Burtt, AY838391`; Anginon verticillatum (Sond.) B.L.Burtt, AY838404 a; Annesorhiza altiscapa Schltr. ex H.Wolff, AY838405 a; Annesorhiza filicaulis Eckl. and Zeyh., AY838407a; Annesorhiza latifolia Adamson, AY838408`; Annesorhiza macrocarpa Eckl. and Zeyh., AY838410a;
Apium graveolens L., AF110545d; Arctopus echinatus L., DQ832351 k ; Berula erecta (Huds.) Coville, AF164819 f; Bupleurum falcatum L. AFI I05664; Bupleurum fruticosum L., AF110569d; Chamaesciadium acaule C.A.Mey., AY838411 a; Chamarea longipedicellata B.L.Burtt, AY838413a; Chamarea snijmaniae B.L.Burtt, AY838414`; Chamarea sp.I AY838415`; Chamarea aff. gracillima (H.Wolff) B.L.Burtt, AY838416`; Dasispermum suffruticosum (Bergius) B.L.Burtt, AY838417 a; Deverra burchellii (DC.) Eckl. and Zeyh., AY838418`; Echinophora tenuifolia L. AF164812f; Ezosciadium capense (Eckl. and Zeyh.) B.L.Burtt, AM982518"; Foeniculum vulgare Mill., AFI 10543d; Helosciadium nodiflorum Koch, AF164820'; Heracleum sphondylium L., AF164800f; Hennas gigantea L.f., AY838420a; Hennas quercifolia Eckl. and Zeyh., AY838421`; Hermas quinquedentata L.f., AY838422"; Heteromorpha involucrata Conrath, AF110577 4; Heteromorpha papillosa C.C.Towns., AY838425`; Itasina filifolia (Thunb.) Raf., AY838426`; Komarovia anisospenna Korov., AF110555d; Lichtensteinia lacera Cham. and Schltdl., AY838427c; Lichtensteinia obscura (Spreng.) Koso-Pol., AY838428C; Molopospennum peloponnesiacum Koch, AY838432a; Notobubon galbanurn (L.) Magee, AY838435 e; Pastinaca sativa L., AF110538d; Pleurospennum foetens Franch., AFI 10559d; Pleurospennum uralense Hoffm., AF110560d; Polemannia montana Schltr. and H.Wolff, AF110570d; Polemannia simplicior Hilliard and B.L.Burtt, AFI 10571 d; Polemanniopsis marlothii (H.Wolff) B.L.Burtt, DQ832374k; Pseudocarum eminii H.Wolff, AY838441`; Pseudocarum laxiflorum (Baker) B.-E.van Wyk, Y838442'; Sanicula arctopoides Hook. and Am., DQ832375 k; Sium lattfolium L., AF110552d; Smyrnium olusatrum L., AF110551 d; Steganotaenia araliacea Hochst., DQ832384 k; Stoibrax dichotomum (L.) Raf., AM982519"; Torilis arvensis Link, AF110548d . b d a Olmstead et al. (1992), Kondo et al. (1996), Plunkett et al. (I 996b), Downie and Katz-Downie (1999), Downie et al. (2000a), f Downie et al. (2000c), g Chandler and Plunkett (2004), h Andersson et al. (2006), ' Calvino et al. (2006), Silvertown et al. (2006), k Calvillo and Downie
(2007), Forest et al. (2007), Lahaye et al. (2008), " present study
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Springer APPENDIX C2
MAGEE, A. R., B.-E. VAN WYK, P. M. TILNEY and S. R. DOWNIE 2009. A taxonomic revision of Capnophyllum (Apiaceae: Apioideae). South African Journal of
Botany 75: 283-291.
Available online at www.sciencedirect.com SOUTH AFRICAN °°;° Science D !red JOURNAL OF BOTANY
ELSEVIER South African Journal of Botany 75 (2009) 283-291 www.elsevier. com/locate/sajb
A taxonomic revision of Capnophyllum (Apiaceae: Apioideae)
A.R. Magee a ' * , B.-E. van Wyk a , P.M. Tilney a , S.R. Downie b
Department of Botany and Plant Biotechnology University of Johannesburg. PO Box 524, Auckland Park 2006, South Africa
b Department of Plant Biology, University of Illinois at Urbana-Champaign, Urbana. Illinois 61801, USA
Received II April 2008; received in revised form 21 October 2008; accepted 5 February 2009
Abstract
The Cape endemic genus Capnophyllum Gaertn. is revised. As a result of valuable recent collections and extensive fieldwork, this hitherto neglected genus was found to comprise four annual species, two of which are newly described, namely C. lutzeyeri Magee and B.-E.van Wyk, and C. macrocarpum Magee and B.-E.van Wyk. The four species are distinguished from one another by their fruit morphology (relative length of the styles, the shape and position of the stylopodium, fruit size, surface sculpturing, and the presence or absence of a sterile apical portion) and fruit anatomy (marginal wings slightly or prominently involute and secondary ribs present or absent). A comprehensive key to the species, their complete nomenclature and typification, together with complete descriptions and known geographical distributions for all the species are presented and illustrated. © 2009 SAAB. Published by Elsevier B.V. All rights reserved.
Keywords: Cape endemic; Fruit anatomy; Kruhera; Morphology; New species; South Africa
1. Introduction nophyllum group). In contrast, Krubera was shown to be only distantly related, occupying a position somewhere between the Capnophyllum Gaertn. is a genus of small annual herbs tribes Apieae and Selineae (Downie et al., 1998, 2000; Winter endemic to South Africa. The name is derived from the Greek et al., 2008; Magee et al., in press). (kapnos=smoke, but is also the Greek name for Fumaria; Until relatively recently, the genus in the strictest sense was phyllon= leaf) and refers to the distinctly glaucous leaves that considered to be monotypic, with two varieties of C. africanum closely resemble those of the genus Fumaria L. (Sonder, 1862; (L.) Gaertn. recognised on the basis of fruit surface sculpturing. Adamson, 1950). Capnophyllum is sometimes extended to Goldblatt and Manning (2000), however, found that the two include the Mediterranean Krubera peregrinum Lowe (e.g. varieties were morphologically and geographically distinct and Tulin et al., 1968; Dyer, 1975; Downie et al., 1998). However, therefore raised C. africanum var. leiocarpon Sond. to the according to Meikle (1977), Krubera Hoffm. can be distin- rank of species, as C. leiocarpon (Sond.) J.C. Manning and guished from Capnophyllum by fruit and floral differences. Goldblatt. As a result of valuable recent collections and Bunt (1991) followed Meikle (1977) in considering Capno- extensive fieldwork, this hitherto neglected genus was found phyllum to be a South African endemic genus. Recently, Magee to comprise four species, two of which are as yet undescribed. et al. (in press) using molecular and morphological data, We present here a detailed taxonomic treatment of the genus, provided strong support for the monophyly of Capnophyllum including a key to the species, their complete nomenclature, and for its placement within the Lefebvrea Glade of the tribe typification, formal descriptions, as well as the known Tordylieae, together with the Cape genera Dasispermum, Son- geographical distributions. derina and Stoibrax capense (therein referred to as the Cap-
2. Materials and methods
* Corresponding author. The complete collections of Capnophyllum from the E-mail address: [email protected] (A.R. Magee). following herbaria were studied: BM, BOL, JRAU, K, NBG
0254-6299/S - see front matter © 2009 SAAB. Published by Elsevier B.V. All rights reserved. doi:10.1016/j.sajb.2009.02.001 284 A.R. Magee et al. / South African Journal of Botany 75 (2009) 283-291
Fig. I. General morphology of Capnophyllum. (a) C. macrocarpum in situ (photo by B—E. Van Wyk), showing the glaucous Fumaria-likc leaves charactcristic of the genus as well as the sparse compound umbels and small stature typical of this species; (b) C. leiocarpon in situ (photo by A.R. Magee), showing the distinctive decumbent branches; (c) typical sympodial growth form; (d) representative lower leaf pinnac; (e) typical petal in ventral and lateral view; (f) umbellule of C. africanum showing the distinct tubcrculcs on the ovaries. Vouchers: (c) Magee et al. 125, JRAU; (d) Rester 6978, JRAU; (e) Williamson 3825, BOL; (f) Magee et al. 124, JRAU. Scale: c=25 mm, d and c=5 mm, f= I mm.
(including SAM and STE), PRE and S (herbarium acronyms as et al. 133 (JRAU). Kruberav peregrinum: Lippert 22969 in Holmgren et al., 1990). Extensive field work was undertaken (PRE); Sandwith 6215 (K). to study all the taxa in situ, thus providing crucial additional To study the three-dimensional structure of the vittae (oil insight into the species concepts proposed in this study. From canals), mature fruit were softened by soaking them in boiling this material, together with geographical information from water for 24 h. The exocarp was then peeled off while keeping Leistner and Morris (1976), the recorded distribution of the the fruit submerged in water to prevent desiccation. species was carefully verified and mapped. The line drawings (Figs. 1 and 2) were made by the first author with the aid of a 3. Results and discussion camera lucida attachment on a Wild M3Z stereomicroscope. FAA and herbarium material were used to study fruit 3.1. Vegetative morphology anatomy. The herbarium material was first rehydrated and then placed in FAA for a minimum of 24 h. The material was The four species of Capnophyllum are all annual herbs subsequently treated according to a modification of the method which exhibit a sympodial growth pattern similar to that found of Feder and O'Brien (1968) for embedding in glycol in the closely related genus Sonderina H. Wolff (Magee et al., in methacrylate (GMA). This modification involves a final infiltra- press). In both these genera, the stem consists of a series of tion of the material in GMA for five days. Staining was done sympodial segments each ending in a terminal umbel which according to the periodic acid Schiff/toluidine blue (PAS/TB) becomes laterally displaced by continued growth from the staining method (Feder and O'Brien, 1968). The terminology axillary bud of the uppermost leaf, which takes over as the main used to describe the fruit anatomy follows that proposed by growth axis (Burtt, 1991). In Capnophyllum the plants are all Kljuykov et al. (2004). Voucher specimens for the fruit anatomical well-branched sprawling herbs in which the branches become study are listed below. prostrate or decumbent (Fig. 1 a—c). As the generic name Capnophyllum africanum: Van der Mere 1767 (NBG); implies, the leaves are always characteristically glaucous. They Winter 110 (JRAU); Anon. sub. NBG 754111. Capnophyllunt are pinnate to bi-pinnate and mostly cauline (Fig. 1), with the leiocarpon: Stirton & Zantovska 11430 (NBG); Taylor 12003 ultimate leaflet segments flat or subterete and less than 1 mm (PRE); Williamson 3825 (PRE). Capnophylhan lutzeyeri: Ma- broad (Fig. 1d). All the species are relatively small in stature gee et al. 106 (JRAU). Capnophyllum macrocarpum: Magee (ranging from 50-500 mm in height). Capnophyllum lutzeyeri A.R. Magee et al. / South African Journal of Botany 75 (2009) 283-291 285
Fig. 2. Variation in fruit morphology (a—d) of Cap/top/Om species with drawings of the stylopodia (c—h): (a, c) C. africanum; (b, f) C. lutzeyeri; (c, g) C. leiocarpon; (d, h) C. macrocarpum. Vouchers: (a 1) Bachmann s.n., BOL: (a2)Bohts 2793, BOL; (a3) Winter 110, JRAU; (a4) Adamson 3081, BOL; (a5) Van der Menve 1767, NBG; (b I ) Lutzeyer 7A, JRAU; (b2) Magee et al. 106, JRAU; (c 1) Williamson 3825, BOL; (c2)Stinon & Zantovska 11432, NBG; (c3) Pillans 7899, BOL; (d, three fruit) Magee et al. 133, JRAU, (c) Winter 110, JRAU; (f) Lutzeyer 7A, JRAU; (g) Stirton & Zantovska 11430, NBG; (h) Magee 133, JRAU. Scale: a—d=3 mm, c—h= I mm. appears to be more robust than the other species, while The shape and position of the stylopodia in mature fruit were C. macrocarpum (Fig. la) is generally much smaller at maturity. found to be useful diagnostic characters (Fig. 2). The stylopodia However, as both these species are known from only a single are either conical and raised above the fruit apex as in locality, the size of the plants may eventually prove to be of C. africanum (Fig. 2a and e), C. leiocarpon (Fig. 2c and g) limited diagnostic value. and C. lutzeyeri (Fig. 2b and 1), or very shortly conical to flattish and diagnostically sunken below the fruit apex in C. macro- 3.2. Reproductive morphology carpum (Fig. 2d and h). The styles are at first erect and relatively short, but may lengthen as the fruit mature. Their relative length The relatively small compound umbels appear to be laterally is an important diagnostic character. In C. macrocarpum (Fig. 2d borne along the stems but are in fact terminal and become leaf- and h) the styles remain erect and relatively short whereas in the opposed due to continued growth from the axillary shoots. The other species they lengthen markedly and become reflexed. The bracts of the involucre and involucel may be either free or styles of C. leiocarpon become reflexed far beyond the base of connate at their bases, and the rays and raylets are invariably the stylopodium (Fig. 2c and g), while in the similar C. lutzeyeri glabrous (Fig. I f). The umbels of C. macrocarpum are always they are reflexed only up to, at most, the stylopodium base very sparse with only two or three rays consistently present (Fig. 2f). In C. africanum there appears to be some variability in (Fig. I a). The other species usually have at least four rays, that the styles may become reflexed either up to or beyond the although two or three may rarely be present in C. leiocarpon base of the stylopodium. and C. lutzeyeri. The fruit of Capnophyllum are dorsally compressed, with the The hermaphroditic flowers are pentamerous, with indistinct marginal ribs expanded into narrow wings (Fig. 2) and the sepals and white, oblong to obovate, papillose petals with commissure extending over the full width of the mericarp (i.e. to acuminate inflexed tips (Fig. I e). The ovaries of C. africanum the very edge of both marginal ribs). The marginal wings are (Fig. I 0 and C. macrocarpum are distinctive in that they are distinct in that they are slightly to prominently involute, so that prominently tubercled, while in the other species they are the commissural surface is very slightly to prominently concave smooth. (Fig. 3). In transverse sections of immature fruit, the commissural 286 A.R. Magee et al. / South African Journal of Botany 75 (2009) 283-291
Fig. 3. Transverse sections through the fruit of Capnophyllum (a—d) and Krubera (c): (a) C. lutzeyeri; (b) C. macrocarpum (almost mature fruit); (c) C. africanum; (d) C. leiocarpon; (c) K. peregrinum with inlay showing enlargement of a commissural vitta. Vouchers: (a) Magee et al. 106, JRAU; (b) Magee et al. 133, JRAU; (c) Winter 110, JRAU; (d) Williamson 3825, PRE; (c) Lippert 22959, PRE. cc=commissural cavity; cv=commissural vitta; Ir=lateral rib; mcdr=median rib; mr=marginal rib; rod—rib oil duct; sr=sccondary ribs; vv=vallecular vitta. Scale: a—c= I mm, inlay=0.2 mm. area between the marginal wings is composed of parenchymatous sural and vallecular vittae unlike those of the species of cells which do not become lignified with maturation of the fruit. Capnophyllum (Fig. 3a—d) where the vittae are relatively Instead these cells disintegrate to create a hollow between the two large and conspicuous. The fruit of Krubera also differ mericarps (Fig. 3a and b). The median and lateral ribs are slightly markedly in the presence of large rib oil ducts. The fruit prominent to prominent and are diagnostically tubercled in C. anatomical data therefore supports the separation of the two africanum (Fig. 2a) and C. macrocarpum (Fig. 2d). Secondary genera as found by Magee et al. (in press). ribs are often present above the vallecular and commissural vittae in C. africanum, C. leiocarpon and C. lutzeyeri (Fig. 3a, c and d). 4. Taxonomic treatment The fruit of C. macrocarpum (Fig. 2d) are the largest in the genus, with even the smallest fruit of this species distinctly larger than the Capnophyllum Gaertn., Fruct. 2: 32 (1792); DC., Prodr. 4: largest fruit of C. africanum (Fig. 2a). In transverse section there 187 (1825); Sond. in Harv. and Sond., Fl. Cap. 2: 562 (1862); are two commissural vittae as well as four solitary vallecular vittae Adamson in Fl. Cape Penins. 625 (1950); Dyer, Gen. S. A fr. (Figs. 3 and 4) in each mericarp of all the species. Flowering Pl. 2: 426 (1975), p.p. maj.; B.L. Burtt in Edinb. J. In Capnophyllum macrocaipum there is a prominent sterile Bot. 48(2): 189 (1991). Type: C. africanum (L.) Gaertn. apical portion on the fruit that can most easily be seen by Abioton Raf., Good Book 56 (1840) reimp. Scad. Gen. Omb. comparing the extent of the commissural vittae (Fig. 4a). In this Pl. 56 (Amer. Midl. Nat. Repr. No. 3, 1913), nom. illegit. Type: species the commissural vittae terminate well below the A. africanum (L.) Raf. stylopodium; in C. lutzeyeri they terminate slightly below the Actinocladus E.Mey. in Ind. Sem. Hort. Bot. Regiomont stylopodium leaving a much smaller sterile portion (Fig. 4c). In (1846) and in Ann. Sci. Nat. Bot. Ser. 3,7: 380 (1847). Type: the remaining species the commissural vittae terminate at the A. cinerascens E.Mey. base of the stylopodium so that there is no conspicuous sterile Sprawling sympodial herbs, 0.05-0.5 m tall, annual. Stems portion (Fig. 4b and d). well-branched at the base, branches usually prostrate or decumbent. Leaves cauline, 15-210 min x 7-90 mm, usually 3.3. Comparison of Capnophyllum and Krubera becoming smaller towards the upper part of the stem, pinnate to bi-pinnate, glabrous, glaucous (and closely resembling those of The fruit of Capnophyllum and Krubera are superficially Fumaria). Petioles 5-110 mm long, angular, basal sheaths similar in having prominent ridges on the dorsal surface, a broad prominent, 3-12 min x 2-9 mm, margins widely membranous. commissure, dorsally compressed mericarps and marginal ribs Ultimate leaflets ovate to broadly ovate or obovate, 7- extended into wings. On close examination of the fruit anatomy, 17 mm x 3-13 mm, pinnatisect, venation not visible or pinnate; however, the two genera are clearly distinct. The fruit of Kru- segments linear-oblong, 1-6 mm long, less than 1 mm broad, bera (Fig. 3e) have extremely small, inconspicuous commis- flat or subterete, apex obtuse to acute. Umbels compound; A.R. Magee et al. / South African Journal of Botany 75 (2009) 283-291 287
or flexuose, smooth or tubercled; marginal ribs narrowly winged, slightly to prominently involute, leaving a small to large commissural cavity; secondary ribs often present above the vallecular and commissural vittae; commissural vittae 2, terminating either directly below or some distance from the base of the stylopodium; vallecular vittae 4; commissure very broad, 100% of mericarp width; carpophore bipartite.
4.1. Diagnostic characters C Species of Capnophyllum are annual, often sprawling, d white-flowered herbs with soft, glaucous leaves resembling those of Fumaria. They differ from other vegetatively similar Sp annual African genera by the dorsally compressed, narrowly- winged fruit with a broad commissure (100% of mericarp width). They are distinct from African peucedanoid genera in their sympodial growth pattern resulting in leaf-opposed umbels. All other African peucedanoid genera have a mono- podial growth pattern with terminal umbels. Furthermore, the fruit differ from those of other annual African peucedanoids in the usually prominent median and lateral ribs, the narrowly- winged marginal ribs which are slightly to prominently involute, the often conspicuous secondary ribs on both the commissural and dorsal surfaces, the slightly to prominently concave commissural surface of the mericarps and the strongly elliptical outline in both dorsal and lateral views as a result of the convex outer surface. a The species often co-occur with the superficially similar and widespread Sonderina hispida (Thunb.) H.Wolff. Although the Fig. 4. Three-dimensional structurc of the commissural vittac of Capnophyllum fruit of Capnophyllum are fundamentally different from those species: (a) C. macrocatpum; (b) C. africanum; (c) C. hazeyeri; (d) C. leiocarpon. of Sonderina (in Sonderina the fruit are isodiametric, lack Vouchers: (a) Magee et al. 133, JRAU; (b) Van der Merwe 1767, NBG; (c) Magee marginal wings and have a narrow commissure), when in flower et al. /06. JRAU; (d) Williamson 3825. BOL. cv =commissural vitta; sp=stcrilc they may easily be confused with one another. However, even portion. Scale: I mm. when in flower Capnophyllum species can be distinguished by their prostrate habit, glaucous leaves, tubercled ovaries in C. cifi•icanum and C. macrocarpunz (glabrous or more usually peduncle short; involucral bracts 2 to 4 (to 6), connate or free, scabrous or pilose in Sonderina hispida), petals appearing lanceolate to ovate, apex acuminate, margins widely membra- truncate or at most shallowly-emarginate in dorsal view nous, glabrous; rays (2 to) 4 to 10 (to 14), 10-45 mm long at (deeply-emarginate in Sonderina), and glabrous umbels (scab- anthesis, glabrous; involucel bracteoles 4 (to 6), connate or free, rous in Sonderina hispida). lanceolate to ovate, apex acuminate, margins widely membra- nous, glabrous; raylets 6 to 25, 2-8 mm long at anthesis, glabrous. Flowers hermaphroditic; pentamerous; sepals indis- 4.2. Distribution and habitat tinct; petals white, ±1 mm long and broad, broadly oblong to The four species are endemic to the Cape Floristic Region of obovate, tips inflexed, acuminate, septum present on inner face, apex truncate to shallowly emarginate, papillose; stamens with South Africa (Figs. 5 and 6) and occur in sandy soil near to the anthers inflexed; ovary inferior, smooth or tubercled, stylopo- coast. The species are generally allopatric in their distributions; however there is some overlap between the ranges of dium shortly conical or flat, raised above or sunken below the around Langebaan and Saldanha. fruit apex; styles at first erect, short, often longer in mature fruit, C. a•icanum and C. leiocarpon either remaining erect or becoming reflexed up to or beyond the base of the stylopodium. Fruit a schizocarp, dorsally com- 4.3. Phenology pressed, elliptic to broadly elliptic or ovate to broadly ovate in dorsal view, 5.5-9.0 min x 2.5-5.5 mm; narrowly elliptic and Seeds germinate during winter after the first autumn rains. convex in lateral view; base obtuse to truncate or shallowly Flowering occurs in spring (September and October) with concave; apex obtuse to truncate; mericarps homomorphic, very mature fruit forming from late spring to early summer slightly to markedly concave on the commissural surface; (November and December). All four species are annuals that median and lateral ribs slightly prominent to prominent, straight wither and die during the dry mid-summer (January). 288 A.R. Magee et al. / South African Journal of Botany 75 (2009) 283-291
HEIGHT ABOVE SEA LEVEL lb. Flowers and fruit with smooth median and lateral ribs = Over 1500 m (without tubercles) E:1 900 - 1500 m 3 ECZI 300 - 900 m I= Under 300 in Styles of mature fruit reflexed and extending±to the base of the stylopodium, short (0.7-0.8 mm long); mericarp markedly concave on the com- missural surface; known only from Stanford in the Western Cape 3 C. lutzeyeri Styles of mature fruit reflexed and extending far beyond the base of the stylopodium, long (1.2- 1.7 mm long),; mericarp only slightly concave on the commissural surface; known from the west coast from Langebaan northwards to Port Nolloth
2 C. leiocarpon
4.5. Capnophyllunt ofricanum
C. africanum (L.) Gaertn., Fruct. 2: 32, tab. 85, f. 6 (1792); DC., Prodr. 4: 187 (1825); Ecklon and Zeyh., Enum. Pl. Afric. Austral. 353 (1837); Sonder in Fl. Cap. 2: 562 (1862); Adamson
HEIGHT ABOVE SEA LEVEL 1=1 Over 1500 m CD 900 - 1500 m 1=3 300 - 900 m = Under 300 m
Fig. 5. Geographical distribution of Capnophyllutn africanum.
