Studies on pollen morphology of selected species of Annonaceae from Thailand

The pollen morphology of 49 species and one variety, representing 18 genera of the family Annonaceae from Thailand, is described and illustrated based on observations using scanning electron microscopy (SEM). The pollen grains of Anaxagorea show double-spheres as a result of the intine extrusion. The palynological evidence confirms the close relationship of Dasymaschalon, Desmos and Asian Friesodielsia. Decussate tetrads in Goniothalamus and coexisting tetragonal, rhomboidal, T-shaped, tetrahedral and decussate tetrads in Mitrephora are reported. Pollen morphology is consistent within Huberantha, Marsypopetalum and Monoon, but is more diverse in Polyalthia sensu stricto Pollen unit, shape, size, ornamentation and aperture number reaffirm the great diversity among and within genera in Annonaceae.     

Cytotaxonomical studies on AsianAnnonaceae

Chromosome numbers of 42 species and 3 varieties from 24 genera of theAnnonaceae have been determined (Table 1); reports for 15 of the genera are new. Among Asian genera 2n = 14 occurs only in the specializedMezzettia, while 2n = 16 is wide-spread and also has been found inAnaxagorea with some primitive characters. 2n = 18 is reported for 11 genera, and tetraploidy (2n = 36) has been observed inPolyalthia. Therefore, an original basic number of x = 8 or x = 9 is suggested at least for the Asian genera of theAnnonaceae.—Cytotaxonomical notes on the critical speciesPolyalthia rumphii andP. affinis are given, and the new combinationNeouvaria parallelivenia (Boerl.)Okada & Ueda is proposed.     

Pollen of Ceratostema (Ericaceae, Vaccinieae): tetrads without septa

The pollen morphology of two species of the Neotropical genus Ceratostema (Ericaceae) was examined by light, scanning and transmission electron microscopy. The Ceratostema species examined have 3-colporate pollen grains united in permanent tetrahedral tetrads that show a common condition encountered in the Ericaceae. But the septal exine was absent between two neighboring grains in each pollen tetrad of Ceratostema. The pollen tetrads without septa are the first report for the Ericaceae as well as other angiosperm families.     

From stowaways to karst‐aways: unloading the Peninsular Malaysia endemic Polyalthia brunneifolia (Annonaceae)

Detailed examination of specimens collected from limestone outcrops in Peninsular Malaysia and previously included in Polyalthia brunneifolia shows that they do not belong in this species. Three new species are described to accommodate these collections: Polyalthia chinii I.M.Turner & Utteridge from Bukit Serdam, Raub, Pahang; P. guabatuensis I.M.Turner & Utteridge from Batu Caves, Selangor, and P. guamusangensis I.M.Turner & Utteridge from Gua Musang, Kelantan. Conservation assessments are included for the new species and the general threats faced by species confined to limestone hills are discussed. In addition, an updated key to all species of Polyalthia known from Peninsular Malaysia is provided.     

Pollen morphology of some selected plant species from Horton Plains,Sri Lanka

The pollen morphology of four species: Gaultheria leschenaultii (Ericaceae), Psychotria zeylanica (Rubiaceae), Rubus leucocarpus (Rosaceae) and Symplocos bractealis (Symplocaceae) from Horton Plains, Sri Lanka was investigated using LM and SEM. Psychotria zeylanica is endemic to Sri Lanka and the other three taxa are common in Horton Plains. Gaultheria leschenaultii has tetrads with 3-colporate, coarsely granular to granulo-rugulate pollen grains; in the polar region and at the colpus margin the exine is smooth with micro-rugulate structure. Psychotria zeylanica has 3- (or 4-) colporate, reticulate-spinulose to granular pollen grains. Rubus leucocarpus has 3-colporate, striate-perforate pollen grains and Symplocos bractealis has 3- (or 4-) colporate pollen grains with a verrucate-perforate exine. Pollen grains of these four species are very indicative of human impact and palaeoclimate.     

