South African Journal of Botany 128 (2020) 326À332

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South African Journal of Botany

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Zulustylis (Abildgaardieae, ) À a new genus from sub-Saharan Africa

A.M. Muasyaa,c,*, P. Goetghebeurb, I. Larridonb,c a Bolus Herbarium, Department of Biological Sciences, University of Cape Town, Private Bag X3, Rondebosch 7701, South Africa b Systematic and Evolutionary Botany Lab, Department of Biology, Ghent University, K.L. Ledeganckstraat 35, 9000 Gent, Belgium c Royal Botanic Gardens, Kew, Richmond, Surrey TW9 3AE, United Kingdom

ARTICLE INFO ABSTRACT

Article History: The generic placement of two African species currently included in the genus , F. hygrophila and F. Received 1 May 2019 variegata, has been contentious. These two species, which have been known for the past half century, are gla- Revised 2 September 2019 brous tufted perennials whose spikelets have semi-distichous to spiral glume arrangement, bisexual flowers, Accepted 22 November 2019 and a pistil with an enlarged and deciduous style base. While the two species fit the gross morphology of Available online 5 December 2019 Fimbristylis, they differ in lacking the characteristic Kranz anatomy. We present phylogenetic inference of the Edited by AR Magee Abildgaardieae based on DNA sequence data and map photosynthetic type. C4 photosynthesis has arisen twice in Abildgaardieae, in the Bulbostylis and the Fimbistylis clades, with a C3 clade (including Arthrostylis) Keywords: sister to rest of tribe and a C3 grade (the two African Fimbristylis species, Trachystylis, Actinoschoenus) sepa- C3 photosynthesis rating the two C4 clades. The new genus Zulustylis is described here, to segregate the two aberrant African Fimbristylis hygrophila Fimbristylis species, and its presented. Fimbristylis variegata Kranz anatomy Crown Copyright © 2019 Published by Elsevier B.V. on behalf of SAAB. All rights reserved. New combinations Phylogenetic inference Taxonomy

1. Introduction 1 G, 1H), and can be distinguished from Scirpeae by the unique embryo types (Fimbristylis-, Bulbostylis-, and Abildgaardia-type) and A number of changes in generic circumscription and classification consistent lack of perianth segments. Goetghebeur (1998) classified in the Cyperaceae have been made within the last two decades. The six genera in Abildgaardieae (Abildgaardia Vahl, Bulbostylis, Crosslan- majority of these have been caused by the shifting paradigm to recog- dia W.Fitzg., Fimbristylis, Nelmesia Van der Veken, Nemum Desv.), all nize monophyletic taxa (Humphries and Linder, 2009). During this sharing presence of multiple florets per spikelet and distinctly thick- period, increased use of molecular data in reconstructing phyloge- ened style bases in the majority of the taxa. At the same time, Goet- netic relationships has revealed the need to revise the classification ghebeur (1998) classified four other genera (Actinoschoenus Benth., at several suprageneric ranks, and the family is currently classified Arthrostylis R.Br., Trachystylis S.T.Blake, Trichoschoenus J.Raynal), shar- into two subfamilies (Cyperoideae and Mapanioideae) and into 18 ing a similarly thickened style base, in tribe Schoeneae due to their tribes (Muasya et al., 2009; Semmouri et al., 2019). Nearly every one spikelets having increasingly longer glumes but only the penultimate of the tribes has had changes in classification, with species and even glume (or rarely 2) bearing a bisexual floret (Figs. 1A, 1E, 1F). The lat- genera moved from tribe to tribe, the most drastic being the reclassi- ter four genera have previously been placed in the segregate tribe fication of the monotypic South African genus Hellmuthia Steud. from Arthrostylideae (Goetghebeur, 1986; Bruhl, 1995). subfamily Mapanioideae (Goetghebeur, 1998) into the Cyperoideae Recent phylogenetic studies (e.g. Ghamkhar et al., 2007; Simpson (Muasya et al., 2009). et al., 2007; Muasya et al., 2009; Semmouri et al., 2019; Roalson et al., Tribe Abildgaardieae is predominantly pantropical, comprising c. 2019a) have shown that Arthrostylis and allied genera are part of the 540 species, with highest diversity in Bulbostylis Kunth and Fimbristy- Abildgaardieae clade. Furthermore, Crosslandia has been demon- lis Vahl (WCSP, 2019). Abildgaardieae was circumscribed (Lye, 1973) strated to be a lineage within Fimbristylis (Ghamkhar et al., 2007; to include taxa that have many-flowered bisexual spikelets (Figs. 1B, Roalson et al., 2019b), and that Nemum (and perhaps Nelmesia) are lineages within Bulbostylis (Muasya et al., 2009; Hinchliff and Roal- son, 2013; Spalink et al., 2016; Semmouri et al., 2019; Roalson et al., * Corresponding author at: Bolus Herbarium, Department of Biological Sciences, University of Cape Town, Private Bag X3, Rondebosch 7701, South Africa. 2018, 2019a). The generic placement of two African species (Fimbris- E-mail address: [email protected] (A.M. Muasya). tylis hygrophila Gordon-Gray, F. variegata Gordon-Gray; Figs. 1C, 1D) https://doi.org/10.1016/j.sajb.2019.11.027 0254-6299/Crown Copyright © 2019 Published by Elsevier B.V. on behalf of SAAB. All rights reserved. A.M. Muasya et al. / South African Journal of Botany 128 (2020) 326À332 327

