Pollen Ultrastructure of the Haemodoraceae and Its Taxonomic Significance

Grana 22: 79-103, 1983

Pollen ultrastructure of the Haemodoraceae and its taxonomic significance

MICHAEL G. SIMPSON

Simpson, hl. G. 1983. Pollen ultrastructure of the Haemodoraceae and its taxonomic significance. -Gram 22:79-103, 1983. Uppsala 31 May 1983. ISSN 0017-3134. The classification of the genera and tribes of the monocot family Haemodoraceae has been variable and uncertain. In order to provide additional taxonomic characters, SEhl and TEhl were used in describing the pollen ultrastructure of seventeen genera and nine- teen species of the family. The Haemodoraceae as a whole, with the exception oflannria and Lophioka, is palynologically united by the absence of a typical tectate-columellate exine architecture. Within the family, all eight genera of the tribe Haemodoreae have boat-shaped, monosulcate, heteropdar pollen grains with a verrucate (foveolate in one genus) exine wall sculpturing. In contrast, all six genera of the tribe Conostylideae (as defined here) possess variously shaped 2-, 3-, or 7-8-porate, isopolar or apolar grains with a characteristic rugulate sculpturing. A distinctive 2-layered exine, lacking a foot- layer, occurs in all members of the Conostylideae and in four genera of the Haemodoreae. The four other genera of the Haemodoreae are linked to the 2-layered members by a hypothesized gradation in wall architecture. Additional ultrastructural evidence supports the delimitation of the tribes Haemodoreae and Conostylideae, the placement of Phlebo- carya within the tribe Conostylideae, and the recognition of subgroups of genera within each tribe. In contrast to the Haemodoraceae, members of the Tecophilaeaceae possess a tectate-columellate exine, arguing against the classification of that family as a third tribe (Conanthereae) of the Haemodoraceae. Lanaria and Lophiola are aberrant within the Haemodoraceae in having tectate-columellate exine mlls with monosulcate, exine-less apertures. The classification of Lanaria or Lophiola within the tribes Haemodoreae or Conostylideae or within the Tecophilaeaceae is refuted based on palynological evidence. The pollen of Pauridia, which also has a tectate-columellate architecture, is otherwise distinct from that of Lanaria and Lophiola in having disulcate apertures and in lacking an ektexinous foot-layer. The inclusion of Pauridia in the Haemodoraceae versus the Hypoxi- daceae is unresolved based on the present study.

hfichael G. Simpson, Department of Botany, Duke University, Durham, NC 27706, USA (hlnniiscript received 14 April 1982, revised version accepted 24 October 1982) Downloaded by [SDSU San Diego State University] at 10:48 25 February 2015 INTRODUCTION 1969, Hutchinson 1973, Robertson 1976). The The Haemodoraceae is a monocot family of 13-17 intrafamilial classification of the Haemodoraceae genera and approximately 80 species with distribu- has varied considerably in past treatments (see tions in southern Africa, northern South America, Geerinck 1968 for a detailed synopsis). The four Central America, Mexico, eastern North .America, family tribes originally proposed by Bentham & Australia, and New Guinea. Members of the family Hooker (1883), Euhaemodoreae (= Haemodoreae), are characterized as perennial, rhizomatous herbs Conostyleae (= Conostylideae), Conanthereae, with equitant, unifacial leaves and trimerous flowers and Ophiopogoneae, have been shifted in various in a terminal, cymose inflorescence (Geerinck 1968, combinations to other families. For example, Pax

Grana 22 80 hi. G. Sittipson

(1888,1930) and Pax & Hoffmann (1930) considered stratification and sexine pattern (Erdtman 1966: the family Haemodoraceae to be equivalent to 198-199). In contrast, five genera of the tribe Con. Bentham & Hooker’s tribe Euhaemodoreae; the ostylideae (sensu Pax & Hoffmann, 1930). Aiigio. Conostylideae and Conanthereae were placed in zotilhos, Blatrcoa, Cotiosfylis, Plrlebocarp, and the Amaryllidaceae (subfamily Hypoxidoideae) and Tribotinnthes, have isopolar, subisopolar, or apolar the Ophiopogoneae in the Liliaceae. Baillon (1894) grains with either 2, 3 or 8 porate apertures (24 viewed the Haemodoraceae as an unnatural as- 53 pm) and with a consistent and characteristic LO semblage. Melchior (1964) treated the family as pattern: “at low adjustment, dark islands (with consisting of three tribes: Haemodoreae, Conos- fainter darker dots = bacula) separated by brighter tylideae, and Conanthereae. However, the most channels” (Erdtman 1966: 46). Erdtman emphasized recent treatments (Hutchinson 1934, 1959, 1973, that this common LO-pattern unites these genera Geerinck 1969) have recognized two family tribes: despite their shifting aperture number. Finally, two Haemodoreae (10-1 1 genera) and Conostylideae other genera of the family, Larinria and LophioIrl, (3-6 genera). Hutchinson, furthermore, grouped comprise a third pollen type of monosulcate grains the Haemodoraceae with five other families in the (29-35 gm) with a thin exine and finely reticulate order Haemodorales, but treated the tribe Conan- sculpturing. thereae as a distinct family, the Tecophilaeaceae, Radulescu (19731, using light microscopy of in the Liliales. In addition to these conflicts of acetolyzed preparations, described twelve species family definition, the circumscription of tribes has of Cotiostylis (tribe Conostylideae), 1 species each varied. Three genera, Larinria, Lophiola, and of Dilatris and Schiekiu (tribe Haemodoreae), and Plrlebocary, have been variably transferred be- 1 species of Cyairellri (tribe Conanthereae of the tween the tribes Haemodoreae and Conostylideae, Haemodoraceae, sensu Melchior 1964). The pollen even by recent authors (see Geerinck 1968, Robert- of the tribe Conostylideae (i.e., the 12 spp. of son 1976). Lanorin, moreover, is sometimes clas- Cotiostylis) was characterized as porate with 0, 2, sified in the Liliaceae or Tecophilaeaceae (De Vos or 3 apertures, having reticulate exine sculpturing 1961, 1963, Willis 1973). The genus Pairridin, placed (verrucate in 2-porate grains of C. setosn) which is in the tribe Haemodoreae by Pax (1888, 1930), is pilate or pilate-tegillate in optical cross-section. included in the family Hfpoxidaceae in most recent Radulescu described pollen of the Haemodoreae as treatments (Geerinck 1969, Hutchinson 1973; see monosulcate and heteropolar with a reticulate- Thompson 1979). foveolate sculpturing and a tegillate-baculate exine. Because of past uncertainties in the classifica- Cyariella of the Conanthereae only differs from tion of the Haemodoraceae and presumed closely the Haemodoreae in having a pilate-tegillate exine. related taxa, evidence has been used increasingly Despite numerous studies in many different fields from the fields of anatomy (Schulze 1893, Stenar of the taxa of the Haemodoraceae and related 1927, 1938, Green 1959, Cheadle 1968, 1969, Simp- families, “the substantial and continuing disagree- son & Dickison 1981), biochemistry (Hegnauer ments concerning the limits of the Haemodoraceae 1963, Edwards, Churchill, & Weiss 1970, Cooke & and the relationships of the genera assigned to it Edwards 1981), cytogenetics (Green 1960, Hilliard indicate that the family merits additional detailed & Burtt 1971, Ornduff 19791, and embryology study” (Ornduff 1979). The purpose of the present (Stenar 1927, 1938, Dellert 1933, De Vos 1956, 1961, study is to provide additional characters from pollen 1963, Simpson 1981 a). However, pollen studies ultrastructure in assessing taxonomic and phylo- Downloaded by [SDSU San Diego State University] at 10:48 25 February 2015 (Erdtman 1966, Radulescu 1973), using light micro- genetic relationships. Specific questions asked in scopy and acetolyzed preparations, have probably this study are: (1) What, if any, pollen ultrastruc- proven most useful in assessing taxonomic rela- tural features characterize the Haemodoraceae as tionships in the family. a whole? (2) Does pollen wall architecture support Erdtman (1966) recognized three pollen types the classification of the Tecophilaeaceae (= tribe within the family. Six genera of the tribe Haemo- Conanthereae) within the Haemodoraceae? (3) doreae (sensu Pax 1930), Hneniodorirni. Palrridia, What similarities and differences, based on wall Dila tris , La cIiir mi th es , Wach eii rlorfia , and Xiph i- structure, are evident between the tribes Haemo- dirrtii, are characterized by monosulcate pollen (20- doreae and Conostylideae? (4) Does pollen ultra- 90pm long) with a “usually not very distinct” exine structure clarify the tribal classification of PlilCbo-