4.4. Key to the species of Capnophyllunt
la. Flowers and fruit with tubercles on the median and lateral ribs (most prominent on ovaries and young fruit) 2 Fruit more than 8.5 mm long; stylopodium sunken below the apex of the fruit, very shortly conical to flat; fruit with conspicuous sterile apical portion (commissural vittae terminating some distance from the base of the stylopodium); rays 2 or 3 per umbel; known only from De Hoop Nature Reserve 4. C. moo -occupant Fruit 8 mm long or less; stylopodium not sunken below the apex of the fruit, shortly conical; fruit without a conspicuous sterile apical portion (commissural vittae terminating directly below the base of the stylopodium); rays more than 3 per umbel; from Gordon's Bay northwards along the coast to Vredenburg Fig. 6. Geographical distribution of Capnophylltun leiocarpon (diamonds), C. 1. C. afi-icanunt lutzeyeri (triangle), C. macrocarpunt (square). A.R. Magee et al. / South African Journal of Botany 75 (2009) 283-291 289 in Fl. Cape Penins. 625 (1950). Conium africanum L., Sp. Pl. has also been collected on Robben Island. It occurs in open 243 (1753); Mant. Alt. 252 (1771); Jacq., Hort. Vindob. 2: tab. Strandveld on deep sandy soils. 194 (1772); Thunb., Fl. Cap. 257 (1823). Caucalis africana (L.) Crantz, Cl. Umbell. Emend. 109 (1767). Cicuta africana (L.) 4.8. Additional specimens examined Lam., Encycl. 2: 4 (1786), saltem quoad basionym. Abioton africanum (L.) Rat:, Good Book 56 (1840); reimp. Scad. Gen. South Africa. —3217 (Vredenburg): 5 km from Vredenburg Omb. 56 (Amer. Midl. Nat. Repr. 3, 1913). Type: "Caucalis to Saldanha (—DD), Stirton 10723 (K, NBG). —3318 (Cape afra, folio minoris Rutae" in Boerhaave, Index Pl. Alt. 1: 63 Town): Elandsfontein west, second ridge W of farmhouse (—AA), (1720), (lecto., designated by Van Wyk and Tilney in Jarvis Thompson 3534 (NBG, PRE); Geelbek Farm, Langebaan (—AA), et al., 2006). Bosenberg & Rutherford 138 (NBG); flats at S end of Langebaan Capnophyllum simplex Lagasca, Gen. and Sp. Nov. 13 lagoon (—AA), Goldblatt 2711 (NBG, PRE); along R27, 14 km (1816), nom. illegit. from Langebaan to Cape Town, near Club Mykonos turnoff(—AA), Capnophyllum jacquinii DC., Prodr. 4: 187 (1830), p.p. min. Magee & Boatwright 114 (JRAU); along R27, resting area near Actinocladus cinerascens E.Mey. in Ind. Sem. Hort. Bot. Swartberg, 33° 13' 12" S 18° 12' 15" E (—AA), Forest et al. 654 Regiomont (1846) and in Ann. Sci. Nat. Bot. Ser. 3, 7: 380 (NBG); along R27, from Velddrif to Cape Town, 33° 07' 38" S 18° (1847). Type: Hort. Region, Zeyher ex herb. Meyer s.n. 07' 53" E (—AA), Boatwright et al. 228 (JRAU); along R27, from (S!, lecto., here designated). [Note: The specimen in S is Velddrif to Cape Town, 33° 10' 33" S 18° 10' 21" E (—AA), chosen here as the label is in Meyer's handwriting and bears the Boatwright et al. 229b (JRAU); sand near Hopefield (—AB), annotation "mihi".]. Bachmann s.n. sub BOL 1627 (BOL, K); 11 km NNE Sprawling or rarely somewhat erect herb, 0.05-0.3 m tall. of Yzerfontein (—AD), Strid & Strid 38062 (NBG); Buffelsrivier Leaves 15-210 mm long x 8-60 mm. Petioles 5-110 mm long, (—CB), Van der Merwe 1767 (K, NBG, PRE); Buffelsrivier, Hardy basal sheaths 3-12 mm x 3-9 mm. Ultimate leaflets ovate to Joubert's sample plots near Bokbaai (—CB), Taylor 4165 (NBG, broadly ovate, 7-17 mm x 4-13 mm, venation not visible or PRE); Melkbosstrand (—CB), Dahlstnind 1064 (PRE); Clifton-on- pinnate; segments linear-oblong, 1-6 mm long, less than 1 mm sea (—CD), Phillips s.n. sub PRE 59930 (PRE); Milnerton (—CD), broad, flat or subterete. Umbels with peduncle short, 5-60 mm Andreae 381 (NBG); Wall 196 (S); Robben Island (—CD), Adam- long; involucral bracts (2 to) 4 (to 6), connate or free; rays 4 to 7 son s.n. (BOL); Sunset Beach, S of Table View (—CD), Van (to 14), 10-45 mm long at anthesis; involucel bracteoles 4 Slageren & Newton 842 (K); Brackenfell, Cape Flats (—DC), Du (to 6), connate or free; raylets 10 to 25, 2-7 mm long at Plessis 108 (PRE); Cape Flats Nature Reserve (—DC), Low 866a anthesis. Flowers with ovary tubercled; stylopodium shortly (PRE); Cape Flats (—DC); Pappe s.n. (K); Wallich s.n. (BM); sandy conical, raised above the fruit apex; styles lengthening as fruit flats near Kuils River (—DC), Anon ex herb. Worsdell sm. (K); high matures, (0.3—) 0.5-0.8 mm long, becoming reflexed up to or dunes W of Sarepta (—DC), Acocks 1112, 1294 (S); Stikland (—DC), beyond the base of the stylopodium. Fruit elliptic to broadly Acocks 2253 (S); Vygekraal (—DC), MacOwan 1751 (BM, NBG, elliptic or ovate to broadly ovate, 6.5-8.0 mm x 2.5-4.0 mm; K), Wolley Dod 706 (K). —3418 (Simonstown): Fish Hoek (—AB), base obtuse or shallowly concave; apex obtuse to truncate; Young 27349 (PRE, fragment labelled A); near Glencaim (—AB), mericarps very slightly concave on the commissural surface; Hutchinson 640 (BM, BOL, K, PRE); Kalk Bay (—AB), Levyns median and lateral ribs prominent, flexuose, tubercled; marginal s.n. sub BOL 1625 (BOL); Kommetjie, Imhoff's Gift (—AB), ribs slightly involute; secondary ribs usually present above the Winter 110 (JRAU); Oaklands (—AB), Jameson son. (K); vallecular and commissural vittae; commissural vittae terminat- Retreat Railway Station (—AB), Bolus 2793 (BOL, PRE); ing directly below the base of the stylopodium. Scarborough (—AB), Adamson 3081 (BOL); Varkensvlei Experi- mental Farm (—BA),Joubert 26147 (NBG); Wynberg Flats (—BA), Anon ex herb Prior sm. (K); Gordons Bay (—BB), Parker 3860 4.6. Diagnostic characters (BOL, K, NBG); Somerset in Hottentots Holland (—BB), Ecklon & Zeyher 2252 (S); Strand (—BB), Parker 4349 (K, NBG). Precise Capnophyllum africanum is vegetatively similar to locality unknown: "Prom. b. Spei", Masson s.n. (BM); " In C. leiocarpon and C. /utzeyeri but differs in that both the ovaries arenosis ad likes marls Sinu False Bay", Bolus 4632 (BM, BOL); and fruit are covered with tubercles on the median and lateral "In solo arenosa prope Capetown", Bolus 2793 (BOL, K, NBG); ribs, as in C. macrocarpum. It differs from the latter species, Burchell 362 (K, PRE); Zeyher 742 (K, two sheets); Zeyher 635 however, in the larger number of rays per umbel (4 or more), the (K); Ecklon & Zeyher 2252 (NBG). smaller fruit (less than 8 mm long) without a sterile apical portion and with the shortly conical stylopodium raised above the fruit 4.9. Capnophyllum leiocarpon apex. C. leiocarpon (Sond.) J.C.Manning and Goldblatt in Gold- 4.7. Distribution and habitat blatt and Manning, Cape Pl. 705 (2000). Capnophyllum africanum var. leiocarpon Sond. in Harv. and Sond., Fl. Cap. Capnophyllum africanum is endemic to the Western Cape 2: 562 (1862); B.L. Burtt in Edinb. J. Bot. 48(2): 189 (1991). Province, occurring from Gordons Bay and the Cape Peninsula Type: South Africa. Near Cape Town, Drege 6243 (S!, lecto., northwards along the coast to Vredenburg (Fi g. 5). This species here designated; BM!, G, K!, two sheets, MO, isolecto.). 290 A.R. Magee et al. / South African Journal of Botany 75 (2009) 283-291
Sprawling or somewhat erect herb, 0.1-0.5 m tall. Leaves Pan Nature Reserve (—CB), Magee & Boatwright 116 50-200 mm x 20-90 mm. Petioles 8-90 mm long, basal (JRAU), Van Rooyen & Ramsey 256 (NBG); Berg River sheaths 4-12 mm x 3-6 mm. Ultimate leaflets ovate to broadly Mouth (—CC), 0' Callaghan 8/11 (NBG). — 3318 (Cape ovate or obovate, 7-16 mm x 5-10 mm, venation not visible or Town): along R27, from Veldrif to Cape Town, 33° 10' 33" S pinnate; segments linear-oblong, 1-6 mm long, less than 1 mm 18° 10' 21" E (—AA), Boatwright et al. 229a (JRAU). broad, flat or subterete. Umbels with peduncle short, (3—) 10- 60 (-105) mm long; involucral bracts 2 to 4, connate, 4.10. Capnophyllum lutzeyeri sometimes free; rays (2 to) 4 to 8, 10-30 mm long at anthesis; involucel bracteoles 4 (to 6), connate; raylets 10 to 25, 2-7 mm C. lutzeyeri Magee and B.-E.van Wyk, species novum, Cap- long at anthesis. Flowers with ovary smooth; stylopodium nophyllo africano et C. leiocarpo stylopodio brevi costis fructus shortly conical, raised above the fruit apex; styles lengthening dorsalibus secondariisque prominentibus similis sed fructu non as the fruit matures, 1.2-1.7 mm long, becoming reflexed far tuberculato stylis brevioribus et supetficiebus commissurialibus beyond the base of the stylopodium. Fruit elliptic to broadly valde concavis differt. Type: South Africa. Caledon district (3419): ovate, 5.5-8.0 mm x 3.0-4.0 mm; base and apex obtuse to Grootbos Nature Reserve, Steynsbos (—CB); 15 December 2006; truncate; mericarps slightly to very slightly concave on the H. Lutzeyer 7A (NBG!, holo.; JRAU!, iso.). commissural surface; median and lateral ribs prominent, Sprawling herb, 0.2-0.5 m tall. Leaves 15-60 mm x 7-25 mm. flexuose, smooth; marginal ribs slightly involute; secondary Petioles 5-15 mm long, basal sheaths 3-8 mm x 2-5 mm. Ulti- ribs usually present above the vallecular and commissural mate leaflets ovate to broadly ovate, 7-15 mm x 4-9 mm, vittae; commissural vittae terminating directly below the base of venation not visible; segments linear-oblong, 1.5-3 mm long, less the stylopodium. than 1 mm broad, subterete. Umbels with peduncle short, 15- 75 mm long; involucral bracts 2 to 4, free?; rays (2 to) 4 to 15, 10- 4.9.1. Diagnostic characters 30 mm long at anthesis; involucel bracteoles 4, free?; raylets 7 to Capnophyllum leiocarpon differs from C. africanum and 10, 2-6 mm long at anthesis. Flowers with ovary smooth; C. macrocarpum in the absence of tubercles on the ribs of both stylopodium shortly conical, raised above the fruit apex; styles the ovaries and fruit. It can be distinguished from C. lutzeyeri lengthening slightly as the fruit matures, 0.7-0.8 mm long, by the longer styles of the mature fruit that become reflexed far becoming reflexed up to or just above the base of the stylopodium. beyond the base of the stylopodium, and the only very slightly Fruit broadly elliptic, 6.0-6.5 mm x 3.5-4.0 mm; base obtuse to concave commissural surfaces of the mericarps. truncate; apex obtuse; mericarps markedly concave on the commissural surface; median and lateral ribs slightly prominent, 4.9.2. Distribution and habitat straight, smooth; marginal ribs prominently involute; secondary This widespread species occurs along the coast from near ribs present above the vallecular and commissural vittae; Langebaan in the Western Cape Province northwards to Port commissural vittae terminating slightly below the base of the Nolloth in the Northern Cape Province (Fig. 6). It occurs in stylopodium. open Strandveld on deep sandy soil. 4.10.1. Diagnostic characters 4.9.3. Additional specimens examined Capnophyllum lutzeyeri differs from C. africanum and
South Africa. — 2916 (Port Nolloth): +15 km E of Port Nolloth C. macrocarpum in the smooth fruit and from C. leiocarpon in
(—BB), Williamson 3508 (BOL), 3825 (BOL, NBG, PRE). — 2917 the shorter styles, which become reflexed only to the base of the (Springbok): Farm Zonnekwa 328, 1 km N of crossroads to stylopodium, and the markedly concave commissural surface. Graafwater and Sonnekwa B on road from Vaalkol (—CD), Le Roux
& Lloyd 5/7 (NBG). —3017 (Hondeklipbaai): Farm Koingaas 4.10.2. Distribution and habitat 475, on road from Hondeklipbaai to Koinghaas on S banks of The species is known only from the private nature reserve Swartlintjies River (—AB), Le Roux & Lloyd 327 (NBG); Grootbos, near Stanford in the Western Cape Province (Fig. 6), Farm Ghaams 492, SE of Spoeg River (—AD), Boucher 5667 where it was collected on acid derived sand in Fynbos (NBG); sand dunes between Wallekraal and Hondeklipbaai (—AD), vegetation that had burnt the previous year. No plants were
Goldblatt 4222 (K, PRE). — 3117 (Lepelfontein): Brand-se-Baai found in subsequent searches by Mr H. Lutzeyer, suggesting
(—BD), Van Rooyen 2154 (PRE). — 3217 (Vredenburg): that this species may be a short-lived fireweed. Paternoster Bay (—DD), Lavranos 11708 (PRE); 5 km from
Vredenburg to Saldanha (—DD), Stirton 10723 (NBG). — 3218 4.10.3. Additional specimens examined
(Clanwilliam): Nortier Experimental Station, Lamberts Bay South Africa. — 3419 (Caledon): Steynsbos, Grootbos (—AB), Boucher 2539 (K, PRE), Liengme 988 (BOL), Van Nature Reserve (—CB), Magee et al. 106 (JRAU, NBG, PRE). Breda 4296 (K, PRE), Van der Merwe 1664 (PRE); Elands Bay (—AD), Metelerkamp 233 (BOL), Pillans 7899 (BOL); 4.11. Capnophyllunz macrocarpum 5 km from Elands Bay on road to Lamberts Bay (—AD), Stirton & Zantovska 11430 (NBG); 2 km from Redelinghuys C. macrocarpum Magee and B.-E.van Wyk, species novum, (—BC), Stirton & Zantovska 11432 (NBG, two sheets); Capnophyllo africano costis dorscdibus tuberculatis ovarii Dwarskersbos (—CA), Taylor 12003 (NBG, PRE); Rocher fructusque similis sed fructu maiori portione apicali sterile A.R. Magee et al. / South African Journal of Botany 75 (2009) 283-291 291
Longo, stylopodio immerso rudimentali et superficiebus com- express gratitude to Dr J.C. Manning (NBG) for assistance with missurialibus valde concavis differs. Type: South Africa. collections and for alerting us to unusual collections. Mr and Mrs Caledon district (3420): De Hoop Nature Reserve, near to De Lutzeyer are thanked for their enthusiasm and hospitality while Mond (-AD); 4 November 2007; A.R. Magee, B.-E. Van Wyk studying the species growing on Grootbos Nature Reserve near and J.S. Boatwright 133 (NBG!, holo.; BOL!, JRAU!, K!, iso.). Stanford. Dr H. Glen is gratefully acknowledged for translating Sprawling herb, 0.04-0.08 m tall. Leaves 25-85 min x 10- the diagnoses. We are grateful to Mr A. Marais and his team from 40 mm. Petioles 10-45 mm long, basal sheaths 5-8 mm x 2- De Hoop Nature Reserve for their kind hospitality. This study 3 mm. Ultimate leaflets ovate to broadly ovate, 9-12 min x 8- was funded by the University of Johannesburg and the National 10 mm, venation not visible; segments linear-oblong, 2-5 mm Research Foundation. long, less than 1 mm broad, flat or subterete. Umbels with peduncle short, 3-8 mm long; involucral bracts 2 or 4, connate References or free; rays 2 or 3, 7-35 mm long at anthesis; involucel bracteoles 4, connate; raylets 10 to 15, 2-5 mm long at anthesis. Adamson, R.S., 1950. Umbcllifcrae. In: Adamson, R.S., Salter, T.M. (Eds.), Flowers with ovary tubercled; stylopodium very shortly conical Flora of the Cape Peninsula. Juta and Co., Cape Town. Burn, B.L., 1991. Umbelliferac of southern Africa: an introduction and to flat, sunken below the fruit apex; styles not lengthening as the annotated checklist. Edinburgh Journal of Botany 48, 133-282. fruit matures, 0.3-0.5 mm long, remaining erect or rarely Downie, S.R., Ramanath, S., Katz-Downie, D.S., Llanas, E., 1998. Molecular becoming somewhat reflexed to just above the base of the systematics of Apiaccae subfamily Apioideac: phylogenetic analyses of stylopodium. Fruit broadly elliptic to broadly ovate, 8.5- nuclear ribosomal DNA internal transcribed spacer and plastid rpoC/ intron 9.0 mm x 4.5-5.5 mm; base obtuse or shallowly concave; apex sequences. American Journal of Botany 85, 563-591. Downie, S.R., Watson, M.F., Spalik, K., Katz-Downie, D.S., 2000. Molecular obtuse to truncate; mericarps markedly concave on the systematics of Old World Apioideae (Apiaccae): relationships among some commissural surface; median and lateral ribs prominent, members of tribe Peucedaneac sensu lato, the placement of several island- flexuose, tubercled; marginal ribs prominently involute; endemic species, and resolution within the apioid superclade. Canadian secondary ribs absent; commissural vittae terminating some Journal of Botany 78, 506-528. distance from the base of the stylopodium. Dyer, R.A., 1975. The genera of Southern African flowering plants, vol. I. Dicotyledons. Botanical Research Institute, Pretoria. Feder, N., O'Brien, T.P., 1968. Plant microtcchnique: some principles and new 4.11.1. Diagnostic characters methods. American Journal of Botany 55, 123-142. As in Capnophyllum africanum, both the ovaries and fruit Goldblatt, P., Manning, J.C., 2000. Cape Plants: A Conspectus of the Cape Flora are covered with tubercles on the median and lateral ribs. of South Africa. Strclitzia, vol. 9. National Botanical Institute, Cape Town. Capnophyllum macrocarpum differs from this species in the Holmgren, P.K., Holmgren, N.H., Barnett, L.C., 1990. Index Herbariorum I: The Herbaria of the World. Ed. 8. Regnum Vcgetabile. New York Botanical larger fruit (more than 8.5 mm long) with a conspicuous sterile Garden, New York. apical portion and the rudimentary stylopodium that is sunken Jarvis, C.E., Rcduron, J.-P., Spencer, M.A., Caffcrty, S., 2006. Typification of below the apex of the fruit. Furthermore, the umbels of this Linnaean plant names in Apiaccae. Taxon 55, 207-216. species appear to have fewer rays (2 or 3). Kljuykov, E.V., Liu, M., Ostroumova, T.A., Pimenov, M.G., Tilncy, P.M., Van Wyk, B.-E., 2004. Towards a standardised terminology for taxonomically important morphological characters in the Umbelliferae. South African 4.11.2. Distribution and habitat Journal of Botany 70, 488-496. This species is known only from De Hoop Nature Reserve in Leistner, 0.A., Morris, J.M., 1976. Southern African place names. Annals of the the Western Cape Province (Fig. 6). It was collected in fynbos, Cape Province Museum 12. on deep sandy soils. A search of the surrounding area did not Magee, A.R., Van Wyk. B.-E., Tilncy, P.M., Downie, S.R., in press. Generic reveal any further populations, indicating that the species may delimitations and relationships of the Cape genera Capnophyllum, Dasi.spernnun and Smulerina, the North African genera Krubera and Stoi- be highly localised. brax, and a new monotypic genus of the subfamily Apioideae (Apiaccac). Systematic Botany. 4.11.3. Additional specimen examined Mcikle, R.D., 1977. Flora of Cyprus, vol. I. The Bcntham-Moxon Trust, London. South Africa. - 3420 (Caledon): De Hoop Nature Reserve, near to De Mond (-AD), Magee et al. 120 (JRAU). Sonder, W., 1862. Umbelliferac. In: Harvey, WH., Sonder, W. (Eds.), Flora Capcnsis. vol. 2. Hodges. Smith and Co., Dublin. Tutin, T.G., Heywood, V.H., Burges, N.A., Moore, D.M., Valentine, D.H., Acknowledgements Walters, S.M., Webb, D.A., 1968. Flora Europaca, vol. 2. Cambridge University Press, Cambridge. The curators and staff of the cited herbaria are thanked for Winter, P.J.D., Magee, A.R., Phcphu, N., Tilncy, P.M., Downie, S.R., Van Wyk, their kind hospitality and assistance during visits and for making B.-E., 2008. A new generic classification for African peucedanoid species (Apiaccac). Taxon 57. 347-364. specimens available on loan. The authors would also like to
Edited by J Van Staden APPENDIX C3
MAGEE, A. R., B.-E. VAN WYK, P. M. TILNEY and S. R. DOWNIE 2009. Generic delimitations and relationships of the Cape genera Capnophyllum,
Dasispermum and Sonderina, the North African genera Krubera and Stoibrax, and a new monotypic genus of the subfamily Apioideae (Apiaceae).
Systematic Botany 34(3): in press. Systematic Botany (2009), 34(3): pp. 1-15 0 Copyright 2009 by the American Society of Plant Taxonomists Generic Delimitations and Relationships of the Cape Genera Capnophyllum, Dasispermum, and Sonderina, the North African Genera Krubera and Stoibrax, and a New Monotypic Genus of the Subfamily Apioideae (Apiaceae)
Anthony R. Magee,'-3 Ben-Erik van Wyk,' Patricia M. Tilney,' and Stephen R. Downie 2 'Department of Botany and Plant Biotechnology, University of Johannesburg, P.O. Box 524, Auckland Park 2006 South Africa 2Department of Plant Biology, University of Illinois at Urbana-Champaign, Urbana, Illinois 61801 U.S.A. 3Author for correspondence ([email protected] )
Communicating Editor: Mark P. Simmons
Abstract—Generic circumscriptions and phylogenetic relationships of the Cape genera Capnophyllum, Dasispermum, and Sonderina are explored through parsimony and Bayesian inference analyses of nrDNA ITS and cpDNA rps16 intron sequences, morphology, and combined molecular and morphological data. The relationship of these genera with the North African genera Krubera and Stoibrax is also assessed. Analyses of both molecular data sets place Capnophyllum, Dasispermum, Sonderina, and the only southern African species of Stoibrax (S. capense) within the newly recognized Lefebvrea Glade of tribe Tordylieae. Capnophyllum is strongly supported as monophyletic and is distantly related to Krubera. The monotypic genus Dasispermum and Stoibrax capense are embedded within a paraphyletic Sonderina. This complex is distantly related to the North African species of Stoibrax in tribe Apieae, in which the type species, Stoibrax dichotomum, occurs. Consequently, Dasispermum is expanded to include both Sonderina and Stoibrax capense. New combinations are formalized for Dasispermum capense, D. hispidum, D. humile, and D. tenue. An undescribed species from the Tanqua Karoo in South Africa is also closely related to Capnophyllum and the Dasispermum- Sonderina complex. The genus Scaraboides is described herein to accommodate the new species, S. manningii. This monotypic genus shares the dorsally compressed fruit and involute marginal wings with Capnophyllum, but is easily distinguished by its erect branching habit, green leaves, scabrous umbels, and fruit with indistinct median and lateral ribs, additional solitary vittae in each marginal wing, and parallel, closely spaced commissural vittae. Despite the marked fruit similarities with Capnophyllum, analyses of DNA sequence data place Scaraboides closer to the Dasispermum—Sonderina complex, with which it shares the erect habit, green (nonglaucous) leaves, and scabrous umbels.
Keywords—ITS, Lefebvrea Glade, morphology, phylogeny, rps16 intron, Scaraboides manningii, South Africa.
A more evolutionary classification of the large and taxo- didyma (Sond.) Adamson, along with the North African spe- nomically complex cosmopolitan family Apiaceae is currently cies, back to the genus Stoibrax, as Stoibrax capense. Burtt (1989, emerging as a result of molecular systematic studies, together p. 145), furthermore, expressed his doubts about the generic with rigorous comparisons of morphological and anatomical concept of Sonderina, stating that the genus was "probably too data. Several small genera of uncertain circumscription and close to Stoibrax for it to be maintained". Burtt (1991), in his affinity are evident in recent checklists of African Apiaceae checklist of Southern African Umbelliferae, treated five spe- (Burtt 1991; Lebrun and Stork 1992; Van Wyk and Tilney 2004). cies within Sonderina. One of these, the Namibian endemic The majority of these genera are either poorly known or have Sonderina streyi Merxm., has subsequently been transferred to not been studied in recent years but may be extremely impor- the early diverging African genus Anginon Raf. (Allison and tant in the understanding of relationships within the family Van Wyk 1997). As a result, only four closely related species as a whole. The South African endemic genera Capnophyllum, are now recognised within the taxonomically difficult genus Dasispermum, and Sonderina were identified as three such taxa. Sonderina. Of these, only two species (Dasispermum suffruticosum and A similar disjunction has also been proposed for Sonderina humilis) have heretofore been included in molecu- Capnophyllum, with some authors (e.g. Tutin et al. 1968; Dyer lar systematic studies (Calvifto et al. 2006; Winter et al. 2008). 1975) extending the genus to include the Mediterranean In the phylogenetic analysis by Winter et al. (2008) using Capnophyllum peregrinutn (L.) Lange. Meikle (1977), however, nuclear ribosomal DNA internal transcribed spacer (ITS) treated the Mediterranean species as distinct under the mono- sequences, both species were shown to be closely related to typic genus Krubera Hoffm., a decision maintained by Burtt a group of recently circumscribed African peucedanoid gen- (1991). A recent taxonomic revision of Capnophyllum (Magee era, here referred to as the Lefebvrea Glade (viz., Afroligusticum et al. 2009b) recognized four species, two of which were newly C. Norman, Afrosciadiunt P. J. D.Winter, Cynorhiza, Lefebvrea described, and excluded Krubera peregrina Lowe on the basis A. Rich., Nanobubon, and Notobubon) within tribe Tordylieae. of important differences in fruit anatomy. The monophyly of Sonderina has not yet been assessed, nor A thorough taxonomic study of the genera Capnophyllum, has its putative relationship with Stoibrax been confirmed. Dasispermum, and Sonderina along with extensive field work Sonderina was described by Wolff (1927) to accommodate has revealed one new monotypic genus (herein described four of five South African species previously included in as Scaraboides manningii) and four new species (Magee et al. Ptychotis Koch by Sonder (1862). Wolff (1927) transferred the 2009b; Magee et al. unpublished). The present study is aimed fifth species, Ptychotis didyma Sond., to Tragiopsis Pomel (now at resolving generic circumscriptions and relationships of Stoibrax), which already included four North African spe- these previously neglected South African endemic genera. As cies. Adamson (1939) considered this geographically disjunct the phylogenetic relationships of African Apiaceae genera are treatment to be unnatural and transferred the South African often hard to predict on the basis of morphological characters species, Tragiopsis didyma (Sond.) H. Wolff, to Sonderina. Burtt alone, analyses of both morphology and anatomy in combi- (1989), however, argued that such a Cape and North African nation with molecular data (specifically, ITS and rps16 intron disjunction was not uncommon and transferred Sonderina sequences) are here presented and explored.
