SMITHSONIAN CONTRIBUTIONS TO BOTANY NUMBER 50

Pollen Morphology and Phylogenetic Relationships of the Berberidaceae

Joan W. Nowicke andJohn J. Skvarla

SMITHSONIAN INSTITUTION PRESS City of Washington 1981 ABSTRACT Nowicke, Joan W., and John J. Skvarla. Pollen Morphology and Phylogenetic Relationships of the Berberidaceae. Smithsonian Contributions to Botany, number 50, 83 pages, 215 figures, 3 tables, 1981.-Pollen from 68 collections repre- senting 14 genera and 40 species of the family Berberidaceae was examined by light microscopy, SEM, and TEM. In part, the pollen data reinforce the traditional view of closely related pairs or small groups of genera. In Berberis and Mahonia the pollen morphology would support separate family status as well as congeneric treatment. The unusual exine structure in Nandina would reinforce its treatment as a monotypic family, Nandinaceae. The distinction of Bongardia from Leontice and of Dysosma from Podophyllum is confirmed by pollen data. The presence of a fundamentally similar tectum in Achlys, Dysosma, Epimedium,Jeffsonia, Podophyllum peltatum, P. hispidun, and Vancouveria suggests closer relationship among these genera than has been previously thought. The _close similarity of the polle-n in Jeffersonia and Plagio- rhegma confirms their congeneric treatment, Palynologically, Bongardia, Caulo- phyllum, and Leontice are more closely related to each other than to any remaining genera. In three taxa, Diphylleia, Podophyllum hexandrum, and Ranta- nia, certain characteristic(s) of the pollen render it unique and for the most part nullify any systematic value within the family. The pollen morphology of the Berberidaceae s. 1. is not similar to that of the Ranunculaceae, Hydrastis excepted, nor to Lardizabalaceae. There appear to be unusual examples of parallelism between the Berberidaceae and Cistaceae, and between Podophyllum and Croton.

OFFICIALPUBLICATION DATE is handstamped in a limited number of initial copies and is recorded in the Institution’s annual report, Srnithsonian Year. SERIESCOVER DESIGN: Leaf clearing from the katsura tree Cercidiphyllurn japonicurn Siebold and Zuccarini.

Library of Congress Cataloging in Publication Data Nowicke, Joan W Pollen morphology and phylogenetic relationships of the Berberidaceae. (Smithsonian contributions to botany ; no. 50) Bibliography: p. 1. Berberidaceae. 2. Palynotaxonomy. I. Skvarla, John J., joint author. 11. Title. 111. Series: Srnithsonian Institution. Srnithsonian contributions to botany ; no. 50. QKl.S2747 no. 50 [QK495.B45] 581s [583’.117] 80-21960 Contents

Page Introduction ...... 1 Acknowledgments ...... 2 Previous Treatments of the Family ...... 2 Materials and Methods ...... 6 Results and Discussion of Pollen Analyses ...... 7 Ektexine Relationships ...... 19 General Discussion ...... 21 Summary ...... 26 Literature Cited ...... 28 Tables and Figures ...... 31

... 111

Pollen Morphology and Phylogenet ic Relationships of the Berberidaceae

Joan W. Nowicke and John J. Skuarla

Introduction 600 species, with as many as 500 of these belong- ing to Berberis L. The geographical distribution of the flowering The genera most commonly regarded as be- plants presents some perplexing problems, but longing to the Berberidaceae are the following. none more so than that of disjunct or discontin- Achlys De Candolle, Berberis L., Bongardia C. A. uous genera that, by definition, occupy widely Meyer, Caulophyllum Michaux, Diphylleia Mi- separated regions. If these taxa are regarded as chaux, Dysosma R. E. Woodson, Epimedium L., monophyletic, then it follows, largely by assump- Jeffersonia Barton (Plagiorhegma Maximowicz) , tion, that either at one time their range must Leontice L., Mahonia Nuttall, Nandina Thunberg, have included intervening areas or the taxa have Podophyllum L., Ranzania Ito, and Vancouveria C. fruits and/or seeds with adaptations for long Morren and Decaisne. These genera are not con- distance dispersal. There are, in fact, floristic sidered to be a single closely related group, and relationships based upon two regions (rarely this is reflected in the fact that few systematists three) having a high number of disjunct genera have included all in one family. common to both, and it is more logical and probable that the high number is an indication Most modern generalists consider the genera as of a previously more continuous range. The floras primitive or at least unspecialized and place them of eastern North America and eastern Asia are with the Rannuculaceae, Menispermaceae, Lar- one of the classic examples, with as many as 80 dizabalaceae, and several very small families as genera having species in both regions. These gen- the Order Ranunculales or even Berberidales. era are not randomly spread among the dicots or This study of pollen morphology in the Ber- monocots, but tend to be concentrated in the beridaceae is part of an extensive and continuing more primitive families, one of which is the Ber- research project on the phylogenetic relationships beridaceae. Even in the widest sense this is a small of the Order Centrospermae. This unusual group family, consisting of 10 to 12 genera and about of families with the unique nitrogen-containing pigments, the betalains, and distinctive sieve tube Joan W. "dowzcke, Department of Botany, .Vatzonal ,2luseum oj plastids (Behnke, 1976) has pollen with an ektex- Natural Histoy, Smithsonian Institution, Washington, ine characterized as spinulose and punctate/ D.C. 20560. John J. Skuarla, Department of Botany and Micro- biology, C'niuersib of Oklahoma, Norman, Oklahoma 730l.Q. tubuliferous. This particular surface pattern,

1 2 SMITHSONIAN CONTRIBUTIONS TO BOTANY found in 85% of the taxa examined, is the pre- Chissoe at the University of Oklahoma. The scan- dominant type in each of the betalain families as ning electron micrographs were taken at the well as the two anthocyanin families, Caryophyl- Scanning Electron Microscope Laboratory at the laceae and Molluginaceae. Both authors have National Museum of Natural History. We thank regarded this ektexine type as unspecialized and Aaron Goldberg and Harold Robinson for their consider that the significance attached to it re- critical review and helpful comments. sides mainly in its high frequency. These results This research was supported in part by a grant have been published in two papers, one based on from the Smithsonian Institution Fluid Research light microscopy and SEM (Nowicke, 1975) and Fund to J. W. Nowicke and in part by National the other emphasizing TEM (Skvarla and Now- Science Foundation Grant DEB 75-19846 and icke, 1976). DEB 80-02585 to J. J. Skvarla. In the first investigation of families outside the Centrospermae, the pollen of the Plumbagina- Previous Treatments of the Family ceae, Polygonaceae, and Primulaceae was exam- ined in a study that combined and integrated The Japanese botanist Masao Kumazawa results from light microsocpy, SEM, and TEM worked on both the Ranunculaceae and Berber- (Nowicke and Skvarla, 1977). Examination of 136 idaceae for a number of years (1930a, 1930b, taxa in these families, considered to be related to 1932a, 1932b, 1935, 1936a, 1936b, 1937a, 1937b, the Centrospermae by various authors, revealed 1937c, 1938a) and published a paper in 1938 a wide range of variation in the ektexines, but not (193813) in which he reviewed and discussed the the common one in the Centrospermae. The relationship within and between the families. Ku- Polygonaceae may be one of the most palynol- mazawa (1938a:g) regarded both families as in- ogically diverse families in the angiosperms, with cluding “extremely heterogeneous types” with the variation in shape, apertures, tecta, and exine systematic affinities of some genera still unde- structure. cided. The Berberidaceae and Ranunculaceae The Order Ranunculales is the second group have peculiar anatomical features not commonly of families to be investigated palynologically for found in dicots: fused cotyledons, trimerous parts, evidence of relationships to Centrospermae. At V-shaped xylem. The developmental mode of the this writing almost 150 species have been exam- pollen grains and endosperm are more character- ined in the Ranunculaceae, 40 species in the istic of the monocots than dicots. Berberidaceae s. l., 14 in the Lardizabalaceae, Although numerous authors have considered seven in the Coriariaceae, four in the Sabiaceae, that the connecting links between the Ranuncu- and three in the Corynocarpaceae. Due to the laceae and Berberidaceae were to be found in the large number of taxa and the general conclusions transitional genera Glaucidium-Hydrastis and Podo- that the palynological data do not support a close phyllum-Diphylleia, Kumazawa (1 938a) regarded relationship among any of the above families, the the above generic pairs as widely separated, citing decision was made to treat each family in a differences in vegetative structures and anther separate publication. dehiscence, as well as the results from his most For purposes of comparison and the reader’s recent investigation of ovular structures in the convenience, we have included electron micro- two families (1938b). graphs of the common pollen type in the Centro- Kumazawa (1938a: 12) considered Nandina as spermae (Figures 1-6) as well as the predominant the “farthest deviated” genus in the Berberida- type in the Ranunculaceae (Figures 7-12). ceae; in fact, he would support family status AcKNowLEDGMENTS.-The authors wish to ac- based on the combination of vegetative and ovu- knowledge the fine technical assistance of J. L. lar characteristics as well as a type of anther Bittner at the Smithsonian Institution and W. F. dehiscence not found in any other berberidaceous NUMBER 50 3 genera. He no longer regarded Diphylleia as closely and Sabiaceae. In his list (1968:365) of orders and related to Podophyllum, the two differing in dehis- families, Podophyllaceae are indicated as belong- cence of the anthers, the pollen, and the type of ing to the Ranunculaceae, but no genera are vernation. He segregated Podophyllum as a mono- listed; Nandinaceae are a part of the Berberida- generic family, but left the fate of Diphylleia “for ceae. consideration” (1938a: 13). For the remaining Takhtajan (1969:208) separated Podophyllum genera, Kumazawa established a Berberidaceae and Diphylleia as the Podophyllaceae and also consisting of two subfamilies: Berberidoideae gave Nandina family status. The remaining genera with only Berberis and Mahonia; and Epimedioi- are assumed to be in the Berberidaceae since they deae with three tribes, Epimedeae with Epimedium do not appear elsewhere in his scheme as families. (Vancouveria?), Leontice (Bongardia?), Caulophyllum, In addition to the above families, he regarded Jeffersonia, and Plagiorhegma, Achylieae with Ach- Lardizabalaceae, Sargentodoxaceae, Menisper- lys, and Ranzanieae with Ranzania. He cited the maceae, Ranunculaceae, Glaucidiaceae, Hydras- pollen of Berberis and Mahonia as the most striking tidaceae, and Circaeasteraceae as belonging to distinction between the two subfamilies, possibly the Order Ranunculales. indicating a large phylogenetic gap between Hutchinson (1959) designated the second order them. in his division Herbaceae as Berberidales, but the Buchheim (1964) maintained the relationship Berberidaceae are very reduced, consisting of only of Diphylleia and Podophyllum by segregating the Berberis and Mahonia; Nandina is placed in a sepa- two genera in a subfamily, the Podophylloideae. rate family in this order. The remaining berberi- The second and much larger subfamily, Berberi- daceous genera are united as the Podophyllaceae, doideae, has the remaining genera, but their sim- but placed in the first order, the Ranales (Ran- ilarities and distinctions are maintained by the unculales). establishment of three tribes: Nandineae with Thorne (1974) designated the first superorder only Nandina; Berberideae with Berberis, Mahonia, in the dicotyledons as the Annoniflorae, which he and Ranzania; and the Epimedieae with Aceran- segregated into three orders: the Annonales, the thus, Achlys, Bongardia, Caulophyllum, Epimedium,Jef- largest with about 23 families; the Berberidales fersonia, Leontice, Plagiorhegma, and Vancouveria. His with six families; and the Nymphaeales with only concept of the Order Ranunculales, which he four families. The significant difference between described for the Syllabus der Pflanzenfamilien, in- Thorne’s concept of the Berberidales and that of cludes the following families: Ranunculaceae, most other generalists is the addition of the Pa- Berberidaceae, Lardizabalaceae, Menisperma- paveraceae whose distinction is partially acknowl- ceae, Nympheaceae, Ceratophyllaceae, and Sar- edged by treating it as a suborder. The other gentodoxaceae (only Sargentodoxa) . suborder, the Berberidineae, includes Lardizaba- Buchheim’s (1964) interpretations of the rela- laceae, Sargentodoxaceae, Menispermaceae, Ber- tionships of the above families are of significance beridaceae, and Ranunculaceae. Thorne main- because he also described the Magnoliales, con- tained a wide family concept, preferring to rec- sidered, at least by Hutchinson (1959), to be the ognize the traditional generic alliances as subfam- woody counterpart of the Ranunculales. Un- ilies or tribes. doubtedly his treatment of each order benefited In a more recent study utilizing comparative from the knowledge and perspective provided by serology, Jensen (1974) concluded: that Berberis the treatment of the other. and Mahonia are very closely related, even con- Cronquist (1968) placed the Berberidaceae in generic; that Podophyllum and Diphylleia are simi- the Order Ranunculales, consisting of Ranuncu- lar in serological aspect; and that Nandina is very laceae, Circaeasteraceae, Lardizabalaceae, Men- similar to the above four genera, but closer to ispermaceae, Coriariaceae, Corynocarpaceae, Berberis and Mahonia. He also suggested that these 4 SMITHSONIAN CONTRIBUTIONS TO BOTANY five genera be segregated as a unit such as rhegma (Jeffersonia), differing only in the area of subfamily, and the distinction between the above elongation at a later stage. Furthermore, the be maintained by establishing three tribes. In ovules of the basilary types are in reality slightly comparing the reactions of other genera to the dislocated towards the lateral walls, which also Berberis vulgaris antiserum and to the Podophyllum supports Kumazawa’s view that parietal placen- emodi (= P. hexandrum) antiserum, Jensen found tation is the original type. that Vancouveria and Jeffersonia indicated at least Not all of Kumazawa’s results (1938a) are rel- some similarities to each system, but Caulophyllum evant here, but those on Nandina are significant and Epimedium had only a very weak reaction to in the later discussion. It was the only genus each. Jensen also noted that Caulophyllum and investigated in which the funiculus was absent; Leontice are very similar in serological respects. in the other genera it ranged from very short to But pending further studies, he would place the prominent. Nandina had an atypical micropyle in remaining genera of the Berberidaceae, Achlys, which the outer integument did not project at the Bongardia, Caulophy llum, Epimedium, Jeffersonia, point of closure. Leontice, and Vancouveria, in a second subfamily. The following review of the size and distribu- Chapman (1936), in a study of carpel mor- tion of the berberidaceous genera is essential to phology in the Berberidaceae, used serial sections the discussion of the results of this study. If a to trace the distribution of the vascular bundles genus has been the subject of a separate study, and proposed that only an original condition of the pertinent conclusion or observations are in- three separate, spirally arranged carpels from cluded. Closely related genera are discussed to- which two distinct lines evolved would satisfac- gether. torily explain the present variation. The two lines Achlys De Candolle has two species, A. triphylla that Chapman suggested would require a fusion (Smith) De Candolle in Pacific North America, of the carpels with the loss or suppression of one and A. japonica Maximowicz from Asia. This is or two. For Berberis, Mahonia, Leontice, and Caulo- the only disjunct genus in the family with a phyllum the distribution of the vascular bundles Pacific North America and Asia distribution. could be the result of suppression of the upper Berberis L. is the largest genus in the family two carpels and an expansion of the lowest one. with as many as 500 species widely distributed in In the other line, the two fused carpels found in both the Old World and the New World. The Epimedium, Vancouveria, Nandina, Achlys, Jeffersonia, closely related Mahonia Nuttall, with approxi- Diphylleia, and Podophyllum could be derived by mately 75 species, is the second largest genus. the loss of one carpel in the process of fusion. She Although Ahrendt (1961) published an exhaus- concluded that the separation of Berberis and tive study of these two genera, it is unfortunate Mahonia with their independent evolution oc- that he did not discuss the relationships of Berberis curred very early; that the lack of a close rela- and Mahonia to the remaining berberidaceous tionship of Nandina to the other genera would also genera. indicate an early separation; that Jeffersonia and Bongardia C. A. Meyer has a single species, B. Achlys are more closely related to Epimedium than chrysogonum (L.) Grisebach, found along the Af- to each other; and that the carpel morphology of ghanistan-Iran-U.S.S.R. border. It was first de- Diphylleia and Podophyllum does not support a close scribed as a species of Leontice by Linnaeus (1 753). relationship to each other. Caulophyllum Michaux is a disjunct genus of two According to Kumazawa’s developmental very similar species, C. thalictroides (L.) Michaux studies (1938a) the basilary placentation ac- in eastern North America as far west as Nebraska corded to Berberis, Mahonia, Caulophyllum, and Ach- and Missouri, and C. robustum Maximowicz from lys is in fact a modification of the parietal placen- China, Japan, and Korea. The latter sometimes tation found in Epimedium, Nandina, and Plagio- has been treated as a variety of the former. NUMBER 50 5