We first inferred the gene trees for each of the four regions sepa- rately to identify potential incongruence. As there was little conflict, the matrices of the four regions were concatenated for the down- stream analyses. PartitionFinder 2.1.1 (Lanfear et al., 2012) was used to determine an appropriate data-partitioning scheme from potential partitions that were defined a priori (in this case, each marker was treated as a separate partition, as was each codon in the two coding markers), as well as the best-fitting model of molecular evolution for each partition, using the Bayesian Information Criterion. The optimal data-partitioning scheme was ITS, ndhF codon 1 + rbcL codon 1, ndhF codon 2, ndhF codon 3, rbcL codon 2, rbcL codon 3, trnL-trnF. Maxi- mum likelihood (ML) analyses of the optimally partitioned data were performed using RAxML 8.2.10 (Stamatakis, 2014) with model set to GTR (gamma) for each partition. The search for an optimal ML tree Fig. 1. Photographic illustration of the inflorescence morphology in the Abildgaar- dieae. Arthrostylis aphylla (A), Bulbostylis atrosanguinea (B), Zulustylis hygrophila (C), was combined with a rapid bootstrap analysis of 1000 replicates. Par- Zulustylis variegata (D), Actinoschoenus repens (E), Trachystylis stradbrokensis (F), Abil- titioned analyses were conducted using Bayesian Inference (BI) in dgaardia ovata (G), Fimbristylis ferruginea (H). Vouchers: A, Brass 18669 (K); B, Luke MrBayes 3.2.6 (Ronquist et al., 2012). Based on the PartitionFinder 4599 (K); C, Venter 2832 (K); D, Huntley 734 (K); E, Robinson 5977 (K); F, Blake 22673 analysis, the GTR (invgamma) model of sequence evolution was set (K); G, Verdcourt 3253 (K); H, Ward 7924 (K). Scale bar = 1 mm. for the ITS partition; the GTR (gamma) model for the ndhF codon 1+rbcL codon 1 partition and the ndhF codon 3 partition; the F81 has remained uncertain. These species were described as part of Fim- model (propinv) for the ndhF codon 2, rbcL codon 2 and rbcL codon 3 bristylis (Gordon-Gray, 1966) and were later transferred into Abil- partitions; and the F81 model (invgamma) for the trnL-trnF partition dgaardia (Lye, 1971). As Abildgaardia has been considered to be a in the concatenated data set. The analysis was allowed to run for synonym of Bulbostylis (e.g. Lye, 1971, 1973; Haines and Lye, 1983), 100 million generations across four independent runs with four these species have by extension also been viewed as belonging to chains each, sampling every 10,000 generations. Convergence, associ- fi Bulbostylis. The uncertainty in af nity is epitomized by the segrega- ated likelihood values, effective sample size (ESS) values and burn-in tion of these two species as the only C3 species of Fimbristylis by values of the different runs were verified with Tracer 1.6 (Rambaut fi Bruhl (1995) and by Gordon-Gray (1995) who classi ed one of the and Drummond, 2014). The first 25% of the trees from all runs were species in Abildgaardia (A. hygrophila (Gordon-Gray) Lye) and the excluded as burn-in before making a majority-rule consensus of the other in Fimbristylis (F. variegata). Molecular phylogenetic studies 30,000 posterior distribution trees using the “sumt” function. All phy- (Besnard et al., 2009; Hinchliff and Roalson, 2013; Bouchenak-Khe- logenetic analyses were run using the CIPRES portal (http://www. ladi et al., 2014; Spalink et al., 2016; Semmouri et al., 2019) have phylo.org/; Miller et al., 2010), and were executed for both full and shown that these sister species are not part of the clade including reduced sampling alignments. Trees were drawn using FigTree 1.4.3 Abildgaardia and Fimbristylis. We investigate the taxonomic place- (http://tree.bio.ed.ac.uk/software/figtree/) and adapted in Adobe ment of these two sub-Saharan African species of Abildgaardieae and Photoshop CS5. revise their taxonomy. 3. Results 2. Materials and methods Since the topologies of the concatenated ML and BI trees were Representative herbarium specimens of tribe Abildgaardieae were entirely congruent, we here provide the BI tree with both the BI pos- studied to identify distinguishing morphological features. In addition, terior probability values (PP) and ML bootstrap values (BS) (Fig. 2). all available specimens of the two target species were studied in Analysis of the individual markers show strong support for the Abil- selected herbaria (BOL, BR, GENT, K, NBG, NU, PRE; codes follow dgaardieae sensu lato. The first diverging lineage has Arthrostylis (sis- Thiers 2019+) as well as consultation of key literature (e.g. Gordon- ter to Fimbristylis composita) as sister to rest of tribe; then strongly Gray, 1995; Goetghebeur, 1998). Gross morphological observations supported lineages observed consistently comprise Bulbostylis were made to enable species descriptions, with the aid of dissecting (including Nemum), a grade with three lineages (Fimbristylis hygro- microscope and measurements made using a ruler or digital calipers. phila sister to F. variegata, Trachystylis, Actinoschoenus) and a Fimbris- We used data from previous studies on the morphology of embryos tylis clade (including Abildgaardia and Crosslandia). But none of the (Semmouri et al., 2019) and the photosynthetic pathway type (based individual markers provides support at the backbone. on anatomy and carbon isotopes; Bruhl and Wilson; 2007). Images of Table 2 summarizes morphological variation in tribe Abildgaar- spikelets and florets were made using a Leica S9i dissecting micro- dieae and the gross morphology of inflorescences is illustrated in scope with an inbuilt digital camera, and plate composed using Fig. 1. The majority of genera in tribe Abildgaardieae share the pres- Adobe Photoshop CS5. ence of a thickened style base, but there is variation on whether this DNA sequence data of 54 taxa including all accepted genera of is persistent or deciduous. Two clades (Bulbostylis, Fimbristylis) have tribe Abildgaardieae (except Nelmesia for which efforts to sequence C4 photosynthesis (characterised by Kranz anatomy), whereas C3 DNA from herbarium specimen were unsuccessful) were sampled for photosynthesis is observed in first diverging lineage and the grade this study. Data of four markers (the chloroplast genes ndhF and rbcL, leading to Fimbristylis clade. Furthermore, the C4 clades, together the chloroplast intergenic spacer trnL-trnF and the nuclear ribosomal with Fimbristylis hygrophila and F. variegata, have spikelets whose marker ITS) generated in previous studies (Ghamkar et al., 2007, glumes do not increase in length from the base, and where more Simpson et al., 2007; Besnard et al., 2009; Hinchliff and Roalson, than two florets are fertile. 2013; Semmouri et al., 2019) were retrieved from GenBank. The sam- ples with species names, voucher information, origin and GenBank 4. Discussion accession numbers for the sequences, are given in Table 1. Sequences were aligned using MAFFT 7 (Katoh et al., 2009; Katoh and Standley, The two aberrant African species placed in Fimbristylis (or Abil- 2013), afterwards, alignments were checked manually in PhyDE dgaardia) are not phylogenetically part of the Fimbristylis clade. Our 0.9971 (Muller€ et al., 2010). results here agree with previous studies (Bruhl, 1995; Besnard et al., 328 A.M. Muasya et al. / South African Journal of Botany 128 (2020) 326À332