Crana 22 Polleii rrlfrastnrcfrrreof the Haeriiodoraceae 81

cnrya, Lanuricr, or Lophiola? (5) Is the classifica- The pollen sections were dehydrated, critical-point dried, tion of Pmrridicr within the Haemodoraceae sup- mounted on a stub, and coated as before for SEhl viewing ported palynologically? (6)What is the pollen ultra- (see Fig. 7 E, 8 E). For TEhl analysis of wall architecture, pollen samples structural basis for the three pollen groups of Erdt- were fixed in 4.2% gluteraldehyde for 1-2 hours, rinsed man? several times in 0.1 hl Sorensen’s phosphate buffer, and post-fixed in 2% OsO, for 1 hour. After two rapid H,O rinses and dehydration to 100% ethanol, the material was infiltrated in a series of increasing concentrations of MATERIALS AND METHODS Spurr’s resin (Spurr 1969). Fully infiltrated grains were Pollen samples were obtained either from herbarium placed in an obconical BEAhI capsule and polymerized sheets or from liquid preserved field collections. Anthers 8-12 hours in a 65°C oven. Sections ca. 500-800 A thick from dried herbarium specimens (“DRIED’) were re- were prepared using a Dupont diamond-knife on a Cam- expanded in Aerosol OT for 1-4 days, followed by several bridge-Huxley ultramicrotome, and mounted on uncoated H,O rinses. Flowers from field collections were fixed 200 mesh copper grids. Preparations were post-stained in either formalin/acetic acid/alcohol (“FAA”) or a soh- with uranyl acetate (I5 min) and lead citrate (7 min) and tion of 4% gluteraldehyde and 4% formalin in 0.1 hl viewed with a Siemens Elmiskop 101 TEhl. Sorensen’s phosphate buffer (“GLUT”). Ifrrgerzbachia brosiliertsis C. Nees & hlart. (Haemo- The following 19 species, in 17 genera, were examined: doraceae, sensu Hutchinson 1973), which recently has Arzigozarirhos flmidits DC. “FAA”-hl. G. Simpson been recognized as a species of Chlorophj!iirn of the 23IX81J (DUKE); Barberetfa aitrea Harv. “FAA“-R. Liliaceae (Ravenna 1977), is not included in the present Ornduff 7661 (UC); Bloncoa cartesceris Lindl. “GLUT”- study. Pollen terminology generally follows that of Walker hl. G. Simpson 181X81AA (DUKE); Corzosrylis bealiarirr & Doyle (1975). F. hluell. “FAA”-Arboretum, U. C. Santa Cruz, 3 Nov. 1980 (UCSC);Di/afris corynbosn Berg. “FAA”-P. Gold- blatt 3242 (hl0);D. piloiisii Barker “DRIED’-E. Werder- mann & H.-D. Oberdieck 754 (US); Hae/mdoritrii spi- RESULTS carrim R. Br. “FAA”-hl. G. Simpson 16IX81C (DUKE); H. simplex Lindl. “GLUT”-M. G. Simpson 20IX81A Tribe Haernodoreae (DUKE); Locltrinrzfhes carolirzinnn (Lam.) Dandy Dilafris (2 of 5 species examined) “FAA”-hl. G. Simpson 14VI80A (DUKE); Lniinria D. pilnrzsii Barker.-Pollen grains monosulcate and loma (L.) Dur. & Schinz (= L. phtriton Ait.)”DRIED’- R. D. A. Bayliss 4369 (US); Lopliiola oitrea Ker-Gawler heteropolar (Fig. 1 A) with verrucate to baculate “FAA”-hl. G. Simpson 14VISOB (DUKE); Macropidin non-apertural sculpturing (Fig. 1 B) and gemmate to frtligirzosa (Hook.) Druce “FAA”-hl. G. Simpson pilate apertural sculpturing (Fig. 1 C).Non-apertural 18IX81DD (DUKE);Paitridia ntinrtra (L.f.)Dur. & Schinz exine 2-layered, a commissural line occuring at the (= Harv.) “DRIED”-P. hlacOwan & P. hypoxidoides junction of the two layers (Fig. 1 E). Outer exine H. Bolus 291 (US);P/ilebocaya ciliara R. Br. “GLUT”- hl. G. Simpson 16IX81A (DUKE); Pyrrorlziza rzeblirine layer composed of irregularly baculate, laterally hlaguire & Wurdack “DRIED”-B. hlaguire, J. J. Wur- fused elements (Fig. 1 D), the distal ends forming dack & G. S. Bunting 37222 (US); Schiekia orinocemis the verrucate sculpturing; inner exine layer com- (Kunth) hleisn. “FAA”-B. hlaguire 41569 (NY); Tri- posed of smaller, distinct, generally papillate ele- bonanrhcs arisrrnlis Endl. “DRIED”-A. J. Eames & A. T. Hotchkiss, 23 Aug. 1953 (US); Wacherzdorfin rhyr- ments (Fig. 1 E). Apertural wall with an exine of siyorn L. “FAA”-R. Ornduff 7691 (UC); Xipliidrrrti scattered, 2-layered, gemmate elements atop a rela- coerrtlettnr Aubl. “FAA”-J. hl. hlacDougal 1043 tively thick, 2-layered intine (Fig. 1 F), the thick- (DUKE). ened outer intine with radially oriented, channel- For SEM studies, whole dehisced anthers containing like structures (Fig. 1 F). mature pollen were placed in a modified capsule between

Downloaded by [SDSU San Diego State University] at 10:48 25 February 2015 two 2 pm hlillipore filters or two 0.1 or 5 pm Nucleopore D. coryiuboso Berg.-Grains monosulcate, heter- filters. If the pollen of a species was later observed to be opolar (Fig. 1 G). Sculpturing of non-apertural wall covered with debris or pollen-kit, the technique of Lynch verrucate (Fig. 1 H) and of apertural wall finely & Webster (1975) was followed and the isolated pollen gemmate (Fig. 1 I); larger wart-like elements scat- placed in the capsules. Anthers or free pollen were dehydrated to 100% ethanol, then infiltrated with 100% tered among the minute gemmate elements near the Freon 113 (intermediate fluid). The material was critical- aperture border (Fig. 1 I). Non-apertural exine point dried in a BOhlAR SPC 900/EX drier with CO, bordering the aperture and at the ends of the grain (transition fluid). Pollen grains were tapped onto a stub smooth and perforated (Fig. 1 H, I). Non-apertural covered by double-stick Scotch tape, sputter coated (ca. 200 A thickness) with goldlpalladium (60/40), and viewed exine 2-layered by presence of a commissural line with a Joel JSM-S1 SEM. Optionally, cross sections of (Fig. 1 J); inner exine layer composed of discon- pollen grains were prepared using a freezing microtome. tinuous to fused baculate elements (Fig. 1 J). Exine