Diphylleia Michaux has three species, one in For Vancouveria, Stearn (1938) recognized three eastern North America, D. cymosa Michaux, and species: V. hexandra (Hooker) C. Morren and De- two in Asia, D. grayii R. Schmidt in Japan and D. caisne, the most common and the most widely sinensis Li from China, and is a disjunct genus. distributed (California, Oregon, and Washing- Dysosma R. E. Woodson was established as a ton), and V. chrysantha Greene and V. planipetala monotypic genus based on Podophyllum pleianthum Calloni, both found in California and Oregon. Hance. At that time (1928), Woodson regarded The report of V. hexandra from Vancouver Island all the following taxa as variations of Dysosma or continental Canada is apparently erroneous; pleiantha (Hance) Woodson: Podophyllum versipelle Stearn did not see any collections from these Hance, P. veitchii Hemsley and E. H. Wilson, P. regions. difforme Hemsley and E. H. Wilson, P. esquirolii Jeffersonia Barton has two species, J. diphylla Leveille, and P. ontoi Hayata. Later: Hu (1937) (L.) Persoon from eastern North America, and J. transferred two species of Podophyllum: P. delavayi dubia (Maximowicz) Bentham and Hooker, which Franchet, now Dysosma delavayi (Franchet) Hu occurs in southeastern Manchuria, adjacent Ko- and P. aurantiocaule Handel-Mazzetti, now rea, and eastern Siberia. The disjunct status of Dysosma aurantiocaule (Handel-Mazzetti) Hu. Jeffersonia depends on the validity of the reduction Epimedium L. and the closely related Vancouveria of Plagiorhegma dubia Maximowicz; most authors C. Morren and Decaisne were treated by Stearn follow Bentham and Hooker in this, Hutchinson in a monograph published in 1938. He reduced excepted (1959). two monotypic genera, Aceranthus C. Morren and Leontice L., an Old World genus of eight to 10 Decaisne, and Vindicta Rafinesque, to Epimedium species, is sometimes given separate family status, since the type species for each was E. diphyllum the Leonticaceae (Airy-Shaw, 1966), including Loddiges; but he maintained the generic status Bongardia and Cau lophy llum. of Vancouveria, citing a group of morphological Nandina Thunberg is a monotypic genus from distinctions that occur only individually in var- China whose distinctions have been acknowl- ious species of Epimedium. edged by Chapman (1936), Ernst (1964), Ku- Stearn (1938) recognized 21 species of Epime- mazawa (1938a), and most generalists. dium, all with an Old World distribution, but remarked on the difficulty of assessing their rela- Podophyllum L. is a disjunct genus with one tionships to each other, the characteristics seem- species, P. peltatum L., very common in the eastern ing to be either on a generic level and applicable half of North America, and an uncertain number, to all, or on a species level and applicable to one. but not more than four or five species, distributed He did segregate the 21 species into eight small in eastern Asia. The present authors follow Soe- groups which he then placed in two sections jarto, Faden, and Farnsworth (1979) in the use of defined by the origin or position of the leaves: Podophyllum hexandrum Royale rather than P. emodi section Rhizophyllum in which all leaves are Wallich for the most common .4sian species. basal with only one “group” of two species; and Rantania Ito is a monotypic genus with the section Phyllocaulon in which all leaves are at- single species, R. japonica (Ito) Ito, restricted to the tached to the flowering stem, with the remaining northern part of Honshu, the largest of the Jap- seven “groups” and 19 species. Stearn (1938:428) anese Islands. Kumazawa (1937~)concluded based the four subsections, some of which include after a detailed study that this species did bear more than one “group,” on the number of leaves some resemblance to Caulophyllum and Epimedium, on the floral stem, but noted that “this is liable to but that it also had in the stamens, petals, and some variation.” After attempting to verify the fruit some of the restricted or peculiar character- identification of some voucher specimens, the istics found in Berberis and Mahonia. He proposed authors would agree. that R. japonica represented an intermediate type 6 SMITHSONIAN CONTRIBUTIONS TO BOTANY and was not as closely related to the Epimedieae bert, 1961). They seem better treated as an intermediate as he had previously thought. group but close enough to the other ranunculads to warrant The cytological data on the genera of the Ber- subfamily treatment in the Ranunculaceae (Kumazawa, 1938; Buchheim, 1964). beridaceae are almost complete and are of signif- icance to “General Discussion” herein. The fol- The most comprehensive study of the pollen of lowing reports were taken from Darlington and the Berberidaceae to date may well be a paper Wylie (1956): Achlys 2n = 12; Berberis x = 14, 2n published in 1936 which also included the Ran- = 28, 56; Bongardia x = 6, 2n = 14; Caulophyllum unculaceae and Lardizabalaceae. Kumazawa x = 8, 2n = 16; Dzphylleia x = 6, 2n = 12; (1936a) provided descriptions and discussions for Epimedium x = 6, 2n = 12; Jeffsonia x = 6, 2n = 46 genera and 230 species with 72 illustrated by 12; Mahonia x = 14, 2n = 28; Nandina x = 10, 2n line drawings. Although his terminology differs = 20; Podophyllum x = 6, 2n = 12; Rantania x = 7, somewhat from those currently in vogue, there is 2n = 14; Vancouveria x = 6, 2n = 12. Leontice no problem in translation, e.g., expansion furrow armeniaca with 2n = 14, and L. leontopetalum with being the equivalent of colpus. He recognized n = 8 + lB, and 2n = 16, were taken from four pollen types based on apertures: Type 0, Ornduff (1968). Six genera, Achlys, Dzphylleia, inaperturate; Type I, 3-zonocolpate, rarely 6-8- Epimedium, Jeffersonia, Podophyllum, and Vancou- zonocolpate; Type 11, pantocolpate; and Type veria, have chromosome number of 2n = 12. 111, pantoporate. Shce Kumazawa (1936a) re- There are two monotypic genera, Glaucidium garded Diphylleia as pantoporate, and Berberis and Siebold and Zuccarini and Hydrastis Ellis ex L., Mohonia as inaperturate, the Berberidaceae, in his that are always included in the Order Ranuncu- estimation, had all four types. lales and are associated with Berberidaceae and In a more recent contribution, Roland-Hey- Ranunculaceae but at different levels of recogni- dacker (1974) described the pollen of Berberis tion. Wettstein (1935) placed both genera in the vulgaris L. and Mahonia aquifolium Nuttall as hav- Berberidaceae; Cronquist (1968) included both ing unique helicoid colpi as well as a compacted in the Ranunculaceae; Hutchinson (1959) di- ektexine, a granular endexine, and a persistent vided the Ranunculaceae s. 1. into the Ranuncu- intine that reacted positively to tests for polysac- laceae S.S. and Helleboraceae with both genera in charides. the latter family; Takhtajan (1969) acknowledged their distinction from the Ranunculaceae, from Materials and Methods the Berberidaceae, and from each other by sepa- rate family status, Glaucidiaceae and Hydrasti- The pollen of 40 species assigned to 14 genera daceae. Thorne (1974: 187) summarized their in the Berberidaceae (Table 1) has been examined characteristics and relationships as follows. in light microscopy (LM) and in scanning elec- tron microscopy (SEM) and a limited number in Hydrastrs and Glaucidium, sometimes placed with the Ber- transmission electron microscopy (TEM). The bericadeae or as a distinct family, do have the berberidaceous pollen of three species from the Order Centro- rhizomatous habit of growth and Gluuczdium frequently has spermae (Table 2) and seven from the Ranun- only one carpel. However, their flowers have numerous stamens, many to two carpels or one carpel, no petals or culaceae (Table 3) have also been included. An- nectaries. and their leaves are simple and palmately lobed. thers were removed from herbarium specimens They are distinguishable from the other Ranunculaceae by and all material acetolyzed according to proce- the absence of nectaries, ovules with longer outer integu- dures outlined in Erdtman (1966). Samples for ment. rather fleshy fruit, and distinctive haploid chromo- SEM were vacuum coated with gold and exam- some numbers of n = 10 and 13. Their chemistry is very similar to that of the other Ranunculaceae (Hammond, ined with a Cambridge Stereoscan MK IIa, and 1955: Jensen, 1968) but they possess several alkaloids in S410, or a Coates and Welter 106B Field Emission common also with the Berberidaceae (Willaman and Schu- Microscope. NUMBER 50 7

Samples for the TEM were incorporated into attempts to evaluate and integrate the pollen agar, dehydrated through increased concentra- data with those from vegetative and floral mor- tions of ethyl alcohol, and subsequently embed- phology (Kumazawa 1930a, 1930b, 1932a, 1932b, ded in araldite-epon resins (Skvarla, 1973). Pollen 1935, 1936a, 1936b, 1937a, 193713, 1938a, 1938b; samples were stained in 0.125% OsO4 in 0.1 M Takeda, 1915; Terabayashi, 1977, 1978), carpel sodium cacodylate buffer for two hours prior to morphology (Chapman, 1936), serology Uensen, agar incorporation. Thin sections were made with 1974), and cytology as reported by Darlington diamond knives, collected on uncoated grids and and Wylie (1956) and Ornduff (1968). stained with uranyl acetate and then lead citrate. Electron microscope observations were made with Results and Discussion of Pollen Analyses a Philips model 200 transmission electron micro- scope. Pollen from three families in the Centrosper- Light slides of all samples are deposited at the mae is illustrated in Figures 1-6: Anacampseros Palynological Laboratory, Department of Bot- fifamentosa Sims (Portulacaceae), Figures 1, 2, has any, Smithsonian Institution. an aperture condition that can be described as The species examined, the collector and num- pantocolpate but the configuration and the shape ber, and country or U.S. state, as well as figure of the individual "colpi" are not always uniform, number(s) if illustrated, are given in Tables 1, 2, the tectum is spinulose and perforate: Tunica and 3. For the most part the names used are stricta (Bunge) Fischer and Meyer (Caryophylla- taken from the herbarium label or the most recent ceae), Figures 3, 4, has a pantoporate aperture annotation; each collection sampled was similar type and a tectum that is spinulose and sparsely to those remaining of a particular species as iden- punctate; Acrodon bellidiflorus N. E. Brown (Aizo- tified. For most of the disjunct berberidaceous aceae), Figures 5, 6, is 3-colpate with a tectum genera, the geographical location, North America that has been described as reticulate (Radulescu, as opposed to Asia, provided sufficient identifi- 1974), but the size of the perforations varies cation for the purposes of this study. within a sample and those illustrated in the high The investigation of the Berberidaceae is part magnification micrographs are, in fact, unusually of a study comparing pollen morphology of the large. Ranunculales with that in the Centrospermae The possibility of a close relationship between and detailed measurements and/or descriptions the Berberidaceae and any families in the Cen- are usually not included. However, within the trospermae is unlikely, but most taxonomists con- Berberidaceae s. 1. a significant difference in size, sider the Berberidaceae and Ranunculaceae to be as indicated by polar length, does distinguish two closely related and for this reason the latter family groups of genera from each other. Size could have is illustrated by seven species: in SEM, Figures been a factor in the transfer of Hydrastis L. from 7-18, in TEM, Figures 115-123, and in the data the Ranunculaceae to the Berberidaceae. For this of Table 3. It should be noted that the following reason the longest dimensions of ten grains in discussion of Ranunculaceae pollen is based on collections of Berberidaceae and Ranunculaceae the examination of almost 150 species (Nowicke were recorded. The high, low, and the mean are and Skvarla, unpublished data). given in Tables 1 and 3, respectively; these figures Ranunculus oreophytus Delile (Figures 7, 8) and should be treated with reserve in view of the small Hepatica transsilvanica Fuss (Figures 15, 16) have a sample size. pantocolpate aperture condition: Clematis heraclei- The results from the pollen investigation of folia De Candolle (Figures 9, 10) has apertures each genus are presented first, along with a dis- best described as pores but neither their shape cussion of the generic relationships as indicated nor their distribution over the surface of the grain by palynology. A general discussion follows and is uniform; Glaucidium palmatum Siebold and Zuc- 8 SMITHSONIAN CONTRIBUTIONS TO BOTANY carini (Figures 1 1, 12), Batrachium aquatile Dumor- and ours do not conflict (Nowicke and Skvarla, tier (Figures 13, 14), and Hamadryas magellanica unpublished data). Lamarck (Figures 17, 18) have a 3-colpate con- In addition to the Ranunculaceae all modern dition. All six species have, in a general sense, a generalists (see “Introduction”) consider the Lar- spinulose and punctate/perforate tectum. dizabalaceae to be closely related to the Berberi- Many of the Ranunculaceae taxa examined in daceae. The results from pollen studies do not thin section have prominent or very large colu- support this contention. For the most part the 14 mellae, e.g., Ranunculus oreophytus (Figure 115), species examined in the Lardizabalaceae have Batrachium aquatile (Figure 1 19), Hamadryas magel- similar pollen: 3-colpate, the tectum can be psi- lanica (Figure 121), and to a lesser extent in late with punctae, or indentations, or finely punc- Clematis heracleifolia (Figure 1 17) and in Hepatica tate-striate; in thin section the tectum is the most transsilvanica (Figure 120). Glaucidium palmatum conspicuous unit of the ektexine, the columellae (Figure 118) illustrates reduced columellae, a con- are diminutive in size and sparsely distributed, dition that is found in Aconitum (Nowicke and the foot layer is very thin, the endexine is well Skvarla, unpublished data) and Adonis (Nowicke developed in the region of the colpus but very and Skvarla, 1980, fig. 158). For the most part reduced in the mesocolpus (Skvarla and Nowicke, the pollen in this family has a well developed unpublished data). endexine layer (Figures 115, 116, 118, 120, 121). We are of the opinion that most of the taxo- The perforations of the tectum, obscure in SEM nomic difficulty associated with the Berberida- (Figures 8, 10, 12, 14, 16, 18), are more conspic- ceae s.1. can be attributed to exceptional and uous in thin section. Hydrastis canadensis L. (Fig- restricted sporophytic characteristics: the per- ures 122, 123), will be discussed with the tribe ianth absent in Achlys, multiseriate in Nandina; Epimedieae. petals spurred and/or retroflexed in Epimedium There is an unusual characteristic, demon- and Vancouveria; stamens tactile in Berberis, Ma- strated only in transmission microscopy, that is honia, and Rantania; filaments connivent in Van- common to some Berberidaceae and to some couveria; the fruits bladder-like in Bongardia and Ranunculaceae: columellae-shaped ektexine Leontice, follicle-like in Epimedium and Vancouveria, units penetrate the endexine in the apertures horizontally cleft in Jeffersonia; seeds large, fleshy, (hereafter referred to as aperture columellae) . and glaucous in Caulophyllum; the habit woody in This condition has been found in all species of Berberis, Mahonia, and Nandina, and semisucculent Ranunculus examined in TEM, illustrated here by in Bongardia and Leontice. R. oreophytus (Figure 116), in Batrachium aquatile Close scrutiny of the entire sections “Results (Figure 119), and in Hepatica transsilvanica (Figure and Discussion of Pollen Analyses” and “General 120). It is also present in some species of Anemone Discussion” will reveal that the authors have not and some species of Clematis (Nowicke and been consistent, from genus to genus, on the Skvarla, 1980, figs. 155, 156). In the Berberida- degree of importance attached to the same paly- ceae s. l., it is most conspicuous in the two closely nological characteristics. The endexine layer is a related genera Epimedium and Vancouveria (Figures case in point: it is taxonomically significant in 149, 154, 155, 159, 160, 161; see discussion of Nandina, but insignificant in all remaining genera; these genera). another, the condition in which the pollen is shed Santisuk (1979) investigated the pollen of 124 (monads versus tetrads) is significant in only one taxa in the tribe Ranunculeae using light and species and insignificant in all the remaining ones. scanning electron microscopy and established 10 The above statements are not intended to justify pollen types “based on the types of columellae or validate any or all of the generic realignments and aperture and on the nature of the tectum” proposed here, but to point out that we have been (Santisuk, 1979:3). For the same taxa his data forced to evaluate irregular pollen data against NUMBER 50 9 an already uneven background of floral and veg- nated as preformed but the configuration of the etative morphology. furrows is not uniform, e.g., Berberis vulgaris L. Within the Berberidaceae the following range (Figure 23) and Mahonia nervosa (Pursh) Nuttall of variation was found in apertures, tectum, and (Figures 27, 28). exine structure. There are three types of aper- Kumazawa, who examined four species of Ber- tures, irregular or spiral found in Berberis and beris and two of Mahonia, made the following Mahonia, 6-pantocolpate in Ranzania, and 3-col- points about their germinating apparatus (1936a: pate, found in all the remaining genera examined. 35). The tectum can be psilate, punctate, punctate- striate, finely reticulate, striate, striate-reticulate, (1) The margin of the furrow expanded is irregularly two layers of striae, “random” rods (small, var- denticulated in surface view, suggesting the breaking of the iously distinct rods with one end projecting out- exine. (2) In some cases no furrows are found on the surface of ward), spinose, or gemmate, with some of the the shrunken and swollen pollens. categories grading into another. The structure of (3) There is no rule concerning the position of the furrow. the exine is typical of angiosperms: foot layer, (4)The shrunken pollens with furrows may represent the columellae, and tectum for all genera except Ber- reshrunken form of the swollen pollens. beris, Mahonia, and Rantania, which have an un- stratified exine. The surface of the exine is unspecialized and Berberis L. (Figures 19-24, 124-132), and Ma- randomly variable: punctate, psilate and punc- honia Nuttall (Figures 25-30, 133-138) are consid- tate, psilate, or punctate-striate occur within a ered to be closely related by generalists and spe- species or even within a sample, e.g., Mahonia cialists alike (see “Introduction”). The pollen of fremontii (Figures 25, 26). all species examined (Table 1) in these two genera In thin section the exine structure of Berberis is very similar (Figures 19-30, 124-138) and can- (Figures 124- 132) and Mahonia (Figures 133- 138) not be distinguished from each ,other using LM, are indistinguishable from each other. The ektex- SEM, or TEM. Of far greater significance than ine is nearly amorphous and not organized into the expected similarity of the pollen is the fact typical foot layer, columellae, and tectum units. that the unifying characteristics are unmistakably Because of this lack of structural organization the primitive and serve to emphasize the extent of pollen of Berberis and Mahonia was examined in the separation of Berberis and Mahonia from the both the acetolyzed and unacetolyzed conditions. remaining genera. Following acetolysis, the ektexine contains nu- Within the taxa examined in Berberis and Ma- merous channels and cavities of a highly pleo- honia the shape and extent of formation of the morphic nature, which for purposes of this dis- apertures, i.e., some grains appear inaperturate, cussion are classified as follows: (1) some com- is highly variable but could be classified as either pletely bisect the ektexine (Figures 128, 130, 136), irregular or spiral. The irregular category applies (2) some partially bisect the exine and occur with to grains with cracks or breaks, e.g., Berberis fendleri sufficient frequency as to resemble highly irregu- A. Gray (Figures 2 1, 22), B. grandflora Turczani- lar columellae (Figures 125, 126, 128, 131-133), now (Figure 24), Mahoniafremontii (Torrey) Fedde and (3) some channels appear as irregular, iso- (Figures 25, 26), M. oiwakensis Hayata (Figure lated, open holes in the ektexine (Figures 125, 29), and M. haematocarpa (Wooton) Fedde (Figure 126, 128, 129, 131, 133). Without acetolysis there 30). Occasionally the “furrows” delimit the sur- appears to be a fourth category of channels: face of the grain into plate-like areas, e.g., M. extremely short, abundant, randomly oriented fremontii (Figure 25) or M. oiwakensis (Figure 29). and “closed” or filled with electron-dense mate- More rarely some grains have areas with pebble- rial (Figure 124). shaped pieces of exine, e.g., B. ilicfolia Forster The primary difference between the examina- (Figure 19). The spiral apertures could be desig- tion of acetolyzed and unacetolyzed ektexine im- 10 SMITHSONIAN CONTRIBUTIONS TO BOTANY ages is that of stain density: the images from ber of 56; this does not deny the existence of acetolyzed grains have considerably greater den- polyploidy in Mahonia. sity than those from unacetolyzed grains. This no Our results agree with those of Roland-Hey- doubt explains the appearance or visibility of the dacker (1974), except for her characterization of fourth category of channels. Whether or not the all(?) apertures as spiral. lack of staining of unacetolyzed pollen was due The presence of irregular apertures, a surface to the dried (i.e., herbarium) nature of the pollen that is psilate or nearly so, and above all the before processing is not known and certainly it unstratified exine, are primitive characteristics would be useful in future work to collect pollen and are in agreement with the phylogenetic po- directly from the field and immediately process sition accorded Berberis and Mahonia by most sys- for TEM. This might clarify the nature of the tematists. thin electron-translucent layer or membrane(?) Ranzania japonica (Ito) Ito (Figures 139- 148), found on the ektexine of unacetolyzed Berberis although widely cultivated in the United States, (Figure 124) pollen. The extent of such a layer or has a very restricted distribution, northern Hon- specificity (it was not observed in Mahonia, Figure shu, Japan. Kumazawa (1936a) described the 138) as well as its equivalence to the fibrous- pollen and illustrated it with line drawings granular extra-ektexinous layer as indicated in (1936a:fig. 67,, 671, 672), but for purposes of the acetolyzed preparation of Berberis ilicifo lia comparison the authors preferred to examine the (Figure 13 1) is not clearly understood at this time. pollen of Ranrania in SEM and TEM. Despite its appeal as an ornamental plant, none In the aperture regions the ektexine is repre- of the major U.S. herbaria had specimens iden- sented as knobs or isolated fragments (Figures tified as such. The material finally obtained (Ta- 127, 132, 134-136). It appears identical in aceto- ble 1) consisted of one pollen sample sent from lyzed and unacetolyzed preparations. Japan by Terabayashi; one flowering specimen, The endexine in Berberis (Figures 124- 127, 130- partially dried, from the garden of a private 132) and Mahonia (Figures 133-137) is prominent individual in New York State, Epstein s.n.; and and consists of a fibrous-granular layer; the inner all material at the Royal Botanic Gardens, Kew, surface is smooth and/or uniform but at the only two collections, one from a cultivated plant, interface with the ektexine the endexine appears the other a sterile specimen collected by Tukeda to partially fill or encroach upon the cavities and s.n., 27 August 1905 in the vicinity of Mount channels. It is present in the apertures and ap- Shirouma, in Honshu. pears to support the ektexine fragments. The The sample from Japan (Terabayashi 154 KYO) endexine is not noticeably altered by acetolysis. was examined in LM and SEM and is illustrated The presence of irregular cavities (Figures 124- in Figures 139 and 140. Although the grains were 132) and the concomitant segregation of material not numerous, they were uniform in size, shape, suggest an early stage of columellae development. aperture type, and the surface of the exine, and In the six species of Berberis, the mean diameter the two in Figure 139 are representative. They ranges from a low of 38.4 pm in B. uulgaris to a also appear to be a dyad (see TEM discussion of high of 50.6 pm in B. grandiflora; in the four species Figure 147). The configuration of the six colpi is of Mahonia, the low is 34.1 pm in M. haematocarpa such that there are four threefold axes delineating and 33.8 pm in M. oiwakensis and the high is 51.2 the surface of the grain into four triangular areas. pm in M. neruosa. This degree of size variation is Most of the colpi have rounded ends and the not unexpected since Berberis is known to have resulting uniform width gives the impression of a tetraploid entities as well as diploid; Mahonia has precise aperture formation. The opening is fre- not, to our knowledge, been reported as having quently blocked by a protruding wedge of exine. either a haploid number of 28 or a diploid num- The surface of the grain is psilate. NUMBER 50 11