Evolution of pollen,stigmas and ovule numbers at the caesalpinioid–mimosoid interface (Fabaceae)

In this study we examine the pollen, stigmas and ovaries from 62 collections of herbarium material representing 16 genera, using light and scanning electron microscopy. The caesalpinioid Dimorphandra group (Burkea, Dimorphandra, Erythrophleum, Mora, Pachyelasma, Stachyothyrsus and Sympetalandra) pollen grains are small, tricolporate monads, with perforate or psilate ornamentation. Dinizia, Pentaclethra and Aubrevillea have morphological characters that have suggested either a mimosoid or caesalpinioid placement. Dinizia pollen is in permanent tetrads with clavate ornamentation. Pentaclethra pollen grains are monads, two species have tricolporate pollen and the third is porate. Aubrevillea has tricolporate, finely reticulate monads. All ten genera have variable, non‐predictable stigma type and ovule number. The mimosoid Adenanthera group (Adenanthera, Tetrapleura, Amblygonocarpus, Pseudoprosopis, Calpocalyx and Xylia) pollen grains are in 8‐ to 16‐grain polyads. In all Adenanthera group species, the stigmatic cavity is only large enough to accommodate one polyad. In addition, the number of ovules present matches the number of pollen units in one polyad. Polyads have porate, operculate apertures that differ in layout, aperture morphology and development when compared with caesalpinioid and other eudicot pollen. © 2010 The Linnean Society of London, Botanical Journal of the Linnean Society, 2010, 162 , 594–615.     

Reproductive biology of Lactoris fernandeziana (Lactoridaceae)

Lactoris fernandeziana, monotypic in its family, is endemic to the cloud forests of Robinson Crusoe Island. Although there has been considerable study of the relationships of Lactoris, as a rare species and as a putative primitive paleoherb, little is known of its reproductive biology. Knowledge of the latter is essential for effective conservation programs. The species is gynomonoecious. The overall proportion of flowers is ∼1 female:1 hermaphrodite. The inconspicuous semipendulous green flowers, usually in mixed-gender inflorescences, do not produce rewards. Hermaphrodite flowers are herkogamous and protogynous. Pollen grains are shed from the extrorse anthers in permanent dry tetrads. There is a mean of 12879 tetrads per hermaphrodite flower. Both flower types bear an average of ∼18 ovules. The P/O (pollen/ovule) ratios imply facultative or obligate xenogamy, but hand pollinations show that Lactoris is self-compatible. No floral visitors were ever observed, but stigmata of open-pollinated flowers bore tetrads, and 64% of such styles had pollen tubes. Flowers enclosed in large mesh (1 mm) bags bore similar numbers of tetrads and pollen tubes. Thus, we conclude that Lactoris is anemophilous, a syndrome perhaps reflected by the P/O ratio. Low genetic diversity (isozymes and DNA) supports selfing and implies limited distance wind pollen dispersal. The small size of the island, the ± 1000 extant Lactoris plants, coupled with anemophily, self-compatibility, and pendant flower position, have yielded a geitonogamous system with high seed set and low genetic diversity. If inbreeding depression is expressed, it is in seed germination and seedling vigor, for Lactoris is very difficult to cultivate. For this species, effective conservation practices need to focus on habitat preservation and promotion of outcrossing.     

New species of Polyalthia (Annonaceae) from Borneo and their relationship to Polyalthia cauliflora

A group of seven Polyalthia species from Borneo allied to P. cauliflora Hook. f. & Thomson is revised. Three of these are described as new. Polyalthia myristica sp. nov. and P. trochilia sp. nov. are similar to P. stenopetala (Hook. f. & Thomson) Finet & Gagnep., particularly in having seeds with a deep circumferential groove. Polyalthia kinabaluensis sp. nov., endemic to Mount Kinabalu, is similar to P. cauliflora, but differs in having petals slightly connate at the base and larger monocarps. Polyalthia ichthyosma sp. nov. is the fourth new species described. It bears some similarity to one of the P. cauliflora group, P. dictyoneura Diels, but differs in not being cauliflorous. Two synonyms of Polyalthia cauliflora, Unona desmantha Hook. f. & Thomson and Unona pycnantha Hook. f. & Thomson, are lectotypified.     

A New Pollen Type in the Winteraceae

Pollen of Zygogynum balansae and Z. pomiferum is described with the aid of the scanning electron microscope. Other members of the primitive ranalean family Winteraceae hitherto investigated have pollen in permanent tetrads, but these two species have solitary grains. Sculpturing is similar on the distal and proximal faces of each grain, excluding the distal apertural zone. The exine pattern resembles that on distal faces of tetrads of other Zygogynum species investigated, including Z. bailloni, the pollen of which is illustrated. Although Zygogynum and Exospermum have rather similar pollen, with sculpturing differing from other Winteraceae in consisting of a less coarse reticulum, pollen of Exospermum stipitatum is clearly distinct from the Zygogynum species investigated, as seen in the scanning electron microscope. It is not clear whether or not the solitary pollen type has evolved from the tetrad form.     