1 2 SYSTEMATIC BOTANY [Volume 34
MATERIALS AND METHODS data. Character transformations were treated as unordered and equally weighted (Fitch parsimony; Fitch 1971). Tree searches were performed Morphological Data—All relevant South African genera and a major- using a heuristic search with 500 random sequence additions, tree bisec- ity of their species were studied and sampled in situ, including two as yet tion-reconnection (TBR) branch swapping, and the MULPARS option in undescribed species of Sonderina (here referred to as S. sp. 1 and S. sp. 2). effect, but saving no more than 5 of the shortest trees from each search. This material was supplemented by a study of specimens from the follow- These equally parsimonious trees were then used as starting trees for TBR ing herbaria: BM, BOL, JRAU, K, LE, MO, NBG, PRE, S, SAM, THUNEi- branch swapping (MULPARS and STEEPEST DESCENT in effect) with UPS. The distribution data for Scaraboides manningii was recorded using the maximum number of trees saved set at 12,000; these trees were permit- Quarter Degree Grid Cells (outlined in Leistner and Morris 1976). In this ted to swap to completion (Downie et al. 1998). Bootstrap percentage val- system the basic unit is the one-degree square of latitude and longitude, ues (BP; Felsenstein 1985) for the separate ITS and rps16 intron data sets which is designated by a degree reference number (i.e. degrees of latitude were determined from 500,000 replicate analyses using fast stepwise addi- and longitude of the north-west corner) and the district name of that square. tion of taxa, while BP values for the morphological and combined data Line drawings were made by the first author with the aid of a camera sets of the Lefebvrea Glade were determined from 1,000 bootstrap replicates, lucida attachment on a Zeiss compound microscope or a Wild M3Z holding 10 trees per replicate and with TBR and MULPARS selected. Only stereomicroscope. values greater than or equal to 50% are reported, and the following scale Preserved (FAA; formaldehyde: acetic acid: alcohol: water) and her- was applied for support percentages: 74%, weak; 75-84%, moderate; barium materials were used to study fruit anatomy. Herbarium mate- and 85-100%, strong. All data sets (except the separate morphological rial was first rehydrated and then placed in FAA for a minimum of 24 h. data set) were subsequently analysed by Bayesian inference (BI; Yang and This material was subsequently treated according to a modification of the Rannala 1997) using MRBAYES version 3.1.2 (Huelsenbeck and Ronquist method of Feder and O'Brien (1968) for embedding in glycol methacry- 2001; Ronquist and Huelsenbeck 2003). An appropriate model of evolu- late (GMA). This modification involves a final infiltration in GMA for five tion was selected for each data partition using the program MODELTEST days. Transverse sections, about 3 pm thick, were cut using a Porter-Bliim version 3.06 (corrected Akaike information criterion; Akaike 1974; Posada ultramicrotome. The sections were examined for the presence of crystals and Crandall 1998). For the combined analysis in which morphological using a light microscope, after which they were stained according to the data were included, the datatype = standard option of MRBAYES for the periodic acid Schiff/ toluidine blue (PAS/TB) method of Feder and O'Brien nonnucleotide data partition was used. For the separate ITS and rps16 (1968). To study the three-dimensional structure of the vittae, mature fruit intron data sets, the analysis was performed for three million generations were softened by soaking in boiling water for 24 h. The exocarp was then of Monte Carlo Markov Chains (MCMC) and a sampling frequency of peeled off while keeping the fruit submerged in water to prevent desicca- 100, while for the morphological and combined data sets of the Lefebvrea tion. The terminology used to describe the fruit anatomical features fol- Glade only two million generations of MCMC and a sampling frequency of lows Kljuykov et al. (2004). 10 was used. The resulting trees were plotted against their likelihoods in A matrix of 23 morphological and anatomical characters was prepared order to determine where the likelihoods converge on a stable plateau. All for 31 species of the Lefebvrea Glade based on examination of herbarium the trees before this convergence were discarded as the 'burn-in' phase. specimens and literature (Appendices 1 and 2; Magee et al. 2008a; Magee et A majority rule consensus tree was produced from the remaining trees al. 2008b; Winter et al. 2008; Magee et al. 2009a; Magee et al. 2009b). These in order to show the posterior probabilities (PP) of all observed biparti- data were also combined with ITS sequences from the same taxa for simul- tions. The following scale was used to evaluate the PPs: 0.5-0.84, weak; taneous phylogenetic analysis (Kluge 1989; Nixon and Carpenter 1996). 0.85-0.94, moderate; 0.95-1.0, strong. Molecular Data—DNA was extracted using the 2 x CTAB method To assess congruency of relationships within the Lefebvrea Glade, as of Doyle and Doyle (1987) from materials collected in the field and from inferred by separate MP analyses of the ITS, rpsl6 intron and morphologi- herbarium specimens and used to assess the generic delimitations and cal data sets, the bootstrap consensus trees from each analysis were com- phylogenetic relationships of the Cape endemic genera Capnophyllum pared. These trees were considered incongruent only if they displayed (12 new accessions), Dasispermum (two new accessions), Sonderina (13 new 'hard' (i.e. incongruencies with strong bootstrap values) rather than 'soft' accessions), the undescribed monotypic genus Scaraboides (two new acces- (i.e. incongruencies with weak bootstrap values) incongruence (Seelanan sions), and the largely North African genus Stoibrax (five new accessions). et al. 1997; Wiens 1998). In addition, a partition homogeneity test (incon- Additional accessions of the rpsl6 intron region for the closely related gruence length difference test, ILD; Farris et al. 1995) was performed in African peucedanoid genera Cynorhiza (two new accessions), Nanobubon PAUP*. This test was implemented with 1,000 replicate analyses, using (two new accessions), and Notobubon (five new accessions) were also the heuristic search option with simple addition of taxa, and with TBR and included. The 45 new accessions for which ITS (18 accessions) and rpsl 6 MU LTREES options selected. intron (27 accessions) sequences were obtained are presented in Appendix 3. To evaluate the significance of differing topologies, we used the Previously published rpsl6 intron accessions are listed in Appendix 4, and Shimodaira-Hasegawa test (SH; Shimodaira and Hasegawa 1999), as previously published ITS accessions are available in Winter et al. (2008). implemented in PAU?' (applying the RELL resampling method with For amplification of the ITS and rps16 intron regions, we used the prim- 1,000 bootstrap replicates). All DNA sequences have been submitted to ers described by Sun et al. (1994) and Oxelman et al. (1997), respectively. GenBank (Appendix 3) and all PAUP* matrices deposited in TreeBASE Amplified PCR products were purified using a QlAquick PCR purifica- (study number S2197). tion kit (Qiagen Inc., Valencia, California) according to the manufacturer's Morphological characters were reconstructed on the MP trees from the instructions and directly sequenced on a 3130 xl Genetic Analyzer (Applied combined ITS/ rps16 intron/morphology data set using parsimony with Biosystems Inc., Foster City, California) using BigDye Terminator version Mesquite version 2.5 (Maddison and Maddison 2008). 3.1 chemistry (Applied Biosystems Inc.). For each molecular data set, com- plementary strands were assembled and edited using Sequencher version 3.1.2 (Gene Codes Corporation, Ann Arbor, Michigan) and aligned manu- RESULTS ally in PAUP' version 4.0b10 (Swofford 2002), with gaps positioned so as to minimize nucleotide mismatches. ITS Data Set—The ITS matrix consisted of 633 unambig- Phylogenetic Analyses—To assess the phylogenetic positions of the uously aligned nucleotide positions with 382 variable and aforementioned genera, the newly obtained ITS sequences were added to 317 parsimony informative characters. Parsimony analyses the 125 taxon ITS matrix of Winter et al. (2008). This matrix represents all tribes and major clades of the apioid superclade plus outgroups from tribes resulted in the preset maximum tree limit of 12,000 trees, each Smyrnieae and Oenantheae (Downie et al. 2001), with those species of the of 2,113 steps (ensemble consistency indices [CI; Kluge and latter used to root the trees. The newly obtained rps16 intron sequences Farris 1969] of 0.33 and 0.31, with and without uninformative were analysed with 27 additional rps16 intron sequences from GenBank characters, respectively; ensemble retention index [RI; Farris (Appendix 4), the latter also representing several relevant major clades of the apioid superclade. The rps16 intron trees were rooted with Slum 1989] of 0.72). MODELTEST selected the GTR + I + G model of latifolium and Berula erects of tribe Oenantheae. To further explore rela- evolution for use in the BI analysis. The MP strict consensus tionships within the Capnophyllum group, combined data sets (ITS/ rps16 tree yielded a similar topology as the BI consensus tree (with intron, ITS/morphology, and ITS/ rps16 intron/morphology) for 31 taxa the differences between these trees summarized in Fig. 1). In EIJ of the Lefebvrea Glade were analysed, with Lefebvrea abyssinica A.Rich. used both analyses, the same groupings as reported previously by as the outgroup. Phylogenetic analyses of all data sets were conducted initially using Winter et al. (2008) were retrieved. The Lefebvrea Glade was the parsimony (MP) algorithm of PAU?' with gaps treated as missing weakly supported in both the BI and MP trees (PP = 0.75, BP
2009] MAGEE ET AL.: CAPE GENERA OF APIACEAE 3
Notobubon galbanum 30 Notobubon capense 37 Notobubon tenuifolium 60 Notobubon tenuifotium 44 Notobubon galbanum 10 1.0 Notobubon capense 43 Notobubon galbanum 2452 Notobubon gummiferum Notobubon tenuifollum 52 Notobubon galbaniopse Notobubon striatum Notobubon striatum 098 Notobubon pungens Notobubon pungens Notobubon ferulaceum Notobubon ferulaceum 98 Notobubon laevigatum Notobubon laevigatum 1.0 1—•-• Nanobubon s.um 99 Nanobubon stectum Nanobubon cap!Ilaceum Cynorhlza typica Cynorfeza ca 1:0 C Afroligusticum petitianum 82 Afroligusticum °Noir DasIspermum suffruticosum 2451 1 Dasispermum suffruticosum 3137 e7 80 Dastspermum suffruticosum 117 oj Sonderfna htspida 107 0.80 10 DasIspermum suffruttcosum 2292 tamq 0 97 E Sondorfna hlspida 115 Sonderfna sp. 1 dep a ep 0.51 1 Stolbrax capense dou 82 Stolbrax capense ep Scaraboldes manning)!' es 0.53 Sonderina tenuls gli Son derfna sp. 2 ni 1.0 tu 92 1=2: ZZig: 6 "Capnophyllum futreyerff
1.0 Capnophylium felocarpon 125 dnoi Capnophyllum lelocaspon 6978 1130 Capnophyllum afrlcanum 087 57 Capnophyllum ahlcanum Capnophyllum macrocarpum - 0.75 Afrosciadium magalismontanum 1.0 Stenosemis caffm se Stenosemis angustifolla 1.0 Notobubon pearson)! 100 Notobubon pearsonfi
098 9 0 LefebvreaLofem gra nts 774 0.99 76 Lefebvre° grantii 89 71 Lefebvre° grantli 1 Lefebvrea abyssinica 0.99 99 Lefebvrea abyssinica 0.99 Ducrosia anethifolia 89 t2t9 4= Kalakia marginate Cymbocarpum anethoides 0.99 0.99 Heradeum dada 7b 1.0 Conium maculatum 100 Conium sphaerucarpurn
0.99 Selineae 94 Ota= Prangos pabularfa 0.99 Ferulago galbanifera Azilia eryngioides Echinophoreee 0.98 Trachyspermum aethusifotium 099 I-- Opopanax hspidusi I-- Smymiopsis ouched e Coriandrum sativum 0 94 Bifora radians 0.53 Kruboro peregrine Levisticum officinal° Seselopsis tianschanica 7 Spheenolobium tianschanicum 1.0 Devena burr:twill 1.0 Deverra denudate ssp. aphyfla 19 Deverra triradiata 93 l°° Sclerosciadium nodiflomm 19.. 10 Apium graveolens 0 85 1 0 99 Apium prostmtum 0.99 88 Naufraga baleanca 0 78 0 Anethum graveolens 1.0 i— Stolbrax dichotomum 100 I— Stolbrax dichotomum 099 Pimpinelieae 100 0.99 ea Pyramidoptereae 99 88 ass 1 Careae 0. Smymieae 100 es Afrocarurn imbricatum 0.99 99 Slum repandum 99 i Berula thunbergli 74 Sium bracteatum Helosciadium n3pens
FIG. 1. Bayesian inference (BI) tree of ITS sequence data. Posterior probability (PP) values are presented above the branches. Bootstrap percentage (BP) values are presented below the branches. BP and PP values below 50% and 0.50, respectively, are not indicated. Branches supported only by BI are indicated by dashed lines, those branches that differ in the parsimony strict consensus tree are indicated alongside as gray lines. 4 SYSTEMATIC BOTANY [Volume 34
< 50). Lefebvrea formed the earliest diverging lineage in the Notobubon formed a weakly supported Glade in the MP MP strict consensus tree, while in the BI tree its position was strict consensus tree, although without N. pearsonii, which in unresolved. both the BI and MP trees was placed as sister (though sup- With the exception of Notobubon pearsonii, the woody Cape ported strongly only in the BI tree with PP = 0.99) to an unre- genus Notobubon was strongly supported as monophyletic in solved Cynorhiza typica. Stoibrax dichotomum was once again the BI tree (PP = 0.98), while in the MP strict consensus tree the shown to be part of tribe Apieae in both BI (PP = 0.94) and latter Glade was not resolved. The Cape genera Capnophyllum, MP trees (BP < 50) and not closely related to its South African Dasispermum, and Sonderina, together with the only South congener, S. capense, or to its putative relative Sonderina. The African species of Stoibrax (S. capense) and the monotypic placement of Stenosemis outside of the Lefebvrea Glade was not genus Scaraboides, all formed a group (hereafter referred to consistent with that found in the ITS studies and may be due as the Capnophyllum group) within the Lefebvrea Glade, which to the low resolution and limited sampling within the tribe was strongly supported in the BI tree (PP = 0.99); however, in Tordylieae. the MP strict consensus tree, this Glade was supported with a Morphological Data Set—MP analysis of 23 morphologi- BP value of less than 50%. Scaraboides manningii was placed cal and anatomical characters from 31 species of the Lefebvrea within a weakly supported polytomy with members of the Glade resulted in 10 minimal length trees each of 50 steps Dasispermum–Sonderina complex (PP = 0.60) in the BI tree, (CI = 0.60, RI = 0.88). The relationships among members of while in the MP strict consensus tree it was weak supported the Lefebvrea Glade were generally better resolved than those to be sister to the Sonderina humilis–S. tenuis group (BP < 50). inferred from the molecular analyses, although with generally Dasispermum suffruticosum and Stoibrax capense, together with lower BP values (Fig. 3A). Notobubon pearsonii was included E3 Sonderina hispida and Sonderina sp. 1 comprise a strongly sup- within a weakly supported Glade of Notobubon (BP = 54), ported Glade (PP = 1.0, BP = 96) that arise from within a para- Nanobubon was moderately supported as monophyletic (BP = phyletic Sonderina. Constraining the Dasispermum–Sonderina 80), and Stenosemis was strongly supported (BP = 93) as mono- complex to monophyly so that Scaraboides manningii was its phyletic. The Capnophyllum group comprising the predomi- sister group resulted in trees that were not significantly dif- nantly annual genera Capnophyllum, Dasispermum, Scaraboides, ferent (p = 0.261). Sonderina, and Stoibrax capense formed a separate lineage as in The North African species Krubera peregrina and Stoibrax the molecular analyses. Capnophyllum was strongly supported dichotomum, sometimes considered to be closely related to as monophyletic (BP = 89), with Scaraboides manningii as its Capnophyllum and Sonderina respectively, were inferred to be sister group. Dasispermum and Stoibrax were again recovered distantly related to each other and to the Capnophyllum group. within a paraphyletic Sonderina (BP = 72). Both accessions of the type species of Stoibrax (S. dichotomum) Combined ITSIrps16 Intron Data Set—The combined ITS were moderately to strongly placed within tribe Apieae (PP = and rps16 intron matrix for 31 taxa within the Lefebvrea Glade 0.99, BP = 76), while Krubera peregrina was resolved in a Glade consisted of 1,593 characters, of which 228 were variable and comprising Coriandrum sativum L., Bifora radians M.Bieb., and 112 parsimony informative. Missing data represented 13% Levisticum officinale Koch. of the entire data matrix, as rps16 intron sequences were rps16 Intron Data Set—The rps16 intron matrix consisted unavailable for eight taxa. Visual inspection of the two sep- of 936 unambiguously aligned nucleotide positions with 191 arate bootstrap consensus trees and results of the ILD test variable and 92 parsimony informative characters. MP analy- suggested that the two matrices were not significantly incon- ses yielded the preset maximum tree limit of 12,000 trees, each gruent (p = 1.0). Parsimony analyses of combined molec- of 282 steps (CI = 0.77 and 0.63, with and without uninforma- ular data yielded 97 trees, each of 359 steps (CI = 0.74 and tive characters, respectively; RI = 0.84). MODELTEST selected 0.59, with and without uninformative characters, respec- the K8luf + G model of evolution for use in the BI analysis. tively; RI = 0.77). The GTR + I + G and the K8luf + G models Overall the MP strict consensus tree yielded a similar topology were retained for the ITS and rps16 intron data partitions, to that of the majority rule consensus tree obtained from the BI respectively. Trees obtained from both the MP and BI analy- analysis (differences between the results of these analyses are ses yielded the same overall topologies (Fig. 3B). As in the a721 presented in Fig. 2). Although the resolution within these trees analyses of partitioned molecular data, a lineage comprising was poor, a lineage comprising Capnophyllum, Dasispermum, the Capnophyllum group (incl. Capnophyllum, Dasispermum, Scaraboides, Sonderina, and Stoibrax capense (Capnophyllum Scaraboides, Sonderina, and Stoibrax capense) was recovered (PP = group) was retrieved in both analyses (PP = 0.89, BP < 50). 1.0, BP = 62). Capnophyllum was again strongly supported as This same Capnophyllum group was resolved in the ITS trees monophyletic (PP = 1.0, BP = 100). Dasispermum suffrutico- with greater taxon sampling (Fig. 1). Similarly, Dasispermum sum and Stoibrax capense were strongly embedded within a suffruticosum and Stoibrax capense were again strongly embed- subclade of a paraphyletic Sonderina, together with Sonderina ded within a subclade of Sonderina (PP 1.0, BP < 50) compris- hispida and Sonderina sp. 1 (PP = 1.0, BP = 100). Scaraboides ing the type species S. hispida. Scaraboides manningii is placed manningii was again strongly supported as part of a Glade within a strongly-supported polytomy (PP = 1.0, BP = 87) comprising Dasispermum, Stoibrax capense, and all species of with members of the Dasispermum–Sonderina complex in the Sonderina (PP = 1.0, BP = 91), although its exact position within BI tree, while in the MP strict consensus tree Sonderina humi- this Glade remained equivocal. In the BI trees Scaraboides man- lis and Sonderina sp. 2 formed the earliest branching lineages, ningii was weakly supported as a sister group to the subclade followed by a polytomy (BP < 50) comprising Sonderina ten uis, comprising Stoibrax-Dasispermum (PP = 0.74), while in the Scaraboides manningii, and the Stoibrax capense–Dasispermum MP strict consensus tree it formed one branch of a trichot- suffruticosum Glade. Constraining the Dasispermum–Sonderina omy. As in the separate ITS and rps16 analyses, constraining complex to monophyly so that S. manningii was its sister the Dasispermum–Sonderina complex to monophyly so that group resulted in trees that were not significantly different S. manningii was its sister group resulted in trees that were not (p = 0.191). significantly different (p = 0.142).
2009] MAGEE ET AL.: CAPE GENERA OF APIACEAE 5
0.99 Sonderina sp. 1 0.64 Sonderina hispida 112 Sonderina hispida 107
Dasispermum suffruticosum 1.0 0.65{ Stoibrax capense 128
Stoibrax capense 131 dep Sonderina hispida 115 1.0
dou 87 "Scaraboides manningii " Sonderina tenuis Ati Sonderina sp. 2 nli 0.89 Sonderina humilis w "Capnophyllum lutzeyeri" al B
— Capnophyllum leiocarpon 125 1 0
0.84 1.0 Capnophyllum leiocarpon 6978 mpi
96 Capnophyllum africanum 124 dn ea
"Capnophyllum macrocarpum " Capnophyllum africanum 654 Nanobubon capillaceum °pep 1.0 Nanobubon strictum 86 C Nanobubon strictum 1.0 Notobubon pearson fi 97 0.95 0.99 Notobubon pearsonii Cynorhiza typica 53 Cynorhiza typica 3372 Notobubon capense 43 Notobubon tenuifolium Notobubon gummiferum Notobubon capense 37 Notobubon pungens Notobubon galbanum 0.93 Notobubon ferulaceum 1.0 Heracleum maximum 1.0 69 Heracleum lanatum 1.0 87 Heracleum sphondylium
0.88 . 0 Pastinaca sativa 1.0 81 C Malabaila sekakul 76 Zosima orientalis Echinophora tenuifolia 0.98 0.79 Aethusa cynapium 1.0 Thaspium pinnatifidum 94 Zizia aurea Stenosemis caffra 1.0 Crithmum maritimum 88 1.0 Conium maculatum 100 Conium sphaerocarpon Naufraga balearica 0.99 0.74 1660 1.0 Apium graveolens 53 99 0.83 Stoibrax dichotomum 94 Deverra burchellii Ammi majus co Foeniculum vulgare Smyrnium olusatrum Berula erecta Sium latifolium Fic. 2. Bayesian inference (BI) tree of rps16 intron sequence data. Posterior probability (PP) values are presented above the branches. Bootstrap per- centage (BP) values are presented below the branches. BP and PP values below 50% and 0.50, respectively, are not indicated. Branches supported only by BI are indicated by dashed lines, those branches that differ in the parsimony strict consensus tree are indicated alongside as gray lines.
6 SYSTEMATIC BOTANY [Volume 34
A. morphology - Notobubon capense C. ITS/morphology 0.98 Notobubon capense 37 Notobubon galbanum 1.0 ,Notobubon tenuifollum 44 99 Notobubon gaibanum 2452 Notobubon tenuifollum 1.0 Notobubon gummiferum 71 87 i gotobubon gummlferum Notobubon galbaniopse 0 97 - Notobubon galbaniopse 0. Notobubon pungens 1.0 Notobubon pungens 50 Notobubon ferulaceum Notobubon striatum 51 Notobubon laevigatum 0.79 or--- Notobubon ferulaceum 2500 Notobubon pearsonii 99 1-Notobubon laevigatum 2510 0.61 Notobubon striatum Notobubon pearsonii 42 Stenosemis caffra 1.0 Stenosemis caffn3 C Stenosemis angustifolia 100 I---Stenosemis angustifolia Nanobubon strictum Nanobubon striatum 58 Nanobubon capillaceum Nanobubon capillaceum Cynorhlza typica Cynorhiza typica 53 Afrosciadium magatismontanum Afrollgusticum petitianum Afroligusticum pelltianum Afrofigusticum °Moth Afroligusticum Motif 0.87 DaSiSpOm10171suffmticosum 117 1.0 Sonderina hlspida 107 .0 ss 67 Sonderina Stoibrax capense hisplda sp. 1 99 0.83 Stoibrax capense 128
Sonderina humus 0.88 62 62 I F Sonderina humilis 72 Sonderina tenuis Dasispermum suffruticosum nderina tenuls doudoo ki 1 doudoo Sonderina sp. 2 nu
Sonderina sp. tli Sonderfna sp. 2 p Scaraboides manningir 8 gu "Scaraboides manningii " wni "Capnophyllum futzeyeri"
43 1.0 Capnophyllum feiocarpon 6978 Capnophyllum Iutzeye& dnoJ 75 100 Capnophyllum africanum 124 Capnophylturn lelocarpon dno Tapnophyllurn macrocarpum" 89 Capnophyllum africanum "Caprtophylturn macmcarpum" Afrosciadium megalismontanum Lefebvre° abysslnlca Lefebvrea abyssinica
- 0.01 B. ITSIrps16 intron D. ITSI rps16 intron/morphology 0 Notobubon capense 37 0.97 Notobubon capense 37 1.0 Q, Notobubon tenuifolium 44 1.0 Notobubon tenulfollum 44 Notobubon galbanum 2452 1.0 160 0 galbanum 2452 6 86 "Notobubon gummiferum .0 NotobubonNotobubon gummiferum Notobubon galbaniopse 0.97 Notobubon galbanlopse 0.99 Notobubon pungens 50 Notobubon pungens 50 Notobubon striatum 51 Notobubon striatum 51 1.0 Notobubon ferulaceum 2500 Notobubon ferulaceum 2500 100 Notobubon laevigatum 2510 Notobubon laevigatum 2510 0.50 11.54 Nanobubon striatum 58 Notobubon pearsonll 42 0.52 I- Cynorhlza typica 53 Stenosemis caffra i.o Afroligusticum petitianum Stenosemis angustifolia 87 I Afroligusticum effiolii Nanobubon strfctum 58 Nanobubon capillaceum Nanobubon capitlaceum 1 0 Dastspermum suffruticosum 117 Cynortliza typica 53 1.o L. Sonderina hispida 107 Afroligusticum petitianum 1.0 100 " Sonderina sp. 1 Afroligusticum Stoibrax capense 128 deo 0.79r- Dasispermum suffruticosum 117 0.79 1.0 "Scaraboides manningr 1 .0 I- Sondertne hispfda 107 dou 91 Sonde:Ina humilis fil Sonderina sp. 1 Sonderfna tenuls Stoibrax capense 128 umip 051 Sonderfna sp. 2 do2
6 Sonderfna humflis i
"Capnophyllum lutzeyeri* dou 0.53 Sonderfna tenuls ki 1.0 dno Capnophyllum lelocarpon 6978 Sonderfna sp. 2 100 "Capnophyllum maaocarpum" nii "Scaraboides manningii" w
Capnophyllum africanum 124 8
"Capnophyttum futzeyeri" , 0.53 Afrosciadium magalismontenum Capnophyllum leiocarpon 6978 1.0 Stenosemis caffra dno 100 Capnophyllum africanum 124 99 L_ Stenosemis angustifoila "Capnophyllum macrocarpum" Notobubon pearsonii 42 Afrosdadium magalismontanum Lefebvre° abyssinica Lefebvrea abyssinlca •••• 0.01 - 0.01 FIG. 3. A. Strict consensus tree of 18 equally most parsimonious trees based on the parsimony analysis of morphological data. B. Bayesian infer- ence (BI) phylogram of the combined ITS/rps16 intron data sets. C. Bayesian inference phylogram of the combined ITS/morphological data sets. D. Bayesian inference phylogram of the combined ITS/rps16 intron/morphological data sets. Posterior probability values are presented above the branches. Bootstrap values from the parsimony analysis are presented below the branches. Bootsrap and PP values below 50% and 0.50, respectively, are not indicated. Branches supported only in the BI tree are indicated by dashed lines, those branches that differ in the parsimony strict consensus tree are indicated alongside as gray lines. 20091 MAGEE ET AL.: CAPE GENERA OF APIACEAE 7
Combined ITS1Morphological Data Set—The combined for the ITS data. Overall, the MP strict consensus tree yielded ITS and morphological matrix for 31 taxa within the Lefebvrea a similar topology as those retrieved from the BI analysis (dif- Glade consisted of 712 characters, of which 206 were vari- ferences between the results of these analyses are presented able and 115 parsimony informative. Visual inspection of in Fig. 3C). As in the morphological analysis, subclades the two separate MP bootstrap consensus trees revealed no comprising the Cape genera Notobubon (PP = 0.79, BP < 50), hard incongruence, however, the ILD test suggested that the Nanobubon (PP = 1.0, BP = 77), and Stenosemis (PP = 1.0, BP = two were significantly incongruent (p = 0.001). Following the 100) were supported as monophyletic (PP = 0.73, BP < 50). suggestions of Seelenan et al. (1997) and Wiens (1998), the The Capnophyllum group was again recovered (PP = 1.0, BP = two matrices were still combined for simultaneous analyses. 81), with Capnophyllum strongly supported as monophyletic Parsimony analysis yielded 200 trees, each of 364 steps (CI = (PP = 1.0, BP = 100). Dasispermum and Stoibrax capense again 0.68 and 0.55, with and without uninformative characters, arose from within a paraphyletic Sonderina. This broadened respectively; RI = 0.76). The GTR + I + G model was retained Dasispermum-Sonderina complex was weakly to moderately
1. Life history 3. Growth pattern 12. Petal vestiture 14. Fruit compression perennial ■ sympodial ■ papillose ■ isodiametric ■ short-lived perennial monopodial leathery platyspermous monocarpic Notobubon capense 31 Notobubon tenuifolium 44 Notobubon galbanum 2452 A B Ci D Notobubon gummiferum i-i p___:Notobubon galbaniopse Notobubon pungens 50 -11' I Notobubon striatum 51 --L_ Notobubon ferulaceum 2500 I Notobubon laevigatum 2510 I Notobubon pearsonii 42 Firc Stenosemis caffra 1_1 - Stenosemis angustifolia r -Nanobubon stnctum 58 FP i. Nanobubon capillaceum rCynorhiza typica 53 Atroligusticum petitianum Afroftgusticum elliolii Dasispermum suffruticosum 117 ° 8 0 Sonderina hispida 107 m Sonderina sp. 1 ■ -c3 1 Stoibrax capense 128 Sonderina humilis . 0-o Sonderina tenuis ■ ...c Sonderina sp. 2 a e-- I"Scaraboides manning? 3 _r 1 Capnophyllum lutzeyeri" ua f-L Capnophyllum leiocarpon 6978 Ei Capnophyllum africanum 124c-0 "Capnophyllum macrocarpunr Afrosciadium magalismontanum - Lefebvrea abyssinica
15. Fruit in lateral view 16. Median and/or lateral ribs 21. Commissure 23. Cells external to vittae ■ very broadly elliptic as developed as marginal rib base to rib base ■ enlarged, upright to rotund < developed than marginal near rib tip to near rib tip square narrowly elliptic O rib tip to rib tip indistinct Notobubon capense 31 Notobubon tenuifolium 44 Notobubon galbanum 2452 E F G H Notobubon gummiferum p D_._ Notobubon galbaniopse Notobubon pungens 50 Notobubon strialum 51 Notobubon ferulaceum 2500 Notobubon laevigatum 2510 11 ■ Notobubon pearsonii 42 r-- Stenosemis caffra = Stenosemis angustifolia Nanobubon stnctum 58 Nanobubon capillaceum l i.i Cynorhiza typica 53 Afroligusticum petitianum Afroligusticum elliotii Dasispermum suffruticosum 117 ll - 0 Trirl__PJ Sonderina hispida 107 1 03 Sonderina sp. 1 -tt _ 1 Stoibrax capense 128 Sondetina humilis O -0 u Sonderina tenuis ■ '.. I fl Sonderina sp. 2 a E- 1 i"Scaraboides manning? 1"Capnophyilum lutzeyerr co Capnophyllum leiocarpon 6978 g Capnophyllum africanum 124 -0 E1-1 "Capnophyllum macrocarpum° _ Afrosciadium magalismontanum 1 Lefebvrea abyss/nice
FIc. 4. Reconstruction of eight morphological characters supporting either the Capnophyllum group or the Dasispermum-Sonderina complex (indicated with a dashed line), when optimized over the ten minimal length trees inferred from MP analysis of combined ITS/rps/6 intron/morphology data. 8 SYSTEMATIC BOTANY [Volume 34
supported (PP = 0.83, BP = 62), with Scaraboides as its sister The Glade comprising Scaraboides and the Dasispermum- group (PP = 0.85), although in the MP strict consensus tree Sonderina group was strongly supported (PP = 1.0, BP = 83). a trichotomy comprising Capnophyllum, Scaraboides, and the As in all prior analyses, Sonderina is rendered paraphyletic by Dasispermum–Sonderina complex was retrieved. the inclusion of Dasispermum and Stoibrax capense. Combined ITS/rps16 Intron/Morphology Data Set—The Morphological Character Evolution—Parsimony-based combined ITS, rps16 intron and morphological matrix for 31 reconstructions of eight morphological characters supporting taxa within the Lefeborea Glade consisted of 1,616 characters, either the Capnophyllum group or the Dasispermum–Sonderina of which 251 were variable and 135 parsimony informative. complex are each summarized onto one of the ten minimal Missing data represented 8.6% of the entire data matrix, as length trees inferred from MP analysis of combined ITS/rps16 rps16 intron sequences were unavailable for eight taxa. Visual intron /morphology data (Fig. 4). Monocarpic life history El inspection of the three separate MP bootstrap trees revealed (character 1, Fig. 4A), sympodial growth pattern (character 3, no hard incongruencies. Parsimony analyses yielded 10 trees, Fig. 4B), and the broadly elliptic to rotund fruit in lateral view each of 425 steps (CI = 0.69 and 0.56, with and without uninfor- (character 15, Fig. 4E) were reconstructed as synapomor- mative characters, respectively; RI = 0.77). Trees obtained from phies for the Capnophyllum group. Petal vestiture (character both the MP and BI analyses yielded similar overall topolo- 12, Fig. 4C) was ambiguously reconstructed at the base of the gies (differences between the results of these analyses are pre- Capnophyllum group; this character can either be interpreted sented in Fig. 3D). The topology resolved from the combined as a synapomorphy for the Capnophyllum group with rever- ITS/rps16 intron /morphology analyses was generally similar sals in Scaraboides manningh and Dasispermum suffruticosum, to those obtained from analyses of the combined ITS/mor- or as a convergent character supporting both Capnophyllum phology dataset. The genera Capnophyllum (PP = 1.0, BP = 100), as well as the Dasispermum–Sonderina complex, with a rever- Nanobubon (PP = 0.97, BP = 68), and Stenosemis (PP = 1.0, BP = sal in Dasispermum suffruticosum. Reconstruction of the short- 100) were each recovered as monophyletic. The Capnophyllum lived perennial habit (character 1) differed slightly among the group (i.e. Capnophyllum, Dasispermum, Scaraboides, Sonderina, 10 minimal length trees depending on the relative position of and Stoibrax capense) was again retrieved (PP = 1.0, BP = 81). Sonderina sp. 1. In eight of the trees, this character was recon-
vv
1 mm
1 mm I mm medr Ir
IIMEMNIMIBISM E 1 mm 1 mm
FIG. 5. Transverse sections through the fruit of A. Stoibrax hanotei, Wall s.n. (S); B. Stoibrax capense, Boatwright et al. 212 URAU); C. Sonderina hispida, Van Wyk 3539 URAU); D. Krubera peregrina, Lippert 22959 (PRE); E. Stenosemis caffra, MacOwen sm. sub South African Exchange Club 904 (GRA); F. Dasispertnum suffruticosum, Winter 78 (JRAU); and G. Capnophyllum africanum, Winter 110 URAU). c - carpophore; cv - commissural vitta; Ir - lateral rib; mr - marginal rib; medr - median rib; rod - rib oil duct; vb - vascular bundle; vv - vallecular vitta. 2009] MAGEE ET AL.: CAPE GENERA OF APIACEAE 9
structed as an autapomorphy for both Sonderina sp. 1 and Dasispermum suffruticosum (as shown in Fig. 4A), while in the remaining two trees where Sonderina sp. 1 and Dasispermum suffruticosum were sister, it was reconstructed as a synapo- morphy. The Dasispermum–Sonderina complex was supported by an isodiametric fruit compression (character 14, Fig. 4D), median and/or lateral ribs as developed as the marginal ribs (character 16, Fig. 4F), commissure extending from at most rib base to rib base (character 21, Fig. 4G), and square or enlarged, upright cells external to the vittae (character 23, Fig. 4H). Flimsy leaf texture (character 6) and concolorously glau- cous leaves (character 7) were both reconstructed as synapo- morphies for Capnophyllum (not shown).