6-838361 Grana 22 82 hf. G. Siiiipsoiz

and intine of apertural wall (Fig. 1 K) similar to (Fig. 4A). Non-apertural wall with a verrucate those in D. pilnizsii. Outer exine layer thin at inter- proximal surface (Fig. 4B-, and a smooth, micro. face between apertural and non-apertural walls, pore dense border surrounding the aperture (Fig. corresponding to the perforated border (Fig. 1 L). 4A, B). Non-apertural exine 2-layered, with a corn. missure between the layers (Fig. 4C, D). Outer Xipliidilriii (monotypic) layer composed either of distinct, closely appressed X. coerrrlerriiz Aubl.-Pollen monosulcate and verrucate elements in the proximal region (Fig. 4 C) heteropolar (Fig. 2A). Sculpturing of the non- or relatively homogeneous in the smooth border apertural wall verrucate with granular material be- region (Fig. 4D). Inner exine layer throughout of tween the major verrucae (Fig. 2B), a relatively appressed, radially elongate elements comprising smooth border encircling the aperture (Fig. 2 A). 1/2-2/3 of the exine thickness (Fig. 4C, D). Aper- Apertural wall sculpturing verrucate (Fig. 2 C). ture with a relatively thick intine and an exine of Non-apertural exine composed of closely spaced, closely spaced, 2-layered, baculate elements, the baculate elements, 2-layered as defined by a median inner exine layer reduced (Fig. 4 E). commissure (Fig. 2E); exine between the major baculate elements granular, particularly in the Pyrrorlzizn (monotypic) lower exine layer. Intine of non-apertural wall 2- P. rzebliizne hlaguire & Wurdack-Grains mono- layered (Fig. 2E). Aperture wall composed of sulcate and heteropolar (Fig. 4F) with verrucate distinct, closely spaced, verrucate to baculate non-apertural and gemmate to verrucate apertural exine elements atop a relatively thick, 2-layered wall sculpturing (Fig. 4G). Non-apertural wall com- intine (Fig. 2D, G). Outer layer of aperture intine posed of a thin intine and an exine with 2-layers: thickened at the interface with the non-apertural a continuous outer layer with a verrucate outer wall (Fig. 2 F). surface and an inner layer of amorphous, discontinuous, somewhat granular exine material (Fig. 1Voclzetzdor-o (1 of 5 species examined) 4 I); inner exine layer along the border of the aper- 1V. thprsifora L.-Pollen monosulcate and heter- ture reduced (Fig. 4 H). Apertural wall composed opolar (Fig. 3A). Aperture wall convex in shape of scattered verrucate exinous elements atop a (Fig. 3 A), consisting ifwidely separated, 2-layered thick, 2-layered intine (Fig. 45). Outer intine layer baculate exine elements (Fig. 3C) over a relatively expanded at the aperture periphery (Fig. 4H) and thick fibrillar intine (Fig. 3 D, F). Non-apertural containing radially oriented, channel-like structures wall proximally verrucate (Fig. 3A, D), a granular (Fig. 4 J). material present between adjacent verrucae (Fig. 3 B). Exine of the proximal region 2-layered, a corn- Scliiekia (monotypic) missural line present between the large, verrucafe S. orinocensis (Kunth) Meism-Grains monosul- elements of the outer layer and smaller, irregular cate and heteropolar (Fig. 5 A), with foveolate- granular elements of the inner layer (Fig. 3G). fossulate non-apertural exine (Fig. 5B) and ver- Aperture wall encircled by a smooth non-apertural rucate to gemmate apertural sculpturing (Fig. 5 C). border, pitted with micropores (Fig. 3A, C, D). Apertural wall of scattered gemmate exine elements Exine of the border continuous with that of proxi- and a thick, homogeneous intine (Fig. 5 D, E). Non- mal wall exine and 2-layered, a prominent commis- apertural exine composed of 3 layers: an outer, per- sure present between the homogeneous, micropore- forated homogeneous layer; a middle layer of dis- Downloaded by [SDSU San Diego State University] at 10:48 25 February 2015 filled outer layer and the irregular, somewhat gran- continuous, amorphous exine deposits; and a con- ular inner layer (Fig. 3H). Intine of non-apertural tinuous, somewhat granular inner layer (Fig. 5 F). wall generally 2-layered (Fig. 3G). Outer intine Middle and inner exine layers reduced at the aper- layer at the aperture interface thickened (Fig. 3 E), ture periphery (Fig. 5 E) and absent along two ap- comprising the bulk of the apertural intine wall parent regions of constriction of the proximal wall (Fig. 3 F). (Fig. 5 D, GI.

Barberetta (monotypic) Haeinorlorrriiz (2 of 20 species examined) B. airrea Haw.-Pollen monosulcate, heteropolar H. spicatirni R. Br.-Grains monosulcate, heter- with verrucate to baculate apertural sculpturing opolar (Fig. 6A) with verrucate non-apertural and

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- - ...-.~, .- 84 bl. G. Sinzpsort

a F

Fig. 2. Xiphidirrnz cocrrrlerrrri Aubl. (Haemodoreae). (A) (e), with commissural line between the exine layers (ar- Whole grain, aperture at right; note raphidesrystal. SEM rows); note granular exine between major elements. TEhl x2 100. (B) Non-apertural wall; note granular exine (ar- X22400 (F) Interface between aperture (N) and non- row) between major verrucae. SEhl ~9000(C) Verrucate apertural wall (right); note thickening of outer intine layer apertural wall. SEhl ~7700.(D) Cross-section of whole in the apertural wall. TEhl X6000. (G) Apertural wall, (0). Downloaded by [SDSU San Diego State University] at 10:48 25 February 2015 grain, showing aperture TEhl x2400. (E) Non-aper- showing 2-layered intine (i)and scattered exine elements tural wall, showing 2-layered intine (i)and 2-layered exine (P). TEhl X30900.

gemmate apertural sculpturing (Fig. 6B). Non- oriented, electron transparent channel-like struc- apertural exine composed of laterally appressed, tures (Fig. 6 F) and thickened at interface between bnculate structural elements basally interconnected non-apertural and apertural walls (Fig. 6 D). by thin exinous material (Fig. 6C, E); commissure H. simples Lind1.-Grains monosulcate, heter- not present. Apertural wall of scattered baculate to opolar with verrucate non-apertural and apertural gemmate elements atop a thick, 2-layered intine wall sculpturing (Fig. 6G). Aperture region in- (Fig. 6C. F). Outer intine layer containing radially distinct, defined only by the slightly more separated

Gram 22 Pollen iiltrtisfnrctrrre of rlie H(~PIIIO~~O~-~ICC~IC85 Downloaded by [SDSU San Diego State University] at 10:48 25 February 2015

Fig. 3. Wnchendorfin ihjrsifloro L. (Haemodoreae). (A) outer layer of the ?-layered intine (i). TEhl x6500. (F) Whole grain, showing convex aperture (above) and flat- Aperture wall. showing ?-layered intine (i) and ?-layered tened proximal surface (below); note smooth aperture exine (e); note junction between exine layers (arrow). border (0).SEhl x 1600. (B) Close-up, proximal surface, TEhl X 1 1200. (G)Non-apertural, proximal wall, showing showing granular exine (arrow) between major verrucae. 2-layered intine (i) and 2-layered exine (e).with commis- SEhl ~5500.(C) Close-up, aperture; note smooth rnicro- sural line between the two exine layers (arrows). TEhl pore-tilled border (b).SEhl ~3600.(D) Cross-section of x2I 300. (H) Non-apertural wall of smooth border region; whole grain, showing proximal wall (p), aperture ((I), and note commissural line between two exine layers (arrows) border (b). TEhl ~2600.(E) Interface between apertural and rnicropores (m).TEhl X20300. (0) and non-apertural (right) walls; note thickening of