THE EVOLUTION OF POLLEN TETRADS IN ONAGRACEAE

In Onagraceae, pollen is shed in mature tetrads in most Epilobieae, many species of Ludwigia (Jussiaeeae), and two closely related species of the large genus Camissonia (Onagreae). Mature tetrads of Camissonia cardiophylla and representative species of Epilobium and Ludwigia were examined with light, scanning, and transmission electron microscopes. Morphological diagnoses of monad units indicated that individual taxa could be readily distinguished. Statistical analyses of tetrads which remained after acetolysis treatment revealed significant differences in the strength of the binding mechanisms. Mechanisms of tetrad cohesion were found to consist of two principal types. Common to all taxa is cohesion of pollen wall surfaces at the aperture margins; this mechanism is well known in many angiosperm groups. With the exception of Camissonia, the remaining taxa also display binding by means of short exine fragments between adjacent pollen units. These fragments, termed bridges and reported here for the first time, are located in the area extending from the aperture margins to near the center of the proximal exine faces. Thin sections reveal that layers of the bridges are identical with those of the exine. Comparisons were made between bridges and viscin threads, both of which occur on the proximal faces of the grains. Viscin threads are present on all pollen grains in Onagraceae and exhibit distinctive morphologies, and bridges were viewed morphogenetically as related to viscin threads but including an endexine layer and occupying a position near the apertures where cohesion of wall surfaces also occurs. In an evolutionary sense, the formation of mature tetrads almost certainly occurred independently in Camissonia and may have done so in Ludwigia and the Epilobieae.     

An Ultrastructural Study of Pollen Wall Ontogeny in Silene Pendula

Pollen of 41 species representing all seven genera of the Neottieae were examined by scanning electron microscopy. Except for Lecanorchis, the genera of the Neottieae constitute a natural group based on pollen morphology. Pollen occurs as single grains in the primitive species and in tetrads in the other species. Most grains are monoaperturate, porate or tenuate, and the tetrads often have irregularly shaped grains. Exine structure varies from tectate-perforate to semitectate. Lecanorchis is anomalous among the Neottieae in that it has 0–5, sunken, relatively small pores. Pollen morphology of this genus indicates that it is probably more closely related to the Gastrodieae than the Neottieae. There are at least four basic phyletic units in the Orchidaceae: the Neottioideae, Apostasioideae, Cypripedioideae, and Epidendroideae. These groups are distinguished by the presence of monads in at least their more primitive members and by their unique pollen types.     

Beschorneria yuccoides and Asimina triloba (L.) Dun: Examples for proximal polar germinating pollen in angiosperms

Beschorneria yuccoides (Agavaceae) microspores are arranged mostly in planar tetrads. Later on, the pollen grains of the tetrad usually fall apart, but sometimes remain loosely connected by ektexine elements. The ektexine consists of a tectum, of short columellae, and of a thin, discontinuous foot layer. An endexine is absent. The bilayered intine is without any additional thickening that would usually indicate an aperture region. From this point of view the pollen grain might be considered as omniaperturate. The pollen ornamentation is reticulate with wide lumina and robust, smooth muri.

The pollen grains show an indistinct sulcus characterised by a loose reticulate ornamentation. The sulcus is not exactly at the distal pole, but shifted towards the equator. No pollen tubes are formed regularly at the sulcus. Instead, pollen tubes are normally formed at the proximal pollen face. The proximal area, indicating a large germination field, is morphologically and functionally clearly an aperture (a germination zone); however, it does not represent a sulcus. The proximal face of all pollen grains appears as ornamented, with some exine lumps.

Asimina triloba (Annonaceae) pollen is shed in permanent planar or decussate tetrads. The distal sides are microreticulate to foveolate, and do not show an aperture; the psilate proximal sides are the germination areas of A. triloba.

The presence of apertures placed at the proximal pole was reported for distinct taxa of several angiosperm families. For Drosera, Dionaea (Droseraceae) and most probably for the diaperturate Cuphea species (Lythraceae) the existence of polar germination areas can be excluded. However, in some Annonaceae taxa with permanent tetrads (Annona cherimola, Asimina triloba) a situation similar to Beschorneria might be present, and indeed a proximal polar pollen tube is formed. Beschorneria yuccoides, Annona cherimola and Asimina triloba are unequivocal examples of angiosperm pollen with an exactly proximal aperture (germination area).     