The cultivated specimen from New York was endexine. The ektexines in the New York State examined in LM, SEM, and TEM, but illustrated collection (Figures 142- 143) are highly channeled here only in TEM, Figures 142, 143, and 146. In and virtually indistinguishable from sections of the SEM preparation about one-third of the acetolyzed pollen from Berberis and Mahonia (Fig- grains appear to be sterile, based on the much ures 125- 136). The endexine, however, in contrast smaller size. The remaining material, frequently to Berberis and Mahonia, is proportionately thicker collapsed or ruptured, etc., is also variable in size, and contains fragments of the ektexine (Figures but intact grains measure 25-35 pm in diameter. 142-143). Further, the outer (i.e., distal) surface The apertures are 6-pantocolpate, and almost all of the endexine appears lamellate in some sections grains, including the smaller ones, manifest this (Figures 142-143). The New York State collection condition to some extent, i.e., less than six fur- apparently contained some heteromorphic or rows, the furrows extended so that some grains anomalous pollen: the oblique section represented have a wide angle L-shaped opening but even in by Figure 146 is of a whole pollen grain and collapsed and ruptured material the furrows are indicates an ektexine that appears more gemmate straight and not irregular cracks or breaks as in or nodular than the amorphous structures illus- Berberis and Mahonia, Figures 19-30. trated in Figures 142-143. The ektexine from Most of the minimal pollen sample from the pollen of the Kew collection (Figures 141, 144- cultivated collection (K) was prepared for TEM 145) differs from the New York State collection (Figures 141, 144, 145, 147, 148) but a small in that it appears to contain extremely short, fraction was acetolyzed and examined in SEM. narrow to broad, irregular columellae and a thin This material consists of a single expanded grain, but distinguishable foot layer (see particularly ca 19 pm in diameter, and a half dozen clusters Figure 145). The sections represented by Figures of collapsed grains, with as few as three and as 147 and 148 include dyad representatives in this many as ten. None of the grains had any evidence collection and indicate that binding of adjacent of apertures, neither the irregular cracks of Berberis grains is by fusion of their ektexine surfaces. Since and Mahonia, Figures 19-30, nor the 6-pantocol- pollen samples of Rantania were so limited we pate type as illustrated in Figure 139. The surface were unable to prepare any samples without ace- of the collapsed grains has a remarkably uniform tolysis. texture, similar to small pebbles with no space Both Kumazawa (1938a) and Buchheim (1964) between adjacent ones. had very similar concepts of a tribe Epimedieae. The pollen morphology in the collections of The former placed Epimedium with Leontice, Cau- Terabayashi and the cultivated one from New lophyllum,Jeffersonia, and Plagiorhegma as the tribe York agrees with that described and illustrated Epimedieae, but it is unclear whether Vancouueria by Kumazawa (1936a). is reduced to Epimedium, and whether Bongardia is The pollen morphology found in the cultivated reduced to Leontice, since neither genus is listed collection from Kew is perplexing due to the under any other subfamily or tribe. The latter apparently complete absence of apertures. None author included Achlys, Bongardia, Caulophyllum, of the grains split or ruptured, nor did the pattern Epimedium, Jeffersonia, Leontice, and Vancouveria in of collapse suggest internal lines of weakness. If the tribe Epimedieae. The only difference is that the pollen is considered to be anomalous or ab- Kumazawa placed Achlys in its own tribe, Ach- normal, the lack of any other aberrations (in the ly ieae. limited material examined in SEM), such as small All of the above genera have grains that are 3- grains, is in itself peculiar. colpate, the colpi long and narrow, and have an The two collections of Rantania examined by incomplete tectum. The structure of the exine in TEM are basically similar in that both show a all of these genera conforms with the common nearly amorphous ektexine and fibrous-granular type in the angiosperms: foot layer, columellae, 12 SMITHSONIAN CONTRIBUTIONS TO BOTANY and tectum, with the endexine present in varying sonia Barton (Figures 43-48, 93, 95, 98, 107, 170- amounts and distribution. 174), and Vancouveria planipetala Calloni (Figures The present authors’ use of the tribe Epime- 37, 38, 101, 102, 165-169) very likely exists in dieae is more restricted and includes Achlys, Epi- Epimedium, but not in the specimens available for medium, Jeffsonia, and Vancouveria. this study. Epimedium L. (Figures 31-36, 94, 99, 104, 105, Epimedium membranaceum K. Meyer (Figure 94) 110, 112, 114, 149-156), the largest genus in the and E. sagittatum (Siebold & Zuccarini) Maxi- Epimedieae with at least 25 species distributed in mowicz (Figure 99) have a perforate tectum in the Old World, has a range of variation in the which only the angular shape of the perforations tectum with phylogenetic implications since it and occasional long ridge (muri or striae?) indi- links various entities of the family. For the most cate that it is a modification of the striate-reti- part we follow Stearn’s (1938) treatment of Epi- culate type, medium and Vancouveria. Four species, Epimedium brevicornu (Figures 35, It should be emphasized that the designation 105, 150), E. cremeum (Figures 114, 151-154), E. applied to the tectum in each of the following diphyllum (Figures 3 1, 32, 149), and E. grandifllorum species is based on the most common form. Al- (Figures 112, 155, 156) were examined in thin most all samples had sufficient variation to indi- section, and found to be almost indistinguishable cate a link to other species. from each other. The endexine is present only in Epimedium diphyllum (Morren & Decaisne) Lod- the apertures and is penetrated by aperture col- diges (Figures 31, 32, 149), E. cremeum Nakai umellae (see previous discussion of Ranuncula- (Figures 1 14, 15 1- 154), E. grandiflorum Morren ceae). This condition is illustrated in Figure 149 (Figures 112, 155, 156), and E. sempervirens Nakai of Epimedium diphyllum and in the higher magni- (Figures 33, 34, 110) all have a tectum consisting fication sections in Figure 154 (E. cremeum) and of small rods with one end projecting as a very especially in Figure 155 (E.grandzjorum). The foot small tip. These rods are illustrated best in the layer is thin, irregular and with notable radial high magnification SEMs (Figures 32, 34, 110, channels (Figures 150, 153, 156). Columellae are 112, 114). Although the distribution of the rods short and fairly regular. The structure of the appears to be random, there are some grains in tectum in Figures 149, 153, 154, and 155 is con- which the free tips are arranged like the vanes of sistent with that depicted in SEM (Figures 3 1-34, a pinwheel. This configuration is also found in 110, 112, 114); the gaps reflect the loose packing Podophyllum peltatum (Figures 77, 78, 109, 11 1, of the rods, and the small peaks reflect their free 113). The similarity of the pinwheel tectum to tips. In Figures 150, 151, and 152 the unattached the crotonoid one (Lynch and Webster, 1975) is dots or circles represent a cross section of the free discussed under “Ektexine Relationships” at the tip. end of the palynological section. The mean polar length calculated for each of Epirnedium alpinum L. (Figures 36, 104), classi- the eleven collections of Epimedium (Table 1) fied here as having a striate-reticulate tectum, ranges from 28.2 pm for E. diphyllum to 33.9 pm nonetheless has lost some of the distinction of the for E. brevicornu and E. sagittatum. individual striae. In Epimedium and closely allied genera-Achlys The tectum of Epimedium brevicornu Maximow- (Figure 177), Jeffersonia, Vancouveria (Figures 157- icz (Figures 35, 105, 150) is also classified as 160, 163)-and in all material of Bongardia (Fig- striate-reticulate, but, of the species of Epirnedium ure 193), Caulophyllum (Figures 184, 188, 189), and examined, it has, at least in some grains, the of Leontice (Figures 180-183) that was examined longest and most distinct striae. A predominantly in thin section, the foot layer has radially oriented striate condition such as that found in Achlys channels. These channels separate the foot layer DeCandolle (Figures 49-5 1, 96, 175- 179),Jeffer- into units that appear to be the result of colu- NUMBER 50 13 mellae expanding laterally at the base. Since the phylla (L.) Persoon (Figures 46-48, 98, 107, 170, foot layer is formed developmentally in this man- 17 1) and the Asian representative,]. dubza (Max- ner, it seems more likely that these channels imowicz) Bentham and Hooker (Figures 43-45, should be interpreted as evidence of incomplete 93, 95, 172-174), have grains that are 3-colpate fusion as opposed to the idea that they have with a striate ektexine. formed in an already solid wall. In each of five collections examined (Table 1) The three species of Vancouveria Morren and there are some grains in which the individual Decaisne (Figures 37-42, 97, 100-102, 157- 169) stria appears to consist of an outer or surface have ektexines in which the variation illustrates section and an inner or subsurface section. In the a near perfect continuum in the degree of distinc- high magnification SEMs of both species (Figures tion of the individual striae: V. hexandra Morren 45,48, 93, 95, 98, 107), at least some of the surface and Decaisne (Figures 39, 40, 100, 160-164) is striae are branched and can be traced to the point finely striate-reticulate; V. chrysantha Greene (Fig- where they sink, alter direction, and become part ures 41, 42, 97, 157-159) is striate-reticulate with of the inner layer. the striae more prominent than V. hexandra; and One logical interpretation of such exine struc- I.’. planipetala Calloni (Figures 37, 38, 102, 165- ture is that it provides a strong wall by the 169) has a & striate ektexine. lamellation of cross grained layers since the long Each species was examined in thin section: axis of the striae in the outer layer is at right Vancouveria chrysantha, Figures 157- 159; I.’. hexan- angle or less to the long axis of the striae in the dra, Figures 160-164; and V. planzpetala, Figures inner layer. 165-169. ,411 are similar to Epimedzum, Figures In Figure 45 ofJeffersonia dubia and especially 149- 156: the endexine is restricted to the aperture in Figure 48 of J. diphylla the wider spacing of the regions and has aperture columellae, and there striae in both layers produces the effect of one are channels in the foot layer. Differences in the layer of slats upon another. In the Pennsylvania structure of the tectum among the three species collection of J. diphylla (Figures 47, 48, 98), the are in agreement with the differences illustrated surface striae are shorter, are not closely packed by SEM: the more open striate-reticulate pattern and are deposited in a patchwork design, whereas of V. chrysantha, Figure 97, is reflected in the in the New York collection (Figures 46, 107) the irregularity and gaps of the tectum, Figures 157- surface striae are longer, more densely packed, 159; the finely striate-reticulate pattern of V. hexandra, Figures 40 and 100, is reflected in the and with a nondescript or interwoven pattern. more continuous tectum illustrated in Figures 160 Most of the grains in each collection of]. dubia and 163: the mostly striate pattern of V. planipe- (Figures 43-45, 93, 95) have tecta in which the tala, Figures 38, 101, and 102, reflects this condi- surface striae are parallel; in Figure 93 of the tion in Figure 166 and in the lower grain in Korean collection some of the distinction of the Figure 169 in which the tectum appears to consist individual striae has been lost. of a “string of beads” due to the mostly parallel This particular tectum type, designated here as striae being cut at right angle. “two layers of striae,” is widely distributed in the Within Vancouveria the difference between the dicots: Aceraceae (Clarke and Jones, 1978; Bies- mean polar length of the two collections of V. boer, 1975), Cneoraceae (Lobreau-Callen et al., hexandra, 34.6 pm for Allen 66 and 39.4 pm for 1978), Cistaceae (Saenz de Rivas, 1979; Nowicke Ebert s.n., was greater than differences among the and Skvarla, unpublished data), Gentianaceae three species; however, both collections were cited (Jonsson, 1973), Leguminosae (Graham and Bar- as V. hexandra by Stearn (1938). ker, in press; Larsen, 1975). The American species of Jeffersonia Barton, In TEM, the pollen ofJe&sonia (Figures 170- (Figures 43-48, 93, 95, 98, 107, 170-174),J. di- 174) has a narrow, smooth to fragmented endex- 14 SMITHSONIAN CONTRIBUTIONS TO BOTANY ine, the foot layer appears very uneven, irregular, 46-48, 107) and Asian (Figures 43-45, 93, 95) and has occasional channels, and the columellae taxa supports the congeneric status of Jeffersonia are not well formed. In J. diphylla (Figure 171) the and Plagiorhegma. This does not, however, deny oblique angle of section has somewhat distorted any palynological relationship between Jeffsonia the characteristics of the various layers but the diphylla and Podophyllum peltatum: there are occa- endexine is present in non-apertural regions. In sional grains in J. diphylla (Figure 107) with a Figure 172 of J. dubia, the median section (at right tectum that is similar to some grains of P. peltatum angle to the long axis) depicts the endexine as in which one of the pinwheel vanes (small rods) thin and fragmented except near the apertures. is predominant (Figure 108). The foot layer is not uniform and has channels, The genus Achlys De Candolle is represented and the tectum consists of almost touching cir- by numerous collections of A. triphylla (Smith) De cular units, which is how the mostly parallel striae Candolle (Figures 49-51, 96, 175-177), but only of J. dubia would appear in cross section (see also by a depauperate type collection of A. japonica Vancouveria planipetala, Figure 169). In Figures 173 Maximowicz (Figures 178, 179). The two collec- and 174 the endexine is more uniform than in tions examined of the American species have Figure 172. In Figure 174 there are small units of grains that are 3-colpate and have a striate tec- 3, 4, 5, or even 6 circles connected below by a tum. The only pollen sample of A. japonica, Tera- solid line. This probably represents a cut that is bayashi 209 (KYO), was unsatisfactory due to a at right angle to the outer striae and parallel with paucity of material and evidence of sterility. part or all of an inner stria. Some of the grains, however, are 3-colpate with The striate tectum of Jeflersonia pollen (Figures a tectum similar to that of A. triphylla (Figure 49), 43-48, 93, 95, 98, 107) is similar to that of Van- i.e., coarse striae mostly parallel to each other. couveria planipetala (Figure 102), Achlys triphylla In TEM (Figures 175-179), the endexine is (Figures 49-51, 96) and especially to that of thin in the mesocolpal regions and thicker near Hydrastis (Figures 52-54). Comparison of the high the apertures (Figure 175), the foot layer uneven magnification SEMs of J. dubia (Figure 45) and J. with occasional channels and is much thicker diphylla (Figure 48) with some whole grains of than the endexine. The columellae are narrow Hydrastzs (Figures 53 and 54) reveals the same and short, the tectum is thick and the striae type of structure: two layers of striae. The size of appear to be closely packed. As in Jeffirsonia the Jeffersonia pollen (the mean of the five collec- (Figures 170- 174) none of the sections of Achlys tions, reported in Table 1, varies from 29.3 pm to have clearly defined aperture columellae. 33.9 pm), would support a closer relationship to In comparing the striate grains of Achlys, Jeffer- Achlys, Epimedium, Vancouveria, and even Hydrastis Sonia, and Vancouveria planipetala, the striae in Ach- t han to Bongardia , Caulophy llum , and Leont ice. lys and in V. planipetala are usually parallel and None of the sections of Jeffsonia (Figures 171- more compacted than in either species of Jeffer- 173) have apertures with ektexinous material that Sonia. All of the above have the same fundamental could be designated, with confidence, as aperture ektexine structure but modifications in the dep- columellae (see discussion of Ranunculaceae, osition of the striae produce variation in the Epimedium, and Vancouveria). tectum. There are, in fact, grains in each of the The pollen morphology of Jeflersonia is taxo- collections examined that have striae similar to nomically significant in the following ways: the that in Figure 43, and are indistinguishable from distinction of J. diphylla (Figures 46-48, 98, 170, each other (Calder and Savile 8323 and Terabayashi 17 1) from Podophyllum (Figures 73-78, 8 1-84, 9 1, 209 of Achlys excepted). 106, 109, 11 1, 113, 198-202) supports the separate Although the two collections of Achlys triphylla generic status accorded by Barton (1 793); the had a high incidence of sterility, which could similarity of the pollen in the American (Figures negate a size characteristic, the longest grains in NUMBER 50 15 the Evert sample are 28.6 pm and in the Calder and the Ranunculaceae, as well as the close relation- Savile sample, 33.0 pm. ship with Glaucidium. The pollen morphology of Hydrastis canadensis In Leontice L. (Figures 61-66, 69-72, 180-183), L. (Figures 52-54, 103, 122, 123) has considerable an Old World genus with as many as 10 species, taxonomic significance: the striate-reticulate tec- the pollen of L. altaica Pallas (Figures 61, 62), L. tum (Figures 52-54, 103, 122 and 123) distin- armeniaca Boivin (Figures 65, 66, 180-182), L. guishes this species from all other Ranunculaceae eversmannii Bunge (Figures 7 1, 72), L. leontopetalum examined, 150 species and 44 genera (illustrated L. (Figures 69, 70, 183), and L. odessana Fischer here by Figures 7-18, 115-121), including Glau- (Figures 63, 64) were examined. All have similar cidium (Figures 11, 12, 118), with which it is grains: the colpi are very long, almost to the poles frequently paired. (Figures 61-63,65,69, 71, 72); the margins some- The two collections (Table 3) examined in times undulate (Figure 61); the tectum is almost SEM have grains that are slightly different, but continuous in L. altaica (Figures 6 1 , 62) and in L. this type of variation is not uncommon in the odessana (Figures 63, 64), but in L. armeniaca (Fig- Berberidaceae (nor Ranunculaceae). The collec- ures 65, 66), L. leontopetalum (Figures 69, 70), and tion from Arkansas (Figures 52 and 103) is similar in L. eversmannii (Figures 71, 72), the tectum is to some grains of Epimedium alpinum (Figure 104), incomplete and the lumina are smaller near the E. breuicornu (Figure 105), and Jeffersonia diphylla colpi, and/or larger in the mesocolpus. (Figure 107). The collection from Ohio, illus- In thin section Leontice armeniaca (Figures 180- trated by a polar and equatorial view (Figures 53 182) and L. leontopetalum (Figure 183) have a well- and 54), has a tectum that closely resembles that developed endexine, a foot layer with radial chan- found in many grains of Jeffersonia diphylla (Figure nels, small irregular columellae, and a thick, per- 48), in which two layers of striae can be seen. for ate t ec t um . In thin section (Figures 122, 123), the structure In the disjunct genus Caulophyllum Michaux of the exine in Hydrastis is also different from that (Figures 55-60, 184-189), two collections of each in the Ranunculaceae (Figures 115-121). The of the two entities have been examined (Table 1). prominent columellae and more or less continu- The results are ambiguous with reference to the ous tectum that characterize many Ranuncula- status of the Asian taxon, either as a species, C. ceae examined in thin section (Skvarla and Now- robustum Maximowicz (Figures 55, 56, 59, 184), or icke, unpublished data) are not found in Hydrastis. as a variety of the American species, C. thalzctroides This conforms with impressions based on SEM (L.) Michaux (Figures 57, 58, 60, 185-189). micrographs. All four collections (Figures 55-60) have very Achlys, Epimedium, Hydrastis,Jeffersonia, and Van- similar pollen, long narrow colpi (Figures 55, 57) couueria have pollen with 3-colpate apertures, a with an incomplete or reticulate tectum (Figures tectum that is striate or striate-reticulate with 55-60), and would be difficult to distinguish from overlapping variation, an endexine (mostly) in each other using light microscopy. the region of the aperture, channeled foot layer, In TEM (Figures 184-189), both species of and a thin tectum. Caulophyllum have a consistent endexine, readily On the basis of all taxa examined to date in defined in the mesocolpal regions (Figures 184, the Order Ranunculales, more than 250 collec- 189) as well as in the colpus (Figures 184, 188), tions in Ranunculaceae, Berberidaceae, and Lar- an irregular foot layer, short columellae, and an dizabalaceae, only five genera, Achlys, Epimedium, incomplete tectum. Channels (Figure 184) or Hydrastis, Jeffsonia, and Vancouveria (four of which even gaps (Figure 189) are present in the foot have three species or less), have the pollen mor- layer. phology described above. As such the palynolog- The pollen of Caulophyllum closely resembles ical data challenge the inclusion of Hydrastis in that of Leontice. The tecta of C. robustum (Figure 16 SMITHSONIAN CONTRIBUTIONS TO BOTANY