Grnno 22 86 hf. G. Sinpsoi1

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Fin. 4. Pollen rrltrrrstrrrctrrre of the Htrettiorlorncene 87

exine elements. Exine composed of laterally ap- Tribe Conostylideae pressed to separated baculate elements (Fig. 6 H); Arigiozarithos (1 of 10 species examined) Some exine elements occasionally 2-layered, the A. flauidirs DC.-Pollen grains isopolar, usually inner layer very thin (Fig. 61). Intine of apertural diporate (Fig. SA), rarely (in the same anther) tri- and non-apertural walls indistinguishable, 2-layered, porate (Fig. 8 C). Apertural wall protrusions with radially oriented, electron transparent channel- hemispheric in shape (Fig. 8 A, B). Non-apertural like structures in the outer layer throughout (Fig. wall with a rugulate sculpturing; apertural wall 6H). psilate to slightly scabrate (Fig. 8B). Non-apertural exine composed of irregular, tangentially elongate ~nchrintithes(monotypic) structural elements (Fig. SD, E); major tangential L. crrrolitriotin (Lam.) Dandy.-Grains monosul- exine elements interconnected by thin, presumably cate, heteropolar (Fig. 7A). Sculpturing the non- of exinous basal strands, demonstrated in acetolyzed apertural wall verrucate; that of the apertural wall grains (Fig. 8D). Inner exine layer composed of gemmate (Fig. 7B). Tetrads of pollen from im- distinctive papillate protuberances (Fig. 8 E, F), a mature anthers decussate, with apertures posi- line of breakage or a commissure often visible be- tioned at the distal pole (Fig. 7C); thus, grains ab- tween papillate protuberances and the major struc- solutely heteropolar. Exine of the non-apertural tural elements (Fig. 8 F). Apertural wall essentially wall composed of Iaterally appressed, apparently free of exine, consisting of a thick, 3-layered fibrillar fused baculate structural elements, the distal ends intine (Fig. 8G). Middle intine layer thickened in forming the verrucate sculpturing (Fig. 7D, E); the interface region of the aperture and containing exine elements homogeneous with commissure numerous, radially extended channel-like struc- absent (Fig. 7D). Apertural wall of distinct pilate to tures (Fig. 8G). gemmate exine elements atop a thick, 2-layered intine; a fibrillar material often (but not 'always) present between the exine elements (Fig. 7F). Macropidin (monotypic) Outer intine layer thickened at interface of non- Al.ficliginosa (Hook.) Druce.-Grains isopolar and apertural and apertural walls (Fig. 7 G), containing diporate with hemispheric to obconical shaped channel-like structures (Fig. 7 F). apertural walls (Fig. 8 H). Non-apertural walls rugulate; apertural walls psilate (Fig. 8 I). Aperture wall lacking exine (Fig. SJ), consisting of a thick, 3-layered intine with prominent channel-like structures in the middle layer (Fig. 8K). Non-apertural Fig. 4. A-E: Burberetfauureu Haw. (Haemodoreae). (A) Whole pollen grain, aperture facing. Note smooth, micro- exine composed of sinuous, tangentially elongate pore-pitted border (b)surrounding aperture. SEhl x2700. elements with papillate protuberances on the inner (B) Close-up of proximal wall, showing large verrucae and surface (Fig. 8 L); commissure present between smooth, micropore-pitted region at end of grain (b). SEhl the outer exine and the inner protuberances, in- x5 600. (C) Non-apertural, proximal wall; note 2-layered dicating a 2-layered nature to the wall (Fig. exine, with cornrnissural line (arrows). TEhl ~22800. 8M). (D) Non-aperturalwall of border region, showing 2-layered exine with cornmissural line (arrows). TEhl ~21200.(E) Cotrostjlis (1 of 24 species examined) Apertural wall. Note junction (arrow) between two layers C. Oenlinrrn F. Muell.-Pollen grains isopolar. of exine elements (e).TEhl X 13800.

Downloaded by [SDSU San Diego State University] at 10:48 25 February 2015 F-J: PJrrorhizu ueblirine hlaguire & Wurdack (Haemo- usually triangular and triporate; aperture walls doreae) (F) Pollen grain. collapsed aperture facing. SEhl hemispheric in shape (Fig. '9 A). Aberrant 3-, 2-, X2500. (G)Close-up of wall, showing apertural (left) and or 4-aperturate grains occasionally present (Fig. non-apertural (right) walls. SEhl ~6800.(H) Interface 9 C, D, E). Non-apertural walls rugulate; apertural between aperture ((1) and non-aperturalwall (lower right); note thickening of outer intine layer and reduction of walls psilate to scabrate (Fig. 9B). Non-apertural inner exine layer. TEhl x8 100. (I) Non-apertural wall wall comprised of an exine of continuous, tangen- showing 2-layered intine (i)and 2-layered exine (e);scat- tially elongate elements with minute papillate pro- tered amorphous elements (arrow) make up the lower tuberances on the inner face (Fig. 9G). Apertural exine layer. TEhl ~21400.(J) Apertural wall; note scat- wall without exine (Fig. 91). composed of a rela- tered exine elements (P) and 2-layered intine (i) with channel-like structures in the outer layer (arrow). TEhl tively thick, obscurely layered intine, with radially x20 100. oriented, channel-like structures (Fig. 9 H).

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a 90 Al. G. Sittipsoti

Downloaded by [SDSU San Diego State University] at 10:48 25 February 2015 Fig. 7. Lnchnnnthes cnrolinionn (Lam.) Dandy (Haerno- baculate elements and apparent micropore channel open doreae). (A) Pollen grain, aperture above; note raphide ings at inner surface (arrow). SEhl x22 300. (F) Aperture crystal. SEhl ~2500.(B) Close-up of wall, with verrucate wall; note 2-layered intine (i), exine elements (e), anc. non-apertural wall (left) and gemmate apertural wall fibrillar deposit between exine elements; outer intine (right). SEhl x5200. (C) Pollen tetrad, showing decussate layer contains apparent channel-like structures (arrow). arrangement of microspores and distal orientation of TEhl X23 000. (G) Cross-section of whole grain. Note apertures. SEhl x I 300. (D) Non-apertural wall; note aperture (n), with greatly thickened outer intine layer. intine (i) and I-layered exine (e). TEhI ~31600.(E) TEhl x4400. Acetolyzed non-apertural exine; note laterally appressed

Grana 22 Polleii iiltr-tistrirctirre oftlic Hne/Iiochr-ricetie 91

\vall composed of a thin intine and a continuous, apertural walls (Fig. 11 D), containing irregular, ?-layered exine, a commissural line evident be- sinuous, channel-like structures (Fig. 11 E). tween the outer homogeneous and inner papillate esine layers (Fig. 9 N). Lophidti (monotypic) L. mre(i Ker-Gawler.-Grains monosulcate and heteropolar (Fig. 12 A). Non-apertural wall re- 7riDor1ririrlics (1 of 3 species examined) 7. austrrilis Endl.-Pollen apolar, globose with 7-8 porate apertures (Fig. 10 A). Non-apertural wall Fig. 8. A-G: cl,iigo~orir/ios~ai,i[~irsD.C. (Conostylideae). sculpturing rugulate, that of the apertural wall ir- (A) Pollen grain, showing two hemispheric apertural walls regular (Fig. IOB). Exine of the non-apertural wall and attached raphide crystal. SEhi x 1200. (B) Close-up 2-layered, a commissure occuring between the con- of wall; note rugulate sculpturing of non-apertural wall tinuous outer layer and the inner layer of discon- (left) and paucity of exine on aperture (right). SEhl x2 600. (C) Rare, 3-porate grain; note raphides. SEhi. ~900.(D) tinuous, generally papillate elements (Fig. IOD). Acetolyzed non-apertural wall, showing basal exine lntine of the non-apertural wall 2-layered, the inner material interconnecting major exine elements. SEhI layer thicker (Fig. IOC, D). Apertural wall com- XS 000. (E) Cross-section of acetolyzed non-apertural posed of closely spaced verrucate exinous elements wall, showing three-dimensional structure; note papillate protuberances (arrow) on inner surface of wall. SEhl atop a thick 2-layered intine (Fig. IOE). Outer aper- ~8000.(F) Non-apertural wall, showing exine with inner tural intine layer contiguous with that of the non- papillate elements and commissural line (arrows). TEhi apertural wall (Fig. IOC) and greatly thickened in x22 700. (G)Interface between aperture (a)and non-aper- the aperture region (Fig. lOC, E), containing promi- tural wall (right); note exine-less, 3-layered apertural nent, radially-oriented, channel-like structures intine, with a thickened middle layer containing channel- like structures (arrow). TEhi X 13000. (Fig. IOE). H-hi: Mncropiclici firligimsn (Hook.) Druce (Conostyl- ideae). (H) Pollen grain; note two pores. SEhl x 1200. (I) Close-up of wall; note rugulate sculpturing of non-aper- Plilebocnr-yn (1 of 3 species examined) tural wall (lower left) and exine-less apertural wall (upper P. cilintn R. Br.-Pollen grains cylindrical, diporate, right). SEhl x3200. (J) Longitudinal-section of grain, and isopolar; sculpturing of non-apertural and aper- showing aperture wall (n),devoid of exine. TEhi X 1 300. turd walls rugulate (Fig. 10 F). Exine of non-aper- (K) Apertural wall, 3-layered; note extensive channel- tural wall composed of two equally thick layers, a like structures of middle layer (arrow). TEhl x21 100. (L) Non-apertural wall; note characteristic papillate elements prominent commissural face occurring at the junc- of inner exine layer. TEhl ~28600.(hl) Close-up of non- tion of the layers (Fig. IOH). Toward the aperture, apertural wall, showing 2-layered exine with a commis- lower exine layer gradually reduced to thinner, sure (arrows) between the layers. TEhi x58700. distinct, papillate structures (Fig. 10 I). Apertural Fig. 9. A-I: Co/iosrjlis bcrilinrio F. hluell. (Conostyl- wall composed of a thick, 3-layered intine, with lideae). (A) Pollen grain, showing 3 hemispheric pores, channel-like structures apparent in the middle layer devoid of exine; note raphide crystals. SEhi X 1600. (B) (Fig. IOG); scattered exine elements present out- Close-up of wall; note rugulate sculpturing of non-apertural wall (upper left). SEhI X3 100. (C) Rare 4-porate side apertural intine wall (Fig. lOG). grain. SEhl x900. (D) Rare 2-porate grain. SEhi X 1000. (E) Aberrant 3-porate grain. SEhi ~900.(F) Slightly oblique section of whole grain, showing 3-apertures (a), Additional taxa which are devoid of exine. TEXI X 1 600. (G)Non-apertural wall; note papillate protuberances on inner surface