Table 1 Voucher table listing the species included in the molecular study with voucher information, distribution of the species (WCSP, 2019) and GenBank accession numbers.

Species Specimen voucher Country of origin ndhF rbcL trnL-trnF ITS

Fimbristylis fusiformis Wangwasit & D.A. Simpson (Abildgaardia) K. Wangwasit 080927-17 (K) Thailand KY652919 F. ovata (Burm.f.) J. Kern (Abildgaardia ovata (Burm. f.) Kral) Tsusaka 19708 (OKAY) Japan AB373097 AB369943 AB250638 F. ovata (Burm.f.) J. Kern (Abildgaardia ovata (Burm.f.) Kral) Muasya et al. 684 (K, EA) Kenya AJ295754 F. oxystachya F. Muell. (Abildgaardia) K.L. Clarke 165 (NE) Australia AY506704 AY506762 F. squarrulosa F. Muell. (Abildgaardia schoenoides R. Br.) K.L. Clarke 70 (NE) Australia AY506706 AY506761 Actinoschoenus thouarsii (Kunth) Benth. (Actinoschoenus L. Gautier 2600 (G) Madagascar FM160509 AM999783 aphyllus (Vahl) ined.) F. composita Latz (Actinoschoenus) K.L. Clarke 213 (NE) Australia AY506702 AY506755 Arthrostylis aphylla R. Br. K.L. Clarke 183 (NE) Australia AY506700 AY506757 Arthrostylis aphylla R. Br. unavailable Australia AY725939 Bulbostylis atrosanguinea (Boeckeler) C.B. Clarke A.M. Muasya 1037 (CAN, K) Kenya KJ513485 Y12992 B. barbata (Rottb.) C.B. Clarke K.L. Clarke 113 (NE) Australia AY506709 AY506764 B. barbata (Rottb.) C.B. Clarke Hoshino et al. 13779 (OKAY) Japan AB261628 AB369944 B. burchellii (Ficalho & Hiern) C.B. Clarke Poilecot & Dulieu 7065 (G) Zimbabwe FM160506 AM999780 B. densa (Wall.) Hand.-Mazz. Hirahara et al. 19651 (OKAY) Japan AB261629 AB369945 AB261663 B. densa (Wall.) Hand.-Mazz. V. Klaphake 1411 (NSW) Australia AY506710 B. hispidula (Vahl) R.W. Haines A.M. Muasya 1025 (EA, K) Kenya GU075448 Y12944 Nemum equitans (Kuk.)€ J. Raynal (Bulbostylis) H. & E. Hess 52/686 (GENT) Angola MH286440 MH286447 N. spadiceum (Lam.) Desv. ex Ham. (Bulbostylis) Baldwin 9766 (K) Guinee (Conakry) Y12945 Trachystylis stradbrokensis (Domin) Kuk.€ K.L. Wilson 8175 (K) Australia AM999970 AM999877 EF178591 F. hygrophila Gordon-Gray (Zulustylis hygrophila Abbott 8841.10 (BOL) South Africa FM160508 AM999782 (Gordon-Gray) Muasya) F. variegata Gordon-Gray (Zulustylis variegata J. Browning 366 (GENT) South Africa AM999929 AM999835 (Gordon-Gray) Muasya) F. crosslandii Roalson, R.L. Barrett, & Larridon K.L. Wilson 10147 (K) Australia EF178538 (Crosslandia setifolia W. Fitzg.) F. crosslandii Roalson, R.L. Barrett, & Larridon K.L. Clarke 185 (NE) Australia AY506716 AY506769 (Crosslandia setifolia W. Fitzg.) F. vaginata (R. Br.) Domin (Crosslandia) J.J. Bruhl 2057 (NE) Australia AY506705 AY506759 F. aestivalis (Retz.) Vahl J. Jung 1008057 (AJOU) South Korea JX644685 JX644785 JX644879 F. arnhemensis Latz K.L. Clarke 177 (NE) Australia AY506722 AY506776 F. autumnalis (L.) Roem. & Schult J. Jung 1009036 (AJOU) South Korea JX644686 JX644780 JX644880 F. bisumbellata (Forssk.) Bubani K.L. Clarke 107 (NE) Australia AY506724 AY506778 F. cephalophora F. Muell. K.L. Wilson 10070 (NSW) Australia AY506720 AY506720 F. cinnamometorum (Vahl) Kunth K.L. Clarke 61 (NE) Australia AY506725 AY506773 F. complanata (Retz.) Link J. Jung 1010290 (AJOU) South Korea JX644687 JX644781 JX644881 F. complanata (Retz.) Link A.M. Muasya 1029 (K) Kenya GU075444 F. cymosa R. Br. K.L. Wilson 10041 (NSW) Australia AY506750 AY506798 F. cymosa R. Br. Abbott 23691 (FLAS) USA KJ773512 F. densa S.T. Blake K.L. Clarke 180 (NE) Australia AY506747 AY506779 F. dichotoma (L.) Vahl J. Jung 1009010 (AJOU) South Korea JX074642 JX644688 JX644787 JX644882 F. ferruginea (L.) Vahl A.M. Muasya 2171 (EA) Kenya AM999927 AM999833 F. ferruginea (L.) Vahl J. Hodgon 445 (NE) Australia AY506744 AY506797 F. lanceolata C.B. Clarke K.L. Wilson 10113 (NSW) Australia AY506730 AY506786 F. laxiglumis Latz K.L. Clarke 106 (NE) Australia AY506736 AY506785 F. microcarya F. Muell. K.L. Clarke 104 (NE) Australia AY506731 AY506787 F. neilsonii F. Muell. K.L. Wilson 10051 (NSW) Australia AY506719 AY506784 F. nutans (Retz.) Vahl J. Hodgon 541 (NE) Australia AY506733 AY506789 F. polytrichoides (Retz.) Vahl S. Jacobs 8688 (NSW) Australia AY506737 AY506795 F. pterigosperma R. Br. K.L. Clarke 118 (NE) Australia AY506729 AY506794 F. quinquangularis (Vahl) Kunth (incl. F. miliacea (L.) J. Jung 909180 (AJOU) South Korea JX644692 JX644783 JX644886 Vahl, nom. rej.) F. rara R. Br. K.L. Clarke 105 (NE) Australia AY506728 AY506780 F. schultzii Boeckeler K.L. Clarke 108 (NE) Australia AY506748 AY506791 F. sericea (Poir.) R. Br. K.L. Wilson 10042 (NSW) Australia AY506743 AY506803 F. sieboldii Miq. ex Franch. & Sav. J. Jung 1010277 (AJOU) South Korea JX644690 JX644788 JX644884 F. squarrosa Vahl J. Jung 807041 (AJOU) South Korea GQ130376 JF313195 GQ130344 F. squarrosa var. esquarrosa Makino (incl. F. velata) J. Jung 808010 (AJOU) South Korea JX644693 JX644786 JX644887 F. squarrosa var. esquarrosa Makino (incl. F. velata) Hoshino et al. 17692 (OKAY) Japan AB261630 F. stauntonii Debeaux & Franch. S.-M. Yun s.n. (AJOU) South Korea JX644694 JX644779 JX644888 F. tetragona R. Br. K.L. Clarke 173 (NE) Australia AY506746 AY506799 F. thermalis S. Watson Hessing s.n. UCR220075 (UCR) USA MF963420 MF964172 F. tristachya R. Br. K.L. Clarke 3 (NE) Australia AY506745 AY506802 F. tristachya var. subbispicata (Nees) T. Koyama J. Jung 1010018 (AJOU) South Korea JX644695 JX644782 JX644889 Eleocharis acicularis (L.) Roem. & Schult. W. Scheibler s.n. (UNIL) uncertain FM160544 GU977050 E. acicularis (L.) Roem. & Schult. Arai & Hirahara 18168 (OKAY) Japan AB369953 E. acicularis (L.) Roem. & Schult. J. Jung 1010110 (AJOU) South Korea JX644718 E. confervoides (Poir.) Steud. C.E. Hinchliff 694 (WS) USA GU075435 GU110789 E. confervoides (Poir.) Steud. M.W. Chase 2706 (K) unavailable FN870956 E. ovata (Roth) Roem. & Schult. J. Jung 1007045 (AJOU) China JX644677 JX644721 JX644871 E. palustris (L.) Roem. & Schult. G. Besnard 34À2007 (G) unavailable FM160551 KM360762 GQ244894 GU977092 E. vivipara Link O. Ueno s.n. Japan FM160554 AM999829 E. vivipara Link J.B. Nelson 732 (RSA) unavailable AF190614 A.M. Muasya et al. / South African Journal of Botany 128 (2020) 326À332 329