55) and C. thalictroides (Figure 57) are almost Typically, the endexine is prominent in the region indistinguishable from L. leontopetalum (Figure 69) of the colpus (Figures 190, 192, 196), and much and L. euersamannii (Figure 71). The exine struc- thinner, almost absent, in the mesocolpus. ture of C. robustum (Figure 184) is almost indistin- Certain characteristics of the pollen support a guishable from that of L. leontopetalum (Figure relationship among Bongardia, Caulophyllum, and 183). Both genera have larger pollen, the mean Leontice. All three have large grains (relative to polar lengths of the five species of Leontice as listed Achlys, Epimedium, Hydrastis, Jeffersonia, and Van- in Table 1 are 48.8 pm, 54.7 pm, 43.7 pm, 55.1 couveria) and a tectum that is reticulate. The pm, and 46.5 pm, and in the two collections of difference between the tectum of Bongardia-an- each species of Caulophyllum, 51.5 pm, 50.2 pm, gular-shaped lumina and uniformly thin “muri” 48.9 pm, and 45.1 pm. without small perforations (Figure 68)-and that The pollen of the monotypic genus Bongardia found in Caulophyllum and in Leontice-circular C.A. Meyer (Figures 67, 68, 190-197) has grains lumina and “muri” that are variable in size but that are 3-colpate with an incomplete tectum of frequently larger than the lumina (Figures 56, an apparently reticulate configuration. The high 58-60, 70, and 72)-suggests different origins. magnification (X 7500) SEM of the surface (Fig- The variation in the size of the lumina in Caulo- ure 68) and the tangential thin section (Figure phyllum and Leontice indicates derivation from a 195) illustrate lumina or perforations (?), angular tectum that was continuous, whereas in Bongardia in shape and not noticeably larger in the meso- the restriction of a tectum to the distal fusion of colpal region, and “muri,” without small perfo- long, narrow, and evenly distributed columellae rations, all of which could indicate a modification indicates that this condition was original, or if it of an originally striate-reticulate tectum. has evolved from a continuous tectum, then a Bongardia chysogonum (L.) Grisebach (Figures very long period of time must have occurred. 67, 68, 190-197) was first described as a species However subtle the above distinctions (long col- of Leontice and in SEM the tectum of Bongardia is umellae, angular lumina, thin muri) may appear remarkably similar to that found in some grains to nonpalynologists, the data reinforce the sepa- of at least one species of Leontice, L. altaica. The rate generic status accorded Bongardia on other larger size (the two collections recorded in Table bases. 1 have a mean of 48.9 pm and 54.5 pm) would None of the material examined in Bongardia, align Bongardia with Leontice and Caulophyllum. Caulophyllum, and Leontice (two, four, and five However, in thin section in TEM (Figures 190- collections, respectively) have any grains that 193, 195-197) and fracture in SEM (Figure 194), might indicate a relationship to the tectum in the the pollen of Bongardia can be distinguished from tribe Epimedieae, and Podophyllum peltatum. Achlys, all other taxa examined in the family, including Epimedium, Jeffersonia, Vancouveria, Bongardia, Cau- L. altaica, by the presence of long columellae LophyLlum, and Leontice do have in common the (Figures 190, 193, 194, 196). which may account channeled foot layer. for 75% of the thickness of the ektexine, whereas There are a number of generic pairs in the in Leontice (Figures 180, 183) or in Caulophyllum Berberidaceae that undoubtedly are closely re- (Figures 184, 188, 189) the columellae are very lated, Berberis (Figures 19-24) and Mahonia (Fig- short, making up less than 20% of the total thick- ures 25-30), Epimedium (Figures 3 1-36) and Van- ness. The magnitude of the difference is such that couueria (Figures 37-42), Caulophyllum (Figures 55- Bongardia could be identified in LM alone, but it 60) and Leontice (Figures 61-66, 69-72), and the is transmission microscopy that reveals the differ- pollen morphology supports these traditional ent structure under a very similar surface. The views. But the distinction of the pollen found in foot layer, with radial channels (Figure 193), is Dzphylleia (Figures 85-88, 203-205) from that of thicker and more prominent than the tectum. Podophyllum (Figures 73-78, 81-84, 91, 106, 108, NUMBER 50 17

109, 11 1, 113, 198-202) appears to challenge their Nunan collection of the thickness of the tectum common association by almost all modern gen- (Figures 200, 201) illustrates the problem. eralists. Buchheim (1964) acknowledged the sup- Although the pinwheel configuration is more posed relationship by treating the two genera as precise in Podophyllum peltatum (Figures 77, 78, the only members of a subfamily Podophyllo- 109, 1 1 1, and 113) than in Epimedium diphyllum ideae. Hutchinson (1959), while not assigning (Figures 3 1, 32), E. cremeum (Figure 114), E. gran- genera to subfamily categories, did have Podo- dijlorum (Figure 1 12), or E. sempervirens (Figures phyllum and Diphylleia key out as the first and 33, 34, 110), the similarity is undeniable. How- second genus. Takhtajan (1969) gave separate ever, all species of Epimedium examined in thin family status to these two genera. section (Figures 149, 150, 153-156) have a well Our investigation of the genus Podophyllum L. defined foot layer with channels, while P. peltatum (Figures 73-78,81-84,91, 106, 108, 109, 111, 113, has perhaps the most reduced foot layer of all 198-202) reveals an unusual range of variation in species examined in the family. the pollen morphology, but the significance of the The mean polar lengths in the nine collections variability remains obscure due to a paucity of of Podophyllum peltatum (Table l), 37.3 pm, 36.0 material. The following discussion is based on pm,35.4pm,33.9pm,36.5pm,35.3pm,35.4~m, 38.5 pm, and 37.7 pm respectively, overlap with one collection of P. hispidum Hao (Figures 8 1, 9 I), some species of Epimedium, of Jeffersonia, and of eight of P. peltatum L. (Figures 73-78, 106, 108, Vancouveria. While the mean length of P. hispidum, 109, 11 1, 113, 200-202), and two of P. hexandrum 38.9 pm, is slightly larger than any of P. peltatum, Royale (Figures 82-84, 198- 199). the high for each of the nine would include the The variation in the tectum of Podophyllum low, 36.4 pm, for P. hispidum. peltatum (Figures 109, 11 1, 113) overlaps with that Podophyllum hexandrum (Figures 82-84, 198, 199) of some species of Epimedium (Figures 110, 112, is the only taxon examined in all the Order 114). Ranunculales that has the pollen shed as tetrads, The pollen (Figures 73-'78, 106, 108, 109, 11 1, with the members arranged in either tetrahedral 113, 200-202) of the widespread and common or rhomboidal configuration. The elongate aper- May Apple, Podophyllum peltatum L., is shed as a tures (Figures 82, 83) can be regarded as colpi monad, 3-colpate, the colpi long, the membrane and their distribution, while not irregular, is not covered with flecks of exinous material. The tec- necessarily consistent from one tetrad to the next. tum could be described as consisting of short, The surface is covered with gemmae of variable flattened "rods" with one end projecting. There size that in turn have a ripplelike surface (Figures is considerable variation in the distinction of the 84, 198, 199). rods or striae and the degree to which one end is The tetrad mechanism is fusion of the gemmae- free and projecting. Figures 74, 76, and 78 illus- producing layer along the common wall (Figure trate this variation. In each of the eight collections 198) and the cytoplasm of each tetrad member there are some grains in which the free tips are would be discrete. Both the endexine and foot arranged like the vanes of a pinwheel, as shown layer are thin and fairly uniform, and delicate in Figures 77, 78, 109, 11I, and 113. columellae support gemmae of widely disparate Podophyllum peltatum (Figures 200-202) is as sizes (Figures 198, 199). It should be noted that variable in TEM as in SEM. For the most part, Figure 199 is somewhat oblique. Sections through all material sectioned has a thin foot layer, short the larger gemmae illustrate the undulate surface slender columellae, and a predominant tectum. characteristic. The tectum is probably repre- However, in each collection there is some evi- sented by a fusion of the small gemmae. dence of radial channels in the foot layer as well Pollen of the above two species of Podophyllum as aperture columellae. The difference within the Figures 73-78, 82-84, 106, 108, 109, 111, 113, 18 SMITHSONIAN CONTRIBUTIONS TO BOTANY

198-202) examined in this study could scarcely posed of irregularly placed rods that form the appear more different. In P. hexandrum it is shed base of stout, blunt spines. The sparse distribution as tetrads and thin section confirms fusion by the of the rods makes the tectum appear punctate gemmae-producing layer between adjacent (Figures 85, 88). grains. The apertures are furrow-like and not In TEM (Figures 203-205) the most striking always in the same position from tetrad to tetrad, feature is the delicate structure that supports and the tectum consists of gemmae of variable massive spines. The tectum in each species ap- sizes. In contrast, the pollen of P. peltatum is shed pears irregular and broken, but it is consistent as a monad, is 3-colpate, and the ektexine or with the surface depicted in SEM; small sparsely tectum has a “pinwheel” configuration or modi- distributed columellae connect the tectum with fications thereof. an equally irregular foot layer. The endexine is A minimal pollen sample from an Arnold Ar- thin and fragmented. boretum collection identified as Podophyllum his- Like the other disjuncts, the two species of pidum (Figures 81, 91) has 3-colpate grains, the Diphylleia have subtle differences in the pollen, colpi long and narrow with flecks of exine mate- i.e., the Asian taxon (Figures 87, 88, 205) has rial on the membrane. The SEMs depict an more numerous and more slender spines than the almost complete or continuous tectum with faint North American one (Figures 85, 86, 203, 204), lines (Figure 91), indicating that it might be a but this distinction might not be maintained if modification of the striate-reticulate type found additional collections were examined. elsewhere in the genus and family. These results Podophyllum has two strikingly different pollen should be treated with reserve. types, the tetrads of P. hexandrum (Figures 82-84, The only material available for the genus Dy- 198, 199) and the monads of P. hispidum (Figures sosma R. E. Woodson was that of D. pleiantha 81, 91) and P. peltatum (Figures 73-78, 106, 108, (Hance) Woodson (Figures 79, 80, 92, 206), the 109, 111, 113, 200-202) and the pollen of Diphyf- type species. The 3-colpate grains (Figures 79, leia has to be compared with each. But the tectum 80), in which the tectum is again regarded as a found in Diphylleia, stout blunt spines, has no modification of the striate-reticulate type, con- counterpart in any of the other taxa examined in tribute little new information regarding the ex- the family. tent of Dysosma’s relationship with Podophyllum. Nandina domestica (Figures 89, 90, 207-21 1) was The monad condition and the surface of the described by Thunberg (1 78 1) ostensibly from a tectum appear much closer to Podophyllum peltatum Chinese collection, but the origin of this subshrub than to P. hexandrum. remains a matter of conjecture since the plant has In thin section (Figure 206), however, Dysosma a long history of cultivation and can be found in could be distinguished from all remaining genera many parts of Asia as an escape. by a prominent tectum and foot layer that has a The distinction of this monotypic genus has complementary, undulating interface bridged by been acknowledged by segregating it as a tribe or delicate, uniform columellae. The endexine is even as a family, Nandinaceae. The pollen mor- thin and uniform in the mesocolpal regions and phology (Figures 89, 90, 207-21 1) supports Ku- noticeably thickened in the colpus. mazawa’s (1938a:12) opinion that it is the “far- Diphylleia Michaux (Figures 85-88, 203-205) thest deviated” if it is included in the Berberida- has at least three species and an eastern North ceae. America and eastern Asia type of disjunction. As demonstrated in SEM (Figures 89 and go), The pollen of the North American species, D. the pollen morphology is among the most com- cymosa Michaux (Figures 85, 86, 203, 204) and mon types in the dicotyledons: the apertures are that of an Asian one, D. sinensis Li (Figures 87, 3-colpate and the tectum is deeply punctate with 88, 205), are 3-colpate and have a tectum com- the punctae evenly distributed. NUMBER 50 19