DISCUSSION Lefebvrea Glade—Winter et al. (2008) reported that the African species previously attributed to Peucedanum and its platysper- mous allies comprise a Glade sister to a small affiance of south- west Asian species (Ducrosia anethifolia Boiss., Kalakia marginata (Boiss.) R. Alava, Cymbocarpum anethoides DC.) within tribe Tordylieae. Within their African group, here referred to as the Lefebvrea Glade, the nonplatyspermous, South African endemic species Dasispermum suffruticosum, Sonderina humilis, Stenosemis caffra, and Stenosemis angustifolia E. Mey. ex. Harv. & Sond. were also included. Our broadened analyses show that the Lefebvrea Glade also includes all species of Sonderina and Capnophyllum, the monotypic genus Scaraboides, and the Cape species Stoibrax capense. Winter et al. (2008) separated Dasispermum, Sonderina, and Stenosemis E. Mey. ex. Harv. & Sond. from the peucedanoid genera by their narrower commissure, not extending to the tips of each marginal rib/wing. Capnophyllum and Scaraboides also have dorsally compressed fruit with prominent marginal wings and a broad commissure. These three character states are all ple- siomorphies based on the distribution of characters states (Fig. 4D, F, G). The fruit of Stenosemis differ from those of Dasispermum, Sonderina, and Stoibrax in that the commissure, though not as broad as found in Capnophyllum, Scaraboides, and the peuce- danoid genera, is much broader and extends to beyond the base of the marginal wings, often somewhere between the tip and the centre of the wing/rib. This feature was reconstructed as a synapomorphy for the genus Stenosemis (Fig. 4G). Capnophyllum and Krubera—Although the genus Capnophyllum has sometimes been extended to include the Mediterranean species Krubera peregrina, Magee et al. (2009b) maintained the two genera as distinct, in agreement with Meikle (1977) and Burtt (1991). The fruit of both gen- 1E5 era (Fig. 5D, G) are superficially similar; they have dorsally compressed mericarps, a broad commissure, marginal ribs FIG. 6. Transverse sections through the fruit of Sonderina showing the extended into wings, and prominent ridges on the dorsal A. square, Sonderina sp. 1 (Magee & Boatwright 105, JRAU), or B. upright surface. However, on close examination of the fruit, Magee Sonderina tennis (Van Wyk et al. 3433, JRAU) cells external to the vittae. Scale: 0.07 mm et al. (2009b) found diagnostic differences between the two genera in terms of the size and prominence of both vit- tae and rib oil ducts and the shape of the marginal wings. Dasispermum, Sonderina, and Stoibrax—Burtt (1989) sug- Krubera and Capnophyllum were widely separated in the ITS- gested that the South African endemic genus Sonderina derived trees (Fig. 1), with Krubera placed in a Glade sister may be insufficiently distinct at the generic level from the to Coriandrum sativum and Bifora radians, and Capnophyllum largely North African genus Stoibrax. Previous molecular placed in the Lefebvrea Glade of tribe Tordylieae. In all trees systematic studies have shown the type species of Stoibrax presented herein, Capnophyllum is strongly supported as (S. dichotomum) to be placed in tribe Apieae (e.g. Downie monophyletic and occupies a position within a broader et al. 2001). Our analyses of both ITS and rps16 intron data lineage comprising other largely annual, sympatric, Cape clearly show Sonderina to form part of the Lefebvrea Glade endemic genera (viz., Dasispermum, Sonderina, Stoibrax cap- together with the only South African species of Stoibrax, ense, and Scaraboides). S. capense. Such a relationship was proposed by Adamson 10 SYSTEMATIC BOTANY [Volume 34
(1939, 1950) who treated Stoibrax capense (then Stoibrax young plants of this species and is therefore not clearly vis- didyma) as Sonderina didyma. ible in Fig. 7A. The species is easily distinguished by the pres- The monotypic genus Dasispermum is strongly supported ence of additional wing vittae (not known in any other genus in all analyses as embedded within Sonderina. The single spe- within the family) and parallel, closely-spaced commissural cies, Dasispermum suffruticosum, is a perennial dune endemic, vittae in the fruit (Fig. 7E–G). distinguished by its usually fleshy leaves and often promi- Separate and combined analyses of ITS and rps16 intron nently winged fruit (Fig. 5F) which can be either homo- or data sets place Scaraboides either within the Dasispermum- heteromericarpic. The genus is plastic with regard to both Sonderina complex (but with weak support values) or are of these characters (Taney and Van Wyk 1995). Leaves may equivocal in its placement. Shimodaira-Hasegawa tests indi- be less fleshy in some individuals within a population, espe- cate that a sister group relationship of Scaraboides to this com- cially those in more shaded sites and the ribs of some fruits plex (as retrieved in the analyses of the ITS/morphology and may not be expanded into wings. Both characters appear to ITS/rps16 intron/morphology data sets) cannot be rejected. In be adaptations to harsh littoral conditions and wind disper- the analysis of morphological data, the genus was moderately sal. Dasispermum shares the sympodial growth habit, a syna- supported (BP = 75) as sister to Capnophyllum. When com- pomorphy for the Capnophyllum group (Fig. 4B), as well as the bined with the molecular data sets, the position of Scaraboides isodiametric fruit, median and/or lateral ribs as well devel- was either unresolved (MP strict consensus tree, Fig. 3C) or oped as the marginal ribs, narrow commissure (Fig. 5A, B, C, F), sister to the Dasispermum–Sonderina complex (Fig. 3C and D). and the square or upright cells external to the vittae of the The genus is morphologically distinct from the Dasispermum- aail fruit (Fig. 6A, B) with Sonderina and Stoibrax. These character Sonderina complex, which has isodiametric fruit with a nar- states are all synapomorphies for the Dasispermum–Sonderina row commissure (Fig. 5B, C, F). The inclusion of Scaraboides complex (Fig. 4D, F, G, H). Furthermore, the close relationship within an expanded circumscription of Dasispermum would between Dasispermum suffruticosum and Stoibrax capense is result in a group that would be impossible to delimit based supported by a chromosome number of n = 9, an unusual on observed morphological characters. Despite the superfi- number for the subfamily (Constance et al. 1976). An expanded cial similarity between the fruits of Scaraboides manningii and circumscription of the genus Dasispermum to include Sonderina Capnophyllum, neither the molecular nor the combined molec- and Stoibrax capense, therefore, seems to be unavoidable. ular/morphological analyses place these taxa together. It is Scaraboides—While revising the genus Capnophyllum therefore clear that Scaraboides represents an independent, (Magee et al. 2009b), the authors were alerted to an unusual easily recognizable lineage. species from the arid Tanqua Karoo region. Although this Capnophyllum Group—All analyses presented herein species shares numerous fruit characters with Capnophyllum, indicate a broader lineage within the Lefebvrea Glade, com- such as dorsally compressed mericarps, broad commissures, prising Capnophyllum, Dasispermum, Scaraboides, Sonderina, concave commissural surfaces, and involute marginal wings and Stoibrax capense. These taxa share a unique combina- El (Fig. 7E–G), it also has morphological characters in common tion of characters, namely the monocarpic or rarely short- with species of Sonderina, such as an erect habit (Fig. 7A), lived perennial life history, the sympodial growth pattern, green ultimate leaf segments, scabrous often sessile umbels the papillose petals, and the broadly elliptic to rotund fruit (Fig. 7A, D), and the absence of involucral and involucel in lateral view. The Glade is therefore defined herein as the bracts (Fig. 7D). The sympodial habit is weakly expressed in Capnophyllum group.
KEY TO GENERA OF THE CAPNOPHYLLUM GROUP 1. Fruit isodiametric; if homomericarpic then with the median and lateral ribs as well developed as the marginal ribs, if heteromericarpic then with either median or lateral ribs as well developed as the marginal ribs, prominent or winged; commissure narrow, extending to the base of each rib Dasispermum 1. Fruit dorsally compressed; homomericarpic with the median and lateral ribs not as well developed as the marginal ribs, marginal ribs prominently winged, median and lateral ribs inconspicuous or prominent but not winged; commissure broad, extending to the tip of each wing 2. 2. Involucral and involucel bracts absent; rays and raylets scabrous; fruit with indistinct median and lateral ribs; additional vittae in the marginal wings; commissural vittae close together; ultimate leaflet segments more than 1.5 mm broad (never subterete), green Scaraboides 2. Involucral and involucel bracts present; rays and raylets glabrous; fruit with prominent median and lateral ribs; additional vittae in the marginal wings absent; commissural vittae widely separate; ultimate leaflet segments less than 1 mm broad (often subterete), glaucous Capnophyllum
TAXONOMIC TREATMENT The concept of the genus Dasispermum, which has nomen- clatural priority, is here expanded to include three species pre- 1. DASISPERMUM Raf., Good Book 56 (1840) emend. Magee & viously treated as Sonderina, as well as the only South African B.-E. van Wyk, emend. nov.—TYPE: Dasispermum mariti- species of Stoibrax, namely S. capense. As a result, this previ- mum Raf., nom illeg. (= Dasispermum suffruticosum (Berg.) ously monotypic genus now consists of seven South African B. L. Burtt). endemic species, two of which are as yet undescribed (Magee et al. in prep.). The genus can be distinguished from all other Sonderina H. Wolff in Pflanzenr. Heft 90: 92. 1927, syn. nov.— genera within the Lefebvrea Glade by a combination of char- TYPE: Sonderina hispida (Thunb.) H. Wolff acters, namely the sympodial growth habit (resulting in leaf- Carum sect. Brachyapium Baill., Hist. Pl. 7: 118. 1879. Brachyapium opposed umbels), papillose petals, isodiametric fruit with the (Bail.) Maire in Bull. Soc. Hist. Nat. Afr. Nord 23: 186. median and/or lateral ribs as prominent as the marginal ribs, 1932, syn. nov.—TYPE: Ptychotis didyma Sond. the narrow commissure extending to, at the most, the base of 2009] MAGEE ET AL.: CAPE GENERA OF APIACEAE 11
Fic. 7. Scaraboides manningii. A. habit, B. lower leaf pinnae, C. petal in ventral and lateral view, D. umbellule, E. fruit, dorsal surface, F. fruit, commis- sural surface, G. transverse section through the mature fruit. A—D: Manning 3010 (NBG); E—G. Manning 3061 (NBG). cv - commissural vitta; vv - vallecular vitta; wv - additional wing vitta. each rib and the presence of square or upright cells external Sonderina caruifolin (Sond.) H. Wolff, Pflanzenr. Heft 90:94. to the vittae of the fruit. 1927, syn. nov.— TYPE: SOUTH AFRICA. Cape, DASISPERMUM SUFFRUTICOSUM (Berg.) B. L. Burtt, Notes Roy. Riebeekkasteel, Zeyher 729 (K!, LE!, NBG!, 5!). Bot. Gard. Edinburgh 45: 93. 1988. Conium suffruticosum Dasispermum humile (Meisn.) Magee & B.-E. van Wyk, Berg., P1. Cap. 77. 1767.—TYPE: SOUTH AFRICA. Cape comb. nov. Petroselinum humile Meisn. in Hook., Lond. J. of Good Hope, Grubb s.n. (STB!). Bot. 2: 531. 1843.—SYNTYPE: SOUTH AFRICA. Natal, Dasispermum capense (Lam.) Magee & B.-E. van Wyk, comb. near Port Natal, Krauss 418 (BM!, K!, MO!); Drege 9545 nov. Caucalis capensis Lam., Encycl. 1: 658. 1785.—TYPE: (not located). SOUTH AFRICA. Cape, Sonnernt s.n. (P-LAM!). Dasispermum tenue (Sond.) Magee & B.-E. van Wyk, comb. Dasispermum hispidum (Thunb.) Magee & B.-E.van Wyk, nov. Ptychotis tenuis Sond., Fl. Cap. 2: 537. 1862.—TYPE: comb. nov. Sium hispidum Thunb., Prodr. 51. 1794.—TYPE: SOUTH AFRICA. Cape, Buffeljagdrivier to Rietkuil, SOUTH AFRICA. Cape, Thunberg s.n. (UPS-sheet 7046!). Zeyher 2672 (LE!, S!). 12 SYSTEMATIC BOTANY [Volume 34
(non angustis glaucis), umbellis scabris (non glabris) saepe sessilibus, bracteis involucralibus involucellaribusque defi- Over 1500 m cientibus, costis dorsalibus fructus indistinctis, vittis additis =J 900 - 1500 m solitariis in quoque alo marginali fructus et vittis paralelis cre- 300 - 900 m bris commissurialibus differt. Under 300 m Erect herb, 0.2-0.4 m tall. Stem single, rarely slightly branched at the base, erect. Leaves 50-120 mm x 20-60 mm, pinnate, glabrous, green. Petioles 20-60 mm long, basal sheaths 7-12 mm x 3-5 mm. Ultimate leaflets broadly ovate, 12-30 mm x 10-28 mm, venation pinnate; segments nar- rowly oblong, 2-9 mm x 1.5-3 mm, flat. Umbels compound; peduncle sessile or rarely short, 0(-30) mm long; involucre absent; rays 4-6, 10-30 mm long at anthesis, slightly sca- brous; involucel absent; raylets 6-9, 5-8 mm long at anthe- sis, scabrous. Flowers pentamerous; petals ± 0.5 mm long and broad, papillose, inflexed tips obtuse, septum absent on inner face, apex truncate; ovary glabrous; stylopodium flat, level with or slightly sunken below the fruit apex; styles not markedly elongated in mature fruit, 0.2-0.4 mm long, remaining erect or rarely becoming somewhat reflexed up to the base of the stylopodium. Fruit broadly elliptic, 5.5- 6.0 mm x 3.0-3.5 mm; base obtuse or shallowly concave; apex obtuse; mericarps strongly concave on the commis- sural surface; median and lateral ribs indistinct; marginal ribs distinctly involute; additional solitary vittae present in the marginal wings. Diagnostic Characters—The fruit is most similar to those of Capnophyllum in the broad commissure, the dorsally com- pressed mericarps with concave commissural surfaces and the involute marginal wings, but differs in the indistinct median and lateral ribs, the additional solitary vittae in each marginal wing and the parallel, closely spaced commis- sural vittae. When in flower, this species may be confused with Dasispermum humile but it is geographically isolated and can easily be distinguished by the flattened stylopodium and the petals which are not keeled on the adaxial face and FIG. 8. The known geographical distribution of Scaraboides manning i. which have only a short, acute tip (not slender and attenu- ate - the typical lobulum inflexum as is found in most other DASISPERMUM sr. 1. Esterhuysen 28979 (BOL!), 34312 (BOL!); Apioideae). Magee & Boatwright 105 (JRAU!), Winter 3850 (JRAU!, Distribution an d Habita t—Scaraboides manningii is restricted PRE!). to the Succulent Karoo of the Western Cape Province, where it is known from only two localities, the Tanqua Karoo National DASISPERMUM sr. 2. Lutzeyer s.n. (JRAU!, NBG!). Park and Mauwerskop near Vanrhynsdorp (Fig. 8). The spe- 2. Scaraboides Magee & B.-E. van Wyk, gen. nov.—TYPE: cies grows in seasonally damp, dolerite or clay soils. S. manningii Magee & B.-E. van Wyk. Etymology—This species is named in honour of Dr. John Manning (NBG) who brought it to our attention and who also A combined generic and specific description (descriptio provided the first complete collection. generico-specifica) is provided under S. inanningii below. Additional Specimens Examined—SOUTH AFRICA. 3118 (Calvinia): Etymology—The generic name is derived from the distinc- Mauwerskop, NW of Matsikammaberg (–DB), Snijman 1056 (PRE, MO). tive dark color and the strongly convex, smooth outline of 3219 (Wuppertal): Top of Boulderkoppie, E of Leeuberg (–BB), Sachse the fruits (making them distinctly beetle-like in appearance), 71 (PRE), Tanqua Karoo National Park. 3220 (Sutherland): NE slopes of Remhoogte, ca. 1 km S from Maansedam, Tanqua Karoo National Park hence Scaraboides [from the Greek, scarabeus (beetle) and -oides (–AA), Bester 7171 (PRE); E foot of Elandsberg, Tanqua Karoo National (like)]. Park (–AA), Manning 3010 (NBG); E foot of Elandsberg, Tanqua Karoo Scaraboides manningii Magee & B.-E. van Wyk, sp. nov.— National Park (–AA), Manning 3061 (NBG); Elandsberg, Wilderness cha- TYPE: SOUTH AFRICA. Sutherland district: Tanqua lets (–AA), Sachse 628 (PRE). Karoo National Park, NE slopes of Elandsberg; 06 August 2006; B. Sachse 116 (holotype: PRE!; isotypes: BOL!, JRAU!, ACKNOWLEDGMENTS. The curators and staff of the cited herbaria are thanked for their kind hospitality and assistance during visits and K!, KMG!, KSAN!, NBG!). for making specimens available on loan. The authors would also like Descriptio generico-specifica: Capnophyllo Gaertn. habitu to express gratitude to Dr. J. C. Manning (NBG) for assistance with col- annuali, commissura lata fructus, mericarpiis dorsaliter corn- lections and for alerting us to unusual collections, Mr. H. and Mrs. E. Lutzeyer for their enthusiasm and hospitality while performing field pressis, superficiebus commissurialibus concavis, alis mar- studies on their reserve in Stanford, Dr. H. Glen (NH) for translat- ginalibus involitis similis sed ramis erectis (non prostratis ing the diagnoses, Dr. J.-N. Labat (P) for providing a digital image of nec decumbentibus), segmentis ultimis foliorum latis viridis Caucalis capense, the Lesley Hill Laboratory, Jodrell Laboratory and Kew 2009] MAGEE ET AL.: CAPE GENERA OF APIACEAE 13
Herbarium for DNA aliquots and the Molecular Systematics Laboratory Leistner, 0. A. and J. M. Morris. 1976. Southern African place names. at the University of Johannesburg for the use of their facilities. This Annals of the Cape Province Museum 12: 1 -565. study was funded by the University of Johannesburg and the National Maddison, W. P. and D. R. Maddison. 2008. Mesquite: a modular sys- Research Foundation. tem for evolutionary analysis. Version 2.5. http:/ /mesquitepreject .org. Magee, A. R., B.-E. van Wyk, and P. M. Tilney. 2008a. A taxonomic revision of the genus Nanobubon (Apiaceae: Apioideae). South African Journal LITERATURE CITED of Botany 74: 713-719. Magee, A. R. B.-E. van Wyk, and P. M. Tilney. 2008b. A taxonomic revision Adamson, R. S. 1939. Some changes in nomenclature II. The Journal of of the genus Cynorhiza (Apiaceae: Apioideae). South African Journal of South African Botany 5: 53-58. Botany 74: 726-734. Adamson, R. S. 1950. Umbelliferae. Pp. 623-625 in Flora of the Cape Magee, A. R., B.-E. van Wyk, P. M. Tilney, and S. R. Downie. 2008c. Peninsula, eds. R. S. Adamson and T. M. Salter. Cape Town: Juta and Ezosciadum (Apiaceae): A taxonomic revision of yet another early Co. diverging South African apioid genus. Plant Systematics and Evolution Akaike, H. 1974. A new look at the statistical model identification. IEEE 276: 167-175. Transactions on Automatic Control 19: 716-723. Magee, A. R., B.-E. van Wyk, and P. M. Tilney. 2009a. A taxonomic revision Allison, I. and B.-E. van Wyk. 1997. A revision of the genus Anginon of the woody South African genus Notobubon (Apiaceae: Apioideae). - (Apiaceae). Nordic Journal of Botany 17: 561 577. Systematic Botany 34: 220-242. Burtt, B. L. 1989. The adoption of Stoibrax for Tragiopsis and Brachyapium Magee, A. R. B.-E. van Wyk, P. M. 'Farley, and S. R. Downie. 2009b. A taxo- (Umbelliferae), and its N-S African disjunction. Pp. 143-147 in nomic revision of Capnophyllum (Apiaceae: Apioideae). South African
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sis for relationships among Epicrates (Boidae, Serpentes). Systematic Biology and Evolution 14: 717-724. Zoology 38: 7-25. Kluge, A. G. and J. S. Farris. 1969. Quantitative phyletics and the evolution APPENDIX 1. Morphological and anatomical characters and states of Anurans. Systematic Zoology 18: 1-32. used in the phylogenetic analysis of the Lefebvrea clade. 'Field observa-
Lebrun, J. - P. and A. L. Stork. 1992. Enumeration des plantes a fleurs d'Afrique tions of Dasispermum suffruticosum and Sonderina sp. 1 indicate that these tropicale, Vol. 2. Geneve: Ville de Geneve Editions. species are not monocarpic annuals but rather short-lived perennials 14 SYSTEMATIC BOTANY [Volume 34 lasting for only a few seasons depending on rainfall, possibly an adap- APPENDIX 3. New accessions of Apiaceae from which ITS and rps16 tation to the dune habitat in which they both occur. Perennials include intron sequences were obtained, with corresponding voucher informa- all shrubs and also species with permanent fleshy roots. 'Some species tion and GenBank reference numbers. The information is listed as fol- of Sonderina are distinct in that their involucre and involucel bracts are lows: taxon — ITS, rps16 intron; voucher information. Taxa where the ITS usually absent or at best strongly reduced and rudimentary. 'Sonderina sequence data has been published previously (Winter et al. 2008) are indi- hispida, Sonderina sp. 1, and Stoibrax capense are unusual in that at least cated with an *. some of the involucre and involucel bracts are pinnately divided, thus resembling the leaves. Capnophyllum Gaertn.: C. africanum (L.) Gaertn. — FM201528, FM201546; Magee 124 (JRAU). C. africanum (L.) Gaertn. — FM201527, 1. Life history' (monocarpic = 0; short-lived perennial = 1; perennial = FM201548; Forest et. al. 654 (NBG). C. leiocarpon (Sond.) J.C.Manning & 2); 2. Habit (herbs = 0; rhizomatous = 1; suffrutices = 2; shrubs or shru- Goldblatt — FM201525, M201545; Bester 6978 (PRE). C. leiocarpon (Sond.) blets = 3); 3. Growth pattern (monopodial = 0; sympodial = 1); 4. Leaf J.C.Manning & Goldblatt — FM201526, FM201544; Magee & Boatwright 125 persistence (one-seasoned or deciduous = 0; permanent, evergreen = 1); (JRAU). C. lutzeyeri Magee & B.-E.van Wyk — FM201524, FM201543; Magee 5. Leaf arrangement (radical or if somewhat cauline then borne on decid- et al. 106 (JRAU). C. rnacrocarpum Magee & B.-E.van Wyk — FM201529, uous branches = 0; cauline, borne on permanent branches =1); 6. Leaf FM201547; Magee et al. 133 (JRAU). Conium L.: C. sphaerocarpum Hilliard texture (coriaceous = 0; flimsy = 1; sclerophyllous =2); 7. Leaf color (con- & Burtt— FM201530, FM201558; Magee at al. 129 (JRAU). Cynorhiza Eckl. colourously green or green above = 0; glacous =1); 8. Inflorescence ves- & Zeyh.: C. typica Eckl. & Zeyh. — *, FM201556; Magee et al. 53 (JRAU). titure (glabrous = 0; scabrous = 1); 9. Ratio of functionally male flowers C. typica Eckl. & Zeyh. —*, FM201557; Van Wyk 3372 (JRAU). Dasispermum (equal ratio of male to female flowers in all raylets of the umbellule = Raf.: D. suffruticosum (Berg.) B. L. Burtt— FM201514, FM201541; Magee & 0; inner raylets of umbellules functionally male =1); 10. Involucre and Boatwright 117 GRAU). Nanobubon Magee: N. capillaceum (Thunb.) Magee involucel bracts' (present = 0; absent or much reduced = 1); 11. Involucre — *, FM201549; Magee & Boatwright 14 (JRAU). N. strictum (Spreng.) and involucel bracts type' (absent or all simple = 0; at least some com- Magee — *, FM201550; Magee et al. 58 (JRAU). Notobubon B.-E.van Wyk: pound, resembling the leaves = 1); 12. Petal vestiture (leathery = 0; papil- N. capense (Eckl. & Zeyh.) Magee — *, FM201551; Magee et al. 43 ()RAU). lose = 1); 13. Fruit length (more than 9 mm = 0; less than 9 mm = 1); 14. N. capense (Eckl. & Zeyh.) Magee — *, FM201555; Magee et al. 37 (JRAU). Fruit compression (platyspermous = 0; isodiametric = 1); 15. Fruit in lat- N. gummiferum (L.) Magee — *, FM201554; Magee et al. 61 (JRAU). N. pear- eral view (narrowly elliptic = 0; broadly elliptic to rotund = 1); 16. Ribs sonii (Adamson) Magee — *, FM201552; Magee et al 42 (JRAU). N. tenuifo- (median and lateral ribs markedly less developed than the marginal ribs lium (Thunb.) Magee — *, FM201553; Magee et al. 44 (JRAU). Scaraboides = 0; median and/or lateral ribs as well developed as the marginal ribs = Magee & B.-E.van Wyk: S. manningii Magee & B.-E.van Wyk — FM201523, 1); 17. Ribs (obtusely tipped = 0; almost trifid with prominent tapering FM201542; Manning 3010 (NBG). Sonderina H.Wolff : S. hispida (Thunb.) tips = 1); 18. Secondary ribs (absent = 0; usually present = 1); 19. Marginal H. Wolff — FM201520, FM201537; Magee & Boatwright 115 GRAU). S. his- wings (absent or flat = 0; involute = 1); 20. Commissural surface (flat = pida (Thunb.) H.Wolff — FM201536; Magee et al. 112 (JRAU). S. hisp- 0; concave = 1); 21. Commissure (100% from rib tip to rib tip = 0; from ida (Thunb.) H.Wolff — FM201521, FM201535; Magee et al. 107 (JRAU). near rib tip to near rib tip = 1; from at most rib base to rib base = 2); 22. S. humilis (Meisn.) H.Wolff — FM201518, FM201538; Van Wyk & Van Wyk Rib vittae (absent = 0; present at base of all ribs = 1; present in marginal 1883 (JRAU). S. sp.l — FM201519, FM201534; Magee & Boatwright 105 wings = 2); 23. Cells external to vittae (indistinct =0; square = 1; enlarged, GRAU). S. sp. 2 — FM201522, FM201540; Lutzeyer s.n. (JRAU). S. tennis upright = 2). (Sond.) H.Wolff — FM201517, FM201539; Van Wyk et al. 3433 (JRAU). Stoibrax Raf.: S. capense (Lam.) B.L.Burtt — FM201516, FM201532; Magee APPENDIX 2. et al. 128 (JRAU). S. capense (Lam.) B.L.Burtt — FM201515, FM201533;
Matrix of morphological character states used in the phylogenetic analysis of the Lefebvrea Glade.