Downloaded by [SDSU San Diego State University] at 10:48 25 February 2015 Cniznr-in (monotypic) of exine (arrows). TEhl x 16200. (H) Fibrillar apertural

I. Iniiotn (L.) Dur. & Schinz.-Grains monosulcate wall, showing channel-like structures (arrow). TEhl nd heteropolar (Fig. 11 A) with a foveolate non- ~19500.(I) Interface between aperture (a) and non- ipertural wall and a psilate apertural wall (Fig. apertural wall (right). TEhl x 10400. J-N: Blnricon rniiesreiis Lindl. (Conostylideae). (J) 1 B). Non-apertural wall exine tectate-columellate Pollen grain; note two hemispheric apertures. SEhl !ectate-perforate) with a thick foot-layer, a com- x 1700. (K) Close-up of wall, showing rugulate non-aper- nissural junction present at the center of the colu- tural wall (right) and exine-less aperture (left). SEhl nellae (Fig. 11C), Non-apertural intine thin, 2- x2 900, (L) Rare 3-porate grain. SEhl X 1 200. (hl) Inter- face between aperture (a)and non-apertural wall (right). 11 yered (Fig. C). Apertural wall composed of a TEhl x9300. (N) Non-apertural wall; note papillate pro- .lick, 3-layered intine (Fig. 11 E). hliddle intine tuberances of inner exine; commissural line (arrows) is layer thickened at junction of aperture and non- faintly visible. TEhl ~62100.

Grana 2; Downloaded by [SDSU San Diego State University] at 10:48 25 February 2015 Polleri iiltrnstriictiire of the Haetiiodorncene 93 Downloaded by [SDSU San Diego State University] at 10:48 25 February 2015 94 hi'. G. Sinipsori

-1 Downloaded by [SDSU San Diego State University] at 10:48 25 February 2015

Fip. I0 Polleii iiltrastriictiire of tlic Hoeinodoracetie 95

ticulate; apcrtural wall scabrate to shallowly ridged Hutchinson 1973) as a whole, with the exception (Fig. 12B). Non-apertural wall composed of a of Lririaria and Lopliiola, is palynologically united tectate-columellate (semitectate) exine with a thick by the absence of a typical tectate-columellate foot-layer and a relatively thin, 2-layered intine architecture. Even Schickia, which has a 3-layered (Fig. 12 C). Apertural wall exineless, consisting of a exine, does not have columellae, possessing in- thick, 3-layered intine (Fig. 12 C). Distinctive radial- stead a discontinuous, amorphous, granular middle ly oriented, electron dense regions present in the layer. Furthermore, in all six genera of the tribe middle intine layer, these regions usually corre- Conostylideae and in four genera of the tribe Hae- sponding to a papilla or ridge of the aperture (Fig. modoreae, the non-apertural exine lacks a foot- 12D). layer and is 2-layered, the difference between genera being primarily in the thickness and continuity Priiiridia (1 of 2 species examined) of the lower exine layer. The remaining four genera P. iiiiiiirtri (L.f.) Dur. & Schinz.-Pollen grains of the Haemodoreae are linked to the 2-layered disculcate and heteropolar (Fig. 12E, G. H). Sculp- members of that tribe by a gradation in exine wall turing of the non-apertural wall foveolate, with structure (Fig. 13). Thus, the occurrence of a com- minute supratectal papillae (Fig. 12 F). Apertural mon pollen wall architecture in the tribes Conos- sculpturing verrucate (Fig. 12 E, F, H). Non-aper- tylideae and Haemodoreae (excluding Loitarici and tural exine tectate-columellate (tectate-perforate) Lopliiola)argues strongly for their close relationship with a thin, electron-dense, apparently endexinous and the monophylesis of the family, sensu Hutchin- basal layer; ektexinous foot-layer absent (Fig. 12 I). son (1973). In contrast to the Haemodoraceae, all Apertural wall with scattered exinous verrucae atop 7 genera of the Tecophilaeaceae (sensu Hutchinson a thick, 2-layered intine (Fig. 12 J). 1973) possess a characteristic tectate-columellate wall stratification with a continuous, ektexinous foot-layer (Simpson 1981 b, work in progress). The DISCUSSION taxonomic treatment by Melchior (1964), which Prilyiologiccil clioracterislics classified the Tecophilaeaceae as the tribe Con- Ultrastructural features of the pollen wall provide anthereae of the Haemodoraceae, is not supported excellent characters for noting similarities and based on evidence from the present study. discontinuities between genera and tribes of the A distinctive commissural plane delimits the Haemodoraceae. The Haemodoraceae (sensu exine layers in the ten genera of the Haemodoraceae with 2-layered exine walls. Although the commissure almost always appears to be a real boundary Fig. 10. A-E: Triborinnthes nirstrnlis Endl. (Conostyl- between layers, it may in some cases represent a ideae). (A) Whole grain, showing three visible apertures region of structural weakness (and therefore break- (a). SEhl x 1900. (B) Close-up, showing rugulate non- age during sectioning). In either case, a structural apertural wall and aperture with irregular exine elements. SEhl ~5200.(C) Section of pollen grain, showing two distinction can be made between outer and inner apertures (0). TEhl ~2400.(D) Non-apertural wall, exine layers in these taxa. However, no difference showing 2-layered intine (i) and 2-layered exine (e); a in staining properties and no endexinelektexine commissural line (arrows) occurs between the 2 exine differentiation can be observed between the two layers; note papillate protuberances of inner exine layer. exine layers. Preliminary studies by the author TEhl X 19600. (E) Apertural wall, showing thick intine using brightfield and fluorescence microscopy Downloaded by [SDSU San Diego State University] at 10:48 25 February 2015 (i)and outer, scattered exine elements; note thickening of outer intine layer, which contains numerous channel-like staining techniques indicate that both exine layers structures (arrow). TEhl ~5500. in these genera have an ektexinous chemical reac- F-I: Plilebocnryn cilintn R. Br. (Conostylideae). (F) tion. Further studies by the author are planned to Whole pollen grain; note 2-porate apertures at ends of grain. SEhl ~2300.(G) Aperture wall, with thick intine determine the exact chemical nature and ontogeny (i) and occasional exine elements (el; note channel-like of the exine in "2-layered" taxa of the Haemodor- structures (arrow)of intine. TEhl ~22000.(H) Non-aper- aceae. tural wall, showing intine (i) and 2-layered exine (e);a Members of the Haemodoraceae also possess a commissural line (arrows) occurs between the two exine layers. TEhl ~45800.(I) Region of aperture border; note common intine wall structure. Non-apertural in- papillate protuberances of inner exine layer and com- tines characteristically have 2-layers, which are missure (arrows). TEhl ~21400. distinguished based on differences in staining pro-