In thin section (Figures 207-21 l), the exine of in Achlys (see page 8), there are palynological Nandina can be distinguished from all members of distinctions that occur in only one or two genera: the Berberidaceae s.1. by the presence of a mas- tetrads only in Podophyllum hexandrum, 6-pantocol- sive endexine. Compared with most members of pate aperture type only in Ranzania, a massive the family, the tectum is thick and almost com- endexine only in Nandina, very long columellae plete (Figure 207), except for the punctae (Figure only in Bongardia, spinose tectum only in Diphyl- 208), and it is uniform in the sense that it is the leia, gemmate tectum only in P. hexandrum, and same thickness in the mesocolpal regions as well aperture columellae only in Epimedium and Van- as near the colpi; the columellae are diminutive couveria. An unstratified exine is found in three (Figures 207, 209), the foot layer is much thinner genera, Berberis, Mahonia, and Razania. than the tectum but recognizable and consistent. EKTEXINERELATIONSHIPS.-The 24 high mag- The massive endexine is lamellar at the interface nification SEMs, Figures 91-114, of tecta found with the foot layer, and uneven and less electron in Achlys, Epimedium, Dysosma, Hydrastis, Jeffirsonia, dense on the inner surface (Figure 21 1). The Podophyllum hispidium, P. peltatum, and Vancouveria continuity of this layer is disrupted by small gaps have been arranged to demonstrate a continuum where the colpus and mesocolpus meet. in variation and the existence of a relationship This study is concerned with generic relation- among these taxa. The relationships exist within ships within the Berberidaceae s. 1 ., and no other and between groubs of figures so that the series taxa examined had a thick punctate tectum sim- should be viewed as a foldout. While the discus- ilar to that of Nandina. Dysosma pleiantha (Hance) sion could start with any one group, that of Woodson (Figure 206) had a thick tectum but Figures 103- 108, representing four genera, Hy- this is the only characteristic that these two spe- drastis, Figure 103, Epimedium brevicornu, Figure cies have in common. 105, E. alpinum, Figure 104, Jeffersonia diphylla, The remarkable development of the endexine Figure 107, and Podophyllum peltatum, Figures 106 layer of Nandina as revealed in thin section (Figure and 108, might serve this purpose best. Palynol- 207) is unique and has not been found in any ogically, Hydrastis, Figure 103, is more closely taxa examined to date in the Centrospermae related to the Epimedium species, Figures 104 and (Skvarla and Nowicke, 1976), or in Plumbagina- 105, and to Jeffersonia, Figure 107, than to any ceae, Polygonaceae, or Primulaceae (Nowicke member of the Ranunculaceae (Nowicke and and Skvarla, 1977). Moreover, the distinction of Skvarla, unpublished data). The similarity of the the pollen of Nandina applies to the Order Ran- striae configuration in Jeffersonia, Figure 107, with unculales as well. Considering each of the various that in the collection of Podophyllum peltatum in families as a whole, the Ranunculaceae does have Figure 108 could suggest a relationship. The dif- the most consistent and well developed endexine ference between the two collections of P. peltatum, (Figures 1 15- 12 1), but at least to date none rivals Figures 106 and 108, is a loss of some of the the one found in Nandina. Palynologically this distinction of individual rods and the fact that species may be more closely related to certain the free tip is no longer free. The difference taxa in the Lardizabalaceae, Akebia trifoliata between the tectum of Hydrastis, Figure 103, and (Thunberg) Koidzumi and Decaisnea fargesii Fran- that of Epimedium alpinum, Figure 104, can also be chet, in that all three have at least the compo- considered as a loss of some distinction. nents of the ektexine represented in the same There is a hypothetical series, Figures 98, 97, proportions, thick tectum, diminutive columellae, 92, 91, in which the tectum from one taxon could and thin foot layer (Skvarla and Nowicke, un- be derived from the preceding one by a loss of published data). some distinction of either rods or striae. The Just as there are exceptional and restricted tectum found in Vancouveria chrysantha, Figure 97, sporophyt ic characteristics, e. g., periant h absent could be derived from Jeffsonia diphylla, Figure 20 SMITHSONIAN CONTRIBUTIONS TO BOTANY

98; the tectum of Dysosma, Figure 92, could be in Figures 48 and 95, in Figure 107 the tectum is derived from Vancouveria chrysantha, Figure 97; and irregularly striate. Podophyllum hispidum, Figure 91, could be derived Hydrastis canadensis may be another example: in from Dysosma, Figure 92, by the loss of all distinc- the collection illustrated in Figures 53 and 54 the tion of the striae or rods. tectum consists of two layers of striae, while in a A second similar series, consisting of Figures second collection, illustrated in Figures 52 and 106, 105, 100, 99, 94, 93, illustrates a continuum 103, the tectum is striate-reticulate. in variation from a striate-reticulate tectum to Another tectal type may be that found in the one that is almost complete, and could link Po- euphorbiaceous genera Croton (Nowicke and dophyllum peltatum, Epimedium brevicornu, Vancouveria Skvarla, unpublished data) and Manihot (Lynch hexandra, E. sagittatum, E. membranaceum, and Jeffer- and Webster, 1975, figs. 1-8; Nowicke and sonia dubia, respectively. Skvarla, unpublished data), in the icacinaceous A third series, Figures 108, 107, 102, 101, 96, genus Platea (Lobreau-Callen, 1973, pl. 3: figs. 1- 95, illustrates the possible stages of the transition 3), in the cistaceous genera Fumana (Saenz de from a striate-reticulate tectum to ones that are Rivas, 1979, figs. 3.4-3C; Nowicke and Skvarla, striate, the striae being mostly parallel to each unpubished data) and Lechea (Nowicke and other. The taxa linked include Podophyllum pelta- Skvarla, unpubished data), in the Myristicaceae tum,Jeffesonia diphylla, Vancouveria planipetala, Ach- (Walker and Walker, 1980), in the Buxaceae lys triphyla, and J. dubia. (Nowicke and Skvarla, unpublished data), in The group consisting of Figures 109-1 14, illus- Aquilaria, Cryptadenza, Lachnaea, Lophostoma, Phal- trates a tectum condition found in some species eria, and Wikstroemia, all members of the Thyme- of Epimedium, Figures 110, 112, and 114, and in laeaceae (Erdtman, 1966; Nowicke and Skvarla, certain collections of Podophyllum peltatum, Figures unpublished data), in Podophyllum peltatum (Fig- 109, 11 1, and 113. All six micrographs have at ures 77, 78, 109, 111, 113), and in some species of least some areas where the free tips are arranged Epimedzum (Figures 110, 112, 114). These tecta like the vanes of a pinwheel. could be classified as a continuous triangular In a review paper (Nowicke and Skvarla, 1980) array. In Scyphocephalium (Walker and Walker, the authors documented the existence of similar 1980, figs. 20, 2 l), Aquilaria, Croton, Lachnaea, Ma- tectum patterns (as illustrated by SEM) in fami- nihot (Lynch and Webster, 1975, fig. 7), and lies or genera that are widely separated on the Wikstroemia the configuration of the triangular or basis of other characters. This phenomenon as prism-shaped subunit is very precise, whereas in well as the existence of very diverse (apparently) Fumana, Podophyllum, and Epimedium it is identifia- morphologies in closely related species raises the ble only in some grains, sometimes only in certain fundamental question of the origin and persist- areas, e.g., along the colpus. ence of such forms. One possible interpretation of these results is Most species of Croton have a tectum as defined that there are a limited number of structurally above-a continuous triangular array-however, defined tectal types, each with a potential varia- C. californicus (Solomon et al., 1973, figs. 46a-46c; tion that may or may not be manifested. The two Nowicke and Skvarla, unpublished data) repre- layers of striae tectum, known to occur in at least sents a variation in this type by having the sub- half a dozen families, could serve as an illustration units rounded or gemmate and with a ripple of such a tectal type. surface. This is more or less paralleled in Podo- Jeffsonia may be an example in which the phyllum: those grains of P. peltatum that have a potential variation is, at least in part, realized: in tectum of uniform “pinwheels” (Figure 78), Figure 48 the tectum consists of two layers of would be equivalent to most species of Croton, striae, in Figure 95 the tectum is striate, in Figure with the “vanes” of the pinwheel equivalent to 45 the tectum appears intermediate between that the traingular subunits; P. hexandrum (Figures 82- NUMBER 50 21

84), with a tectum of ripple-surfaced gemmae tinuous triangular array are closely related and would be equivalent to C. californicus. may be variants of, or derived from, a major Of far greater interest is the remarkable par- structural type. allelism of the pollen morphology in the family Cistaceae with that of the Berberidaceae. Saenz General Discussion de Rivas (1979) examined 36 species representing five of the eight genera in the Cistaceae. He The following discussion considers data from classified the exine sculpturing (1979, table 1) other sources-gross morphology, carpel mor- into five types, rugulose, retipilate, reticulate, phology, serology, and cytology-together with striate, and reticulate-granular, all of which were the implications of pollen analyses. documented by SEM micrographs, and most of That Berberis and Mahonia are closely related is which have a close counterpart in the Berberi- beyond question. Of more fundamental interest daceae s. 1. is the extent of their separation or isolation from The close similarity between Saenz de Rivas’ the remaining genera traditionally aligned as the rugulose type (1979, figs lE, IF), and the reduced Berberidaceae. To regard the woody habit of striae distinction type in the Berberidaceae, as Berberis and Mahonia as secondarily derived from illustrated by Vancouveria chrysantha (Figure 97) a herbaceous one is a situation where Occam’s and Dysosma (Figures 79, 80, 92), would be diffi- Law should be applied: there is no reason to cult to refute. assume that Berberis and Mahonia were ever any- thing but woody in habit. Certainly the distin- The retipilate exine in some species of Fumana guishing characteristics of their pollen are une- (Saenz de Rivas, 1979, figs 3A-3C), in another quivocally primitive. Carpel morphology (Chap- cistaceous genus Lechea (Nowicke and Skvarla, man, 1936) indicates the Berberis-Mahonia line unpublished data), and the pinwheel pattern separated very early from the ancestral stock. found in Podophyllum peltaturn (Figures 77, 78, 109, Both genera have tactile stamens, chromosome 11 1, 113) and some species of Epimedium (Figures numbers of 2n = 28 or 56 and their interfertility 110, 112, 114) are probably derivations of a con- as well as their mutual susceptibility as the alter- tinuous triangular array as discussed above. nate host for wheat rust clearly supports Jensen’s The striate type in the Cistaceae as illustrated (1974) proposal that they are congeneric. All by Halimium atriplicfolium (Saenz de Rivas, 1979, available evidence indicates that Berberis and Ma- fig. 2A) is very similar to the striate-reticulate honia are primitive, isolated genera, and the pres- type in some species of Epimedium: E. alpinum ent authors would agree with Hutchinson’s (1959) (Figure 104) and E. brevicornu (Figure 105). Other restricted view of the Berberidaceae as consisting examples of striate types in the Cistaceae (Saenz of only Berberis and Mahonia. de Rivas, 1979, figs. 2B-2F) are equivalent to the In a family that is said to consist of groups of two layers of striae as illustrated in Jeffersonia genera not closely related to each other, the de- (Figures 45, 48), and in Hydrastis (Figures 53, 54). gree to which Ranzania is separated from the The reticulate-granular type in the Cistaceae remaining genera may be exceeded only by Nan- (Saenz de Rivas, 1979, figs. 1A-1D) has its coun- dina. Ranzania japonica, found in an area of three terpart in the Berberidaceae in those species of degrees of latitude on northern Honshu, has one Epimedium (Figures 31-34, 110, 112, 114) that of the more restricted distributions in all of the have a tectum of small rods with one tip project- dicots. This species possesses floral characteristics ing. that align it with Berberis-Mahonia: petals with The parallel pollen morphology in the Berber- fleshy nectaries at the base, sensitive stamens, an idaceae and Cistaceae suggests that the tecta unstratified exine, and baccate fruit. Certain veg- described by the present authors as two layers of etative characteristics, however, align it with Cau- striae, “randomly” placed small rods, and a con- lophyllum-Leontice: the habit, rhizome, leaf and 22 SMITHSONIAN CONTRIBUTIONS TO BOTANY petiole morphology. The type of aperture, 6-pan- advanced type, especially in regard to the irreg- tocolpate (Figure 139), is unique in the family ular type found in Berberis and Mahonia. s. l., a status that Kumazawa (1937~)also applied The similar floral morphology and exine struc- to the anther dehiscence. Kumazawa recognized ture would support the view that Ranzania, Ber- four modes of dehiscence in the Berberidaceae: berzs, and Mahonia are monophyletic, but the great three were monogeneric, Nandina, Podophyllum, disparity in vegetative morphology and aperture and Ramania, and the fourth included all remain- type strongly suggests that the ancestral stock of ing genera. However, the distinctions of the four Ranzania separated very early in time from that types as illustrated by Kumazawa (1937c:59, fig. which produced Berberis and Mahonia. The present 3) do not, at least to the present authors, seem authors would maintain Rantania as a taxon in- very great. certae. Terabayashi’s first publication in his studies of The almost complete agreement on the close floral morphology of the Berberidaceae was on relationship of Epimedium and Vancouveria is not Ranzania (1977) and the second one on Berberis surprising since the latter genus has been treated and Mahonia (1978). If the flower parts of Ranzania as a section of the former (Baillon, 1871). The as depicted in line drawings by Terabayashi range of variation found in the tectum of Epime- (1977, fig. 1) are compared with those of Berberis dium and Vancouveria would indicate that Stearn’s and Mahonia by the same author (1978, figs. 1, 2), (1938) treatment as closely allied but separate the close similarity would make it difficult to genera is the most valid. Both genera have a deny a relationship between the three genera. chromosome number of 2n = 12, the basic one Certainly the presence of an unstratified exine in for the Berberidaceae. Chapman (1936) placed both the Berberis-Mahonia alliance (Figures 124- Epimedium and Vancouveria with the other predom- 138) and in Ranzania (Figures 141-145) supports inantly two-carpellate genera (Achhs, Diphylleia, this contention. Epimedium , Jeffersonia , Nandina , Podophy llum), bu t Herbarium specimens of Ranzania, consisting of emphasized their special relationship by her state- the only two collections at Kew, were obtained in ment (1936:344), “The structure of the ovaries of April 1979. Unfortunately, one was cultivated, the species commonly placed in the genus Vancou- the other sterile. Nevertheless when these two veria helps clarify the interpretation of conditions collections (Figures 2 12,213) were compared with in Epimedium.” Comparison of herbarium speci- a Japanese species of Berberis, B. amurensis Ru- mens leaves little doubt as to their close associa- precht (Figure 2 14) and one of Mahonia, M. japon- tion. ica (Thunberg) De Candolle (Figure 215), the Comparison of Bongardia, Caulophyllum, and contrast was striking: Ranzania is a slender, deli- Leontice collections, unfortunately, leaves consid- cate herb with a single stem arising from a small erable doubt as to the extent of their affinities to rhizome, and at a height of six to eight inches the each other. Vegetatively, Bongardia and Leontice stem divides producing two petioles and the pe- are almost succulent, a sharp contrast to the thin duncle, which supports a single flower (the culti- leaves and slender stems of Caulophyllum. All three vated, partially dried New York State specimen genera have fruits described as bladder-like; but is f identical). Caulophyllum has this condition only in the earliest The difference in aperture type may be more stage of fruit development, and the large (ca. 1 significant than the similarity in exine structure. cm in diameter), exposed seeds with a dark blue, An unstratified exine is a generalized condition glaucous coat are “easily mistaken for fruits” found in many primitive angiosperms, and in (Ernst 1964: 19). Moreover, each of the two seeds Berberis, Mahonia, and Ranzania it most likely rep- is attached to an erect, conspicuous funiculus, as resents an original, unchanged state. However, much as 7 mm long and 2 mm in diameter, a the 6-pantocolpate aperture of Ranzania is an structure lacking in the other two genera. It could NUMBER 50 23 be argued that the fundamental difference be- early separation or a differential rate of evolution tween the fruits of Caulophyllum and those of Leon- among the characteristics investigated. Neverthe- tice and Bongardia is the rapid and extensive less, Caulophyllum, Leontice, and Bongardia appear growth of the seeds in the former genus. But the to be more closely related to each other than to disparity of the mature fruits and/or seeds as well any remaining genera and could be segregated as as the unusual funiculus would scarcely support the tribe Leon ticeae. a close relationship. The similarity of pollen morphology in Achlys Results from carpel morphology (Chapman, and Jeffersonia, a more or less striate tectum, con- 1936) and serology (Jensen, 1974) support Airy- sistent endexine, and small size range, prompted Shaw’s (1966) viewpoint of a close relationship a comparison of the two genera on other charac- between Caulophyllum, Leontice, and Bongardia, but teristics. There are, however, differences of such not necessarily separate family status since the magnitude as to preclude the possibility of any same results also indicate some affinity to other close relationship: the compact, spicate infloresc- berberidaceous genera. ence of Achlys with numerous, small flowers which The palynological data alone are paradoxical: lack a perianth contrasts sharply with the scapose of all the 3-colpate taxa, Bongardia, Caulophyllum, inflorescence of Jeffersonia with a single, showy and Leontice have the largest individual grains, flower. The fruit of Jeffersonia is as unusual as the averaging 49-35 pm long; all three have an in- inflorescence of Achlys: the oblong capsule opens complete tectum, and in eleven collections there by a horizontal cleft to release the numerous were no variants that indicated a relationship to seeds. The fruit of Achlys is small, one-seeded, and the tectum found in the tribe Epimedieae and either dehiscent (Hutchinson, 1959) or indehis- Podophyllum peltatum. The type of tectal perfora- cent (Rickett, 1971). tion in Bongardia differs sufficiently from that of According to Chapman (1936:346), “The fre- Caulophyllum and Leontice to suggest different quent coupling of the genera Achlys andJeffersonia origins of the “incomplete” characteristic. In thin does not seem to be especially justified when the section and LM, the pollen of Bongardia can be carpel morphology of the two is considered.” She distinguished from that of Caulophyllum, Leontice, regarded them as no more related to each other and all remaining genera by the predominance than each is to Epimedium. of the columellae, which account for 80% of the In a morphological and systematic study of exine. Achlys, Takeda (1915) also dismissed the possibil- In a discussion following Jensen’s presentation ity of a close relationship to Jeffersonia, citing the of the serological results (1974), A. Takhtajan much greater specialization of the latter genus. refers to his study of seed coat characteristics in He regarded Achlys as being related to Epimedium Bongardia, Caulophyllum, and Leontice, and the fact and Leontice, and in fact characterized Achlys as a that Bongardia differs considerably from the other much reduced form of the latter genus. two. Both genera have a chromosome number of 2n All three genera have been reported as having = 12, the basic one for the Berberidaceae, which chromosome numbers other than, or in addition contributes little to the clarification of the rela- to, the basic x = 6: Bongardia n = 6, 7, 2n = 12; tionships of Achlys and Jeffersonia to each other or Leontice n = 7, 8, 2n = 14, 16; Caulophyllum has to to any of the remaining genera. our knowledge only been reported as 2n = 16. Any discussion of the relationships and place- Cltimately any statement regarding their re- ment of Podophyllum must first attempt to resolve lationship depends on the value attached to each the vague circumscription of this genus. of the above characteristics: habit, fruit, seeds, Woodson (1928) elevated Podophyllum pleianthum carpel morphology, serology, pollen morphology, Hance to separate generic status as Dysosma based and cytology. The paradoxical results indicate an primarily on the following characteristics: very SMITHSONIAN CONTRIBUTIONS TO BOTANY large leaves with regular, equal lobes and a finely from the monads of P. peltatum or any other dentate margin, as opposed to smaller leaves with member of the Berberidaceae. However difficult irregular lobes and an entire or uneven margin in it may be to reconcile the distinction of the pollen Podophyllum; an inflorescence of four to 19 flowers, in these two species, they are very similar in as opposed to one; and introrse anthers as op- vegetative and floral morphology and the present posed to extrorse in Podophyllum and all the rest of authors would maintain Podophyllum as including the Berberidaceae. He also cited differences in the both species until samples and/or other data from pollen, that of Dysosma being spherical and rela- other species of Podophyllum or Dysosma become tively small, while that of Podophyllum is lobed and available. relatively large. The latter description can only The pollen found in the two species of Diphylleia mean that Woodson used P. hexandrum pollen as (Figures 85-88, 203-205), D. cymosa from North representative of Podophyllum and interpreted the America and D. sinensis from Asia, is unique and tetrad as a large, lobed grain. reinforces their close relationship to each other Kumazawa (1935: 274) re-examined Podophyl- and distinguishes them from all other members lum plelanthum in view of Woodson’s elevation and of Berberidaceae, s. 1. concluded that “the anthers are quite laterally According to Chapman (1936:347), “the taxo- situated on the connective and the dehiscence is nomic association of Diphylleia and Podophyllum is extrorse.” He also commented that “the size of one which the study of carpel structure does not the pollen grain is larger in P. pleianthum than in confirm.” Furthermore, “it seems likely that the P. peltatum, and this is quite contrary to his two may be derived from the complex which gave [Woodson’s] description.” Our data in Table 1 rise to the two carpel forms, but the separation confirm this, although the difference is not very between them can probably be extended back to great: Dysosma has a mean length of 42.9 pm, and an early representative in that evolution.” Her in eight collections of P. peltatum, the means are views of a remote connection between Diphylleia 37.3 pm, 36.0 pm, 35.4 pm, 33.9 pm, 36.5 pm, and Podophyllum agree with those of Kumazawa 35.4 pm, 35.3 pm, 38.5 pm, and 37.6 pm. (1938a). Woodson (1928) did not, in the opinion of the Li (1947) recognized three species of Diphylleia, present authors, need a pollen difference to con- mostly on the basis of the origin of the infloresc- firm the validity of Dysosma. However, the pollen ence and the extent of the lobing in the leaves: D. of Dysosma (Figures 79, 80, 92) appears to be cymosa Michaux from eastern North America, D. much closer to that of Podophyllum peltatum and P. sinensis Li from China, and D. grayi R. Schmidt hispidurn than that of P. hexandrum is to these two from Japan. Examination of the collections at the species. Examination of the collections (US) seem US indicates that Diphylleia can be separated into to justify his treatment. Hu (1937) apparently three entities using Li’s criteria. agreed with Woodson since he transferred at least Diphylleia and Podophyllum have a chromosome three other species of Podophyllum to Dysosma. number of 2n = 12, basic in the family and If the genus Dysosma and Hu’s (1937) subse- therefore of limited value as an indicator of rela- quent transfers (see page 5) are accepted as valid, tionships. Podophyllum S.S. still includes two very dissimilar According to Jensen (1974:223), Diphylleia and entities regarding pollen morphology: the Podophyllum have “the possibility of great serolog- common and widely distributed P. peltatum L. in ical similarity,” but the two genera have the North America and,judging from the material at reality of widely divergent (apparently) pollen US, a similarly common and widely distributed morphologies (Figures 73-78, 8 1-88, 198-205). P. hexandrum Royale from Asia. The tetrads of P. In considering the variation in pollen mor- hexandrum, in which the tectum is covered with phology of the Berberidaceae s. l., the distinction ripple surfaced gemmae, are radically distinct of the grains in Diphylleia relative to the remaining NUMBER 50 25 members of the family is surpassed only by that In considering the disparity of the palynologi- of the tetrads in Podophyllum hexandrum. cal results with Jensen’s (1974) preliminary data Nandina domestica Thunberg is one of the few from serology (see page 3), it should be noted that berberidaceous genera that has a woody habit conclusions from the latter source are re- and in this respect, as well as articulated leaves, stricted to comparisons among the taxa investi- it is similar to Berberis and Mahonia. Chapman gated. While pollen morphology also relies heav- (1936) placed Nandina with the 2-carpellate gen- ily on comparison, this discipline has reached a era, but cited earlier work that recorded some 3- stage where a number of characteristics have carpellate specimens among the predominantly values attached, primitive or unspecialized, and 2-carpellate ones. She (1936:346) considered the advanced or specialized. Most palynologists variation in Nandina “as an index to the history of would designate the unstratified exine, irregular the forms in the other genera rather than indi- apertures, and undifferentiated surfaces found in cating the genus as ancestral in the evolution of Berberis and Mahonia as primitive without any the family.” knowledge of the rest of the family‘s pollen mor- Kumazawa (1938b: 12) made a strong case for phology. elevating Nandina to separate family status. Reference has been made in the Introduction that the Berberidaceae have an unusually high The outer integument of Nundznu is strongly developed and the micropyle is not observed from outside; moreover, the number of disjunct or discontinuous genera, five nucellus is absorbed before the flower comes into bloom and of the 12 have species that are widely separated the external epidermis and the internal one change into the geographically. Achlys is the only one with a thin walled columnar tissue. These vegetative and ovular Pacific North America and eastern Asia distri- characters, as well as the dehiscing type of anther described bution. Unfortunately, the paucity of material of before (Kumazawa, 1937c), are quite unique among the the Asian taxon as well as the high incidence of berberidaceous genera. sterility in both taxa compromises any statement The pollen of Nandina can be easily distin- about their relationships. guished from all remaining members of the family Caulophyllum, Diphylleia, andJeffersonia have the by the massive endexine. more common eastern North America and east- Nandina has a chromosome number of 2n = 20, ern Asia type of disjunction. In each of these three unique in the Berberidaceae. genera, the pollen of the American species and Jensen’s (1974:225) serological study of the that of the Asian species are fundamentally sim- Berberidaceae does not support the separation of ilar with only subtle differences in the tectum. Nandina since “it features serological similarities The range of variation found within a sample of a high degree with Berberis and Mahonia and and between collections of some species makes with Podophyllum and Diphylleia.” He drew a par- such differences suspect, and the paucity of Asian allel of the distinctive characteristics of Nandina specimens curtailed any further investigation relative to the Berberidaceae with certain groups Podophyllum is another disjunct genus with a in the Ranunculaceae relative to that family distribution similar to that of Caulophyllum, Di- (1974:225): “The fact that the deviating chro- phylleia, and Jeffersonia, but if the pollen of P. mosome number (n = 10) is by no means decisive hexandrum is arbitrarily dismissed due to unparal- is proved by the related family of Ranunculaceae, leled distinction (in all of the Order Ranuncu- in which the Thalictreae and Coptideae also lales), and if the treatment of P. pleianthum as exhibit a completely different karyotypus.” The Dysosma pleiantha is considered valid, then for present authors are of the opinion that variation purposes of comparison the Asian possibilities are in a family as large as the Ranunculaceae is to be reduced to one pollen sample from a collection expected; but in a family of 12 genera, such identified as Podophyllum hispidum at Gray Her- variation may indicate a lack of relationship. barium. The differences in the tectum between P. 26 SMITHSONIAN CONTRIBUTIOKS TO BOTANY