Fig. 2. 50% majority consensus multiple-locus Bayesian Inference tree with the associated Posterior Probability values and the Bootstrap values of the multiple-locus Maximum Likelihood tree. Only bootstrap values above 75% and posterior probabilities above 0.75 are shown.

2009; Hinchliff and Roalson, 2013; Bouchenak-Kheladi et al., 2014; example, if these were to be retained as Fimbristylis, then other gen- Spalink et al., 2016; Semmouri et al., 2019), which have shown these era (Abildgaardia, Actinoschoenus, Trachystylis) would need to be taxa to be closer to the taxa formerly included in the Arthrostylideae. merged with Fimbristylis. We therefore propose to erect a new genus These taxa share the presence of C3 anatomy with the Arthrostyli- for the two species. deae (Bruhl and Wilson, 2007). Furthermore, F. hygrophila is more The genera in tribe Abildgaardieae differ greatly in their species similar to species in Actinoschoenus (e.g. A. repens) in morphology numbers and ecological variability. The C3 lineages are depauperate, (capitate inflorescence, distichous glumes arrangement; Figs. 1C, 1E) comprising under 20 species whereas the C4 clades each have over and in ecology (wetlands). The main differences from Actinoschoenus 200 species. While the C3 lineages are restricted to wetlands, the being in the spikelet bearing more than two fertile florets and glumes other (C4) clades have diversified since mid-Miocene (Besnard et al., not increasing in length from spikelet base towards the apex. We find 2009; Hinchliff and Roalson, 2013; Bouchenak-Kheladi et al., 2014) that these two species cannot be added to any of the recognized gen- into a variety of non-wetland habitats and span a wide distribution era without greatly upsetting the current generic concepts. For range. Perhaps the C3 Fimbristylis species have remained in the 330 A.M. Muasya et al. / South African Journal of Botany 128 (2020) 326À332

ancestral habitats and retained ancestral morphology of tribe Abil- dgaardieae. mbriate fi 5. Taxonomic treatment

d, often 5.1. Genus description fi many larged, deciduous 2 À

> Zulustylis Muasya, gen. nov. New genus similar in gross morphol- ogy to Abildgaardia and Fimbristylis, differing in having C3 anatomy. Type species: Zulustylis hygrophila (Gordon-Gray) Muasya Tufted perennial, rhizome inconspicuous, woody, vertical or semi- vertical. Culms to 600 mm long, wiry, terete, grooved, glabrous. Leaves: sheath 22À165 £ 0.9À1.9 mm, usually pale yellow, spongy, minutely speckled brown, rim smooth and sloping away from leaf blade; blade reduced to a lobe < 4mmlong.Inflorescence a single head of 1 to 4 ses- mbriate whole length 2 & 3- fi

rm their placement in the tribe. Embryo data are based sile spikelets or occasionally with additional 1 to several pedicellate spi- d, fi 3 (sessile or stalked) 1 fi 2 À kelets; pedicel to 13 mm long; bracts 2 to 4, glume-like, 3.7À4.2 mm > long, shorter than the spikelet. Spikelets terete or appearing slightly compressed at base, 5.0À19.0 £ 2.1À4.5 mm, oblong to oblong-lanceo- late. Glumes elliptic to lanceolate, golden yellow or reddish brown with

7 stalked 1 a yellowish midrib tip, lowest 1 or 2 sterile, glabrous, 3.4À9.7 mm long, À margins membranous, midrib prominent and excurrent into a mucro of mbriate 3- fi 0.3À1.2 mm long. Anthers 1.8À3.7 mm long, yellowish to dark brown. d, fi Style 3-branched, ciliate to villous, base shortly pyramidal, 0.5À0.6 mm long. Nutlets 10À1.7 £ 0.5À1.1 mm, trigonous, broadly obovate to pyri- form, surface faintly reticulate to transversely ribbed.