Paleobotanical remains from the Paleocene–lower Eocene Vagadkhol Formation, western India and their paleoclimatic and phytogeographic implications

A small assemblage of macro- and micro floral remains comprising fossil leaf impressions, silicified wood, spores, and pollen grains is reported from the Paleocene–lower Eocene Vagadkhol Formation (=Olpad Formation) exposed around Vagadkhol village in the Bharuch District of Gujarat, western India. The fossil leaves are represented by five genera and six species, namely, Polyalthia palaeosimiarum (Annonaceae), Acronychia siwalica (Rutaceae), Terminalia palaeocatapa and T. panandhroensis (Combretaceae), Lagerstroemia patelii (Lythraceae), and a new species, Gardenia vagadkholia (Rubiaceae). The lone fossil wood has been attributed to a new species, Schleicheroxylon bharuchense (Sapindaceae). The palynological assemblage, consisting of pollen grains and spores, comprises eleven taxa with more or less equal representation of pteridophytes, gymnosperms, and angiosperms. Angiospermous pollen grains include a new species Palmidites magnus. Spores are mostly pteridophytic but some fungal spores were also recovered. All the fossil species have been identified in the extant genera. The present day distribution of modern taxa comparable to the fossil assemblage recorded from the Vagadkhol area mostly indicate terrestrial lowland environment. Low frequency of pollen of two highland temperate taxa (Pinaceae) in the assemblage suggests that they may have been transported from a distant source. The wood and leaf taxa in the fossil assemblage are suggestive of tropical moist or wet forest with some deciduousness during the Paleocene–early Eocene. The presence of many fungal taxa further suggests the prevalence of enough humidity at the time of sedimentation.     

Pollen Morphology of the Periplocaceae from Madagascar

The pollen morphology of eleven (Baroniella Constantin & Galland, Baseonema Schltr. & Rendle, Camptocarpus Decne., Cryptolepis R. Br., Cryptostegia R. Br., Gonocrypta Baill., Harpanema Decne., Ischnolepis Jum. & H. Perrier, Menabea Baill., Pentopetia Decne., and Tanulepis Balf. f.) of the 13 genera of the Periplocaceae from Madagascar have been examined using light-, scanning-, and transmission electron microscopy. All of the genera are characterized by pollen grains arranged in tetrads. The arrangement of the grains may be rhomboidal, decussate or tetragonal. The 4–6 pores present are restricted to the junction area of adjacent grains. Cryptostegia differs from the other genera in that the arrangement of pollen grains is only decussate. In Menabea the tetrads are united into a pollinium. The exine is smooth and consists of a distal stratum (tectum), subtended by a granular stratum consisting of granules of unequal size. Towards the base larger granules are present. In Camptocarpus, Harpanema and Tanulepis the exine is stratified into a distal stratum (tectum), a thin granular stratum and an almost continuous basal stratum (foot layer). The inline is well developed. The pollen grains of tetrads are connected by wall bridges (cross-wall cohesion). The internal walls in Camptocarpus, Harpanema and Tanulepis differ from the other genera in the absence of a tectum. The pollen morphology of the taxa investigated is very similar and of little value for distinguishing the species and genera investigated. The distinctive difference in exine structure between the above mentioned three genera and other genera investigated emphasizes the importance of exine ultrastructure in the Periplocaceae.     

Aperture types withinSapranthus andPolyalthia (Annonaceae)

Within the large palaeotropical genusPolyalthia so far five different pollen types are described (two monosulcate types, two disulcate types, and a single omniaperturate type). One of the disulcatePolyalthia types is similar to the disulcate neotropical genusSapranthus. All the newly described types differ predominantly not in exine but rather in intine characters. From this it can be concluded that seemingly inaperturate grains are disculcate in function, generally with two clearly recognizable germination zones, or omniaperturate, with the whole sporoderm capable of germination. The functions of the intine layers within the germination zones and within the nongerminating areas are very different and cooperate during the first germination steps in bursting the exine. It is difficult clearly to define the germination zones of pollen grains if the exine exhibits no modifications. Based on our and other results a more precise definition is suggested.     

Aperture variation in the pollen of Nelumbo (Nelumbonaceae)

The evolutionary and developmental origin of tricolpate pollen is of great interest because pollen of this kind defines a major clade of angiosperms (eudicots), a clade that is also well supported by molecular data. We examined evidence that tricolpate and monosulcate pollen types are produced alongside each other in the anthers of Nelumbo flowers, as has previously been reported. Observations of pollen in situ within individual anthers revealed mainly tricolpate pollen produced in tetrahedral tetrads, but also a small percentage of clearly aberrant pollen grains that have a great variety of aperture configurations. Previously published evidence for tetragonal tetrads is not supported, and previously reported monosulcate grains are part of a continuum of variation among the aberrant grains in aperture number, position and form. Other eudicots show similar variability in their pollen apertures. The variation in the pollen of Nelumbo is not exceptional, and may not be more significant than variation seen in the other taxa with regard to the origin of the tricolpate and tricolpate‐derived pollen characteristic of eudicots. Nevertheless further studies of aberrant pollen in Nelumbo and other eudicots, together with comparisons of pollen development in “normal” eudicots and closely related species that show radical, and developmentally fixed, reorganization of apertures and pollen polarity, may be helpful in understanding the processes that controlled the transition from the monosulcate to the tricolpate condition.     