hispidum and P. peltatum are more pronounced spicuous aril. Both have a chromosome number than in the other disjuncts, but the type found in of x = 6. Dysosma pleianthum could be interpreted as inter- The close similarity of the pollen in the Amer- mediate and link the two entities. ican Jeffersonia diphylla and the Asian Plagiorhegma dubia supports their congeneric treatment as Jef fersonia and allies them with Achlys, Epimedium, Summary Podophyllum peltatum, and Vancouveria. While Jeffer- sonia has the typical chromosome number of x = The pollen diversity in the Berberidaceae con- 6, and an anther dehiscence by narrow valves firms, for the most part, previous characterization attached at the apex found in at least six other as “groups of genera” not closely related to each berberidaceous genera, no one genus seems to be other, especially regarding the distinction of Ber- especially allied with this Asian-American dis- beris and Mahonia, and of Nandina. Howevx, the junct. Certainly the fruit, which resembles a moss pollen would suggest greater conformity or closer capsule, is unique within the taxa examined. relationship among Achlys, Dysosma, Epimedium, Achlys has pollen similar to Epimedium, Jeffer- Jeffersonia, Podophyllum species, and Vancouveria, sonia, Podophyllum peltatum, and Vancouueria, and a than has been previously thought. chromosome number of x = 6. The highly re- For Berberis and Mahonia the characteristics of duced flower (sepals and petals are absent) sug- the pollen, an unstratified exine with a random gest no other generic relationships. or unspecialized surface, irregular and/or spiral Achlys, Epimedium, Jeffersonia, and Vancouueria apertures, as well as their woody habit, indicate are similar in pollen morphology, chromosome that they are primitive genera, additionally dis- number, anther dehiscence, and habit, and could tinguished by a chromosome number of n = 14 be segregated as a tribe. (28 for some species of Berberis), susceptibility to Palynologically the Old World genus Leontice, Puccinia graminis, and articulated leaves. Hutch- the monotypic Bongardia (based on L. chrysogonum), inson’s (1959) concept of the Berberidaceae as and the Asian-American disjunct Caulophyllum, consisting of only Berberis and Mahonia could be are more closely related to each other than to any justified. remaining taxa examined. Although the unifying The pollen of the monotypic genus Nandina can characteristics are quantitative (larger size and be distinguished from those of all other taxa thicker walled), the fact that their tecta are not examined, which in combination with evidence related to the type found in Achlys, Epimedium, from floral morphology (carpel morphology, de- Jeffersonia, Podophyllum peltatum, and Vancouveria velopment, anther dehiscence) and, above all, the may be more significant. All three have chromo- unique chromosome number of 2n = 20, support some numbers other than, or in addition to, n = the case for separate family status. 6: Bongardia n = 6, 2n = 14, Caulophyllum 2n = 16, Palynologically, Hydrastis is more closely re- and Leontice 2n = 14, 16. Caulophyllum and Leontice lated to Achlys, Epimedium, Jeffersonia, and Vancou- have very similar stamen morphology, and all ueria than to any genus examined thus far in the three have anthers opening by valves. Their dis- Ranunculaceae. tinction could be acknowledged by treating them Pollen morphology reinforces the special rela- as a tribe, Leonticeae. tionship between Epimedium and Vancouveria: spe- The structure of the exine supports the separate cies assigned to these two genera were the only generic status of Podophyllum pleianthum as Dysosma members of the Berberidaceae s. 1. to have aper- Woodson, accorded on other bases. The tectum, ture columellae. They are additionally distin- aperture type, and size are similar to P. peltaturn, guished by the presence of saccate petals or nectar and Achlys,Jeffersonia, Epimedium, and Vancouveria. spurs, a follicle-like fruit, and seeds with a con- The pollen of Podophyllum hispidum and P. pel- NUMBER 50 27 tatum is similar in the tectum, apertures, and size Mahonia. The chromosome number of 2n = 14 to that found in Achlys, Epimedium, Jeffersonia, and has been reported only from Bongardia and one Vancouueria. Podophyllum peltatum has a 2n = 12, species of Leontice. characteristic of the above genera, but this species The pollen found in two species of the disjunct has longitudinal anther dehiscence as opposed to genus Diphylleia is unique, so much so that no valves. The pollen morphology of Podophyllum clear relationships within the Berberidaceae are hexandrum is unique within the Order Ranuncu- suggested, while the chromosome number, 2n = lales: tetrads with a tectum supporting ripple- 12, is found in at least six other genera of the surfaced gemmae. In leaf and floral morphology family. Evidence from floral morphology is incon- P. hexandrum is closely related to P. peltatum. The clusive except to deny a close relationship with boundaries of Podophyllum have yet to be estab- Podophyllum. Of all taxa examined in this study, lished. Diphylleia is indeed a genus of uncertain affinities. The characteristics of the pollen of Rantania are Pollen morphology does not support a close ambiguous: the 6-pantocolpate aperture type is relationship between the Berberidaceae and Ran- unique within the Berberidaceae s. l., but the unculaceae nor between the Berberidaceae and unstratified exine is similar to that in Berberis and Lardizabalaceae. Literature Cited

Ahrendt, L.W.A. Ernst, W. R. 1961. Berberis and Mahonia: A Taxonomic Revision. Jour- 1964. The Genera of the Berberidaceae, Lardizabalaceae nal of the Linnaean Society of London, Botany, 57: 1- and Menispermaceae in the Southeastern United 410. States. Journal ofthe Arnold Arboretum, 45: 1-35. Airy-Shaw, H. K. Graham, A,, and G. Barker. 1966. A Dictionary of the Flowering Plants and Ferns. 7th In press. Palynology and Tribal Classification in the Cae- edition, xxii + 1214 + liii pages. Cambridge: salpinoideae. In R. Poitiill and P. Raven, editors, University Press. Advances in Legume Systfmatics. Baillon, H. Hu, H. H. 187 1. Monographie des menispermacees et des berberi- 1937. Notulae Systematicae ad Florem Sinensium. Bul- dacees. Histoire des Plantes, 3:1-76, letin of the Fan Memorial Institute, Botany series, 8: Barton, 0. S. 3 1-46. 1793. A Botanical Description of the Podophyllum Diphyl- Hutchinson, J. lum of Linnaeus, in a Letter to Charles Peter 1959. Dicotyledons. In Families of Flowering Plants, 1: xi Thunberg . . . . Transactions of the American Philo- + 510 pages. London: Oxford University Press. sophical Societ).., 3 :334-347. Jensen, U. Behnke, H. D. 1974. The Interpretation of Comparative Serological 1976. Delimitation and Classification of the Order Car- Results: Nobel Symposium 25. In G. Bendz and yophyllales (Centrospermae) according to Ultra- J. Santesson, editors, Chemistry in Botanical Classi- structural Data from Sieve-Element Plastids: A fication, pages 217-227. New York: Academic Survey Based on 146 Species. Plant Systematics and Press. Evolution, 126:31-54. Jonsson, L. Biesboer, D. D. 1973. Pollen Morphology in African Species of Swertia 1975. Pollen Morphology of the Aceraceae. Grana, 15: L. (Gentianaceae). Grana, 13: 119-128. 19-27. Kumazawa, M. Buchheim, G. 1930a. Studies on the Structure of Japanese Species of 1964. Engler’s Syllabus der PfZanzenfamilien. 12th edition, Ranunculus. Journal of the Faculty of Science, Imperial volume 2. Berlin: Gebruder Borntraeger. [Mag- University of Tokyo, section 3, 2(3):297-343, 2 plates. noliales, pages 108-131; Ranunculales, pages 131- 147.1 1930b. Morphology and Biology of Glaucidium palmatum Chapman, M. Sieb. and Zucc. with Notes on Affinities to the 1936. Carpel Anatomy of the Berberidaceae. American Allied Genera Hydrastis, Podophyllum and Diphylleia. Journal of Botany, 23:340-348. Journal of the Facult).. of Science, Imperial University of Clarke, G.C.S., and M. R. Jones Tokyo, section 3, 2(4):346-380. 1978. Aceraceae. In G.C.S. Clarke, C.R. Janssen, and 1932a. Morphological Studies of Anemonopsis, Actaea, and W.J. Punt, editors, The Northwest European Pol- Cimicijiuga. Journal of the Faculty of Science, Imperial len Flora, 17. Review of Paleobotany and Palynology, Cniuersity of Tokyo, section 3, 2(6):413-454. 26:181-193. 1932b The Medullary Bundle System in the Ranuncu- Cronquist, A. laceae and Allied Plants. Botanical Magazzne (To- 1968. The Evolution and Classification of Flowering Plants. kyo), 46(544) :327-332. x + 396 pages. Boston: Houghton Mifflin Co. 1935. The Structure and Affinities of Paeonia. Botanical Darlington, C. D. and A. P. Wylie Magazine (Tokyo), 49(581):306-315, 1 plate. 1956. Chromosome Atlas of Flowering Plants. xix + 519 1936a. Pollen Grain Morphology in Ranunculaceae, Lar- pages. New York: The Macmillan Co. dizabalaceae, and Berberidaceae. Japanese Journal Erdtman, G. of Botany, 8(1):19-46, 5 plates. 1966. Pollen Morphology and Plant Taxonomy: Angiosperms. 1936b. Podophyllum pleianthum Hance: A Morphological xii + 553 pages. Stockholm: Almquist and Wiksell. Study, with Supplementary Notes on Allied 28 NUMBER i0 29