Etymology

mbriate bi The name Zulustylis recognizes the locality of the two species fi d,

many 1 sessile, 3 which is predominantly in KwaZulu Natal Province of South Africa, 1 cm long well developed short to well developed short to well developed fi À

< and honours the iconic Zulu community.

5.2. Key to the species of Zulustylis

1. Spikelets laterally compressed at base; nutlet pyriform 1. Z. hygrophila 0 mbriate 3- 1 . Spikelets terete; nutlet obovate 2. Z. variegata fi 3) 1 , have not been included here due to lack of phylogenetic studies to con À d, 1 cm long fi 2 1 (2) 1 < > 5.3. Species treatments

1. Zulustylis hygrophila (Gordon-Gray) Muasya, comb. nov. Fim- Trichoschoenus bristylis hygrophila Gordon-Gray, J. S. African Bot. 32: 129 (1966). Abil- and dgaardia hygrophila (Gordon-Gray) Lye, Mitt. Bot. Staatssamml. Munchen€ 10: 547 (1971). Type: South Africa. KwaZulu Natal, Port Nelmesia . Edward (3130): Port Edward (AA), Jan 1951, Huntley 701 (NU holo.!; BM!, BOL!, E! [3 sheets], K! [3 sheets], L, NBG!, PRE!, PRU, iso.).

d, glabrous 3- fi

fi Tufted perennial, resprouting after re; rhizome inconspicuous,

many 1 (2 woody, vertical or semi-vertical. Culms 170À510 £ 0.4À0.8 mm, 2 À > wiry, terete, grooved, glabrous. Leaves: sheath 22À65 £ 0.9À1.9 mm, usually pale yellow, spongy, minutely speckled brown, rim smooth and sloping away from leaf blade; blade reduced to a lobe < 3 mm.

Bruhl and Wilson (2007) Inflorescence a 1 (2) terminal spikelet or occasionally with additional 1 to several pedicelled spikelets; pedicel up to 13 mm long; bracts 2 to 4, glume-like, 3.7À4.2 mm long, shorter than the spikelet. Spikelets mbriate to wooly 2 & 3-

fi appearing slightly compressed with glume arrangement distichous at

d, À £ À 1 cm long short to well developed fi base and spiral toward apex, 10.0 19.0 3.0 4.5 mm, oblong to Arthrostylis< Bulbostylis Zulustylis Actinoschoenus Trachystylisoblong-lanceolate. AbildgaardiaGlumes Fimbristylis elliptic to lanceolate, golden yellow or red- dish brown with a yellowish green midrib tip, lowest 1 or 2 sterile, glabrous, 5.0À9.7 mm long, margins membranous, midrib prominent ; photosynthetic data from and excurrent into a mucro of 0.5À1.2 mm long. Stamens 3, filaments ribbon-like; anthers 2.2À3.7 mm long, yellowish to dark brown. Style ce glabrous hairy, some glabrous glabrous glabrous glabrous glabrous glabrous fi 3-branched, ciliate to villous, base shortly pyramidal, bearing a flap covering the nutlet apex and 0.5À0.8 mm long. Nutlets oret 1

fl 1.2À1.7 £ 0.8À1.1 mm, trigonous, broadly obovate to pyriform, sur- orescence type capitate capitate, anthelate capitate capitate, anthelate anthelate capitate, anthelate capitate, anthelate

fl face faintly reticulate and transversely ribbed. Figs. 1C, 3A. Semmouri et al., 2019 Style 3- Leaf blade HabitLeaf sheath ori perennial annual & perennial perennial perennial perennial perennial, rarely annual annual & perennial Glume arrangementFertile spiral Spiral, rarely distichous distichous to spiral distichous & spiral spiral distichous to spiral spiral, rarely distichous In Style-baseEmbryo typePhotosynthetic systemDistribution C3 enlarged, deciduous Carex-type NE Australia mostly enlarged & persistent Bulbostylis- enlarged, deciduous & Abildgaardia-type C4 enlarged, deciduous Fimbristylis-type pantropical enlarged, deciduous Carex-type enlarged, deciduous Carex-type en S & SE Africa C3 Abildgaardia-type paleotropical Fimbristylis-type E Australia C3 pantropical C3 pantropical C4 C4 Involucral bract (lowermost)Spikelet glume-like leaf-like or short many glume-like 1 leaf-like or shortDistribution Leaf-like and glume-like Ecology: This species leaf-like or short is widespread in eastern and Table 2 Distinguishing features among the genera of tribe Abildgaardieae. Two taxa, on southern Africa, distributed from Tanzania to South Africa. It grows A.M. Muasya et al. / South African Journal of Botany 128 (2020) 326À332 331