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 Afroligusticum elliotii 2 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 Afroligusticum petitianum 2 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Afrosciadium magalismontanum 2 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 Capnophyllum africanum 0 0 1 0 0 1 1 0 0 0 0 1 1 0 1 0 0 1 1 1 0 0 0 Capnophyllum leiocarpon 0 0 1 0 0 1 1 0 0 0 0 1 1 0 1 0 0 1 1 1 0 0 0 Capnophyllum lutzeyeri 0 0 1 0 0 1 1 0 0 0 0 1 1 0 1 0 0 1 1 1 0 0 0 Capnophyllum macrocarpum 0 0 1 0 0 1 1 0 0 0 0 1 0 0 1 0 0 0 1 1 0 0 0 Cynorhiza typica 2 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Dasispermum suffruticosum 1 2 1 1 0 0 1 0 0 0 0 0 1 1 1 1 0 0 0 0 2 0 1 Lefebvrea abyssinica 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Nanobubon capillaceum 2 1 0 1 0 2 0 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Nanobubon strictum 2 1 0 1 0 2 0 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Notobubon capense 2 3 0 1 1 2 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 1 0 Notobubon ferulaceum 2 3 0 1 1 2 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 1 0 Notobubon galbaniopse 2 3 0 1 1 2 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 1 0 Notobubon galbanum 2 3 0 1 1 2 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 1 0 Notobubon gumrniferum 2 3 0 1 1 2 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 1 0 Notobubon laevigatutn 2 3 0 1 1 2 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 Notobubon pearsonii 2 3 0 1 1 2 0 0 0 0 0 0 1 0 0 1 0 0 0 0 0 0 0 Notobubon pungens 2 3 0 1 1 2 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 1 0 Notobubon striatum 2 3 0 1 1 2 0 1 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 Notobubon tenuifoliutn 2 3 0 1 1 2 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 1 0 Scaraboides manningii 0 0 1 0 0 0 0 1 0 1 0 0 1 0 1 0 0 0 1 1 0 2 0 Sonderina hispida 0 0 1 0 0 0 0 1 0 0 1 1 1 1 1 1 0 0 0 0 2 0 1 Sonderina humilis 0 0 1 0 0 0 0 0 0 1 0 1 1 1 1 1 0 0 0 0 2 0 2 Sonderina tenuis 0 0 1 0 0 0 0 0 0 1 0 1 1 1 1 1 0 0 0 0 2 0 2 Sonderina sp. 1 1 2 1 0 0 0 0 0 0 0 1 1 1 1 1 1 0 0 0 0 2 0 1 Sonderina sp. 2 0 0 0 0 0 0 0 0 0 0 0 1 1 1 1 1 0 0 0 0 2 0 2 Stenosemis caffra 2 1 0 1 0 2 0 1 0 0 0 1 1 1 1 1 1 0 0 0 1 0 0 Stenosemis angustifolia 2 1 0 1 0 2 0 1 0 0 0 0 1 1 1 1 1 0 0 0 1 0 0 Stoibrax capense 0 0 1 0 0 0 0 1 0 0 1 1 1 1 1 1 0 0 0 0 2 0 1 2009] MAGEE ET AL.: CAPE GENERA OF APIACEAE 15
Magee et al. 131 GRAD). S. dichotomum (L.) Raf. — FM201531, –; Sanchez- (DC.) Eckl. & Zeyh. AY8384184; Echinophora tenuifolia L. AF164812'; Mata & Molina Abril s.n. (K). Foeniculum vulgare Mill. AF110543'; Heracleum lanatum Michx. AF1105371; Heracleum maximum Bartr. EF426691 6; Heracleum spondylium L. AF1648002; Malabaila sekakul Boiss. AF1648022; Nanobubon strictum (Spreng.) Magee APPENDIX 4. Previously published rps16 intron accessions of Apiaceae AY8384384; Naufraga balearica Constance & Cannon AF164816 2; Notobubon obtained from GenBank.' Downie and Katz-Downie (1999); 2 Downie ferulaceum (Thunb.) Magee AY8384344; Notobubon galbanum (L.) Magee et al. (2000)' Sun and Downie (2004); 6 Calvin() et al. (2006);' Magee et al. AY8384354; Notobubon pearsonii (Adamson) Magee AY8384364; Notobubon (2008c);6 McNeill and Kemper (unpubl.). pungens (E.Mey. ex Sond.) Magee AY8384374; Pastinaca sativa L. AF110538'; Sium latifolium L. AF110552'; Smyrnium olusatrum L. AF110551'; Stenosemis Aethusa cynapium L. AF110539'; Ammi majus L. AF1648142; Apium gra- caffra Sond. AY8384446; Stoibrax dichotomum (L.) Raf. AM9825185; Thaspium veolens L. AF110545'; Berula erecta (Huds.) Coville AF1648192; Conium mac- pinnatifidum (Buckley) A.Gray AY372896 3; Zizia aurea Koch AF110535'; ulatum L. AF110546'; Crithmum maritimum L. AF110540'; Deverra burchelli Zosima orientalis Hoffm. AF1648062. APPENDIX C4
MAGEE, A. R., B.-E. VAN WYK, P. M. TILNEY, F. SALES, I. HEDGE and S. R.
DOWNIE. Billburttia, a new genus of Apiaceae (tribe Apieae) endemic to
Madagascar. Plant Systematics and Evolution. In press. Billburttia, a new genus of Apiaceae (tribe Apieae) endemic to Madagascar
A. R. Magee', B.-E. van Wyk', P. M. Tilney I , F. Sales", I. Hedge3, and S. R.
Downie
'Department of Botany and Plant Biotechnology, University of Johannesburg,
Johannesburg, South Africa
2 Departamento de Botanica, Faculdade de Ciencias e Tecnologia, Universidade de Coimbra, 3001-456, Portugal
3 Royal Botanic Garden, Edinburgh EH3 5LR, Scotland, United Kingdom
4 Department of Plant Biology, University of Illinois at Urbana-Champaign,
Urbana, Illinois 61801, United States of America
Abstract. The genus Billburttia is described to include two new Malagasy endemic species, B. capensoides and B. vaginoides. Although the species are superficially similar to those of the southern African endemic peucedanoid genus
Notobubon, they differ markedly in their fruit anatomical characters, notably a narrower commissure, six commissural vittae, vascular tissue in the tip of the ribs, and sphaerocrystals distributed in and around the epidermis. The two last- mentioned characters are proposed as generic apomorphies for Billburttia. The non-peucedanoid affinity of the genus was confirmed using ITS and rps16 intron sequence data. Both parsimony and Bayesian analyses of these data place
Billburttia within the tribe Apieae of subfamily Apioideae, and not closely related Magee et al.: Billburttia, a new genus of Apiaceae to either Peucedanum (Selineae) or the African peucedanoid genera (Lefebvrea
Glade of Tordylieae).
Keywords: Billburttia capensoides - Billburttia vaginoides - Fruit anatomy — ITS —
New genus - New species - Peucedanum - rps16 intron - sphaerocrystals
Although the family Apiaceae is relatively poorly represented in Madagascar [15 genera (11 endemic) and 31 species (10 endemic)], the Malagasy contingent represents an important component particularly within the early diverging lineages of subfamily Apioideae (Van Wyk et al. 1999; Calvin° et al. 2006;
Calvin° et al. in prep.). These Malagasy species display a diverse range of unusual habit and fruit anatomical characters (Sales et al. 2004). Earlier treatments under-estimated the total number of endemic genera, largely because of inadequate material, with many of the isolated Malagasy species included in
African genera. Humbert (1956) placed eight of the woody Malagasy species within the arboreal African Heteromorpha Cham. and Schltdl. However, after extensive morphological and anatomical studies, the Malagasy species were later excluded from Heteromorpha (Winter et al. 1993; Winter and Van Wyk
1994, 1996) and accommodated largely in four new Malagasy endemic genera
(Van Wyk et al. 1999), with one of the species transferred to the tropical African genus Pseudocarum C.Norman. Sales and Hedge (in press) recently described five new Malagasy species. Two of these species were tentatively placed within the problematic Peucedanum L. (Sales et al. 2004; Peucedanum sp. A and
2 Magee et al.: Billburttia, a new genus of Apiaceae
Peucedanum sp. B). The former species, described here as Billburttia capensoides, though relatively well-collected has consistently been misidentified as part of the Peucedanum capense (Thunb.) Sond. complex (Burtt 1991), due to surprising morphological similarities. Recent detailed studies on African
Peucedanum have shown that all the African species are unrelated to Eurasian
Peucedanum and, therefore, were accommodated in six African endemic genera
(Magee et al. 2008a, Magee et al. 2008b, Magee et al. 2009a, Winter et al.
2008). As a result, the species in Madagascar represented the last remaining challenge to the so-called "Peucedanum problem" in Africa. Using fruit anatomical and molecular sequence data, we show here that the two new species (described herein as "B. capensoides" and "B. vaginoides") are unrelated to Peucedanum and other peucedanoid taxa but represent a distinct new genus described herein as "Billburttia".
Material and methods
Taxonomic study. Herbarium specimens from the collections at BM, E, K,
MO, P and S were studied. Line drawings were made by the first author with the aid of a camera lucida attachment on a Zeiss compound microscope or a Wild
M3Z stereomicroscope.
Fruit anatomy. Fruit from two specimens of "Billburttia capensoides"
(Humbert 3634, P; Rakotozafy 631, P) and one specimen of "B. vaginoides"
(Decary 7583, P) were rehydrated for anatomical study. Following rehydration, the fruit were fixed in FAA for a minimum of 24 h and subsequently embedded in
3 Magee et al.: Billburttia, a new genus of Apiaceae glycol methacrylate (GMA) according to a modification (final infiltration in GMA of five days) of the method of Feder and O'Brien (1968). Transverse sections of about 31.1m thick were made using a Porter-Blum ultramicrotome. These sections were examined for the presence of crystals using a light microscope, after which they were stained according to the periodic acid Schiff/toluidine blue (PAS/TB) staining method of Feder and O'Brien (1968). To study the three-dimensional structure of the vittae, boiling water was poured over the fruits and they were then left to cool and soak for 24 h. The exocarp was peeled off while keeping the fruit submerged in water to prevent desiccation.
Molecular data. Total DNA was extracted from herbarium specimens of
"B. capensoides" (3 specimens) and "B. vaginoides" (1 specimen) using the
PureLinkTM Plant Total DNA Purification Kit (Invitrogen, Carlsbad, California,
USA). For amplification of the nuclear ribosomal DNA internal transcribed spacers (ITS) and chloroplast DNA rps16 intron region we used the primers of
Downie and Katz-Downie (1996) and Downie and Katz-Downie (1999), respectively. Successfully amplified PCR products were purified according to the
ExoSAP protocol of Werle et al. (1994) using 5 units of Exonuclease I (New
England Biolabs, Ipswich, Massachusetts, USA) and 0.5 units of Shrimp Alkaline
Phosphatase (Promega, Madison, Wisconsin, USA). Sequencing reactions were carried out using the BigDye Terminator version 3.1 Cycle Sequencing Kit
(Applied Biosystems Inc.) and sequenced using either an ABI (Applied
Biosystems) 3130 XL or 3730 XL sequencer. Complementary strands were assembled and edited using Sequencher version 3.1.2 (Gene Codes
4 Magee et al.: Bifiburttia, a new genus of Apiaceae
Corporation) and manually aligned in PAUP* (Swofford 2002), with gaps positioned so as to minimise nucleotide mismatches. Sources of material used in the study are listed in Appendix 1. All newly obtained sequences have been deposited in Genbank.
Phylogenetic analyses. To assess the phylogenetic positions of the two species, the newly obtained ITS (three accessions of "B. capensoides" and one accession of "B. vaginoides") and rps16 intron (two accessions of "B. capensoides") sequences were incorporated into the respective ITS and rps16 intron data matrices of Magee et al. (2009b). These matrices include most of the relevant major clades of the "apioid superclade," including a representative sampling of the Lefebvrea Glade (Magee et al. 2009b) which comprises many
African peucedanoid genera (Winter et al. 2008). Preliminary analyses indicated an affinity between the two new species and the tribe Apieae, therefore all available accessions of the latter were also included. The trees were rooted with taxa from tribes Smyrnieae and Oenantheae (Downie et al. 2001). In total, 150 accessions of ITS and 60 accessions of rps16 were considered for the phylogenetic study. Data sets are available on request from the corresponding author. Parsimony analyses (MP) were conducted using PAUP* with character transformations treated as unordered and equally weighed (Fitch parsimony;
Fitch 1971). Tree searches were performed using the strategies employed by
Downie et al. (1998). Bootstrap percentage values (BP; Felsenstein 1985) were determined from 500,000 replicate analyses using fast stepwise addition of taxa.
Only values greater than 50% are reported, and the following scale was used to
5 Magee et al.: Billburttia, a new genus of Apiaceae evaluate support percentages: 50%-74%, weak; 75%-84%, moderate; and
85%-100%, strong. Appropriate models of evolution were selected for each dataset using MODELTEST version 3.06 (corrected AKAIKE information criterion; Posada and Crandall 1998). Bayesian inference (BI) was performed for four million generations of Monte Carlo Markov Chains with a sampling frequency of 100 using MRBAYES version 3.1.2 (Huelsenbeck and Ronquist 2001;
Ronquist and Huelsenbeck 2003). Suboptimal trees were discarded as the 'burn- in' phase (25%), and from the remaining trees a majority rule consensus was produced using the "sumt" command of MRBAYES. Only posterior probability
(PP) values greater than 0.5 are reported, and the following scale was applied:
0.50-0.84, weak; 0.85-0.94, moderate; 0.95-1.0, strong.
Results and discussion
Morphology. "Billburttia capensoides" and "B. vaginoides"are glabrous shrubs or subshrubs up to 2 m tall. Their leaves are cauline and glaucous. The species are easily separated from each other by their ultimate leaf segments. In
"B. capensoides" these are linear-elliptic to elliptic (Fig. 1 a) and closely resemble those of Notobubon laevigatum (Aiton) Magee (Fig. 1i), from which it can be distinguished by the lack of a prominent midrib and well-developed lateral venation. The ultimate leaf segments of "B. vaginoides", in contrast, are filiform to linear with only the main vein visible (Fig 1b). Each stem terminates in a much branched inflorescence borne on a long peduncle. The relatively small fruit (Fig.
1g) are superficially similar to those found in species of Notobubon B.-E.van Wyk
6 Magee et al.: Billburttia, a new genus of Apiaceae
(Fig. 11), but differ in the prominent dorsal ribs and several other important details, as described below.
Fruit anatomical description. In transverse sections (Fig. 1 c-e) the fruit of "B. capensoides" and "B. vaginoides" are slightly dorsally compressed and homomericarpic. The marginal ribs are expanded into narrow or scarcely developed wings and the dorsal ribs are very prominent. The commissure differs notably from that of species of both Eurasian Peucedanum (Fig. 1k) and the
African peucedanoid taxa (e.g. Notobubon, Fig. 1j) in that it extends only to the base of each marginal wing (and not to the tip of each wing, as in the latter genera). The epidermal surface is striate and the epidermal cells are periclinally elongated. There is some lignification of the mesocarp in the ribs. The cells of the endocarp are parenchymatous and periclinally elongated. The vascular tissue is located at the tip of the dorsal and marginal ribs (Fig1c-e) and as such is an unusual and diagnostic character for both species. Rib oil ducts were not observed in the mature fruit. The mericarp has 10 to 12 regular vittae. Along the commissural length there are six vittae of unequal length, while in each vallecula there may be one or two vittae present. Crystals were observed in groups around the epidermis and vittae in unstained sections of both species (Fig. 1f). These crystals were initially thought to be calcium oxalate druses, as reported in some taxa of the Apioideae-Saniculoideae Glade (Rompel 1895, Drude 1897-1898, Van
Wyk and Tilney 2004; Lui et al. 2006, 2007; Magee et al. 2008c). However, as the crystals did not dissolve during staining with PAS/TB, it became apparent that they were not composed of calcium oxalate. On closer examination of both
7 Magee et al.: Billburttia, a new genus of Apiaceae stained and unstained transverse sections they were identified as sphaerocrystals of unknown composition. This appears to be the first report of sphaerocrystals in Apiaceae and, as such, represents a potentially useful diagnostic character at the generic level.
ITS and rps16 intron data. The ITS dataset included 634 unambiguously aligned nucleotide positions of which 384 were variable and 321 parsimony informative. MP analyses resulted in the preset maximum tree limit of 12,000 trees, each of 2,207 steps [consistency indices (CI) of 0.33 and 0.30, with and without uninformative characters, respectively; retention index (RI) of 0.73].
MODELTEST selected the GTR+I+G model of evolution for use in the BI analysis. The MP strict consensus and BI majority-rule consensus trees (Fig. 2a) were largely topologically congruent and retrieved the same clades as those reported by Magee et al. (2009b). The Glade comprising the two new species
(three accessions of "Billburttia capensoides" and one accession of "B. vaginoides") was strongly supported (PP 1.0, BP 100) and formed part of the tribe Apieae (PP 1.0, BP 84). The two species were sister group to a Glade comprising Sclerosciadium nodiflorum Coss., Ammi majus L. and Petroselinum crispum (Mill.) Fuss (PP 0.91, BP 61).
The rps16 intron dataset included 865 unambiguously aligned nucleotide positions of which 179 were variable and 98 parsimony informative. MP analyses resulted in the preset maximum tree limit of 12,000 trees, each of 277 steps (CI of 0.75 and 0.63, with and without uninformative characters, respectively; RI of
0.86). MODELTEST selected the K81uf+G model of evolution for use in the BI
8 Magee et al.: Billburttia, a new genus of Apiaceae analysis. The MP strict consensus tree and the BI majority-rule consensus trees
(Fig. 2b) were largely topologically congruent and retrieved the same clades as those reported by Magee et al. (2009b), although the topologies recovered within the Glade comprising the tribe Apieae differed slightly as discussed below. The position of the Malagasy species, "B. capensoides", within the tribe Apieae (PP
0.97, BP <50) was confirmed (Fig 2b), although rps16 intron data for the closely related "B. vaginoides" was unavailable due to amplification difficulties. In both the BI and MP consensus trees, the two accessions of "B. capensoides" formed a
Glade which was sister group to Stoibrax dichotomum (L.) Raf. (PP 0.85, BP
<50). However, in the BI consensus tree "B. capensoides" and Stoibrax dichotomum formed part of a larger, weakly supported Glade (PP 0.82) including
Deverra DC., Ammi majus, Petroselinum crispum, Apium graveolens L. and
Naufraga balearica Constance and Cannon (BI 0.82), while in the MP strict consensus tree "B. capensoides" and Stoibrax Raf. formed part of a Glade (BP
<50%) comprising Anethum graveolens L. and Foeniculum vulgare Mill.
Phylogenetic position. The two species are clearly unrelated to both
Eurasian Peucedanum and the African peucedanoid taxa. Analyses of both ITS and rps16 intron data strongly place "Billburttia" within the tribe Apieae (Fig 2).
Although "B. vaginoides" was not included within the analyses of the rps16 intron dataset, this species is clearly closely related to "B. capensoides" based on morphological, anatomical and ITS sequence data. It is unclear what the direct sister group of "Billburttia" is, as either Stoibrax or Ammi, Petroselinum and
Sclerosciadium as retrieved as sister groups by either cpDNA or nrITS sequence
9 Magee et al.: Billburttia, a new genus of Apiaceae data, respectively. This may be due to the low resolution within the rps16 intron dataset or the result of hybridization and introgression of the genome. Although the fruit of "Billburttia"are superficially similar to the previously mentioned peucedanoid genera, anatomically they are markedly dissimilar. The position of the vascular bundles in the tips of the ribs and the presence of sphaerocrystals are both unusual fruit anatomical characters which lend support to both the isolated position of the two species and their recognition as a distinct and monophyletic new genus.
Taxonomic treatment
Billburttia Magee and B.-E.van Wyk, gen. nov. Forma foliorum, mericarpiorum et habitu Peucedanum L. et praecipue Notobubon B.-E. van Wyk similis sed ab eorum vittis vallecularibus 6, margine mericarpiorum indistincto, commissuris angustis, sphaerocrystallis in regione epidermidis bene differt. TYPE: B. capensoides Sales and Hedge.
Shrub or subshrub, clump-forming, glabrous, 0.5-2.0 m tall. Rootstock thick to very thick, woody, vertical or oblique. Stems erect, branched in region of inflorescence, terete, clearly and finely ridged, solid, blue-green or reddish.
Leaves cauline, persisting, 1- to 3-pinnate. Ultimate leaf segments filiform—linear or linear-elliptic to elliptic; margins entire; base narrow to broad and cuneate; apex acute to mucronulate; firm to thick-textured; concolorous, glabrous to glaucous; midrib and lateral veins equally developed or only midrib prominent, flush with or sunken below the lamina. Petioles terete, sheathing almost along
10 Magee et al.: Billburttia, a new genus of Apiaceae their entire length. Inflorescence of 1 to 6, lax hermaphrodite and smaller male compound umbels; terminal umbels hermaphrodite; peduncle long, more than two times longer than the diameter of the primary umbel; striate. Primary umbel rounded; rays 15 to 30,15-40 mm long, ± equal, glabrous; involucre present; bracts numerous, linear-oblong, unequal, apex acute to acuminate, glabrous, papery; raylets glabrous, glaucous; involucel present; bracteoles numerous, linear-oblong, unequal, apex acute to acuminate, ± fused at base, glabrous, papery, shorter than raylets; umbellule many-flowered. Flowers pentamerous, predominantly hermaphrodite, those in the lateral umbels entirely or in part functionally male; sepals minute, apex truncate to acuminate, glabrous; petals
1.5 mm long, greenish yellow, yellow or white, elliptic to obovate, acuminate, with inflexed tips, glabrous; stylopodium broadly conical; styles 2, short; ovary bilocular. Fruit slightly dorsally compressed, narrowly ellipsoid or oblong-ellipsoid,
3.5-4.5 mm x c. 1.5 mm, glabrous; mericarps homomorphic; median and lateral ribs prominent; marginal ribs narrowly-winged, not saccate at base; styles becoming strongly reflexed up to or beyond the base of the stylopodium; commissural vittae 6; vallecular vittae 4 to 6, solitary or in pairs; commissure broad, extending to the base of each wing; carpophore bipartite. Endosperm flat.
Etymology. The genus is named in honour of Brian Laurence ("Bill") Burtt
(1913-2008) of the Royal Botanic Garden Edinburgh (Weber and Noltie 2008) whose paper on southern African Apiaceae (Burtt 1991) has contributed greatly to our understanding of the family in that region. He was also a co-author of the account of Gesneriaceae for Flore de Madagascar.
11 Magee et al.: Billburttia, a new genus of Apiaceae
Diagnostic characters. The species of Billburttia are superficially similar to some species of Notobubon. They are small shrubs or subshrubs with a woody rootstock and have relatively small, slightly dorsally compressed fruit. Billburttia is easily distinguished from Notobubon and members of the tribe Apieae by a combination of distinct fruit characters, namely the narrower commissure (not extending beyond the base of each wing, as in Notobubon), the six commissural vittae (vs. two commissural vittae), ribs with the vascular bundles located near the tips, and the presence of sphaerocrystals around the epidermis and vittae.
Distribution. The species are endemic to Madagascar.
1. Billburttia capensoides Sales and Hedge, sp. nov. Herba suffruticosa vel suffrutex, glabra. Caudices verticales. Caules 1-2 m, foliati, erecti, solidi, tenuiter striati. Folia 1-2-pinnata, segmentis ellipticis vel lineari-ellipticis, 5-15 mm latis; vaginae amplexicaules ad 2-6 cm longae. Bracteae 4-8 (-12) mm longae; bracteolae 5-3 x 1-1.5 mm basin versus plus minusve connatae; radii 12-25, 15-
40 mm longi. Mericarpia dorsaliter compressa, basi non saccata jugis aequalibus haud alatis; stylopodium conicum; styli valde reflexi applanati stylopodiis 1.5- duplo longior. TYPE: Madagascar. Perrier de la Bathie 6807 (P!, holotype).
Shrub or subshrub, aromatic. Rootstock very thick, vertical. Stems 1-2 m, leafy, c. 0.5 cm wide at base; blue-green or reddish. Leaves 1 to 2-pinnate.
Ultimate leaf segments linear-elliptic to elliptic, 20-40 x c. 5-15 mm; apex acute to mucronulate; thick-textured; somewhat or clearly glaucous; midrib and lateral veins equally developed. Petioles of lower leaves 20-60 mm long. Primary umbel
12 Magee et al.: Billburttia, a new genus of Apiaceae
(60) 100 to 120 mm wide; rays 12 to 25, 15-40 mm long; bracts 4-8 (-12) mm long; raylets 15 to 20, 4-8 mm long; bracteoles 1.5-3 x 1-1.5 mm. Fruit narrow ellipsoid, 3.5-4.5 x c. 1.5 mm; styles reflexed beyond the base of the stylopodium. Illustrated in Sales and Hedge (in press).