Plants. Botanical ,Magazine (Tokyo), 50(593):268- Rickett, H. W. 276. 197 1. Wildflowers of the United States, Northwestern. Volume 1937a. On the Morphology and Anatomy of Achlysjaponica 5, part 1. xii + 363 pages. New York: McGraw- Maxim. Botanical Magazine (Tokyo), 5 1 (607):660- Hill Book Company. 668. Roland-Heydacker, F. 193% Comparative Studies on the Vernation in the 1974. Caracteres ultrastructuraux et cytochimiques par- Ranunculaceae and Berberidaceae. Journal J ap- of ticuliers du sporoderme des pollens de Berberis aneJe Botany. 13(10): 7 13-726. vulgaris L.. et de Mahonia aqufolium Nutt. Comptes- 1937~.Kanianra japonica (Berberidaceae): Its Morphology, Rendus Academie des Sciences (Paris), 278(D):1475- Biology and Systematic Affinities. JapaneseJournal 1477. qfh’otany. 9.i.i -70. 1938a. Systematic and Phylogenetic Consideration of the Saenz de Rivas. C. Ranunculaceae and Berberidaceae. Botanical Mag- 1979. Pollen AMorphology of Spanish Cistaceae. Grana, azine (Tokyo).5 2(6 1 3) : 9- I 5. 18:91-98. 1938b. On the Ovular Structure in the Ranunculaceae Santisuk. T. and Berberidaceae. Journal of Japanese Botany, 14: 1979. A Palynological Study of the Tribe Ranunculeae. 10~25. Opera Botanica. 48: 1-76. Larsen. S. S. Skvarla, J,J. 197.5. Pollen Morphology of Thai Species of Bauhinia 1973. Pollen. In P. Gray. editor, The Enc);clopedia ofMi- (Caesalpiniaceae). Gram, 14: 114-131. croscopj and .Microtechnique, pages 456 -459. New Li, H. L. York: Van Nostrand Reinhold Co. 1947. Sotes on Asiatic Flora. Journal of the Arnold Arbore- Skvarla. J. J,,and J. 12.’. Nowicke (um. 28:44‘-444. 1976. Ultrastructure of Pollen Exine in Centrospermous Linnaeus. C. Families. Plant Sjstematics and ELsolution, 126355-78. 17.53. SperieJ Plantarum. 1200 pages. Stockholm. Soejarto. D. D., R. B. Faden. and N. R. Farnsworth Lobreau-Callen. D. 1979. Indian Podophjllum: Is It Podoph,llurn emodi or Po- 1953. Le Pollen des Icacinaceae, 11: Observations en dophyllum hpxandrum? Taxon, 28(5/6) :i49-55 1, microscopie electronique, correlations, conclusions (I). Pollen SSpores. 15:47-89. Solomon, A. M.. J. E. King. P. S. Martin. and J. Thomas Lobreau-Callen. D.. S. Nilsson. F. Albers. and H. Straka 1973. Further Scanning Electron Photomicrographs of 1978. Les Cneoraceae (Rutales): Etude taxonomique, Southeastern Pollen Grains. Journal q/ the Arizona palynologique et systematique. Grana, 17: 125- 139. Academ) of Science, 8:133-157. Lynch. S. P.. and G. L. LVehster Stearn. u’. T. 1975. A New Technique of Preparing Pollen for Scan- 1938. Epimedium and I’unrouveria (Berberidaceae), a Mon- ning Electron Microscopy. Grana, 15: 127-136. ograph. Journal of the Linnaean Soriel>. Botany. 5 1: Nowicke. J,\Y. 409-535. plates 24-3 1. 197.5. Pollen Morphology in the Order Centrospermae. Takeda, H. Grana. 15: 5 1-7 7. 1915. On the Genus Achhs: A Morphological and Sys- Sowicke. J. W.. and J.J. Skvarla tematic Study. Botanical Magazine (Tokyo), 1977. Pollen Morphology and the Relationship of the 29(346):169-185. 1 plate. Plumbaginaceae. Polygonaceae, and Primulaceae Takhtajan. A. to the Order Centrospermae. Smithsonian Contribu- 1969. Flowering Plants. Origin and Dispersal. x + 3 10 pages. tronc tu Butany. 37: 64 pages. Washington: Smithsonian Institution Press. 1980. Pollen Morphology: The Potential Influence in Terabavashi, S. Higher Order Systematics. Annals of the Missouri Studies in Morphology and Systematics of Berber- Boianical Garden. 66:633-700. 1977. Ornduff. R., editor idaceae. I: Floral Anatomy of Kanzania japonica. 1968. Index to Plant Chromosome Numbers for 1966. Acta Phytotaxonomica et Geobotanica. 28( 1-3) 145-57. Regnum I)gegetabile, 55:54. 1978. Studies in AMorphologyand Systematics of Berber- Radulescu, N.M. idaceae, 11: Floral Anatomy of Mahonia japonica 1974. Cercetari morfopolinice asupra familei Aizoaceae (Thunb.) DC. and Berberis thunbergii DC. Acta Phy- I. Analele I’nirNersitatii Bucuresti, 22:29-39. totaxonomica et Geobotanica, 29(1-5): 106-1 18. SMITHSONIAN CONTRIBUTIONS TO BOTANY

Thorne, R. F. Coeloraryon, Pycnanthus. and Scyphocephalium). Amer- 1974. A Phylogenetic Classification of the Annoniflorae. ican Journal of Botany. 67(5):603-611. Aluo 8: 147-209. Wettstein. R. Thunberg, C. P. 1935. Handbuch der Systematischen Botanik. 4th edition, x 1781. .Voca Genera Plantarum. Part 1, 28 pages, 1 plate. 1152 pages. Leipzig and Wein: Franz Deuticke. Upsaliae: J. Edman. + Walker, J. \V., and A. G. Walker Woodson. R. E. 1980. Comparative Pollen Morphology of the Mainland 1928. Dysosma: A New Genus of Berberidaceae. Annals oj African Genera of Myristicaceae (Cephalosphaera, the 'Missouri Botanical Garden, 15:335-340, 1 plate. Tables and Figures w N TABLE1 .-Berberidaceae specimens examined, alphabetically by genus (grain size in pn; asterisk indicates longest dimension for tetrad member)

Longest dimension of ten grains Species Collect ion Location Low Mean High Figure numbers

Achlys japonica Maximowicz Terabayashi 209 KYO Japan 22.1 24.3 26.0 178, 179 A. triphylla (Smith) De Candolle Calder & Savile 8323 US Canada 27.3 30.2 33.0 49, 51, 175-177 Evert s.n. 1920 US Washington 24.7 26.3 28.6 50,96 BerberiJ.fendleri A. Gray Eastwood 5272 US Colorado 37.7 39.9 42.9 21, 22, 124, 125 B. grandtflora Turczani now Arbelaez & Cuatrecasas 5908 US Colombia 39.0 43.2 45.5 24 Mexia 7628 US Colombia 48.1 50.6 53.3 126, 127 B. ilicifolia Forster Goodall 834 US Argentina 45.5 49.9 53.3 19, 20, 128-132 B. lischleri C. K. Schneider Rock 17544 US China 36.4 41.9 49.4 B. uulgaris L. Soper & Dale 3950 US Canada 32.5 38.4 41.6 23 Bongardia chrysogonum (L.) Spach Balls 776 US Turkey 44.2 48.9 54.6 194 Sintenis 35 US U.S.S.R. 50.7 54.5 58.5 67,68, 190 193, 195-197 Caulophyllum robustum Maximowicz Kirino 713 US Japan 49.4 51.5 54.6 55, 56 Moran 5204 US Korea 46.8 50.2 53.3 59, 184 C. thalictroides (L.) Michaux Harper3864 US Alabama 44.2 48.9 52.0 60, 185-189 Henry 571 US Pennsylvania 42.9 45.1 48.1 57, 58 Diphylleia cymosa Michaux Braun s.n. 25 Apr 27 US Tennessee 32.5 36.4 39.0 86 Pollard s.n. 16 May 01 US North Carolina 36.4 40.4 44.2 85,203, 204 D. sinensis Li Rock 4230 US China 35.1 39.5 44.2 87,88, 205 Dysosma pleianthu (Hance) R. E. Woodson Steward & Cheo 210 GH China 39.0 42.9 50.7 79, 80, 92 Wilson 3203 GH China 206 Epimedium alptnum L. Porta s.n. Jun 1894 US Italy 29.9 31.5 33.8 36, 104 E. breuicornu Maximowicz Rock 12302 US China 29.9 33.9 37.7 35, 105, 150 E. cremeum Nakai Togasi 1281 US Japan 32.5 33.2 33.8 151-154 Kirino 618 US Japan 26.0 28.7 31.2 114 E. diphyllum (Morren & Decaisne) Loddiges Anon. 1314887 US Japan 26.0 28.2 29.9 31, 32, 149 E. grandiflorum Morren Togasi 1031 US Japan 28.6 3.1.3 33.8 112, 155, 156 E. membranaceum K. Meyer Forrest 25471 US China 29.9 33.2 35.1 94 E. sagittatum (Siebold & Zuccarini) Maximowicz Anon. 1314880 US Japan 32.5 34.1 36.4 99 E. sempervirens Nakai Masayuki Oue 33 US Japan 31.2 33.1 36.4 33, 34, 110 Jeffersoniu diphylla (L.) Persoon Baxter s.n. 15 Apr 11 US New York 29.9 32.9 36.4 46. 107 Chandler s.n. 30 Apr 16 US Michigan 28.6 31.9 33.8 Shafer & Miller 180a US Pennsylvania 31.2 33.9 36.4 Table 1.-Continued z C 5 Longest dimension qf M ten grains P Species z Collection Localion Low Mean Hiph Fipure numbers J. dubia (Maximowicz) Bentham & Hooker Palczevsky 3616 US U.S.S.R. 27.3 29.3 31.2 43, 95, 172-174 Mrs.R. K. Smith s.n. 30 May 38 US Korea 29.9 32.5 35.1 44, 45, 93 Leontice altaica Pallas Anon. 597803 US U.S.S.R. 42.9 48.8 53.3 61, 62 L. armenaica Boivin Koelz 14701 US Iran 52.0 54.7 57.2 65, 66, 180-182 L. euersmannii Bunge Sintenis 121 MO U.S.S.R. 39.0 43.7 48.1 71, 72 L. leontopetalum L. Zillett & Rawi 10275 US Iraq 52.0 55.1 58.5 69, 70, 183 L. odessana Rogowicz 4. Dojcz s.n. 5 Apr 08 MO U.S.S.R. 42.9 46.5 49.4 63, 64 Mahonia fremontii (Torrey) Fedde Hope 9396 US Arizona 31.2 35.2 41.6 25, 26 M. haematocarpa (Wooton) Fedde McKelvey 1225 US Arizona 31.6 34.1 39.0 30 M. neruosa (Pursh) Nuttall Belton s.n. 26 May 43 US Oregon 49.4 51.2 53.3 27, 28, 133-135 M. oiwakensis Hayata Wilson 10844 US Formosa 29.9 33.8 37.7 29, 136-138 Nandina domestica Thunberg zheng 2098 US China 41.6 44.2 48.1 207-211 rai & Class 4034 US China 35.1 39.4 42.9 89, 90 Wilson 2379 US China 35.1 38.9 41.6 Podophyllum hexandrum Royale Rock 12259 US China 31.5* 33.3* 35.8* 83, 84 Rock 12259 GH China 32.5* 33.3* 34.2* 198, 199 Rock 12424 GkI China 27.3* 28.9* 31.3* 82 P. hispidum Hao r. T. yu, 15977 A China 36.4 38.9 41.6 81, 91 P. peltatum L. Braun 391 1 US Kentucky 33.8 37.3 40.3 109, 202 Zrampton 76 US Illinois 29.9 36.0 40.3 77, 78, 113 lones s.n. 21 Mar 62 US Alabama 32.5 35.4 39.0 McDougall 1231 US Mississippi 31.2 33.9 36.4 75, 76, 108, 111 \Tease 3 10 US Tennessee 32.5 36.5 41.6 Vurian 1914 US Georgia 32.5 35.3 39.0 200, 201 Ricksecker s.n. 20 May 95 US Ohio 32.5 35.4 39.0 Rowell, York & Tharp 47 I7 1 US Texas 35.1 38.5 41.6 73, 74, 106 rhieret 10356 US Louisiana 35.1 37.7 41.6 Ranzania japonica (Ito) Ito Zult. 12 Apr 47 K England 141, 144, 145, 147, 148, 212 Epstein s.n. Cult. US New York 36.0 38.0 142, 143, 146 rakeda s.n. 27 Aug 05 K Japan 213 rerabayashi 154 KYO Japan 30.0 32.6 35.0 139, 140 Vancouueria chrysantha Greene rhompson 4605 US Oregon 32.5 34.7 37.7 41, 42, 97, 157-159 V. hexandra Morren & Decaisne illen 66 US Washington 32.5 34.6 35.4 Ebert s.n. Aug lY20 US Washington 33.8 39.4 41.6 39, 40, 100, 160 164 V. planipetala Calloni ;onstance 2514 US California 33.8 36.5 40.3 101 w Yoover 5046 US California 35.1 38.1 41.6 37, 38, 102. 165 169 w TABLE2.-Centrospermae specimens examined, alphabetically by family (grain size in pm)

~ Longest dimension of ten grains Species Figure Collection Location Low Mean Hi,gh numbers AIZOACEAE Acrodon bellidijlorus N. E. Brown Van der Bijl 33 K S. Africa 26.9 27.0 28.6 5, 6 CARYOPHYLLACEAE Tunica stricta (Runge) Fischer & Meyer Goloskokov s.n. 20 Jun 59 US U.S.S.K. 22.1 23.1 24.7 3, 4 PORTULACACEAE Anacampseros jilamentosa Sims Rose & Stewart s.n. 1912 US S. Africa 51.0 54.3 58.5 I, 2

TABLE3.-Ranunculaceae specimens examined, alphabetically by genus (grain size in pm)

~ Longest dimension of ten grains Species _____ Collection Location Low Mean High Figure numbers _____ Batrachium aquatile Dumortier Larsen & Pedersen s.n. 12 Jun 67 US Denmark 29.9 33.4 36.4 13, 14, 119 Clematis heracleifolia De Candolle Mrs. R. K. Smith s.n. 11 Aug 34 US Korea 22.1 24.7 26.0 9, 10, 117 Glaucidium palmalum Siebold & Zuccarini Takeda s.n. 23 May 07 US Japan 24.7 26.3 27.3 11, 12, 118 Hamadryas magellanica Lamarck Goodall 1073 US Argentina 26.0 28.7 29.9 17, 18, 121 Hepatica transsiluanica Fuss Richter s.n. 2 Apr 02 US Hungary 33.8 38.0 41.6 15, 16, 120 Hydrastis canadensis L. Hardin 610 US Arkansas 19.5 22.8 26.0 52, 103, 122, 123 Ricksecker s.n. 12 May 95 US Ohio 19.5 22.6 26.0 53,54 Kanunculus oreophytus Delile Mearns 1402 US Kenya 36.4 37.8 39.0 7, 8, 115, 116

v1 f =i

z F z n 0 z 4 E m C =1 0 FIGIJRES7 12.-Ranunculaceae pollen, SEM: 7, Rununcu1u.r orrophytus Delile, pantocolpate, polar z (II view, X 2260 (Mearns I402 US, Kenya); 8, K. orrophytus, ektexine surface, X 7560 (same collection); 9, Clemutzs heruclezjdzu De Candolle, irregularly (panto)poratq X 3000 (Mrs. R. K. Sniilh s.11. 11 Aug 34 US, Korea); 10, C. heruclez~?liu,cktexirie surface, X 7.500 (same collection); 11, G'luuczdiumpulmu~umSiebold & Zuccarini, 3-colpate, polar view, X 4500 (Takeda s.11. 23 May 07 US, Japan); 12, G. palmatum, ektexine surface, X 10,000 (same collection). (Micrographs reduced to 76'%).) FICIJRES15 III.-Ranunculaceae pollen, SEM: 13, Batruchium aqua/zk Dumortier, kolpate, cquatorial view, X 242.5 (1,arseri c(r Pederseri s.n. 12 Jun 67 US, Denmark); 14, B. aquatilt-, ektexirie surface, X 7500 (same collection); 1.5, Hqmcn trcln.r.riluanicu Fuss, pantocolpate, polar view, X 2220 (liichter s.n. 2 Apr 02 US, Hungary); 16, H. ~mns.riluanicu,ektcxine surfacc, X 7500 (same collection); 17, Hamadryas rnagellanzca Lairiarck, :3-colpate,equatorial view, X 3300 (Goodall 1073 US, Argentina); 18, H. ma.yellanica, ektexine surface, X 7500 (same collection). (Microsraphs reducrd to 7.5%.) FIGURES19 24.-Berberidaceae, Berberis pollen, SEM: 19, H. iliczjiiliu Forster, irregular apertures, X 1700 (Goodall 83.1- US, Argentina); 20, B. zlic@liu, ektexirie surface, X 3000 (same collection); 2 I, B.,fundlrri A. Gray, irregular apertures, X 2000 (Eastwooti ,5272 US, Colorado); 22, B. fendlerz, five grains (onc at upper left may be sterile) illustrating variation in size and apertures, X 9.1-0 (sanir collection); 23, H. uu(yuris Id.,spiral apertures, X 2050 (Soper Kr Dale 3950 US, Ontario); 24, H. grrmu’zflora Turczaninow, four grains illustrating variation in apertures, X 790 (Arbelaez 8r Cuatrecasas 5908 US, Colombia). (Micrographs reduced to 7 l%.) G; '9 n z0 -I E m C =! FIGURES3 I 36. -Berberidaceae, Epirnedium pollen, SEM: 3 I, Epirnedium dipilyllurn (Morren & 0 Ikcaisrre) Loddiges, :%-colpate,polar view, X 3400 (Anon. 131.2887 US, .Japan); 32, E. dzphyllurn, 5 ektexine surface, X 7.500 (same collection); 33, E. sernperuirens Nakai, 3-colpate, equatorial view, X 3000 (Masayuki Oue 33 US, Japan); E. .~m~i~ruirms,ektexine surface, X 7480 (same collection); 35, E. hreozcurnu Maximowicz, 3-colpate, equatorial view, X 2700 (Rock I2302 US, China); 36, 1;. nlpinurn L., kolpate, equatorial view, X 2.500 (Porta s.n. Jim 18!)4 US, Italy). (Micrographs reduced to 75'%.) FIGURES37~42.~--Berberidaccac, Vuncouveria pollen, SEM: 37, V. plunipetulu Calloni, :$-colpate, equatorial view, X 2400 (Hoover 5046 US, California); 38, I:. plr~rizpetulu,ektexine surface, X 7500 (same collection); 39, V. hexundru Morren & Decaisne, :l-colpate, equatorial view, X 2320 (Ebert s.11. Aug 1920 US, Washington); 40, V. hexundru, ektcxine surface, X 7.500 (same collection); 4 1, I:. chrysunhi Greene, 3-colpate, equatorial view, X 2700 (Thonlpson 4605 US, Oregon); 42, C'. chrysnnthu, 3-colpate, polar view, X 3000 (same collect ion). (Micrographs reduced to 77'%,.) FrcirRes 43 48.~. Bcrberidaccae, JeffPrJonzn pollen, SEM: 43, ,/pfler.ronza duhiu (Maxiniowicz) I~c111ham8 I looker, 3-colpate, equatorial view, X 3000 (I’alczevsky 3616 US, U.S.S.K.);44,J. dub, kolpatc, equatorial view, X 2880 (Mrs. K. Sriiith s.11. 30 May 38 CIS, Korea); 4.5,J. K. 4 dubru, ektexirir surface, X 7470 (same collection); Mi,,/. di,thy//a (I,.) Persoori. 3-colpatr, equatorial 0 view, X 2.500 (Raxtcr s.11. 1.5 Apr 11 US, Ncw York); 47, J. diphyllu, :$-colpatc, polar view, X 3200 (Sharer Kr Miller 180a US, Pennsylvania); 48,J.dip/!y/ln, ektexirie surface, X 5280 (smie collcctioii). (Micrographs reduced to 75‘X~) ., kolpate, equatorial view, X 1800 (Gillett Pr Rawi 1027.5 US, Iraq); 70, L. /eontopelalzrm, ektexine surface, X 7500 (sanic collection); 7 1, L. euersmannii Bunge, 3-colpate, equatorial view, X 2300 (Sintenis 121 MO, U.S.S.K.);72, I,. evrrsamannii, ektexine surface, x 7500 (same collection). (Micrographs reduced to 77'h.) F~c:riws 73~78.~Herberidaceae,I’orfophyllurn pellakun pollen, SEM: 73, 3-colpate, equatorial view, X 2500 (Kowell, York & ‘l’harp 47171 CIS, ‘Iexas); 74, cktcxiric surface, X 7530 (same collection); 7.5, :kolpate, rquatorial view, X 2910 (Mcl)ougaII 1231 US, Mississippi); 76. rktexiric surface, X 7.500 (sanic collection); 77, 3-colpate, equatorial view, X 2600 (Crampton 76 US, Illinois); 78, ektexine surface, X 7500 (same collection). (Micrographs reduced to 79‘X .) FIGIJRES79-84.-Berberidaceae, Dysosma arid Podophyllum pollen, SEM: 79, Dysosmu pleianthu (Hance) R. E. Woodson, kolpate, polar view, X 2000 (Steward & Cheo 210 GII, China); 80, I). pleiaritha, 5-colpate, equatorial view, X 2000 (same collection); 8 1, I’odophyllurn hz,\/lzdurri Hao, :kolpate, equatorial view, X 2000 (T. T. Yu 15977 A, China); 82, f’. hexundrum Royale, rhomboidal tetrad, X 1500 (Rock 12424 GH, China); 83, 1’. hexandrum, tetrahedral tetrad, X 141.5 (Rock 122.59 US, China); 84, f’. hexandrum, ektexirie surface, X 7.500 (same collectiori). (Microzraptis rc:duced to 74‘%.) Fic:rrues t(,~~!)O.~Berberidaceac,Diphylleiu arid Nundinu pollen, SEM: 8.5, II$il$h cymosu Michaux, S-colpatc, equatorial view, X 2200 (Pollard s.n. 16 May 01 US, North Carolina); 86, I). cymo.\a, ektexine surface, X 5000 (Brauri s.11.25 Apr 27 US, Teiinessee); 87, I). .rznun~irLi, kolpate, equatorial view, X 2250 (Rock 4230 US, China); 88, D. .sinmsz.v, cktexirie surface, x 5300 (same collection). 89; Nundznu domestzcu Thuriberg, 3-colpate, equatorial view, X 2500 (Tai I(r Class 4034 US, China); 90, N. domeJtzcu, ektexine surfacc, X 8400 (same collection). (Micrographs reduced to 73'K.) (r 0