Grana 22 96 hi. G. Siriipsori

Fig. 11. Larznria lomm (L.) Dur. & Schinz. (A) Whole mellae (arrows). TEhl x24 100. (D) Longitudinal-section pollen grain, aperture at left. SEhl ~4000.(El) Close-up of pollen grain; note aperture (a),devoid of exine. TEM of wall, showing foveolate non-apertural wall (left) and ~3500.(E) Interface between apertural (left) and non-

Downloaded by [SDSU San Diego State University] at 10:48 25 February 2015 psilate aperture wall (right). SEhl x8 100. (C)Non-aper- apertural wall (right); note 3-layered intine of aperture turd wall, showing 2-layered intine (i) and tectate-colu- with numerous, sinuous, channel-like structures (arrow). mellate exine (e);note commissural line at middle of colu- TEhl X21500.

perties or fibrillar structure. Initial chemical in- 1982). Progressing from the non-apertural to the vestigations of some members of the Haemodor- apertural wall, the outer intine layer becomes sub- aceae indicate that the inner layer is primarily cel- stantially thickened and is traversed with narro\v, lulosic and the outer composed of pectic com- radially oriented channel-like structures. lop hid^ pounds, as has been found for numerous angio- lacks these channel-like structures, possessing sperms, both monocots and dicots (Kress & Stone instead curious undulating, electron dense regions

Grana 22 Pollen iiltrastrirctirre of the Hnenioriorncene 97

in the apertural intine. In some taxa, mainly of the rorliiza has a third type of wall architecture, pos- tribe Conostylideae, there is an additional thin, sessing a 2-layered exine, the inner layer being dis- rather ill-defined layer of the apertural intine exter- continuous, amorphous, and somewhat granular. nal to the other two intine layers. Finally, a fourth type is present in Schiekin, which The tribes Haemodoreae and Conostylideae can has a 3-layered exine, the middle layer of which be differentiated based on pollen characters. As resembles the inner exine layer in Pyrrorhiza. originally noted by Erdtman (1966), members of the Unlike that of other members of the tribe, the Haemodoreae have monosulcate, heteropolar exine sculpturing of Schiekiri is foveolate-fossulate, grains while those of the Conostylideae have 2 or resembling that of tectate-columellate taxa, such as more porate, isopolar, subisopolar, or apolar grains. Latinria of the Haemodoraceae (present study) or This trend, supported by Radulescu (1973) for Cyanella and Odorzrostoniirrn of the Tecophila- Schiekia, is here confirmed for Barberetta, Macro- eaceae (Simpson 19816). In fact, the three exine pidia and Pyrrorhiza (Table I). (It should be noted layers of Schiekin show resemblances to a typical that only in Lachrzriiithes is pollen polarity absolute- foot-layer, columellae, and tectum and may re- ly determined, based on observations of tetrads in present an evolutionary modification from the typi- the present study.) The only ultrastructural char- cal tectate-columellate architecture. The wall of acter which diagnostically separates the two tribes Scliiekin differs, however, in that the middle exine is sculpturing. In the Haemodoreae non-apertural layer is structurally discontinuous with the inner walls are verrucate, except for Schiekici which is and outer layers and is amorphous, not columnar, somewhat foveolate-fossulate. In contrast, all mem- in shape. Pyrrorliiza, which has a verrucate sculp- bers of the Conostylideae have a characteristic turing and lacks a third, inner exine layer, is some- rugulate non-apertural wall sculpturing (Table I). what intermediate between Schiekia and 2-layered Additionally, all members of the Haemodoreae members of the Haemodoreae. Thus, a structural have exine as a component of the apertural wall, gradation can be hypothesized between Schiekin, whereas in most members of the Conostylideae, Pyrrorliiza, and the 2-layered members of the tribe apertural exine is lacking. Haemodoreae (Fig. 13). It should be pointed out, Within the tribe Haemodoreae four basic types however, that suggestions of an evolutionary grada- of wall structures can be distinguished (Table I). tion in pollen wall structure presupposes the mono- Four genera of the tribe, Barberetfa, Dilatris, phylesis of the Haemodoraceae. Such a gradation Wachericlorfin, and Xiphidiirriz, possess a distinctive can only be substantiated based on comparative 2-layered exine, evident by the presence of a com- analyses with other taxa using a wide range of char- missural “line” in electron micrographs. Further acters. similarities can be noted between Wnchendorfia Subgroups of pollen morphologies can be noted andXiphidiirnz, which both have a partially granular in the tribe Conostylideae. Four genera of the tribe, lower exine layer, and between Barberetta and Arzgiozanthos, Blnncoa, Coiiostylis, and Macro- Wncliendorfia, which have a similar proximal wall pidia, are very similar in having relatively smooth, sculpturing and identical psilate, micropore-pitted hemispheric to obconical aperture wall protrusions. apertural borders (differing from the smooth, per- Conostylis has 3-porate) triangular grains, while the forated border ofDilatis corymboso). A second wall other three genera have 2-porate, fusiform pollen structure type of the tribe occurs in Haeniodoriirii grains. However, the fact that occasional 2- or 4-

Downloaded by [SDSU San Diego State University] at 10:48 25 February 2015 and Lachrzarithes. These two genera possess an porate grains are found in Coizostylis and 3-porate exine composed of laterally appressed, basally grains in Ariigozarithos is indicative of an inter- fused, baculate structural elements. These elements gradation of pollen morphology between these are not 2-layered, except very rarely in isolated genera. (SEM observations of pollen from several portions of the pollen wall of H. simplex; the occa- other species of both Aizigozarithos and Conostylis sional presence of this thin, inner exine layer in H. indicate morphologies very similar to the species sirriplex may be evidence of a structural homology described here; palynological studies may trove with 2-layered members of the family. Thus, the useful in resolving relationships of species com- wall structure of Hnemodoriini and Laclinnntlies plexes within the two genera). Phlebocarya differs may be homologous with the upper exine layer of in the tribe in having cylindrical pollen grains with the 2-layered members of the tribe (Fig. 13). Pyr- two circular, flattened aperture walls. Triboiinrithes

7-838361 Grana 22 98 hf. G. Sirlzpsorl

Downloaded by [SDSU San Diego State University] at 10:48 25 February 2015 .a

Fig. 12 Pollen iiltrastriictiire of the Haernodoraceae 99

LACHNANTHES

rnQ. .,b WACHENDORFIA XlPHlDlUM

SCHlEKlA PYRRORHIZA €zxZz BARBERETTA DlLATRlS \€3= PHLEBOCARYA

Fig. 13. Hypothetical morphocline of exine wall architec- - ture in the non-tectate-columellate members of the Hae- - TRIBONANTHES modoraceae. The indicated gradation between Schiekia, Pjrrorliizn, and genera with a commissural, 2-layered ANIGOZANTHOS BLANCOA exine is unclear. CONOSTYLIS MACROPlDlA

is unique in the whole family in possessing spherical cause of the small size of the inner exine papillae). pollen with 7-8 pores. =_. A gradation is evident between the exine structure All genera of the Conostylideae have a 2-layered in the Conostylideae and the 2-layered wall struc- exine, the lower layer being composed of discon- ture of four genera of the Haemodoreae. Phlebo- tinuous papillate elements (a 2-layered exine was cnryn especially corroborates this gradation in hav- not observed in Conosiylis: however, perhaps being a thick, continuous inner exine (resembling that of 2-layered members of the Haemodoreae), which, in the aperture region, becomes thin, discontinuous Fig. 12. A-D: Lophiolo mren Ker.-Gawler. (A) hlonosul- and papillate (resembling the exine of the rest of the cate pollen grain, aperture at upper right. SEhl x4000.03) Close-up of wall, showing reticulate non-apertural wall Conostylideae) (Fig. 13). (lower left) and ridged aperture wall (upper right); note apparent “fusion” of exine elements at aperture border. Tctwotiotiiic itiiplicniiotis SEhl x15200. (C) Interface between apertural (n) and The taxonomic position of Phlebocaryn is clarified non-apertural (left) wall; note tectate-columellate non- apertural exine (e)and 3-layered intine (i); middle intine with reference to pollen wall ultrastructure. Plilebo- layer. is expanded at aperture border and contains radial- cnryn has been classified in either the tribe Conos- ly-oriented, electron dense regions (arrow). TEhl X32 800. tylideae (Pax 1888, Pax & Hoffmann 1930) or Hae- (D) Aperture wall, showing general correspondence of modoreae (Bentham & Hooker 1883, Hutchinson electron dense regions with the ridged apertural wall 1973, Geerinck 1969). However, the pollen of Phle- sculpturing (arrow). TEhl x55 700.