Diagnostic characters. Billburttia capensoides is readily distinguished from the closely related B. vaginoides by the broader, linear-elliptic to elliptic ultimate leaf segments with a prominent midrib and well-developed lateral venation. This much-collected Madagascar species has always been previously identified as the superficially similar southern African Peucedanum capense
(Thunb.) Sond. [=Notobubon laevigatum (Aiton) Magee]. The epithet of this new
Malagasy species indicates its superficial similarity to the southern African taxon.
Distribution and habitat. Granite rocks, quartz, dry hillsides, pine woods, dry pine wood, scrubby forest and roadside, sunny places, stream bank in open grassland, damp places; locally abundant, 1000-2658 m.
Phenology. Flowering (August) October to March; fruiting June to
September.
Vernacular name: tsilaninosy (Rakotovao 52).
Selected specimens examined:
Baron 340, s. loc. (K, P); 2021, central Madagascar (BM, K). Bosser 7563,
Faratsiho, Ankaratra (P). Clement, Phillipson & Rafamantanantsoa 2009,
Fianarantsoa, Ambositra to Ambatofinandrahana on R.N. 35, 1600 m (E, MO, P).
Croat 28892A, station forestiere de Manjakatompo near sommet Hosiarivo, in
13 Magee et al.: Billburttia, a new genus of Apiaceae massif l'Ankaratra (MO); 29892, vicinity of col de ('Itremo, massif de ('Itremo (MO,
P). Decary 13035, Ambatofinandrahana, 1700 m (P); 13400, massif du
Tsiafajavona, 2200-2600 m (P); 17248, Ambositra, dans les bois de Tapia (P);
17356, Ambatofinandrahana (P). Du Puy, Labat & Andriantiana M660, west of
Ambositra, 20° 34 S, 46° 35' E, east margin of Itremo massif, 1290 m (K, P).
Guillaumet 3588, plateau d'Andohariana, Andringitra, c. 2000 m (P). Hildebrandt
3571, nord Betsileo, Antsirabe (BM, K, P). Hodgkin & Stansfield s.n. (K). Humbert
3634, env. de Miarinarivo, SE d'Ambalavao, c. 1200 m (P). Humbert & Capuron
28129, env. d'Ambatofinandrahana, c. 1450 m (P). Jard. Bot. Tananarive 221, pentes nord de Vohitra, Antsirabe (P); 3644, Tsiafajavona (P). Keraudren 218, env. d'Ambatofinandrahana (P). Moeller, Andriantiana & Haevermans 01-26,
Fianarantsoa, Itremo, col d'Itremo, 20° 34' S, 46° 34' E (E). Perrier de la Bathie
6792, env. d'Antsirabe, 1500 m (P); 6807, W du massif d'Andringitra, 1000 m, type de l'espece (P). Rakotovao 52, Reserve Naturelle Integrale no. 5, pres de Ia montagne rocheuse Vangomena, plateau de Sonindrana, 2010 m (MO, P).
Rakotozafy 631, Itremo (P). Razafindraba 193, Fianarantsoa, Ambalavao,
Sendrisoa, Antanifotsy, pic Boby d'Andringitra, 2658 m (MO, P). Reserves
NatureIles RN 2278 Razafindrakoto, distr. Ambalavao, canton Sendrisoa (P).
Viguier & Humbert 1357, distr. Betafo, sommet du pic de Vohimalaza pres
Betafo, 1700 m (P); 1386, distr. Betafo, dans Ia coulee de laves de I'lantsifitra, c.
1450 m (P); 1617, Betafo, monts Vararata, c. 2000m (P); 1727, prov. Itasy, distr.
Kitsamby, sur le flanc ouest de l'Ankaratra, entre Ambatofotsy et le Tsiafajavona, c. 2200 m (P). Waterlot s.n., Antsirabe (P).
14 Magee et al.: Billburttia, a new genus of Apiaceae
2. Billburttia vaginoides Sales and Hedge, sp. nov. Herba perennis glabra.
Caudices obliqui. Caules c. 60 cm, tenuiter striati solidi rubentes. Folia 3-pinnata segmentis filiformibus vel linearibus, c. 30 x 1-2 mm, vaginae prominentes amplexicau/es, 1.5-6 cm longae prope basin rubentes. Bracteae 3-9 mm longae, inaequales; bracteolae 3-5 mm ad basim plus minusve connatae, radii 15-30, 20-
40 mm, aequa/es. Mericarpia (immaturae) dorsaliter vix compressa haud a/atis, basi non saccata, jugis prominentibus, stylopodium conicum, styli va/de reflexi teretes, stylopodio 1-1.5 longior. Affinis P. capensoides. TYPE: Perrier de la
Bathie 13557 (P!, holotype).
Subshrub. Rootstock thick, oblique. Stems c. 60 cm, 4-5 mm wide at base; reddish. Leaves 3-pinnate. Ultimate leaf segments filiform to linear, c. 30 x 1-2 mm; apex mucronulate; firm-textured; glabrous; only midrib prominent. Petioles of lower leaves 15-20 (-60) mm long. Primary umbel 60-100 mm wide; rays 15 to 30, 20-40 mm long; bracts 3-9 mm long; raylets 15 to 22, 4-7 mm long; bracteoles, 3-5 mm long. Fruit [immature] oblong-ellipsoid, c. 3.5 x 1.5 mm; styles reflexed up to or beyond the base of the stylopodium.
Diagnostic characters. This species has a very different facies from its closest relative B. capensoides because of its narrower, linear-filiform ultimate leaf segments. The two species grow in similar areas and although in the field they must look clearly different, B. vaginoides being a much smaller plant, in most characters they are surprisingly similar. Mature fruit of B. vaginoides is a
15 Magee et al.: Billburttia, a new genus of Apiaceae desideratum. Humbert originally recognised this taxon as a possible new species and pencilled on some sheets the name "Peucedanum emirnense".
Distribution and habitat. Ravines, edge of streams, damp rocks, gneiss;
2000-2200 m. The field-note of Perrier de la Bathie 13557 (of 1921) states
"Espece en voie d'extinction". Although there were some gatherings after 1921, there are apparently no collections of the species after 1939.
Phenology. Flowering April to June; fruiting March.
Selected specimens examined: Bojer s.n., prov. Emirina (P). Decary 7564, vallee de I'lkopa NO d'Ankazobe (P); 7583, Tampoketsa, au NE de Fenoarivo
(BM, P); 13893, Behenjy (P). Jard. Bot. Tananarive 4261S, Ambohimasimbola
(P). Perrier de la Bathie 6795, Ankaratra, vers 2200 m (P); 6799, mt
Tsiafajevona, 2000 m (P); 13557, flancs est de mt Tsiafajevona, vers 2200 m, type de l'espece (P). Rousson s.n., Ankaratra (P).
The most useful diagnostic characters are included in the following key.
Key to the genus Billburttia and its species:
1. Mericarps isodiametric or laterally compressed in transverse section
All other Malagasy Apiaceae
Mericarps slightly dorsally compressed Billburttia:
Ultimate leaf segments broad, more than 3 mm wide, linear-elliptic to elliptic; shrub or subshrub, 1-2 m tall B. capensoides
16 Magee et al.: Billburttia, a new genus of Apiaceae
2. Ultimate leaf segments narrow, less than 1 mm wide, linear-filiform; subshrub, c. 60 cm tall B. vaginoides
The authors would like to thank the curators and staff from the cited herbaria who kindly made their specimens available for study, Prof. A.E van Wyk for valuable advice, the University of Johannesburg, the South African
Biosystematics Initiative and the National Research Foundation of South Africa for funding.
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21 Magee et al.: Billburttia, a new genus of Apiaceae
Appendix 1. Voucher information of taxa added to the ITS and rps16 intron matrices of Magee et al. (2009a). The information is listed as follows: taxon —
ITS, rps16 intron; voucher information (for new accessions). Those regions not available for a taxon are indicated with a dash.
New accessions Billburttia capensoides Sales and Hedge, FM986437,
FM986441, Du Puy et al. M660 (P); Billburttia capensoides Sales and Hedge,
FM986438, FM986442, Keraudren 119 (P); Billburttia capensoides Sales and
Hedge, FM986439, —, Razafindrabe 193 (P); Billburttia vaginoides Sales and
Hedge, FM986440, —, Bojer s.n. (P). Published GenBank accessions. Ammi majus L., U78386a & U78446a, —; Anethum graveolens L., —, AF110542b;
Deverra denudata (Viv.) R.Pfisterer and Podlech, —, AY838419a; Deverra triradiata Hochst. ex Boiss., —, AF164815 b; Diplolophium somaliense Verdc.,
DQ368843e, —; Foeniculum vulgare Mill., AY581806d, —; Foeniculum vulgare
Mill., EF421428f, —; Petroselinum crispum (Mill.) Fuss, U78387 1 & U78447a, —;
Ridolfia segetum Moris, U783841 & U78444a, —.
a Downie et al. (1998), b Downie and Katz-Downie (1999), b Downie et al. (2000), d Tabanca et al. (2005), e Calvino et al. (2006), f Kersten and Knoess
(unpublished).
22 Magee et al.: Billburttia, a new genus of Apiaceae
Figure legends
Fig. 1 Diagnostic morphological and anatomical characters of Billburttia (a—h) in comparison to the vegetatively similar Notobubon laevigatum (i, j, I, m) and the type species of Peucedanum (P. officinale) (k). a and b ultimate leaf segment of
B. capensoides (a) and B. vaginoides (b), c—e transverse sections through the fruit of B. vaginoides (c) and B. capensoides (d, e), f marginal wing of B. capensoides showing the presence of sphaerocrystals (indicated with arrows) in and around the epidermis, g and h mature mericarp of B. capensoides in dorsal view (g) and in commissural view (h) showing six commissural vittae of unequal length, i ultimate leaf segment of N. laevigatum, j and k, transverse sections through the fruit of N. laevigatum (j) and P. officinale (k), I and m mature mericarp of N. laevigatum in dorsal view (I) and in commissural view (m) showing two commissural vittae. a Du Puy et al. M660 (P), b Perrier de la Bathie 6799 (P), c Decary 7583 (P), d Rakotozapy 631 (P), e and f Humbert 3634 (P), g and h
Humbert 1727 (P), i, I and m Magee 7 (JRAU), j Botha 3547 (PRE), k PE
328993. Scale: a, b, g, h, I = 1mm; c—e, j, k= 0.5 mm; f= 0.2 mm.
Fig. 2 Portions of Bayesian inference trees, showing the position of Billburttia within the tribe Apieae, based on ITS (a) and rps16 intron (b) sequence data.
Posterior probability (PP) values are presented below the branches. Bootstrap percentage (BP) values from parsimony analysis are presented above the
23 Magee et al.: Billburttia, a new genus of Apiaceae branches. BP values below 50% are not indicated. Branches supported only in the BI are indicated by dashed lines.
Addresses of the authors: A. R. Magee ([email protected] ), B.-E. van Wyk and P. M. Tilney, Department of Botany and Plant Biotechnology, University of
Johannesburg, P.O. Box 524, Auckland Park 2006, South Africa. F. Sales,
Departamento de Botanica, Faculdade de Ciencias e Tecnologia, Universidade de Coimbra, P-3000, Portugal. I. Hedge, Royal Botanic Garden Edinburgh EH3
SLR, Scotland, United Kingdom. S. R. Downie, Department of Plant Biology,
University of Illinois at Urbana-Champaign, Urbana, Illinois 61801, U.S.A.
24 Magee et al.: Billburttia, a new genus of Apiaceae
Figure 1
25 Magee et al.: Billburttia, a new genus of Apiaceae
A. ITS Deverra burchellii B. rpsl 6 intron 100 10 Deverra denudata 1.0 Deverra triradiata 79 62 Sclerosciadium nodiflorum 1.0 93 Ammi majus 90 Naufraga balearica 1 0 0.77 Petroselinum crispum 10 Apium graveolens 61 Bilburtia capensoides 0.91 _ 94 Petroselinum crispum r Billburtia capensoides 100 1.0 Ammi majus 85 1.0 Billburtia capensoides 0.74 D Deverra triradiata 1.0 Billburtia vaginoides 52 CD Deverra denudata 991 Apium graveolens 0.95 78 CD cm ' 97 1.0 Apium prostratum 0.82 1.0 Deverra burchellii a) 1.0 Naufraga balearica Billburtia capensoides CD 84 97 Anethum graveolens 1.0 Billburtla capensoides 1.0 0.97 59 70 Foeniculum vulgare 0.97 0.85 93 Stoibrax dichotomum 0.61 Foeniculum vulgare 1.0 0199 Anethum graveolens Ridolfia segetum 77 1.0 Foeniculum vulgare 100 Stoibrax dichotomum 1.0 Stoibrax dichotomum
Figure 2
26 APPENDIX C5
MAGEE, A. R., C. I.CALviNo, M. Liu, S. R. DOWNIE, P. M. TILNEY and B.-E. VAN
WYK. New tribal delimitations for the early diverging lineages of Apiaceae subfamily Apioideae. Taxon. Submitted. New tribal delimitations for the early diverging lineages of Apiaceae subfamily Apioideae
Anthony R. Magee 1'2, Carolina I. Calvitio 3, Mei (Rebecca) Liu4, Stephen R.
Downie5, Patricia M. Tilney 1 & Ben-Erik van Wye
'Department of Botany and Plant Biotechnology, University of Johannesburg,
P.O. Box 524, Auckland Park 2006, Johannesburg, South Africa
*[email protected] (author for correspondence)
2South African National Biodiversity Institute, Compton Herbarium, Private Bag
X7, Claremont 7735, South Africa
3 INIBIOMA, CONICET- Universidad Nacional del Comahue, Bariloche, Rio
Negro 8400, Argentina
4Department of Biology, Harbin Normal University, Hexing Road 50, Harbin
150080, People's Republic of China
5Department of Plant Biology, University of Illinois at Urbana-Champaign,
Urbana, Illinois 61801, U.S.A.
Phylogenetic analyses of the cpDNA trnQ-trnK 5'exon region for 27 genera and 42 species of Saniculoideae and early diverging lineages of
Apioideae were carried out to assess or confirm the tribal placements of the following anomalous genera: Annesorhiza, Astydamia, Chamarea,
Choritaenia, Ezosciadium, ltasina, Lichtensteinia, Marlothiella,
Molopospermum and Phlyctidocarpa. To accommodate these unique early Tribal delimitations in Apiaceae
diverging members of the Apiaceae and to reflect their relationships, a new tribal classification system has become necessary. Many of the early diverging genera (herein referred to as the protoapioids) can readily be distinguished from the euapioids (the remaining apioids) by the presence of scattered druse crystals in the mesocarp. The major morphological discontinuity within the family, however, lies between the combined protoapioids and euapioids
(representing an expanded Apioideae s.l., including the Saniculoideae) and the subfamilies Azorelloideae and Mackinlayoideae. The broadened subfamily
Apioideae is diagnostically different from the other subfamilies in the absence of rhomboidal crystals, the presence of druse crystals scattered throughout the mesocarp (subsequently lost in the euapioids), and the non-woody endocarp.
No such diagnostic characters are available to support the traditional or recently expanded concept of Saniculoideae. The broadened concept of
Apioideae is also supported by the sporadic presence of true wings. This character can be variously interpreted from a phylogenetic point of view, but nevertheless has considerable diagnostic value. A new tribal classification system for the protoapioids is proposed on the basis of molecular, morphological and anatomical evidence. This new system is intended to be practical and non-hierarchical to allow for future realignments amongst the tribes, as more evidence becomes available. It makes provision for hitherto poorly known African taxa and comprises the following eight tribes, five of which are newly described: Annesorhizeae, Choritaenieae, Heteromorpheae,
Lichtensteinieae, Marlothielleae, Phlyctidocarpeae, Saniculeae and
Steganotaenieae.
2 Tribal delimitations in Apiaceae
KEYWORDS: Annesorhizeae, Choritaenieae, euapioids, Lichtensteinieae,
Marlothielleae, Phlyctidocarpeae, protoapioids, trnQ - trnK 5'exon.
INTRODUCTION
The Apiaceae are a large and taxonomically complex family comprising
463 genera and ca. 3,500 species (Plunkett & al., in press), with a near cosmopolitan distribution. The classification system of Drude (1897-98) made little provision for the comparatively small sub-Saharan African and
Madagascan contingent (80 genera and 354 species), many of which were unknown or poorly studied at the time. As suggested by Burtt (1991) and later by molecular systematic studies (Calvin° & Downie, 2007; Calvin° & al., 2006;
Downie & al., 1996, 1998, 2000; Downie & Katz-Downie, 1999; Magee & al.,
2008a, 2008b; Plunkett & al., 1996a, 1996b) the African genera are crucial to an understanding of the evolutionary history of the family and its full range of morpholocial diversity. Many African taxa have indeed been found to occupy early diverging positions within Apiaceae, suggesting a southern African origin for both Apioideae (Calvino & al., 2006) and Saniculoideae (Calvin° & Downie,
2007). In recent years, tribal delimitations within the largest and most complicated subfamily, the Apioideae, have been the focus of intensive phylogenetic research (see Downie & al., 2001; Downie & al., 2009). The most recent treatment of the "core" apioid lineages by Downie & al. (2009), based largely on nrDNA ITS sequence data, recognized 14 tribes and 12 major clades. In this treatment the earliest diverging lineages of the subfamily were
3 Tribal delimitations in Apiaceae
excluded due to their highly divergent ITS sequences and consequent alignment difficulties. Although there has been much progress on the generic placement of several anomalous taxa within these early diverging lineages
(Calvino & al., 2006; Calvin° & Downie, 2007; Magee & al., 2008a) the correct placement of several critical African taxa has remained uncertain. This paper is aimed at resolving the early diverging lineages of the Apioideae, assessing their relationship to their sister subfamily Saniculoideae, and proposing a new tribal classification for these lineages using anatomical, morphological and
DNA sequence data.
MATERIAL AND METHODS
Morphological data. — Populations of several taxa were studied and sampled in situ in South Africa and Namibia over a period of several years.
This material was supplemented by a study of all relevant taxa from the following herbaria: BM, BOL, JRAU, K, LE, MO, NBG (including SAM and
STE), P, PRE, S, THUNB-UPS and WIND.
Fruit from herbarium specimens and formaldehyde-acetic acid-alcohol
(FAA) preserved material were used in the anatomical study. Herbarium material was first rehydrated and then placed in FAA for a minimum of 24 h.
This material was subsequently treated according to a modification of the method of Feder & O'Brien (1968) for embedding in glycol methacrylate
(GMA). Transverse sections, about 3 ion thick, were made using a Porter-
Blum ultramicrotome. The sections were examined for the presence of crystals using a light microscope, prior to staining using the periodic acid
4 Tribal delimitations in Apiaceae
Schiff/toluidine blue (PAS/TB) method of Feder & O'Brien (1968). To study the three-dimensional structure of the vittae, mature fruit were softened by soaking in boiling water for 24 h. The exocarp was then peeled off while keeping the fruit submerged to prevent desiccation. All illustrations were made by the first author with the aid of a camera lucida attachment on a Zeiss compound microscope or a Wild M3Z stereomicroscope.
Taxon sampling. — The cpDNA trnQ-trnK region for 46 accessions, representing 27 genera and 42 species of Saniculoideae and early diverging lineages of Apioideae and two accessions of the South African azorelloid genus Hermas L. were examined. Sources of material, together with their corresponding taxonomic authorities and GenBank accession numbers, are listed in Appendix 1. Data from the trnQ-trnK region for 27 of these 46 accessions were available from previous studies (Calvin° & Downie, 2007;
Calvino & al.,.2008a, 2008b, in prep.; Downie & al., 2008; Tilney & al., 2009).
Sequence data from the rps16 intron region were available for a further ten accessions (Ca'vino & al., 2006; Downie & Katz-Downie, 1999; Magee & al.,
2008a). Rps16 intron data for the remaining nine accessions and data from the tmQ-rps16 and rps16-tmK intergenic spacers for 15 and 17 accessions, respectively, were obtained specifically for this study. The monotypic southern
African genera Choritaenia Benth., Marlothiella H. Wolff and Phlyctidocarpa
Cannon & Theobald were included as previous authors had suggested their placements within the early diverging lineages of the Apioideae-Saniculoideae
Glade (Calvin° & al., 2006; Calvin° & Downie, 2007; Liu & al., 2003, 2007a,
2007b; Nicolas & Plunkett, 2009; Tilney & al., 2009). The genus Hermas
5 Tribal delimitations in Apiaceae
(Azorelloideae) was used to root the trees, based on results from previous molecular phylogenetic analyses (Calvin° & al., 2006).
Molecular data. — Total DNA was extracted from herbarium or silica material using either the 2X CTAB method of Doyle & Doyle (1987) or the
DNeasy Plant Mini Kit (Qiagen). For amplification of the chloroplast gene rps16 and its flanking intergenic spacer regions (i.e., tmQ-rps16 5' exon and rps16 3' exon-trnK 5' exon) we used the primers of Downie & Katz-Downie (1996) and
Lee & Downie (2006), respectively. Successfully amplified PCR products were purified using either the QlAquick PCR purification kit (Qiagen Inc.), according to the manufacturer's instructions, or the ExoSAP protocol of Werle & al.
(1994) using 5 units of Exonuclease I and 0.5 units of Shrimp Alkaline
Phosphatase. Sequencing reactions were carried out using the BigDye
Terminator version 3.1 Cycle Sequencing Kit (Applied Biosystems Inc.) and sequenced on either an ABI (Applied Biosystems) 3130 XL or 3730 XL sequencer.
Phylogenetic analyses. — Complementary DNA strands were assembled and edited using Sequencher version 3.1.2 (Gene Codes
Corporation) and aligned initially using the default pairwise and multiple alignment parameters in the computer program Clustal X (gap opening cost
=15.00, gap extension cost = 6.66, DNA transition weight = 0.50; Jeanmougin
& al. 1998). This alignment was then checked and adjusted manually where necessary, with gaps positioned so as to minimise nucleotide mismatches.
Unambiguous gaps were scored as presence/absence characters using the simple indel coding method of Simmons & Ochoterena (2000).
6 Tribal delimitations in Apiaceae
For analyses of the entire cpDNA trnQ-trnK region, the portion of the matrix representing the tmQ-rps16 5' exon region was scored as missing in three accessions (Polemannia grossulariifolia, Chamarea snijmaniae and
Steganotaenia commiphoroides) and the portion representing the rps16 3' exon-trnK region was scored as missing in one accession (Polemanniopsis sp.
1) because of difficulties in amplifying these regions in these taxa. In addition, portions of the tmQ-rps16 5' exon region for one accession of Choritaenia capense (1618 bp) and of the rps16 3' exon-trnK region for one accession of
Phlyctidocarpa flava (580 bp) were also unobtainable despite our best efforts.
In Molopospermum peloponnesiacum we were unable to amplify both intergenic spacer regions. Overall, missing data represented approximately
10% of the entire matrix.
Phylogenetic analyses were conducted initially using the parsimony
(MP) algorithm of PAUP* version 4.0b10 (Swofford, 2002). Character transformations were treated as unordered and equally weighted (Fitch parsimony; Fitch, 1971). Tree searches were performed using a heuristic search with 1,000 random sequence additions, tree bisection-reconnection
(TBR) branch swapping, and the mulpars option in effect. Bootstrap percentage values (BP; Felsenstein, 1985) were determined from 1,000 bootstrap replicates, holding 10 trees per replicate, with TBR and mulpars selected. Only values greater than or equal to 50% are reported, and the following scale was applied to evaluate support percentages: 74%, weak;
75%-84%, moderate; and 85%-100%, strong. After model selection using
Modeltest version 3.06 under the corrected Akaike information criterion
7 Tribal delimitations in Apiaceae
(Akaike, 1974; Posada & Crandall, 1998), Bayesian inference (BI; Yang &
Rannala, 1997) was implemented using MrBayes version 3.1.2 (Huelsenbeck
& Ronquist, 2001; Ronquist & Huelsenbeck, 2003). We employed the K81uf+G model for the nucleotide data and the 'standard' model (using default parameters) for the indel data (Lewis, 2001). The analysis was performed for
2,000,000 generations of Monte Carlo Markov Chains (MCMC) with trees saved every 100 generations. The analysis was judged to have reached stationarity when the standard deviation between the split frequencies stabilised below 0.009. The initial one-fourth of the trees were discarded as the
'burn-in' phase. A majority rule consensus tree was produced from the remaining trees in order to show the posterior probabilities (PP). The following scale was used to evaluate the PP's: 0.5-0.84, weak; 0.85-0.94, moderate;
0.95-1.0, strong.
Fourteen morphological and anatomical characters considered important for defining early diverging lineages within the Apioideae-
Saniculoideae Glade were scored for the 46 taxa included in the molecular analyses (Appendices 2 & 3). Character data were taken from field observations of the South African taxa, herbarium specimens and literature
(Winter & al., 1993; Vessio, 2001; Liu, 2004; Liu & al., 2003, 2007a, 2007b,
2009; Magee & al., 2008a, 2008b; Tilney & Van Wyk, 2001; Tilney & al., 2009;
Yembaturova & al., 2009, Van Wyk & al. 1999). These morphological data were then reconstructed on the MP trees using Mesquite version 2.5
(Maddison & Maddison, 2008).
8 Tribal delimitations in Apiaceae
RESULTS
Phylogenetic analyses. — The matrix, representing the entire trnQ- trnK region, consisted of 5,072 unambiguously aligned positions and 587 binary scored indels, and resulted in 2,045 variable and 1,285 parsimony informative characters. MP analyses of these data yielded two minimal length trees, each of 2,113 steps [ensemble consistency indices (CI; Kluge & Farris,
1969) of 0.75 and 0.66, with and without uninformative characters, respectively; ensemble retention index (RI; Farris, 1989) of 0.86]. The MP strict consensus tree was identical in topology to that inferred from the BI analysis
(Fig. 1). In both analyses, the same clades and relationships as reported previously (Calvin° & al., 2006; Calvirio & Downie, 2007; Magee & al., 2008a) were retrieved. The subfamily Saniculoideae sensu Calvin° & Downie is monophyletic (PP 0.99, BP 74), with tribes Saniculeae (i.e., Saniculoideae s.s.) and Steganotaenieae strongly supported (PP 1.0, 1.0, BP 100, 99, respectively). Phlyctidocarpa is sister group to Polemanniopsis and
Steganotaenia (tribe Steganotaenieae) with strong or weak support depending on the analysis (PP 0.97, BP 71). Subfamily Apioideae sensu Calvin() &
Downie is also supported as monophyletic (PP 1.0, BP 76) and is sister group to subfamily Saniculoideae sensu Calvin° & Downie (PP 1.0, MP 100). Within the apioid Glade a strongly supported subclade comprising Choritaenia,
Lichtensteinia Cham. & Schltdl. and Marlothiella (PP 1.0, MP 100) is sister group to all remaining Apioideae lineages. The accessions of Bupleurum L.
(tribe Bupleureae), Physospermum Cusson (tribe Pleurospermeae) and Slum
L. (tribe Oenantheae), representing the upper lineages of the subfamily, are a
9 Tribal delimitations in Apiaceae
sister group to a strongly supported tribe Heteromorpheae (PP 1.0, BP 100).
Successively sister to this group is a strongly supported Annesorhiza Cham. &
Schltdl. Glade (PP 1.0, BP 89), comprising Annesorhiza, Astydamia DC.,
Chamarea Eckl. & Zeyh., Ezosciadium B.L. Burtt, ltasina Raf. and
Molopospermum W.D.J. Koch.