vl 4

n 0

E'IcuREs 9lp96.--Berberidaceae pollen, view of ektexine surface, SEM: 91, Podophyllurn hispidurn Hao, X 5000 (T. T. Yu 15977 A, China); 92, Dysosmuplezunlhu (Hance) R. E. Woodson, X 5000 (Steward & Cheo 210 GH, China); 93, JeSfersoniu dubzu (Maximowicz) Bentharn & Hooker, X 7520 (Mrs. R. K. Smith s.n. 30 May 38 US, Korea); 94, Epimediurn membranuceurn K. Meyer, x 7500 (Forrest 25471 US, China); 95, JeJfersonzu dubzu (Maxirnowicz) Bentham & Hooker, x 7500 (Palczevsky 3616 US, U.S.S.R.);96, Achlys lriphyllu (Smith) De Candolle, x 7500 (Evert s.n. 1920 US, Washington). (Micrographs reduced to 75%.) I;IC;~IRES07 102. -1krl)eritlace;re pollcri, view ofektexine surface, SEM: 97, Vuncouvrriu chry.\arithu (irwnr. X 7500 (’I‘honipsoii 460.5 IJS, Orcgon); 98, Jrflrrtonru dipliyllu (L.) I’t.rsoon, X 7500 (Shafrr Kr Millcr 180a US, l’erinsylvania); 99, E/~i,lmrdium cqi//ulum (Siebold Kr Zuccaririi) Maxiriiowicz, X 7500 (Anon. 13148180 US, Japan); 100, Vunrouueriu hexundru Morreii Kr Decaisne, X 7.500 (t

w0, 64 SMITHSONIAN CONTRIBUTIONS TO BOTANY I;ICIJRES 149- I56-Epzrnedium pollen, 1EM: 149, R dzphyllum, low magnification of section near mcdian, rriesocolpus shows foot layer with channels arid without an endexinc whilc the colpus shows a lamellate eridexirie penetrated by columellae (ser also Vancouuerzu chrymntha, Figure 159, V. hexandra, Figures 160, 161, and V. planzpetulu, Figure lW), X 1400; 150, E. hrruzcornu, section through mesocolpus illustrating highly channeled foot layer, X 3800; 151, E. cremeum, tangential section at and slightly beneath exinc surface, the arrows indicate cross sections of free tips of tectal rods illustrated by SEM (see Figure 114), X 4930; 1.52, E. crrmeum, oblique section including eridexirie in rriesocolpal region, X 3060; 153, E. cremeum, radial section through mesocolpus, small peak in tectuni probably represents longitudinal section through a free tip of a rod, X 12,880: 154, E. cremeum, colpus showing lamellate endexine perforated by columellae, X 10,080; 155, E. cqrandzflorum, colpiis similar to Figure 1.54, X 13J1.0; 156, E. grandz/lorum, section of mesocolpus with samc: structiirr as in Figures 1.50 arid 1.53, X 36,400. (Scales equal 1 pni.) 66 SMITHSONIAN CONTRIBUTIONS TO BOTANY FI(;llRk:S 1.57- 164.- Vanouuena pollen, M: 1.57, V. chrysunlha, low magnification oblique section near median showing endexine development in colpal regions and absence in mesocolpal regions as well as abundant channels in the thick irregular foot layer, X 7380; 158, V. chrysanlhu, angle of section is less oblique than in Figure 157, and illustrates foot layer channels and paucity of endexinc characteristic of the rrir:socolpus, X 9430; 159, V. chrysunthu, lower graiii shows thick endexinc with columellae (arrows) in colpus, upper grain shows an oblique section of a mesocolpus, X 9430; 160, V. hrxandru, low magnification cross section at median showing similar niorphology as V. chrysuntha (Figure 157), foot layer channels are abundant but less conspicuous because of low magnification, X 4000; 161, V. hexandru, radial section through colpus showing lamellar nature of endexine arid apparent penetration by colurnellae (also sec Figure 159), X 15,580; 162, V. hpxandrn, oblique view showing cross sections of foot layer channels and endexinc accumulation in colpr~s,X 4.500; 163, V. hexandru, section parallel with colpus showing reduced columellae, foot layer channels arid eridexine, X 11,890; 161, V. hrxnndra, tangrntial section illustrating reticulate arid sonirwhat striatc nature of exine surface, X 5.510. (Scales equal 1 pi.) 68 SMITHSONIAN CONTRIBUTIONS TO BOTANY Fic:rrwics 165 169.- Vanconueria pianipetala pollen, M: 16.5, view of mesocolpus, eridexine absent, the foot layer is thick, irregular and with nunierous channels, the inconsistent and uneven tecrum is the result of both section plane and striate pattern, the circular or club-like appearance of the distal ends of the columellac represent cross sections of these striae, X 26,650; 166, oblique sectioii clearly showing endexine development in the colpus and absence in niesocolpns; the foot layer channels are present bul low magnification and dark printing tend to obscure them, X ,5460; 167 and 168, tangential sections at and slightly beneath exirie surface, Figure 167 agrees with thc mostly parallel exine striae depicted in SEM, Figures 37, 38, 101, arid 102, just as the tangential section of V. iiuxandru (1:igure 164 ) agrees with the tectiim deipictcd in SLM, (Figures 39, 40, and loo), X 5760, in Figure 168 cross sections of columellae arc ~urroundedby exine striae, x 5290; 169, upper grain highly oblique section erriphasizing striate exine surface and foot layer with barely visible cross sections of channels; lower grain shows a colpus wir h thickened endexine and colurriellae, as described previously (see V. ~h~.tu~ilh~~,Figure 159, and V. hexandra, Figiirc Ifl), X ,5460. (Scales equal 1 pin.)

o? W 70 SMITHSONIAN CONTRIBUTIONS TO BOTANY FI(XJRI~S170 179. JeJersoniu arid Achlys pollen, TEM: 170, J. dzphyllu, rnesocolpal view, note the well dcvrhped arid essentially uniform endexine which contrasts with the highly irregular foot layer, X 10,080 (Shafer Nr Miller 180a US, Pennsylvania); 171,J. diphyllu, low magnification view including portions of each mesocolpal region, X 5000 (same collection); 172,J. dubiu, low magnification, median section, the endexirie is present in rnesocolpus but is irregular; the beaded appearance of the tecturri is due to the mostly parallel striae being cut perpendicular to the long axis, X 3780 (I’alczevsky 3616 US, U.S.S.R.); 173,,/. dubia, section in colpus region indicates cktexirie elernents do not penetrate endexine as is commonly noted for Epimpdzum and Vuncouuerzu, X 6200 (same collection); 174, J. dubzu, mc:socolpal-colpal (at bottom) view, arrow indicates “inner” striae, note the regular and consistent foot layer arid eridexine, X 9120 (same collection); 175, Achlys lrzphyllu, low magnification median section, the endexine is very narrow in the mesocolpus and thicker in the colpirs, the foot layer is thicker than the endexine and highly irregular, X 4300 (Calder Kr Savile 8 US, Canada); 176, A. lriphyllu, mesocolpal section, the numerous openings in the tectum, reflect packing of striae as discussed in text, X 4600 (same collection); 177, A. triphyllu, section in colpus showing reduced ektexine over endexine (see corririierits to Figure 173 above), X 8410 (same collection); 178, A. juponicu, low rnagnificat ion of a near equatorial section (approximately perpendicular to section in Figure 175), X 4000 (Terabayashi 209 KYO, Japan); 179, A. pponzcu, in this tangential view the openings (see legend to Figure 176) are abundant in the ektexine, X 4060 (same collection). (Scales equal I p.) 72 SMITHSONIAN CONTRIBUTIONS TO BOTANY E’IGIJRES 180- 189.- Leonticr and Caulophylhm pollen, TEM: 180, L,. nrmeniaca, section of nieso- colpus showing a thick, uniform, and lamellate endexine, irregular foot layer with ctiannrls. short columellae and thick, irregular, incomplete tectum, X 10,120; 181, L,. armenzacci, at rriargirr of colpus emphasizing channeled foot layer and thickening of endexirir (to the left), X 5880; 182, L. urrncnzucu, section includes colpus arid shows ektexine fragments on surface of endexine, these fragments appear joined by a narrow foot layer, X 5880; 183, L. lPO7llO/Jdfhm, low magnification, median section, the endexirie is well developed in non-apertural regions as well as in the colpus, X 3010; 184, Chulophyllurn robusturn, low niagriification median section illustrating same basic structure as in Figure 183: incomplete tectiim, short columellae, channels in foot layer, endexirre present in [ion-aperture arid apcrt lire regions, note trace of foot layer across colpus as suggested in Figure 182, X 2800; 18.5, C. thalictrotd~.r,tangential scction illlistrating circular lumina of exine surface as shown in SEM (see ligure 60), X 2990; 186, C. thalzchzh, tangential view but including cross sections of columrllae, X 2990; 187, C. thalzctrozd~s,oblique cross section indicating iiriiforni endexine, X 1400; 188, C’. thnlictrozdes, colpal view showing thickened lanrellate endcxirie supporting fragments of ektexine, X 6480; 189, C. thalictrozdur, nicsocolpal view similar to I,. urmenzuca in ligurr 180. I.. lron~oprtalurn in Figure 189 arid C robusturn iri Figure 181, X 11,310. (Scales equal I pi.) 74 SMITHSONIAN CONTRIBUTIONS TO BOTANY FIC~JRES190- 197.--Hnn~ardin chrysogonum pollen, TEM arid SEM: 190, low niagnification median section showing portions of the three apertures, note that the endexine is only developed in the colpal rcgions (arrows), X 2730; 191, tangential section just beneath tectum and showing cross sections of coluniellae, X 1400; 192, section of colpus emphasizing thinning of foot layer and presence of endexine, X 4370; 193, mesocolpal view showing thick irregular foot layer containing numerous channels, an endexine is not present in this area of the exine, X 15,960; 194, SEM of fractured grain, the long, slender coluniellae in the mesocolpus and thick, irregular foot layer correlate well with TEM (Figures 190 and 193), X 6060; 195, tangential section showing reticulate exine surface, note identical correlation with SEM surface in fractured grain (Figurc 194, exine fragment to the left of the scale), X 2.530; 196, section parallel with the colpus showing well developed and uniform endexine, although not clearly shown the foot layer has channels (see Figure I%), note that in the coipus the columellae are reduced (as in Figures 190 and 194 which show both colpal and mesocolpal regions), X 11,310; 197, highly oblique section that includes a substantial area of a colpus (bottom half of TEM), X 3700. (Scales equal 1 pm.) 76 SMITHSONIAN CONTRIBUTIONS TO BOTANY z C

m5

FIMJRE~198-206.-- I’odophyllum, Diphylleza arid Dywsmu polleri, M: 198, l’od~ip~~llurnhexundrum, portion of two members of mature tetrad, some gemmac on thc distal or free exine surfaces confirrii the ripple-likc morphology depicted in SEM (see Figure 84), note thickened, undulatc. structure of the foot layer in the oblique section of distal surfacc of tetrad member at the right (arrow), the tectum between tetrad members is shared, resulting in a solid layer that is attached to each individual grain by it’s columellae, the endexine is uniform, riarrow and disrupted (at left), X 3800; 199, P. hexandrum, view of external or distal surface, which, in contrast to Figure 198, shows a slightly thicker endexine, X ,5320; 200, P. pvl/ulurn (Nunan /9/4 US, Georgia), the obliquc angle of section has enhanced the thickness and irregularity of the tectuni produced by ttie “flattened rods” (see SEM Figures 109, 11 1, 1 13), the foot layer is thin and irregular and decreases toward the colpus (at the right), the endexine is fairly iinifornm, slightly thicker than the foot layer and increases in the colpus, X 12,180; 201, 1’. peltutum (same collection), in contrast to Figure 200 this section is less oblique and more parallel to the colpus, the eridexine is smooth, uniforrn arid considerably thicker than thc narrow, somewhat intermittent foot layer, thc colurnellae are short and well developed, the tectiim is thick and fairly regular, the protrusions from [he tecturri probably represent sections through free tips of rods, X 12,180; 202, 1’. pellaturn (Hruun .39// US, Kentucky), section includes colpus, note that exine area to the left of the colpus resembles Figure 201 while exine to ttie right of thc colpus resembles Figure 200, in thc colpus flecks of exine appear to be supported by a narrow foot layer, the endexirie is not clearly differcritiated from the foot layer but is thickened, lamellate and somewhat disrupted, X 9430; the pleomorphic nature of the exirie of 1’. peltalum as shown in Figures 200 202 supports results from SEM (see text discussion); 203, Dzphyllezu cymosa, radial section through niesocolpus, the cndexine is narrow, uniforrn and about equal to the irregular foot layer; the short, regular columellae support a thin tectum interrupted by massive spines, X 11,020; 204, I). rymoui, section through colpus illustrating endexine thickening, X 9660; 205, D.sznensis, slightly oblique section through mesocolpus, X 8050; 206, I)ysoJrna plezuntha, ripper grain shows slightly obliqiic section through mesocolpus and lower one shows section through colpus, note that undulating outer surface of foot layer corresponds with undulating inner surface of tectum, the two layers are separated by narrow colurnellae, the colpus shows thickened, highly lamellate ciidexirie. X 6300. (Scales equal 1 pm.) 7% SMITHSONIAN CONTRIBUTIONS TO BOTANY FIGURES207 21 I.-Nundznn dompstzm pollen, TEM: 207, the most striking aspcct of the central grain and parts of three surrounding grains is the remarkably developed endcxinc, which in most of thcse sections is thicker than the entirc cktcxinc, a condition unparalleled for Berberi- daceae pollen; the tecturii is considered to be thick, hiit thc columcllae are very reduced and the foot layer thin; note traces of ektexirie in the aperture as well as the gap between nieso and colpial endexine (arrows), X 4200; 208 and 20!), tarigential sections. in Figure 208 the section is at the cxinc surface and shows the punctate morphology illustrated by SLM (Figures 89, YO), X 6670, in Figure 209 part of the section (i.c., thc surrounding margins) shows the punctate surface morphology (as seen in Figure 208) whilc the ccntral arca represents the region beneath the tecturn surface and distinguishes the narrow coluniellae depicted in Figure 207, X 4620; 210 and 21 1, oblique sections that include the three apertures, in Figure 210. which is highly oblique and close to a pole, the massiveness of thc cndcxinc is cxaggerated, X 6300, in Figure 21 I, which is less oblique, the thick cndexine is again emphasized arid may also be sorncwhat lamellar (arrows), x 4900. (Scales equal 1 pm.) 80 SMITHSONIAN CONTRIBUTIONS TO BOTANY

FIGURE2 12.-Herbarium specimen, Ranranza pponzca (Ito) Ito, cultivated, Royal Botanic Gardens at Kew, 12 April 1947 (K). NUMBER 50 81

FIGURE2 13.-Herbarium specimen. Ranzaniu japnoica (Ito) Ito, Takedu s.n., 27 .4ugust 190.5 (K),Japan. 82 SMITHSONIAN CONTRIBUTIONS TO BOTANY

FIGURE2 14.-Herbarium specimen, Berbens arnu~ensisRuprecht, Korshznsky s.n., 22 May 1891 (US), U.S.S.R. NGMBER 50 83

FIGURE2 15 -Herbarium specimen, Mohonia japontca De Candolle Mtrushtma s.n, 22 March 1954 (US), Japan.