Msikaba gorge, Mateku River (-BD), 1 Apr 2002, de Wett, L. 145 (PRE); Pondoland, Mtentula, above Msikaba gorge (-BD), 29 Sep 2005, Victor, J.E., 3099 (PRE); Lupatana (-BD), 13 Nov 1970, Strey, R.G., 10238 (PRE); Mkambati Game Reserve (-BD), 7 Sep 1985, Shackleton, C., 302 (PRE); Usikisiki District, S edge of Umsikaba Gorge (-BD), 13 Jan 1947, Acocks, J.P.H., 13266 (PRE); 3130 (Port Edward): Mzamba River mouth south of Port Edward (-AA), Oct 1973, Arnold, T.H., 791 (PRE). UNCERTAIN PROVINCE, Claremont, 20 m, 22 Aug 1893, Schlechter, R. 3132 (GRA, K). Tanzania. Tunduru District, 15 km E of R. Mawese, Milne-Redhead & Taylor 7830 (GENT, K); Tabora District, 19 km on Tabora-Nzega Road, 14 Jun 2008, Bidgood, S., Leliyo, G., Vollesen, K., 7348 (K). Zambia. Central Province, Chakwenga Headwaters, 100À129 km east of Lusaka, 1 Dec 1963, Robinson, E.A., 5877 (PRE); Luanshya, Chin- Fig. 3. Photographic illustration of the floret morphology (glume, anther, style, nutlet) gola, 2 Feb 1960, Robinson, E.A., 3316 (K, NU, SRGH); (1528BC), Lusaka in Zulustylis hygrophila (A) and Zulustylis variegata (B). Vouchers: A, Huntley 701 (K); B, District, Nyanshishi River near Chinkuli, 10 Dec 1972, Kornas, J., (Plan- Huntley 734 (K). Scale bar = 1 mm. tae Africanae) 2792 (GENT, K). Mozambique. San Sebastian Peninsula, east of Vilanculos. 28 Aug in waterlogged habitats (vlei, swamp, lake edges, old rice fields, dam- 2002, Jacobsen, N.H.G., 6084 (PRE); Beira District, Lower Chiniziua saw bos in miombo woodlands) at a wide elevation from sea level to mill, 13 Jul 1972, Ward, C.J., 7889 (K, PRE); Bazaruto Isle, 7 Nov 1958, 1300 m. Resprouting after fire. Mogg, A.O.D., 23346 (PRE). Diagnostic characters: Zulustylis hygrophila is a robust tufted 2. Zulustylis variegata (Gordon-Gray) Muasya, comb. nov. Fim- perennial, unique in having leafless scapes terminating in 1 or few bristylis variegata Gordon-Gray, J. S. African Bot. 32: 134 (1966). Abil- spikelets, and spikelets appearing laterally compressed. This species dgaardia variegata (Gordon-Gray) Lye, Mitt. Bot. Staatssamml. is most similar to Abildgaardia ovata in glume arrangement and nutlet Munchen€ 10: 547 (1971). Type: South Africa. KwaZulu Natal, Port (pyriform) morphology, but A. ovata has well developed leaves and Edward (3130): Port Edward (-AA), Jan 1951, Huntley 734 (NU holo.!; C4 anatomy. BM!, BOL!, E!, K! [2 sheets], L!, NU!, PRE!, PRU, iso.). Tufted perennial, resprouting after fire, rhizome inconspicuous, Additional selected specimens examined woody, vertical or semi-vertical. Culms 150À400 £ 0.4À0.8 mm, wiry, terete, grooved, glabrous. Leaves: sheath 40À60 £ 0.6À1.2 mm, South Africa. KWAZULU-NATAL. 2732 (Ubombo): Sileza Forest usually pale yellow, spongy, minutely speckled brown, rim smooth Reserve, Northern boundary area (-BA), 18 Feb 1993, Matthews, W.S., and sloping away from leaf blade; blade reduced to a lobe < 4mm. 1165 (PRE); Manzingwenya; Vazi Swamp; just E. of Vazi Swamp Inflorescence a single head of 1 to 4 sessile spikelets or occasionally (-BA), 28 Nov 1969, Moll, E.J., 4765 (PRE); Ingwavuma District, Muzi with additional 1 to several pedicelled spikelets; pedicel to 13 mm swamp system near Phelandaba cash store (-BA), 27 Mar 1985, Reid, long; bracts 2 to 4, glume-like, 3.7À4.2 mm long, shorter than the C., 1056 (GENT, K, PRE); Ingwavuma District., Sileza Forest (-BA), 10 spikelet. Spikelets terete but appearing slightly compressed at base, Oct 1985, Ward, M.C., 1079 (NU); Ingwavuma District., Sileza (-BA), 5.0À10.0 £ 2.1À4.0 mm, oblong to oblong-lanceolate. Glumes elliptic 14 Nov 2000, Zietsman, P.C., 4262 (PRE); Ubombo (-BC), 7 Dec 1979, to lanceolate, golden yellowed or reddish brown with a yellowish MacDevette, 58 (NU); Sodwana State Forest, Ozabeni (-CD), 16 Oct midrib tip, lowest 1 or 2 sterile, glabrous, 3.4À7.2 mm long, margins 1986, Van Wyk, G.F., 944 (NU); Eastern side lowermost Mkuze membranous, midrib prominent and excurrent into a mucro of Swamps, Ukhovu Pan area (-DC), 9 Nov 1972, Ward, C.J., 8105 (K, 0.3À0.5 mm long. Anthers 1.4À3.7 mm long, yellowish to dark brown. PRE). 