Reconstruction of morphological and anatomical characters. —
Parsimony-based reconstructions of 14 morphological and anatomical characters (Appendix 2) considered important for defining early diverging lineages within the Apioideae-Saniculoideae Glade are each summarised onto one of the two minimal length trees inferred from MP analysis of trnQ-trnK nucleotide substitution and scored indel data (Fig. 2). True wings/ribs on the fruit (character 7, state 1; Fig. 2G; Fig. 3E, H–K), parenchymatous or lignified endocarp (character 10, state 1; Fig. 2J; Fig. 3C, D, F–K), absence of rhomboidal crystals (character 13, state 1; Fig. 2M), and the presence of druse crystals scattered throughout the mesocarp (character 14, state 1; Fig. 2N) were reconstructed as synapomorphies for the Apioideae-Saniculoideae Glade.
The base of the trees was reconstructed as ambiguous for each of the aforementioned characters. However, if a further outgroup of either
Azorelloideae or Mackinlayoideae had been included, this node would certainly be reconstructed for the plesiomorphic state as in Hermas. As shown by
Calvirio & al. (2008a), Saniculoideae s.s (i.e. tribe Saniculeae sensu Ca'vino &
Downie) is supported by the presence of simple umbels (character 3, state 1;
Fig. 2C), sessile or subsessile fructiferous flowers (character 4, state 1; Fig.
2D), large involucral bracts forming a prominent pseudanthium (character 5,
10 Tribal delimitations in Apiaceae
state 1; Fig. 2E), and fruit with surface outgrowths (character 6, state 1; Fig.
2F). No characters were found to support the clades comprising either
Saniculoideae sensu Calvin° & Downie (Phlyctidocarpa—Petagnaea) or
Apioideae sensu Calvin° & Downie (Sium—Lichtensteinia). The presence of regular vittae (character 11, state 1; Fig. 2K; Fig. 3I—K) was reconstructed as synapomorphies for the lineages Sium—Molopospermum and two species of
Alepidea La Roche (Fig. 3D), and as an apomorphy for Phlyctidocarpa (Fig.
3G). Large rib oil ducts (character 12, state 1; Fig. 2L; Fig. 3C, D, F—H), traditionally a character for Saniculoideae s.s., was ambiguously reconstructed as either a synapomorphy for the Apioideae—Saniculoideae Glade with secondary reversals in the upper lineages (from the Annesorhiza Glade upwards), Arctopus, Sanicula europaea, and Petagnaea or as a convergent character state in Saniculoideae and in Lichtensteinia and Marlothiella. The woody habit (character 1, state 1; Fig. 2A) was reconstructed as convergent for both Marlothiella and the tribe Steganotaenieae, and was ambiguous for the ancestor to tribe Heteromorpheae. Although Dracosciadium Hilliard & B.L.
Burtt is a rhizomatous herb, character 1 (Appendix 2) was coded as polymorphic to reflect the missing Madagascan taxa with which it is allied
(Calvin° & al., in prep.). As this genus forms a Glade with the woody
Madagascan genera, it is likely that with more accessions the woody habit may be reconstructed as a synapomorphy for the Heteromorpheae, with the herbaceous habit of Dracosciadium reconstructed as a reversal. Bifurcate ribs
(character 8, state 1; Fig. 2H), surface vesicles (character 6, state 1; Fig. 2F), and regular vittae (character 11, state 1; Fig. 2K) seem to have evolved
11 Tribal delimitations in Apiaceae
independently in Phlyctidocarpa (Fig. 3G) and other lineages. The proteranthous leaves (character 2, state 1; Fig. 2B) in Lichtensteinia and
Steganotaenieae were reconstructed as synapomorphies for each lineage.
Steganotaenieae was also supported by the presence of heteromericarpic fruit
(character 9, state 1; Fig. 21), where only the sepaline ribs are well-developed
(Fig. 3H). This character was also found in the genus Heteromorpha Cham. &
Schltdl. (Fig. 3K).
The number of parsimony informative indels and reconstructed morphological apomorphies are indicated on a BI phylogram (Fig. 4), above and below branches, respectively. The branch leading to the Apioideae-
Saniculoideae Glade was relatively long and supported by 31 parsimony informative indels (although until more outgroup taxa from the Azorelloideae are included, we cannot be certain how many of these indels are synapomorphic for Hermas) and four morphological synapomorphies. In contrast, the branches leading to Saniculoideae sensu Calvin° & Downie
(Phlyctidocarpa–Petagnaea) and Apioideae sensu Ca'vino & Downie (Slum-
Lichtensteinia) were both relatively short, supported by only three and two parsimony informative indels, respectively (Fig. 4), and without any reconstructed morphological synapomorphies.
DISCUSSION
"Euapioids" and "protoapioids". — The early diverging lineages (a paraphyletic assemblage herein referred to as the protoapioids) of the
Apioideae-Saniculoideae Glade are readily separated from the remaining
12 Tribal delimitations in Apiaceae
apioid lineages (the euapioids) by the presence of scattered druse crystals of calcium oxalate in the mesocarp of the fruit (character 14, Fig. 2N).
Optimization of this character by Calvin° & al. (2008a) suggested that the scattered druse crystals in the mesocarp were either absent or ambiguously reconstructed in the ancestor of the Apioideae-Saniculoideae Glade. This was, however, the result of the incorrect scoring of this character as absent in some of the protoapioid taxa (viz. Anginon Raf., Polemanniopsis, Steganotaenia and several species of Lichtensteinia) based on the reports of Liu & al. (2003,
2006) and Liu (2004). Druse crystals are often dissolved and therefore easily overlooked in PAS/TB stained anatomical sections. Further confounding this problem, they may be only sparsely distributed throughout the fruit mesocarp, so that many sections need to be studied. After careful examination of both stained and unstained (where necessary) fruit sections, we are now able to confirm the presence of scattered druse crystals in almost all protoapioid taxa.
Druse crystals were absent only in Arctopus L., Choritaenia and Petagnaea, and restricted to the commissure in Ezosciadium. These results confirm the suggestion by Liu & al. (2006) that the presence of scattered druse crystals in the mesocarp is a synapomorphy for the Apioideae-Saniculoideae Glade.
Phlyctidocarpa. — In both the BI and MP trees the two accessions of
Phlyctidocarpa flava comprised a strongly supported monophyletic group (PP
1.0, BP 100), weakly to strongly supported as a sister group to the tribe
Steganotaenieae (PP 0.97, BP 71). This monotypic Namibian endemic genus had previously been placed within Drude's (1897-98) subfamily Apioideae, tribe Ammineae, by Theobald & Cannon (1973), based on the presence of
13 Tribal delimitations in Apiaceae
crystals in the commissure and at the base of the fruit ribs, as well as the arrangement of the vittae. These authors acknowledged the similarity of the scattered druse crystals with members of the Saniculoideae s.s. but the significance of this similarity remained unclear to them. It has subsequently become evident that the presence of scattered druse crystals in the fruit is not restricted to the Saniculoideae s.s. (Liu & al., 2006; character 14, Fig. 2N), but that this character extends also into the early diverging lineages of Apioideae s.s. Calvin() & Downie (2007) suggested the placement of Phlyctidocarpa within Saniculoideae based on a preliminary molecular phylogenetic study but the genus was not yet formally included in their broadened circumscription of
Saniculoideae. The isolated position of Phlyctidocarpa within the Glade comprising both Steganotaenieae and Saniculeae, as found in this study, has subsequently also been confirmed by Nicolas and Plunkett (personal communication) using trnT-trnD and rp116 intron data. However, the results from these authors place Phlyctidocarpa successively sister to
Steganotaenieae and Saniculeae, as opposed to the direct sister relationship with Steganotaenieae recovered in our analyses. Phlyctidocarpa is unusual in that, like many of the other protoapioid lineages, it combines characters traditionally used to define both Apioideae s.s and Saniculoideae s.s. The fruit
(Fig. 3G) have unusual surface blisters on the ribs and large rib oil ducts (as in most Saniculoideae s.$), together with prominent regular vittae (as in most
Apioideae s.$). Although the surface blisters are superficially similar to those found in the saniculoid genus Astrantia L. (Fig. 3C), they differ in that the ribs on which they are borne are bifurcate in transverse section (Fig. 3G) due to a
14 Tribal delimitations in Apiaceae
longitudinal division or groove. As a result, both the vallecular vittae and rib oil ducts appear, in transverse section, to be located in the valleculas between the ribs. Somewhat bifurcate ribs are also found in Alepidea woodii and A. cordifolia (Yembaturova & al., 2009).
Choritaenia, Lichtensteinia and Marlothiella. — Previous analyses of molecular sequence data (Ca'vino & al., 2006; Calvino & Downie, 2007) placed the South African endemic genus Lichtensteinia as the earliest diverging lineage within Apioideae sensu Calvin() & Downie. Recently, in an assessment of the subfamily Azorelloideae, Nicolas & Plunkett (2009) using molecular data showed that the anomalous genus Choritaenia was not related to the other genera of the Azorelloideae but rather formed a Glade together with Lichtensteinia, that was weakly supported as sister group to
Saniculoideae sensu Calvin() & Downie. In the present study, both the MP and
BI trees recover a strongly supported Glade (PP 1.0, BP 100) comprising
Choritaenia, Lichtensteinia and Marlothiella, with this Glade as sister group to the remaining members of Apioideae sensu Calvin° & Downie. Choritaenia is an annual herb endemic to the dry interior of southern Africa (Liu & al., 2007a) and clearly represents a highly adapted lineage. Traditionally placed within the
Hydrocotyloideae (now largely included in Azorelloideae and Mackinlayoideae) due to the presence of a woody endocarp, Choritaenia differs from the other members of Azorelloideae in the conspicuous oil vesicles located in its wings
(an autapomorphy for the genus), absence of rhomboidal druse crystals, marginal wings composed entirely of the mesocarp, and further lignification of the mesocarp (Fig. 3E). Oil vesicles, although rare in Apioideae, have also
15 Tribal delimitations in Apiaceae
been recorded in Bilacunaria Pimenov & V.N. Tikhom. and Smyrniopsis Boiss.,
(Tamamschan, 1945; pers. obs.); however, in these taxa they are dispersed in the mesocarp of the fruits. Amongst Apiaceae included in this investigation, wings may develop in one of two ways (Calvino & al. 2008a). In the first, wings are due to the compression or folding of the carpel and are as such composed of both the mesocarp and endocarp with the vascular bundle located at the margin (Fig. 3A & B). This type (type 3 of Calvin° & al. 2008a, hereafter referred to as pseudo-wings) seems to be the plesiomorphic state, present in wing-fruited members of Azorelloideae, Mackinlayoideae, and even in the closely related families Araliaceae and Myodocarpaceae. In the second wing configuration type, the wings are due to an expansion of the mesocarp and are as such composed only of mesocarp, with the vascular bundle usually located at its base (Fig. 3E—J). This type (type 2 of Calvin° & al. 2008a, hereafter referred to as true wings) is restricted to winged or prominently ribbed members of the Apioideae-Saniculoideae Glade and was reconstructed in this study as a synapomorphy for this group (character 7, Fig. 2G). As this character was not directly applicable to non-winged members of the
Apioideae-Saniculoideae Glade, it was scored in the analysis as missing for these taxa (Appendix 3). Calvin° & al. (2008a) also described a third wing type composed entirely of exocarp in some lineages of Eryngium L., but this state was not observed in any of the fruits investigated in this study.
Tilney & al. (2009) suggested that the Namibian endemic genus
Marlothiella may be the closest relative of Lichtensteinia, due to the shared presence of concentric rings of cells around the large rib oil ducts and the
16 Tribal delimitations in Apiaceae
marked differences that may be present in the size of the rib ducts (Fig. 3F) so that the fruit may appear heteromericarpic in transverse section. Indeed, our analyses of the trnQ-trnK region support a close relationship between these two otherwise very different genera. Marlothiella was strongly supported (PP
1.0, BP 100) as a sister group to Choritaenia, with Lichtensteinia sister group to this Glade. Liu & al. (2004) suggested that both Lichtensteinia and
Marlothiella be placed within an expanded Saniculoideae based on the presence of large rib oil ducts and the absence of regular vittae (Fig. 3F).
Calvin° & al. (2008a) considered the presence of rib oil ducts in the fruit as a plesiomorphy in Saniculoideae due their presence in both Azorelloideae and
Mackinlayoideae. Moreover, they considered the loss (or reduction) of rib oil ducts to be a synapomorphy for the upper lineages of Apioideae (from the
Annesorhiza Glade upwards). Large rib oil ducts (traditionally a character for
Saniculoideae s.$) when reconstructed onto the MP trees (character 12, Fig.
2L) could most parsimoniously be considered as either a synapomorphy for the Apioideae–Saniculoideae Glade with a secondary reversal in the upper lineages of Apioideae (upwards from the Annesorhiza Glade), or the result of convergence in the Phlyctidocarpa–Petagnaea and Marlothiella–Lichtensteinia clades. Since relatively large rib oil ducts also occur in some members of the
Azorelloideae, there are various ways of interpreting the evolution of the character (see Calvin° et al. 2008a).
Annesorhiza Glade. — Initially, the Annesorhiza Glade was recognised by Calvin() & al. (2006) as comprising Annesorhiza, Chamarea and Itasina.
Although Astydamia and Molopospermum were weakly recovered as being
17 Tribal delimitations in Apiaceae
closely related, the authors excluded them from the Annesorhiza group until their placements had been confirmed with further data. Magee & al. (2008a) subsequently showed that the annual, South African endemic genus
Ezosciadium was also related to this Glade. In both the MP and BI analyses, all accessions of these six genera were strongly supported to comprise a monophyletic group (PP 1.0, BP 89), successively sister to the tribe
Heteromorpheae and subsequent lineages of the Apioideae. The Annesorhiza
Glade comprises herbaceous members largely typical of the euapioid tribes with regular vittae and compound leaves, but differs most prominently in the presence of scattered druse crystals [except in Ezosciadium, where they are restricted to the commissural area (Magee & al., 2008a)], the thick and highly lignified vascular bundles in the fruit (Fig. 31), and the proteranthous or deciduous leaves.
Paraphyly of Apioideae. — The subfamily Apioideae s.s. as circumscribed by Drude (1897-98) is clearly not monophyletic when the
African taxa are considered (Downie & Katz-Downie, 1999; Downie & al.,
2001; Liu & al., 2003; 2006; Calvin° & al., 2006; Calvin° & Downie, 2007), and the characters traditionally used to segregate this subfamily are reconstructed to be either plesiomorphic (e.g., compound umbels) or synapomorphic and restricted to the upper lineages of the subfamily (e.g., inconspicuous rib oil ducts, presence of regular vittae, absence of scattered druse crystals); Calvin°
& al. (2008a) and this study. Calvin° & Downie (2007) based on phylogenetic studies, accommodated many of the conflicting African taxa in Apioideae or
Saniculoideae resulting in two recircumscribed monophyletic subfamilies.
18 Tribal delimitations in Apiaceae
However, as shown by Calvin° & al. (2008a), the subfamilies Apioideae sensu
Calvin° & Downie and Saniculoideae sensu Calvin° & Downie are each not supported by any morphological or anatomical synapomorphies studied to date. The nodes supporting Apioideae sensu Calvin° & Downie (Slum-
Lichtensteinia) and Saniculoideae sensu Calvin° & Downie (Phlyctidocarpa-
Petagnaea) are both strongly to moderately supported with a low rate of nucleotide substitutions per site in the BI trees (Fig. 6 in Calvitio & Downie
2007; Fig. 4 in this study). Calvin° & al. (2008a) proposed that
Steganotaenieae and Saniculeae (sister tribes of subfamily Saniculoideae sensu Calvin° & Downie) have evolved independently for a long time or have accumulated many independent morphological changes, thus masking any shared morphological feature that was once present in their common ancestor.
Synapomorphies of the latter, however, persist at the molecular level. This hypothesis explains the lack of evident morphological or anatomical synapomorphies in Saniculoideae sensu Calvin° & Downie. The same reasoning can be applied to subfamily Apioideae sensu Calvin° and Downie.
Despite the lack of morphological or anatomical synapomorphies for each of these subfamilies, Calvin° and Downie (2007) decided to maintain the two as distinct, monophyletic taxa. The spirit for their classification was basically threefold: 1) to maintain the identity of two of the major lineages of Apiaceae that commonly originated in South Africa but later independently dispersed to, and diversified in, other parts of the world; 2) to be able to readily reflect a sister group relationship between subfamilies Apioideae and Saniculoideae; and 3) to prevent the loss of long-accepted taxonomic concepts through the re-
19 Tribal delimitations in Apiaceae
accommodation of problematic taxa to adequately reflect phylogeny. The controversy around this classification lies in the absence of any morphological or anatomical synapomorphies that define each subfamily. In order to define subfamilies in Apiaceae based on morphological or anatomical synapomorphies, an alternative would be to incorporate the two tribes of
Saniculoideae sensu Calvino and Downie within a more widely delimited subfamily Apioideae (Apioideae s.l. as defined herein).
The Saniculoideae s.s have traditionally been defined by the presence of simple spinescent or setiferous leaves, simple umbels, showy involucral bracts forming a pseudanthium, sessile or subsessile fructiferous flowers, fruit with exocarp outgrowths, distinct rib oil ducts, scattered druse crystals and the absence of regular vittae. In contrast, Apioideae have been circumscribed by the usually compound leaves, prominent compound umbels, fruit with inconspicuous rib oil ducts and regular vittae. Many of the protoapioid genera, however, share characters with both subfamilies. Lichtensteinia, Marlothiella,
Polemanniopsis and Steganotaenia, have prominent compound umbels typical of Apioideae but saniculoid-like fruit with large rib oil ducts and without regular vittae (Fig. 3F & H). The fruit of Phlyctidocarpa (Fig. 3G) share the blistered exocarp and large prominent rib oil ducts with members of Saniculoideae, together with a compound umbel and regular vittae as in Apioideae. It is interesting to note the recent discovery of regular vittae in some species of the most early diverging saniculoid genus Alepidea (Yembaturova & al., 2009; Fig.
3D) which further narrows the taxonomic distance between the two subfamilies. Due to the presence of equally isolated and morphologically
20 Tribal delimitations in Apiaceae
divergent lineages within the protoapioids, the rank of subfamily for
Saniculoideae no longer seems to be appropriate. Drude's (1897-98) classification system reflects incomplete knowledge of several other lineages
(mostly African) that are morphologically as distinct as the Saniculoideae. The only logical way to delimit a morphologically congruent Apioideae would be to treat the Saniculoideae s.s. as a tribe (the Saniculeae) within a more widely delimited subfamily Apioideae. The union of Apioideae and Saniculoideae into a single subfamily was also proposed by Koso-Poljansky (1916) who included both subfamilies within his Ligusticoideae, based on the parenchymatous endocarps. Optimization of morphological and anatomical data onto one of the two minimal length trees obtained from the MP analyses of the trnQ-trnK region (Fig. 2) shows that such an expanded Apioideae s.l. is well-supported by four synapomorphies (Fig. 4), namely the presence of true wings/ribs (wings or prominent ribs consisting only of mesocarp and exocarp – Fig. 2G), the parenchymatous or sometimes lignified endocarp (becoming secondarily woody in Choritaenia – Fig. 2J), the absence of rhomboidal crystals (Fig. 2M), and the presence of druse crystals of calcium oxalate scattered throughout the mesocarp (subsequently lost in the euapioids – Fig. 2N).
Tribal delimitations. — While morphological and anatomical characters support the lineage comprising the Apioideae s.l., as well as many of the terminal clades within the protoapioids (herein recognized as tribes), there are no available morphological or anatomical characters to support the hierarchical relationships among the tribes (i.e., the clades comprising the
Lichtensteinieae–Oenantheae, Lichtensteinieae–Choritaenieae,
21 Tribal delimitations in Apiaceae
Steganotaenieae—Phlyctidocarpeae or Saniculeae—Ph lyctidocarpeae).
Therefore, in order to reflect the available morphological, anatomical and molecular data, the protoapioids are here segregated into eight small and morphologically isolated tribes (each well-supported by several morphological and/or anatomical characters and/or molecular sequence data), which probably represent relicts from a once more numerous African apioid flora. A taxonomic key to the subfamilies of Apiaceae and tribes of the protoapioids is provided below.
TAXONOMIC TREATMENT
Key to the subfamilies of Apiaceae and tribes of the protoapioids
(Apioideae s.I.):
Endocarp woody, adjacent mesocarp usually parenchymatous; rhomboidal
crystals usually present, druse crystals absent; true wings absent; wing-like
structures (pseudo-wings) or prominent ribs comprising endocarp and
mesocarp with vascular tissue at the tip 2.
2 Fruits laterally compressed; mericarps not separating at maturity; carpophore
absent Mackinlayoideae
Fruits isodiametric or dorsally compressed; mericarps separating at maturity;
carpophore present Azorelloideae
1 Endocarp parenchymatous or sometimes becoming lignified, if somewhat
woody then with adjacent mesocarp lignified; rhomboidal crystals absent;
druse crystals present or absent; true wings often present; wings or
22 Tribal delimitations in Apiaceae
prominent ribs comprising only mesocarp, with vascular tissue almost
always at the base Apioideae s.I.:
3. Fruit with oil vesicles in the wings Choritaenieae
3. Fruit without oil vesicles in the wings 4
4. Inflorescence a pseudanthium; regular vittae absent, if rarely present then
with larger rib oil ducts and amphi-seminal druse crystals in the fruit
Saniculeae
Inflorescence umbellate, if rarely a pseudanthium then with regular vittae,
poorly developed rib oil ducts, and without amphi-seminal druse crystals in
the fruit 5
Druse crystals of calcium oxalate absent or commissural only; without
tanniniferous epidermal cells euapioids
5. Druse crystals of calcium oxalate amphi-seminal, if reduced to commissure
only, then with tanniniferous epidermal cells and only partly bifid carpophore
6
6. Regular vittae absent 7
6. Regular vittae present 9
7. Fruit winged; wing cavities present; woody shrubs or trees
Steganotaenieae
7. Fruit not winged; large rib oil ducts surrounded by a concentric ring of cells;
herbs or small woody shrublets 8
8. Rhizomatous herbs; leaves large, coriaceous Lichtensteinieae
8. Woody shrublets; leaves small, succulent Marlothielleae
23
Tribal delimitations in Apiaceae
9. Fruit surface blistered; ribs bifurcate; vittae and rib oil ducts equal in size
Phlyctidocarpeae
Fruit surface smooth; ribs not bifurcate; vittae larger than rib oil ducts 10
Perennial or annual herbs; leaves proteranthous or deciduous
Annesorhizeae
10. Woody trees, shrubs, suffrutices or lianas; if rarely herbaceous then leaves
persistent Heteromorpheae
1. Annesorhizeae Magee, C.I. Calvin°, M. Liu, S.R. Downie, P.M. Tilney & B.-
E. van Wyk trib. nov. — Type: Annesorhiza Cham. & Schltdl.
Heteromorpheis similis umbello multiplexo, fructu crystallis drusaceis
omnino in mesocarpio dispersis et vittis regularibus, sed habitu herbaceo, fohis
proteranthis vel deciduis et fructu fasciculis vascularibus valde lignosis differt.
Annesorhizeae is similar to the Heteromorpheae in that the species
have compound umbels, fruit with druse crystals scattered throughout the
mesocarp and regular vittae. However, they differ from members of the
Heteromorpheae in the herbaceous habit (rarely annual in Ezosciadium), the
proteranthous or deciduous leaves and fruit with thick and strongly lignified
vascular bundles. The tribe includes Annesorhiza, Astydamia, Chamarea,
Ezosciadium, Itasina and Molopospermum. Although most of the genera are
southern African endemics, Molopospermum is a European genus and
Astydamia is restricted to North Africa and the Canary Islands.
24 Tribal delimitations in Apiaceae
Choritaenieae Magee, C.I. Calvino, M. Liu, S.R. Downie, P.M. Tilney & B.-
E. van Wyk trib. nov. — Type: Choritaenia Benth.
Tribubus multis Apioidearum umbello multiplexo alisque marginalibus veris similis, sed unicus vesiculis olei in alis marginalibus, carpophoro hygroscopico, endocarpio lignoso cum lignificatione adiuncta mesocarpii.
Choritaenieae shares the compound umbels and true marginal wings with many other tribes of the Apioideae, but is unique in that the fruit have oil vesicles in the marginal wings, a hygroscopic carpophore as well as a woody endocarp with further lignification of the mesocarp. It also differs from most other protoapioids in the annual and ephemeral habit and the absence of druse crystals scattered throughout the mesocarp of the fruit. The tribe is monogeneric and comprises the southern African genus Choritaenia.
Lichtensteinieae Magee, C.I. Calvin°, M. Liu, S.R. Downie, P.M. Tilney &
B.-E. van Wyk trib. nov. — Type: Lichtensteinia Cham. & Schltdl.
Marlothiellieis similis umbeffis multiplexis, ductis olei maximis costalibus annulis concentricis cellularum circumcinctis, crystaffis drusaceis omnino in mesocarpio dispersis, sed habitu herbaceo deciduo, foffis non succulentis saepe proteranthis marginibus dentatis setaceis, fructu sine pilis stellatis va/de differt.
Lichtensteinieae is similar to Marlothielleae in the compound umbels, very large rib oil ducts surrounded by concentric ring of cells and druse crystals scattered throughout mesocarp, but differs markedly in the herbaceous deciduous habit, non-succulent leaves (often proteranthous, with dentate,
25 Tribal delimitations in Apiaceae
setaceous margins) and fruit without stellate hairs. It is also similar to the tribe
Annesorhizeae in the herbaceous and deciduous habit but differs markedly in the large ribs oil ducts and the absence of vittae in the fruit. The tribe is monogeneric and comprises the South African genus Lichtensteinia.
Marlothielleae Magee, C.I. Calvirio, M. Liu, S.R. Downie, P.M. Tilney & B.-
E. van Wyk trib. nov. — Type: Marlothiella H. Wolff.
Lichtensteinieis similis umbellis multiplexis, crystafiis drusaceis omnino in mesocarpio dispersis et ductis olei maximis costalibus annulis concentricis cellularum circumcinctis, sed habitu lignoso, foliis succulentis et fructu pilis stellatis differt.
Marlothielleae is similar to Lichtensteinieae in the presence of compound umbels, druse crystals scattered throughout the mesocarp, and the very large rib oil ducts surrounded by concentric rings of cells, but differs in the woody habit, succulent fleshy leaves and fruit with stellate hairs. Although it shares the woody habit and heteromericarpic fruit with members from both
Heteromorpheae and Steganotaenieae, it differs from the former in the large rib oil ducts and the absence of regular vittae in the fruit and from the latter in the isodiametric fruit, ribs that do not develop into wings and that are without wing cavities. The tribe is monogeneric and comprises the Namibian endemic genus Marlothiella.
Phlyctidocarpeae Magee, C.I. Calvirio, M. Liu, S.R. Downie, P.M. Tilney &
B.-E. van Wyk trib. nov. — Type: Phlyctidocarpa Cannon & Theobald.
26 Tribal delimitations in Apiaceae
Annesorhizeis similis habitu herbaceo, vittis regularibus et crystallis drusaceis omnino in mesocarpio dispersis, sed ductis olei costalibus magnis, costis bifurcatis vesiculis superficialibus magnis et fasciculis duobus ventralibus non evolutis differ`..
Phlyctidocarpeae is similar to Annesorhizeae in the herbaceous habit, regular vittae and druse crystals scattered throughout the mesocarp, but differs in the large rib oil ducts, bifurcate ribs with large surface vesicles and the two weakly developed ventral bundles. Although it shares the large rib oil ducts, weakly developed ventral bundles and surface vesicles with members of the tribe Saniculeae it differs markedly in the absence of a pseudanthium, the pedunculate compound umbels, the presence of regular vittae and the bifurcate ribs. Phlyctidocarpa has previously been associated with superficially similar members of the tribe Apieae and other euapioid tribes but differs in the presence of druse crystals scattered throughout the mesocarp of the fruit as well as large rib oil ducts, bifurcate ribs with large surface vesicles and weakly developed ventral bundles. The tribe is monogeneric and comprises the
Namibian endemic genus Phlyctidocarpa.