Downloaded by [SDSU San Diego State University] at 10:48 25 February 2015 E-J: Pairridin niinufa (L.f.) Dur. & Schinz. (E) Pollen bocnrya has at least three structural features shared grain, apparent polar view, showing two sulcate apertures only with genera of the tribe Conostylideae: (1) (a).SEhl ~4000.(F) Close-up of aperture (left) and non- grains with 2 or more porate apertures (2) rugulate apertural wall (right). SEhl x9400. (G) Apparent equa- sculpturing (3) a papillate inner exine layer (found torial view of pollen grain showing two apertures (a). SEhl ~2700.(H) Cross-section of grain; note two aper- only near the aperture in Plilebocnrya). It is pro- tures (a) with outer exine elements, thickened intine, and posed that these are shared derived characters and plasmolyzed regions beneath. TEM ~2700.(I) Non- that the classification of Phlebocaryn in the tribe apertural wall showing intine (i) and tectate-columellate Conostylideae, as done only by Pax (1888) and Pax exine (e); note thin, electron dense foot-layer (arrow). & TEhI x33 500. (J) Interface between aperture (n)and non- Hoffmann (1930), is supported. The placement of apertural wall (right); note thick 2-layered intine (i). TEhl the genus in the tribe Haemodoreae has been based X20 400. primarily on the common occurrence of an im-

Grana 22 100 M. G. SinlpsoJl

Table I. Polleri characters of the Haeniodoraceae Apo. = apolar; Circ. =circular; Fov. = foveolate; Foss. = fossulate; Hemisph. = hemispheric; Het. = heteropola,; Iso. = isopolar; Pap. = papillate; Por. = porate; Psil. = psilate; Retic. = reticulate; Rugul. = rugulate; Sulc. = suIcate; Verr. = verrucate. * Lachnanthes is known to be absolutely heteropolar; polarity in all other genera is assumed ~ ~~ - Aperture Non-apertural wall - No. Type Shape Polarity Exine Sculpturing Exine stratification

Tribe Haemodoreae Bnrberet to 1 Sulc. Furrow Het. VerrJPsil. 2-layered, inner baculate Dilotris . 1 Sulc. Furrow Het. Verr. 2-layered, inner baculate-papillate Hoemodorrrtti 1 Sulc. Furrow Het. Verr. 1-layered Lochtintithes 1 Sulc. Furrow Het.* Verr. 1-layered Pyrorhiza I Sulc. Furrow Het. Verr. 2-layered, middle amorphous Schiekio 1 Sulc. Furrow Het. FOV.-FOSS.3-layered, middle amorphous Xiphidiirnt 1 Sulc. Furrow Het. Verr. 2-layered, inner baculate-granular Wocheridorfin 1 Sulc. Hemisph. Het. VerrJPsil. 2-layered, inner granular Tribe Conostylideae Anigozotithos 2 (3) Por. Hemisph. Iso. Rugul. 2-layered, inner papillate Bloticoo 2 Por. Hemisph. Iso. Rugul. 2-layered, inner papillate Cotlostylis 3 (2,4) Por. Hemisph. Iso. Rugul. 2-layered?, inner papillate hfacropodio 2 Por. Hemisph. Iso. Rugul. 2-layered, inner papillate Plilebocarp 2 Por. Circ. Iso. Rugul. 2-layered, inner baculate-papillate Tribotlarlfhes 7-8 Por. Circ. Apo. Rugul. 2-layered, inner papillate Latinria 1 Sulc. Furrow Het. Fov. Tectate-columellate, foot-layer thick Lopliiola I Sulc. Furrow Het. Retic. Tectate-columellate, foot-layer thick Pniiridin 2 Sulc. Furrow Het. Fov.lPap. Tectate-columellate, basal layer thin, endexinous

bricate perianth (Benthim & Hooker 1883, Hutchin- Tecophilaeaceae, sensu Hutchinson 1973), whose son 1934, Geerinck 1969), which is almost certainly members have monosulcate grains with operculate a primitive character for the group and should not sulci. The wall ultrastructure of the seven genera be used in delimiting hypothesized monophyletic of the Tecophilaeaceae is, like Lariaria and Lophi- taxa (sensu Hennig 1966). oh, tectate-columellate (Simpson 1981 b, work in In contrast to all other members of the Haemo- progress), thus seemingly supporting Erdtman's doraceae, Latinria and Lophiola possess a tectate- contention. However, other families presumed to columellate wall architecture, with a thick, con- be closely related to the Haemodoraceae (sensu tinuous foot-layer (interestingly, however, the Cronquist 1981, Dahlgren 1980, Hutchinson 1973, columellae of Lariaria often appear to be 2-layered). Takhtajan 1980, Thorne 198l)also possess a tectate- Apertural exine is absent in the two genera, whereas columellate pollen wall architecture similar to that all genera of the similarly monosulcate Haemo- of Lariaria, Lopliiola, and the genera of the Teco doreae have a prominent cover of apertural exine philaeaceae. The taxa investigated to date include elements. Lariaria and Lopliiola are also aberrant Barbaceriia of the Velloziaceae (Ayensu & Skvarla Downloaded by [SDSU San Diego State University] at 10:48 25 February 2015 within the family with respect to anatomy (Schulze 1974), Ciirciiligo & Hypoxis of the Hypoxidaceae; 1893, Simpson & Dickison 1981), chemistry (Ed- Helniholtzia & Philydrella of the Philydraceae; wards et al. 1970), and embryology (De Vos 1963, Tncca of the Taccaceae; and Liriope of the Lili- Simpson 1981 a). These data, applied to a cladistic aceae, tribe Ophiopogoneae (Simpson, work in analysis of the complex (Simpson, unpublished), progress). Although additional studies are needed, argue strongly against the classification of the two it is hypothesized here that the tectate-columella!e genera in either the tribe Haemodoreae or Conosty- wall structure is ancestral, based on outgroup corn lideae. Erdtman (1966) proposed that Larinrin and parison (Simpson, unpublished), for the complex Lophiola are palynologically more similar, in size as a whole, as it probably is for the angiosperms S and sculpturing, to the tribe Conanthereae (= a whole (Van Campo 1971). If this interpretation