An attempted analysis of the tetrads in the complex heterozygote species of the sub-genus Oenothera

Summary The statistical parameters of a pollen population of Oenothera purpurata in which the mendelian factor of pollen lethality segregates, showed that the pollen population analysed at anthesis in the Oenothera sub-genus quite faithfully conserves the segregation indicated in the tetrads. Following on this fact, the segregation active grains, inactive grains, empty grains was analysed in the pollen of the complex heterozygotes. The large standard deviation of the percentages of the three pollen classes and the value of the correlation between the percentages of grain classes taken two by two signify that the segregation of the three potential classes of microspore differs from one tetrad to another. By superimposing the correlation coefficient estimated between the percentage of the three grain classes of a bifactorial segregation of corn pollen on the correlation coefficient of the three classes of Oenothera pollen, it is possible to know with certainty the distribution of the potentially active, inactive and empty microspores in the tetrads. This distribution, which is different depending on whether the species is heterogamous or isogamous, allows us to confirm that the empty grains carry the recombined complexes which become lethal as a result of this recombination.     

STERILITY BARRIERS OF SOME ARTIFICIAL F1 ORCHID HYBRIDS: MALE STERILITY. I. MICROSPOROGENESIS AND POLLEN GERMINATION

Microsporogenesis, pollen germination and fertility of males gametes were studied in 24 artificial intergeneric and interspecific F₁ hybrids of orchids. Although parental species had the same chromosome number (2n = 40), microsporogenesis of the hybrids was irregular due to the lack of homology of the chromosomes of the parental species. This led to formation of tetrads of microspores without micronuclei, tetrads with 1–8 micronuclei, triads, dyads with and without micronuclei, and monads. Chromosomes numbers found in haploid microsporocytes ranged from 7 to 40; in micronuclei the chromosome number varied between 1 and 5. In terms of pollen germination, three situations were observed: 1) hybrids whose pollen grains did not germinate in the stigma; 2) hybrids in which the pollen tubes grew down in the style, but did not penetrate into the ovary; 3) hybrids in which the pollen tubes grew down normally through the ovary, reaching the ovules. When the pollen tubes did not penetrate the ovary no fruit was formed. Therefore germination tests carried out in vitro may not indicate pollen fertility, because pollen tube growth in the style of the flower may be insufficient to induce fruit formation or to accomplish fertilization.     

Aperture Orientation in Laurelia Pollen (Atherospermataceae Syn. Subfamily Atherospermoideae of Monimiaceae)

The development of the encircling aperture of pollen grains or the New Zealand species Laurelia novae-zelandiae is described by means of light and scanning electron microscopy. Some observations are also reported from pollen development and structure in the South American species Laurelia sempervirens, mature pollen of which can be distinguished from that of L. novae-zelandiae. The aperture begins to develop while pollen of L. novaezelandiae is still in tetrads, and passes through the distal and proximal poles of each grain. The widest parts of the mature aperture are at and near what are morphologically the distal and proximal poles of each grain. No such meridionosulcate type of pollen was recorded in a recent survey of pollen aperture types of primitive angiosperms and contradicts a recent report that pollen in the Atherospermataceae had equatorially aligned apertures.     

Anther development, microsporogenesis and microgametogenesis in Heliconia (Heliconiaceae, Zingiberales)

Anther development, microsporogenesis and microgametogenesis in several species of Heliconia were investigated as part of a complementary embryological study of the Heliconiaceae. All studied Heliconia species present bithecate and tetrasporangiate anthers with fertile pollen grains; only H. rivularis, a natural hybrid, presented sterile pollen grains of variable size and no content. The anther wall has an uniseriate epidermis and endothecium, the latter with helicoidal thickenings, although some cells of the middle layers also showed thickenings; the biseriate tapetum is of amoeboid non-syncytial type, since the tapetum cells did not fuse together forming a true plasmodium. The microsporogenesis is successive leading to isobilateral tetrads. The inaperturate pollen grains had a very reduced exine consisting of a thin, more or less continuous layer with small spines upon; the pollen grain shape is variable among the species, all of them presenting heteropolar pollen, except H. angusta with isopolar ones. Most of these characteristics were shared with other studied Zingiberales, although more studies need to be done.