ACKNOWLEDGEMENTS
Funding from the South African National Research Foundation, the University of Johannesburg, the USA National Science Foundation (grant number NSF
DEB 0089452 to SRD) and the China National Science Foundation (30870148 to ML) is gratefully acknowledged. We thank Dr H. Glen (NH) for translating the Latin diagnoses, Prof. A.E. van Wyk, Prof. G.M. Plunkett and Mr A.N.
27 Tribal delimitations in Apiaceae
Nicolas for providing us with material of Choritaenia, and Mr C. Danderson for sequencing some of the material. The authors also wish to thank the curators and staff of the cited herbaria.
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34 Tribal delimitations in Apiaceae
Appendix 1. Accessions of Apiaceae from which the cpDNA trnQ-trnK sequences were obtained, with corresponding voucher information, taxonomic authorities, and GenBank reference numbers.
Actinolema macrolema Boiss. — Coode & Jones 2425 (E); tmQ-rps16
DQ832386b, rps16 DQ832337b, rps16-tmK DQ832473b. Alepidea amatymbica Eckl. & Zeyh. — Mohle 480 (MO); tmQ-rps16 DQ832387b, rps16
DQ832338b, rps16-tmKDQ832474 c. Alepidea capensis (Berg.) R.A. Dyer —
Hilliard & Burtt 16645 (E); trnQ-rps16 DQ832389b, rps16 DQ832340b, rps16- tmK DQ832476b. Alepidea peduncularis Steud. ex A. Rich. — Gereau &
Kayombo 4094 (MO); tmQ-rps16 DQ832386c, rps16 DQ832337c, rps16-trnK
DQ832473b. Anginon difforme (L.) B.L. Burtt — Magee 130 (JRAU); tmQ-
rps16 FM986461 h , rps16 FM986476h, rps16-tmK FM986485h. Anginon paniculatum (Thunb.) B.L. Burtt — Downie 2458 (ILL); trnQ-rps16
DQ832400b, rps16 AY838397b, rps16-tmK DQ832487b. Annesorhiza altiscapa H. Wolff — Magee 97 (JRAU); tmQ-rps16 FM986462h, rps16
FM986477h, rps16-tmK FM986486h. Annesorhiza macrocarpa Eckl. & Zeyh.
— Downie 2454 (ILL); tmQ-rps16 DQ832400b, rps16 AY838397b, rps16-tmK
DQ832487b. Arctopus echinatus L. — Ornduff 7380 (UC); tmQ-rps16
DQ832402b, rps16 DQ832351 b, rps16-tmK DQ832489b. Astrantia major L. —
Schilling 2937 (E); trnQ-trnK DQ832443b. Astrantia minor L. — Vautier,
Mermoud & Bersier s.n. (ILL); trnQ-trnK DQ832444c. Astydamia latifolia (L.f.)
Kuntze — Hildenbrand s.n. (ILL); tmQ-rps16 FM986463h, rps16-tmK
FM986487h. Bupleurum angulosum L. — Younger 2565 (E); tmQ-rps16
35 Tribal delimitations in Apiaceae
FM986464h, rps16 AF110568a, rps16-tmK FM986488h. Bupleurum fruticosum L. — McBeath 2592 (E); tmQ-rps16 FM986465h, rps16
AF110569a, rps16-tmK FM986489h. Chamarea snijmaniae B.L. Burtt -
MacGregor s.n. (NBG); rps16 AY838414b, rps16-tmK FM986490h.
Choritaenia capensis Benth.— Zeitman 271 (PRE); tmQ-rps16 FM986467h, rps16 FM986479h, rps16-tmK FM986492h. Choritaenia capensis Benth.-
Van Wyk 545 (PRU); tmQ-rps16 FM986466h , rps16 FM986478h, rps16-tmK
FM986491 h . Dracosciadium italae Hilliard & B.L. Burtt — Ngwenya & Singh
1279 (PRE); EU4346579, EU4346529 . Eryngium giganteum M. Bieb. — Mc
Neal 472 (UC); tmQ-rps16 EU070440d , rps16 EU070502d, rps16-tmK
EU070564d. Eryngium maritimum L. — Medina et al. MP1656 (TEX); trnQ- rps16 EU070454d, rps16 EU070516d, rps16-tmK EU070578d. Eryngium palmatum Panda & Vis. — Constance C-99 (UC); tmQ-rps16 DQ832416c, rps16 DQ832365c, rps16-tmK DQ832504c. Eryngium planum L. — Downie
191 (ILL); trnQ-trnK DQ832456c. Ezosciadium capense (Eckl. & Zeyh.) B.L.
Burtt — Goldblatt & Porter 12578 (NBG); trnQ-rps16 FM986468h, rps16
AM982518f rps16-trnK FM986493h . Hacquetia epipactis DC. — M. F. & S. G.
Gardner 2590 (E); tmQ-rps16 DQ832423 c, rps16 DQ832372c, rps16-tmK
DQ832511 c. Hermas gigantea L. — McDonald 1769 (NBG); tmQ-rps16
DQ832425c, rps16 AY838420b, rps16-tmK DQ832513c. Hermas quinquedentata L.f. — Burman 1080 (BOL); tmQ-rps16 DQ832426c, rps16
AY838422b, rps16-tmK DQ832514c. Heteromorpha arborescens Cham. &
Schltdl.— Van Wyk 4122 (JRAU); tmQ-rps16 FM986469h, rps16 AY838424b , rps16-trnK FM986494h. ltasina filifolia (Thunb.) Raf. — Downie 2453 (ILL);
36 Tribal delimitations in Apiaceae
tmQ-rps16 FM986470 h , rps16 AY838426b, rps16-tmK FM986495h.
Lichtensteinia globosa B.-E. van Wyk & Tilney — Downie 2462 (ILL); tmQ- rps16 DQ832429c, rps16 AY838429b, rps16-tmK D0832517C. Lichtensteinia lacera Cham. & Schltdl. — Downie 2464 (ILL); tmQ-rps16 DQ832427c, rps16
AY838427b, rps16-tmK DQ832515c. Lichtensteinia obscura (Spreng.) Koso-
Pol.— Downie 2457 (ILL); tmQ-rps16 DQ832428c, rps16 AY838428b, rps16- tmK DQ832516c. Lichtensteinia trifida Cham. & Schltdl. — Downie 2460
(ILL); tmQ-rps16 DQ832430c, rps16 AY838430b, rps16-tmK DQ832518c.
Marlothiella gummifera H. Wolff — Manheimer 2987 (JRAU); tmQ-rps16
FM986471 h , rps16 FM986480h, rps16-tmK FM986496h. Molopospermum peloponnesiacum (L.) W.D.J. Koch — Argent ML2 (E); rps16 AY838432b. —.
Petagnaea gussonei (Spreng.) Rauschert — Donila 2005 (PAL); trnQ-trnK
DQ832466c. Phlyctidocarpa flava Cannon & Theobald — Merxmueller &
Giess 30626 (WIND); tmQ-rps16 FM986473h, rps16 FM986482h, rps16-tmK
FM986498h. Phlyctidocarpa flava Cannon & Theobald — Manheimer 2889
(JRAU); tmQ-rps16 FM986472h, rps16 FM986481 h, rps16-tmK FM986497h.
Physospermum cornubiense (L.) DC. — Pimenov et al. s.n. (MW); tmQ- rps16 FM986474h, rps16 AF110556a, rps16-tmK FM986499h. Polemannia grossulariifolia Eckl. & Zeyh.— De Castro 274 (JRAU); rps16 AY838439b, rps16-tmK FM986500h. Polemanniopsis marlothii (H.Wolff) B.L. Burtt —
Downie 2459 (ILL); trnQ-rps16 DQ832432c, rps16 DQ832374c, rps16-tmK
DQ832520c. Polemanniopsis sp. 1 — Manheimer 2769 (JRAU); tmQ-rps16
FM986475h, rps16 FM986483h. —. Sanicula canadensis L. — Downie 737
(ILL); trnQ-trnK DQ832467c. Sanicula europaea L. — Catalan 1896 (JACA);
37 Tribal delimitations in Apiaceae
trnQ-trnK DQ832468c. Sium suave Walter — Downie 12 (ILL); trnQ-trnK
EF185274e. Steganotaenia araliacea Hochst.— Downie 2456 (ILL); tmQ- rps16 DQ832442c, rps16 DQ832384c, rps16-tmK DQ832530c. Steganotaenia commiphoroides Thulin — Friis et al. 4889 (K); rps16 FM986484h, rps16-tmK
FM986501 h .
Downie and Katz-Downie (1999)a; Calvino et al. (2006)h; Calvin° and Downie
(2007)c; Calvin° et al. (2008b) d; Downie et al. (2008)e, Magee et al. (2008a)f;
Tilney et al. (2009)9; present studyh.
38 Tribal delimitations in Apiaceae
Appendix 2. Morphological and anatomical characters and states examined during this investigation and optimised onto the molecular trees.
1. Habit (herbaceous = 0; woody = 1); 2. Leaf persistence (not proteranthous
= 0; proteranthous = 1); 3. Umbel (compound = 0; simple = 1); 4. Flowers
(pedicellate = 0; sessile or subsessile = 1); 5. Involucre bracts (not forming a
pseudanthium = 0; forming a prominent pseudanthium = 1); 6. Fruit surface
(glabrous = 0; surface vesicles = 1; spines/bristles = 2; stellate hairs =3); 7.
Fruit wing/rib development (pseudo-wings/ribs = 0; true wings/ribs = 1); 8.
Fruit ribs shape (simple = 0; bifurcate = 1); 9. Fruit symmetry (homomericarpic
= 0; heteromericarpic = 1); 10. Endocarp (woody = 0; parenchymatous/
lignified = 1); 11. Regular vittae (absent = 0; present = 1; oil vesicles = 2); 12.
Rib oil ducts (small = 0; large = 1; forming cavities = 2); 13. Rhomboidal
crystals (present = 0; absent = 1); 14. Druse crystals (absent = 0; scattered
throughout mesocarp = 1; restricted to commissure = 2).
39 Tribal delimitations in Apiaceae
Appendix 3. Matrix of morphological and anatomical character states
optimised onto the molecular trees. Dashes represent non-applicable
character states that were scored as missing in the analysis.
1 2 3 4 5 6 7 8 9 10 11 12 13 14 Hermas gigantea 0 0 0 0 0 0 0 0 0 0 0 0 1 0 Hermas quinquedentata 0 0 0 0 0 0 0 0 0 0 0 0 1 0 Sium suave 0 0 0 0 0 0 1 0 0 1 1 0 0 0 Physospermum cornubiense 0 0 0 0 0 0 0 0 1 1 0 0 0 Bupleurum angulosum 0 0 0 0 0 0 0 0 1 1 0 0 0 Bupleurum fruticosum 1 0 0 0 0 0 - 0 0 1 1 0 0 0 Polemannia grossulariffolia 1 0 0 0 0 0 1 0 0 1 1 0 0 1 Dracosciadium italae 0/1 0 0 0 0 0 - 0 0 1 1 0 0 1 Anginon paniculatum 1 0 0 0 0 0 - 0 0 1 1 0 0 1 Anginon difforme 1 0 0 0 0 0 1 0 0 1 1 0 0 1 Heteromorpha arborescens 1 0 0 0 0 0 1 0 1 1 1 0 0 1 Annesorhiza altiscapa 0 0 0 0 0 0 1 0 1 1 1 0 0 1 Annesorhiza macrocarpa 0 1 0 0 0 0 1 0 1 1 1 0 0 1 Chamarea snijmaniae 0 1 0 0 0 0 0 0 1 1 0 0 1 Itasina filifolia 0 0 0 0 0 0 0 0 1 1 0 0 1 Ezosciadium capense 0 0 0 0 0 0 0 0 1 1 0 0 2 Astydamia latifolia 0 0 0 0 0 0 1 0 0 1 1 0 0 1 Molopospermum peloponnesiacum 0 0 0 0 0 0 1 0 1 1 1 0 0 1 Choritaenia capensis 0 0 0 0 0 0 1 0 0 0 2 0 0 0 Choritaenia capensis 0 0 0 0 0 0 1 0 0 0 2 0 0 0 Marlothiella gummifera 1 0 0 0 0 3 - 0 1 1 0 1 0 1 Lichtensteinia obscura 0 1 0 0 0 0 - 0 0 1 0 1 0 1 Lichtensteinia trifida 0 1 0 0 0 0 - 0 1 1 0 1 0 1 Lichtensteinia globosa 0 1 0 0 0 0 - 0 0 1 0 1 0 1 Lichtensteinia lacera 0 1 0 0 0 0 - 0 1 1 0 1 0 1 Phlyctidocarpa flava 0 0 0 0 0 1 - 1 0 1 1 1 0 1 Phlyctidocarpa flava 0 0 0 0 0 1 - 1 0 1 1 1 0 1 Polemanniopsis sp. 1 1 1 0 0 0 0 1 0 1 1 0 2 0 1 Polemanniopsis marlothii 1 1 0 0 0 0 1 0 1 1 0 2 0 1 Steganotaenia araliacea 1 1 0 0 0 0 1 0 1 1 0 2 0 1 Steganotaenia commiphoroides 1 1 0 0 0 0 1 0 1 1 0 2 0 1 Alepidea amatymbica 0 0 1 1 1 0 - 1 0 1 1 1 0 1 Alepidea capensis 0 0 1 1 1 2 - 0 0 1 1 1 0 1 Alepidea peduncularis 0 0 1 1 1 2 - 0 0 1 0 1 0 1 Arctopus echinatus 0 0 1 1 1 2 - 0 0 1 0 0 0 0 Actinolema macrolema 0 0 1 1 1 2 - 0 0 1 0 1 0 1 Astrantia major 0 0 1 0 1 1 0 0 1 0 1 0 1 Astrantia minor 0 0 1 0 1 1 - 0 0 1 0 1 0 1 Hacquetia epipactis 0 0 1 1 1 2 - 0 0 1 0 1 0 1 Petagnaea gussonei 0 0 1 1 1 0 0 0 1 0 0 0 0 Sanicula canadensis 0 0 1 1 1 2 0 0 1 0 1 0 1 Sanicula europaea 0 0 1 1 1 2 0 0 1 0 0 0 1 Eryngium maritimum 0 0 1 1 1 2 1 0 0 1 0 1 0 1 Eryngium giganteum 0 0 1 1 1 2 1 0 0 1 0 1 0 1 Eryngium palmatum 0 0 1 1 1 2 0 0 1 0 1 0 1 Eryngium planum 0 0 1 1 1 2 - 0 0 1 0 1 0 1
40 Tribal delimitations in Apiaceae
FIGURE CAPTIONS.
Fig. 1. Bayesian inference (BI) tree of trnQ-trnK sequence data with the previous subfamilial circumscriptions indicated alongside.
Phlyctidocarpa flava was not included in the study of Calvino & Downie, but presumed to be included within subfamily Saniculoideae based on preliminary evidence (Calvirio & Downie, 2009). Posterior probability (PP) values are presented above the branches. Bootstrap percentage (BP) values (>50%) are presented below the branches.
Fig. 2. Reconstruction of morphological characters 1 to 14 from
Appendix 2, when optimized over one of the two minimal length trees inferred from MP analysis of the trnQ-trnK sequence data. The proposed new tribal and subfamilial classification is indicated alongside Fig. 2N.
Fig. 3. A, Myodocarpus involucratus; B, Hermas villosa; C, Astrantia major; D, Alepidea cordifolia; E, Choritaenia capensis; F, Lichtensteinia trifida; G, Phlyctidocarpa flava; H, Polemanniopsis marlothii; I,
Annesorhiza macrocarpa; J, Anginon difforme; K, Heteromorpha arborescens. Vouchers: A, Lowry & Oskolski 4639 (MO); B, Anon s.n.
(JRAU); C, Pauca 165b (PE); D, Van Wyk 4232 (JRAU); E, Hanekom 1834
(PRE); F, Winter & Tilney 4164 (JRAU);G, Gress et al. 6075 (PRE); H,
Taylor 11269 (PRE); I, Rourke 1700 (NBG); J, Van Wyk 2944 (JRAU); K,
Greenway 12558 (PRE). br, bifurcate rib; Ir, lateral rib; medr, median rib;
41 Tribal delimitations in Apiaceae
mr, marginal rib; ov, oil vesicle; rod, rib oil duct; sv, surface vesicle; vb, vascular bundle; vv, vallecular vitta; wc, wing cavity; we, woody endocarp. Scale: A—E, l—K=0.8 mm; F=0.5 mm; G=1 mm H=0.4 mm.
Fig. 4. Bayesian inference phylogram of trnQ- trnK sequence data.
Numbers above the branches indicate the distributions of parsimony informative indels and those below the branches the number of apomorphies as constructed from character optimisation. The scale indicates the number of expected substitutions per site.
42 Figure 1 100 1.0 0.99 74 100 76 1.0 1.0 0.87 50 100 100 1.0 1.0 0.97 100 71 100 1.0 53 89 1.0 1.0 1.0 90 0.93 0.71 100 79 1.0 100 loo 100 loo 100 1.0 72.69 1.0 1 1.0 1.0 . 100 0 0.84 1.0 100 100 1.0 95 1.0 100 92 1.0 1.0 1.0 98 1 99 '.82 95 96 . 0 100 1.0 100 r. 96 0.69 67 100 100 1.0 1.0 1.0 1.0 1.0 1.0 1 1.0 63 99 71 70 - 0 _ . I 1- Phlyctidocarpaflava r 1
I - Anginon Anginon paniculatum Annesorhiza altiscapa Annesorhiza macrocarpa Polemannia grossulariifolia Bupleurum fruticosum Astydamia latifolia Bupleurum angulosum Physospermum comubiense Dracosciadium italae Heteromorpha arborescens Sium suave Itasina filifolia Molopospermum peloponnesiacum Ezosciadium capense Alepidea peduncularis Alepidea capensis Alepidea amatymbica Lichtensteinia globosa Lichtensteinia lacera Lichtensteinia trifida Lichtensteinia obscura Marlothiella gummifera Chamarea snijmaniae Astrantia major Actinolema macrolema Arctopus echinatus Polemanniopsis sp.1 Phlyctidocarpa flava Choritaenia capensis Astrantia minor Polemanniopsis marlothii Steganotaenia araliacea Choritaenia capensis Steganotaenia commiphoroides Hacquetia epipactis Hermas quinquedentata Petagnaea gussonei Eryngium palmatum Eryngium giganteum Eryngium planum Eryngium maritimum Sanicula europaea Hermas gigantea Sanicula canadensis Tribal delimitationsinApiaceae difforme
_I 1 Azorelloideae
s •s eeeppinopes s•s eeepo!dy
43
d oi pi A su C su sen eae D & ° alvin e e owni S oid cul ani sensu C sensu eae D & o n vi a! e e wni o
Tribal delimitations in Apiaceae
1. Habit 2. Leaf persistence 3. Umbel o herbaceous o not proteranthous o compound woody proteranthous ■ simple Hernias gigantea Hernias quinquedentata Sium suave —_I-1 PhysosperMum comublense Bupleurum angulosum Bupleumm fruticosum Polemannia groaSuladifalia Heteromorpha arborescens Anginon paniculatum Anginon difforme Li Dracosciadium kola° AnnesoMiza altiscapa LJ Annesorhiza macrocarpa Chamarea snijmaniae Itasina filifolia Astydamia latifolia Ezosciadium capense Molopospennum pelopomesiacum I..._t_i_ Chontaenia capensts Chontaenia capensis Martothiella gummifera L 1L .I. Lichtensteinia obscure Lic htensteinia trifida Lichtensteinia lacers • Lichtensteinia globosa I_____I •I Phlyclidocarpa (lava Phlycliclocarpa (lava Polemanniopsis sp. 1 _I---' Polemanniopsis martothii Steganotaenia arallacea Steganotaenia commiphoroides I Alepidea amatymbica Alepidea capensis Alepidea peduncularis Arctopus echinatus Actinolema macrolema Astrantia major Astrantta minor Hacquetia epipactis Sanicula eumpaea Sanicula canadensis Erynglum maritimum Eryngium plenum B C Etyngiurn giganteum A Eryngium palmaturn Petagnaea gussonet 4. Flowers 5. Involucre bracts 6. Fruit surface o pedicellate o not forming a pseudanthium o glabrous o surface vesicles ■ sessile or subsessile forming a prominent o spines/bristles pseudanthium ■ stellate hairs Hennas gigantea Hermes quinquedentata Sium suave Physospermum comubiense Bupleurum angulosum Bupleurum fruticosum Polemannia grossutadifolia Heteromorpha arbomscens 1j Anginon paniculatum Anginon diffonne Dracosciadium italae Annesorhiza altiscapa L rte; Annesorhiza macrocarpa Chamarea snijmaniae ltasina filifolia Astydamia latifolia Ezosciadium capense Molopospermum peloponnesiacum Chorftaenia capensis Chorttaenia capensis Marlothiella gummifera Lichtensteinia obscure Lichtensteinia bifida Lichtensteinia lacera ter: Lichtensteinia globosa Phlyctidocarpa (lava Phlyctidocarpa (lava Potemannirmsis sp. I suanotaeaamamoPolemanniopsis ma thii
• Steganotaenia commiphomides Alepidea amatymbica Alepidea capensis Alepidea peduncularis Arctopus echinatus Actinolema macrotema Astrantia major Astrantia minor Hacquetia epipactis Sanicula eumpaea Sanicula canadensis Eryngium maritimum ' Eryngium plenum D E F Eryngium giganteum
EPrraangiUaMnaParr.saontumei
Figure 2
44 -
Tribal delimitations in Apiaceae
7. Fruit wing/rib development 8. Fruit ribs shape 9. Fruit symmetry pseudo-wings/ribs ❑ simple homomericarpic ■ true wings/ribs ■ bifurcate heteromericarpic
Hermes gigantea Hennas quinquedentata Slum suave Physospormum comublonso a Bupleurum angulosum Bupleurum fruticosum Polemannia grossularafolia Heteromorpha arborescens Anginon paniculatum Anginon Mame Dracosciadium italae Annesorhiza altiscapa a Annesorhiza macrocarpa o Chamarea snijmaniae o Itasina filifolia Astydamia !caddie Ezosciadium capense Molopospermum peloponnesiacum .., Choritaenia capensis 11:1 Choritaenia capensis II.7 Martothiella gummifera ., Lichtensteinia obscum Lichtensteinia trifida Lichtensteinia lacera o Lichtensteinia globosa 4a Phlyctidocarpa Have Phlyctidocarpa !lava • Polemanniopsis sp. 1 a Polemanniopsis martothii a Steganotaenia araliacea a Steganotaenia commiphoroides Alepidea amatymbica o Alepidea capensis I r Alepidea peduncularis Arctopus echinatus Actinolema rnacrolerna Astrantia major Astrantia minor Hacquetia °alpacas Sanicula europaea Sanicula canadensis Eryngium mariiimum Eryngium planum Eryngium giganteum H Eryngium palmatum Petagnaea gussonei
10. Endocarp 11. Regular vittae 12. Rib oil ducts woody o absent o small parenchymatous/lignified present large ■ oil vesicles forming cavities Hermes gigantea Hennas quinquedentata Sium suave Physospermum comubiense Bupleurum angulosum Bupleurum frutkosum Polemannia grossulartifolia scens etng _nor% panhi:ualatum Anginon difforme Dracosciadium italae 1=1 Annesorhize altiscapa Annesorhiza macrocarpa Chamarea snijmaniae Rosins Mitotic) Astydamia latifolia Ezosciadium capense Moiopospermum peloponnesiacum r - Choritaenia capensis Choritaenia capensis Madothiella gummifera Lichtensteinia obscura r—, Lichtenstainia trifida Lichtensteinia lacers Lichtensteinia globosa Phlyctidocarpa Save Phlyctidocarpa Sava Polemanniopsis sp. 1 Polemanniopsis marlothil Steganotaenia eraliacea Steganotaenia commiphoroides Alepidea amatymbica Alepidea capensis Alepidea peduncularis Arctopus echinatus Actinolema macrolema Astrantia major Astrantia minor Hacquetia epipactis Sanicula europaea Sanicula canadensis Eryngium maritimum I Eryngium plenum J L Eryngium giganteum r Eryngium palmatum Petagnaea gussonei
Figure 2 (Cont.)
45
Tribal delimitations in Apiaceae
13. Rhomboidal crystals 14. Druse crystals present absent m absent o scattered throughout mesocarp ■ restricted to commissure Hermas gigantea Azorelloideae Hermas quinquedentata m _ Sium suave Oenantheae Physospermum comubiense Pleurospermeae Bupleurum angulosum Bupleurum fruticosum Bupleureae a:: Polemannia grossulartifolia - Heteromorpha arborescens Anginon paniculaturn Heteromorpheae Anginon diffonne Dracosciadium italae Annesorhiza altiscapa 1 Annesorhiza macrocarpa Chamarea snijmaniae ltasina fllifolia Annesorhizeae J Astydamia latifolia I Ezosciadium capense Molopospermum peloponnesiacum Choritaenia capensis Choritaenieae L . Choritaenie capensis Martothiella gummifera 7 Marlothielleae Lichtensteinia obscura Lichtensteinia tda Lichtensteinieae Lichtensteinia lacers Lichtensteinia globosa 0 Apioideae s.l. 0 Phlyctidocarpa flava Phlyctidocarpeae Phlyctidocarpa flava -o. Polemanniopsis sp. 1 0 Polemanniopsis marlothii Steganotaenieae a Steganotaenia araliacea Steganotaenia commiphoroides 4=1 Alepidea amatymbica 1 Alepidea capensis 1 Alepidea peduncularis Arctopus echinatus Actinolema macrolema Astrantia major Astrantia minor Hacquetia epipactis Saniculeae Sanicula europaea Sanicula canadensis Eryngium maritimum _F—r-111 7—'11 Eryngium planum N Eryngium giganteum Eryngium palmaturn Petagnaea gussonei
Figure 2 (Cont.)
46 Tribal delimitations in Apiaceae
_r„," • do v#4.0, y„a;,..••••: -..,„,„,,,,iii.• . .kia''' die'
"••••••:,....::::::::"" . '
Figure 3
47 CD V
Apioideae sensu Calvin° & Downie Saniculoideaesen su CaNino & Downie 48 Sium suave Sium suave altiscapa Eryngium maritimum Eryngium planum Eryngium palmaturn Bupleurum fruticosum Bupleurum Ezosciadium capense Ezosciadium capense Bupleurum angulosum Bupleurum Eryngium giganteum Petagnaea gussonei Sanicula canadensis lepidea amatymbica Sanicula europaea Tribal delimitations in Apiaceae in Apiaceae delimitations Tribal Physospermum cornubiense cornubiense Physospermum Alepidea capensis Astrantia major Chamarea snijmaniae Chamarea snijmaniae Alepidea peduncularis Astrantia minor nnesorhiza Annesorhiza macrocarpa Annesorhiza macrocarpa 11 2 Cacquetia epipactis A Astydamia latifolia Astydamia latifolia 1 14 E-Actinolema macrolema 2 5 LL — Itasina filifolia — Itasina filifolia Molopospermum peloponessiacum Molopospermum ___ Marlothiella gummifera Anginon difforme Anginon difforme Anginon paniculatum Anginon Dracosciadium italae Dracosciadium Choritaenia capensis iChoritaenia capensis -Heteromorpha arborescens -Heteromorpha — Polemannia grossularifolia grossularifolia — Polemannia 10 Phlyctidocarpa flava Phlyctidocarpa flays 3 1 5 8 I 3 4 27 Arctopus echinatus Lichtensteinia trifida Lichtensteinia globosa 3 Steganotaenia araliacea
7 ' Lichtensteinia lacera 3 Steganotaenia commiphoroides Steganotaenia commiphoroides Polemanniopsis sp. 1 - 15 —Liatensteinia obscura 1 r 4 2 L-- Polemanniopsis marlothii €1 _ 4 7 4 31 -Hermas gigantea - Hermas quinquedentata Figure 4