2831 Nkandla: Ngoye Forest Reserve, Eastern Ridge (-DC), 25 Style 3-branched, ciliate to villous, base shortly pyramidal and Oct 1966, Venter H.J.T. 2832 (K, PRE). 2832 (Mtubatuba): Hlabisa Dis- 0.5À0.6 mm long. Nutlets 1.0À1.3 £ 0.5À0.6 mm, trigonous, broadly trict, St. Lucia, eastern shores (-AB), 31 Nov 1959, Feely & Ward, C.J., obovate (not pyriform), surface faintly reticulate (not transversely 14 (K, PRE); Hlabisa Dukuduku (E-corner V. 175) (-AD), 20 Nov 1964, ribbed) (Figs. 1D, 3B). Ward, C.J., 5072 (PRE); Dukuduku Forest Reserve (-AD), 4 Jun 1972, Distribution and Ecology: This species is restricted to southern Ward, C.J., 7728 (PRE); 4 km from St. Lucia Estuary on road to Cape Africa, occurring from Eastern Cape to Mpumalanga. It was previously Vidal (-BA), Sep 1973, Arnold, T.H., 469 (PRE); St. Lucia System, Bhan- described from Eastern Cape and KwaZulu Natal coastline, but several gazi Lake (-BA), 16 Apr 1982, Ward, C.J., 9497 (PRE); Lake Nhlabane, high elevation collections (Smook 8143 & Haines 7051) expand the western shores (-CB), 17 Sep 1991, Ward, C.J., Begg, G.W. 11351 (PRE); range, elevation 100 À 1700 m. All localities are in wetlands (margins 2930 (Pietermaritzburg): Merebank East (-DD), 10 Sep 1965, Ward, of streams, pools) located in sandstone habitats. C.J., 5130 (PRE). 2931 (Stanger): Merebank East (-CC), 20 Sep 1965, Diagnostic characters: This species is similar to Z. hygrophila in Ward, C.J., 5182 (PRE). 3030 Port Shepstone: Umlazi District; main anatomy and morphology, differing in glume arrangement (spiral Umbumbulu-Winklespruit road. (near Adams Mission) (-BB), 23 Mar versus distichous at spikelet base) and nutlet shape (broadly obovate 1966, Ward, C.J., 5519 (PRE); Umtamvuna Nature Reserve, vlei above versus pyriform). the Cleft (-CC), 22 Sep 1984, Abbott, A.T.D., 2129 (NU); Umtavuna Nature Reserve (-CC), 10 Jan 2008, Muasya & Stirton in Abbott 8841.14 Additional specimens examined (BOL). 3130 (Port Edward): Port Edward (-AA), 25 Oct 1975, Stirton, C.H., 5650 (PRE); Port Edward, Umtamvuna River (-AA), 21 Oct 1976, South Africa. MPUMALANGA. 2531 (Komatipoort): KaNgwane, Nicholson, H.B., 1560 (PRE). Uncertain. Mhlambanyati pan, sandy vlei, Songimvelo Game Reserve, Mendon boundary with Schultzenhorst after fire, 27 Mar 1941 Michelmore, A.P.G., 49 (PRE). EASTERN CAPE. Farm (-AA), 9 Dec 1992, Smook, L., 8143 (PRE). KWAZULU-NATAL. 3129 (Port St. Johns): Pondoland, Inland of Mkambati near tree plan- 3030 Port Shepstone: Umzinto District. Pennington, near turn off tation (-BB), 26 Sep 2005, Victor, J.E., 3006 (PRE); Pondoland, Near to from main Durban - South Coast Road (-BC), 5 Jan 1969, Gordon-Gray, Mnyameni River, and near to Amadiba (-BB), 12 Oct 2005, Victor, J.E., K.D.H., 6234 (NU); Oribi flats (-CB), Apr 1937, McClean, A.P.D., 380 3163 (PRE); Tracor; Slopes above KwaDlambu E.-tributary (-BD), 15 (NH, NU); Paddock; Higlands Farm, above Umzimkulwana River Nov 2000, Abbott, A.T.D., 7784 (PRE); Pondoland, Mtentula, near (-CB), 1 Jan 1972, Nicholson, H.B., 1142 (PRE); Umtamvuma (-CB), 23 332 A.M. Muasya et al. / South African Journal of Botany 128 (2020) 326À332

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