Grana 22 Pollen rtltrastrrictrire of the Haemodoraceae 101

is correct, the exine of members of the Haemo- ological (De Vos 1961) and anatomical (Schulze doraceae may conceivably represent the loss of a 1893) characters, no clear taxonomic conclusions foot-layer from an ancestral tectate-columellate can be drawn based on pollen wall architecture. condition. Additionally, the common occurrence of Further palynological studies are needed to verify a tectate-columellate wall in Latzarin, Lophiola, the taxonomic placement of Paiiridia within the and members of the Tecophilaeaceae cannot pro- Hypoxidaceae. vide evidence for their classification in one taxon. In fact, the operculate aperture of members of the Coniparisons with post stirdies Tecophilaeaceae is almost certainly, for that family, Clarifications regarding the three pollen groups in a shared derived character based on outgroup com- the Haemodoraceae recognized by Erdtman (1966) parison (Simpson, unpublished). The absence of can be made with reference to the present ultra- an operculum in Lowria and Lopliiola argues structural study. Erdtman’s description of wall against their classification in the Tecophilaeaceae. structure in six genera of the tribe Haemodoreae However, whether Lanoria and Lopiola should be as “not very distinct’’ is probably a reflection of included within their own tribe of the Haemodor- the atectate architecture found in all members of aceae or transferred (perhaps not together) to some that tribe. The characteristic LO-pattern he ob- other taxon will remain unclear until additional served for five genera of the Conostylideae can be characters are found and comparative analyses with explained ultrastructurally. The “dark islands. . . other families are made. Anatomical evidence has separated by brighter channels” probably cor- recently been presented which supports the classifi- respond to the rugulae separated by fissures in the cation of Lophioln in the Liliaceae, tribe Melanthi- wall. Also, the “fainter darker dots = bacula” al- oideae (Ambrose 1980). most certainly correspond to the papillate pro- The genus Poirridia is palynologically distinct tuberances of the inner exine layer characteristic of from members of the Haemodoraceae. Although the tribe. Finally, the reticulate exine wall of Latin- the pollen of Paiiridki resembles that of Latinria rin and Lopliiolo corresponds with a tectate-colu- and Loplriola in having an apparent tectate-columel- mellate architecture as seen with the electron micro- late exine stratification, it differs in having: (1) two scope. apertures with a cover of ape‘rtural exine elements, Erdtman’s (1966: 199) description of a “coarsely (2) a supratectal papillate sculpturing, and (3) a very granular, distinctly convex operculum” in Wachen- thin, apparently endexinous basal layer with ekt- c/or$a is incorrect. This is not an operculum, but exinous foot-layer absent. The observation in the simply an aperture wall with an outer layer of close- present study of disulcate pollen in Paiiridia con- ly spaced exine structural elements. Radulescu’s firms a similar report by Thompson (1979) but con- (1973) description of granules or verrucae on the flicts with Erdtman’s (1966) description of the genus apertures of Dilatris and Schiekia corresponds to as monosulcate. Although Erdtman treated Paiiridia these exine elements. Similar aperture walls are within the tribe Haemodoreae, he did report resem- found in all other genera of the tribe Haemodoreae; blances in size and sculpturing between Paiiriclio the apertures often disintegrate during acetolysis, and members of the Hypoxidaceae (sensu Hutchin- probably because of the lack of continuous exine son). Preliminary studies by the author indicate that material between the elements. The use of the term two genera of the Hypoxidaceae, Cirrcirligo and reticulate by Radulescu (1973) for the sculpturing of

Downloaded by [SDSU San Diego State University] at 10:48 25 February 2015 Hjpoxis, are similar to Pairridin in having a tectate- Conostjlis and Dilotris is in disagreement with columellate architecture. However, pollen grains Erdtman (1966) and the present study. Radulescu’s of the former two genera differ from Pairridin in report of a pilate-tegillate wall structure in Co- having a monosulcate aperture and in possessing a tiostjlis and a tegillate-baculate exine for Dilatris thicker, continuous, ektexinous foot-layer, similar and Scliiekia inadequately describe the major struc- to that of Lorinria, Lophiolo, or members of the tural elements observed here. Tecophilaeaceae. In fact, the absence ‘of a continuous ektexinous foot-layer in Parrridio indicates similarity to the 2-layered members of the tribe CONCLUSIONS Haemodoreae. Although the placement of Pairridin Observations of pollen ultrastructure are useful in in the Hypoxidaceae is supported based on embry- characterizing and delimiting the Haemodoraceae,

Grana 22 102 hf. G. Siriipsori

family tribes, and groups of genera within tribes. Cronquist, A. 1981. An integrated system of classification Suggestions regarding the classification of family of flowering plants. - Columbia University press N.Y. taxa and the intergradation of wall types have been Dahlgren, R. 1980. A revised system of classification of made based on these pollen characters. Of especial the angiosperms. - Bot. J. Linn. SOC.80:91-124. interest in this study is the general occurrence in Dellert, R. 1933. Zum systematischen Stellung the family of a 2-layered exine wall lacking a foot- Wachendorfia. - Oesterr. Bot. Z. 82: 335-345. layer. Based on comparisons with taxa of several Edwards, J. hl., Churchill, J. A. & Weiss, U. 1970. A chemical contribution to the taxonomic status of other, presumably closely related families, it is Lophiola americana. - Phytochemistry 9: 1563-1563. proposed that the exine structure of the Haerno- Erdtman, G. 1966. Pollen morphology and plant taxo- doraceae is derived and the tectate-columellate nomy. Andosperms. Corrected reprint and ne\v ad- architecture is probably primitive for the complex. dendum. - 553 pp. Hafner Publ. Co., New York. Geerinck, D. 1968. Considerations taxonomiques au sujet However, comparative analyses, such as cladistic des Haemodoraceae et des Hypoxidaceae (hlono. techniques, using all known characters should be cotyledones). -Bull. SOC.Bot. Belg. 101:265-278. made to test hypotheses regarding homology and Geerinck, D. 1969. Genera des Haemodoraceae et des character polarity. The pollen of many more mono- Hypoxidaceae. - Bull. Jard. Bot. Natl. Belg. 39: cotyledonous taxa need to be studied before broad 47-82. Green, J. W. 1959. The genus Conostylis R. Br. 1. Leaf taxonomic comparisons can be noted. The addi- anatomy. - Proc. Linn. SOC.N. S. W.84: 194-206. tional families of the Haemodorales, sensu Hutchin- Green, J. W. 1960. The genus Conostylis. 11. Tax. son (1973) and Dahlgren (1980), are under current onomy. - Proc. Linn. SOC.N.S.W. 85: 334-373. investigation by the author. Hegnauer, R. 1963. Chemotaxonomie der Pflanzen. Band 2. hfonocotyledoneae. - Birkhauser Verlag, Basel und Stuttgart. ACKNOWLEDGEMENTS Hennig. W. 1%6. Phylogenetic systematics. - 263 pp. This study was supported in part by United States Na- University of Illinois Press, Urbana. tional Science Foundation grant DEB-81-09909 and is part Hilliard, 0. hl. & Burtt, B. L. 1971. Noteson some plants of a Ph.D. dissertation submitted to the Department of of southern Africa chiefly from Natal: 11. - Notes R. Botany, Duke University. I wish to thank Patricia G. Bot. Gard. Edinburgh 31: 1-33. Gensel for conceiving the project and making initial ob- Hutchinson, J. 1934. The families of flowering plants, ed. servations; Donald E. Stqne and Richard A. White for 1. vol. 2, hlonocotyledons. - Oxford Univ. Press, reviewing the manuscript; Gwen C. Roney for substantial London. editorial help; Robert Omduff, Bassett hlaguire, and Peter Hutchinson, J. 1959. The families of flowering plants, ed. Goldblatt for supplying critical plant material; Greg J. 2, VOI. 2, Monocotyledons. - pp. 512-792. Oxford Keighery, Steven D. Hopper, and P. G. Wilson for their Univ. Press, London. help in field collection in Australia; herbaria at US, UC, Hutchinson, J. 1973. The families of flowering plants, ed. hlO, DUKE, UCSC, PERTH, and NY for supplying plant 3. -Oxford Univ. Press, London. samples or housing vouchers and Sue D. Dickerson for Kress,W. J. &Stone, D. E. 1982. Nature ofthe sporoderm typing the manuscript. in monocotyledons, with special reference to the pollen of Canna and Heliconia. - Grana 21: 129-148. Lynch, S. P. & Webster, G. L. 1975. A new technique of REFERENCES preparing pollen for scanning electron microscopy. - Ambrose, J. D. 1980. A re-evaluation of the hlelanthio- Grana 15: 127-136. idede (Liliaceae) using numerical analysis. - In: hfelchior. H. 1964. Haemodoraceae. - In: Engler's Petaloid hlonocotyledons (ed. C. D. Brickell, D. F. Syllabus der Pflanzenfamilien, ed. 12, vol. 2. - p. Cutler and hi. Gregory - Linn. SOC.Symposium 527. - 666 pp. Gebrtider Bomtraeger, Berlin. Series No. 8: 65-82. Ornduff, R. 1979. Chromosome numbers and relationships

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