Variation and similarities in pollen features in some basal angiosperms, with some taxonomic implications

Species within three families of basal angiosperms (Trimeniaceae, Winteraceae, Monimiaceae) illustrate differences and similarities in pollen within a species, between species and between genera. Trimenia papuana (Trimeniaceae) has dimorphic pollen (inaperturate, polyforate), each confined to different individual plants. Other species have either disulculate or polyforate pollen. Evolution seems to be from disulculate to inaperturate to polyforate. Present-day Winteraceae have pollen in permanent tetrads except four species of Zygogynum with monads. Why? Did such monads appear as fossils before tetrads in Winteraceae? Molecular studies of Takhtajania perrieri indicate it is basal but its unique bicarpellate unilocular gynoecium seems derived. Although Hedycarya arborea and Kibaropsis caledonica have near-identical permanent pollen tetrads, many other features are very different. Hedycarya species have permanent tetrads or inaperturate monads with spinulose, `starry' or other sculpturing, and it is suggested this and recent molecular data indicate further studies are needed to determine generic limits.     

Embryology ofMalaxis saprophyta,with comments on the systematic position ofMalaxis (Orchidaceae)

InMalaxis saprophyta, anther wall development corresponds to the Monocotyledonous type. The uninucleate tapetum is of secretory type and the endothecium develops U- and V-shaped thickenings on the inner tangential and radial walls. Cytokinesis is simultaneous; tetrahedral, isobilateral and T-shaped tetrads are formed which are compactly aggregated in pollinia. At anthesis the microspore tetrads are 2-celled. The ovule is anatropous, bitegmic and both integuments are dermal in origin. A single hypodermal cell develops directly into a megaspore mother cell. Embryo sac development is predominantly monosporic and less often bisporic. Irrespective of the type of development, the mature embryo sac is 6-nucleate. Although double fertilization occurs, the primary endosperm nucleus degenerates. Embryogeny is of the Onagrad type. The mature embryo lacks differentiation into cotyledon, plumule and radicle. The reticulate seed coat is formed entirely by the outer layer of outer integument. There are three sterile and three fertile valves in the ovary. Although initially parenchymatous, the entire three sterile valves in the ovary and the upper half of the three fertile valves become sclerified after fertilization. The embryological characters support the disputed systematic position ofMalaxis within subtribeMalaxidinae ofEpidendreae.     

Extra divisions and nuclei fusions in microspores from Brassica allohexaploid (AABBCC) × Orychophragmus violaceus hybrids

Abnormal meiosis and microspore development and related defective mutants have often been reported in plants and wide hybrids. Here extra divisions and nuclei fusions were observed to occur in microspore nuclei of partial hybrids between synthetic Brassica hexaploid (2n=54, AABBCC) and another crucifer Orychophragmus violaceus (2n=24). Abnormal spindle were formed and chromosomes were separated into several nuclei of variable sizes after bi-, or multi-polar divisions in the four cells of tetrads. As a consequence, more than eight mini-microspores of different sizes were produced by one tetrad. Genomic in situ hybridization results indicated that no chromosome replication occurred during such divisions. In some tetrads, the four nuclei were fused to form one large cell with increased chromosome number. The extra divisions or fusions appeared only in some flower buds of one plant, some anthers in the same buds, or even in individual cells of tetrads. The possible mechanisms behind these cytological phenomena are discussed.     

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.     

Ontogenesis of monad pollen inPterostylis plumosa (Orchidaceae,Neottioideae)

Pterostylis plumosa (Neottioideae) is an orchid with monosulcate monad pollen. Tetrads may be isobilateral, decussate, tetrahedral, rhomboid or T-shaped, but all pollen grains have a similar shape. Those belonging to the same tetrads are contiguous from microspore release to opening of the anther, with the furrow oriented inwards. Sporophytic proteins are present outside the furrow. The tapetum is of the parietal type without orbicles. The increase in pollen grain size between meiosis and maturity is only three-fold. The generative cell is spherical when the pollen is mature. These features are discussed in relation to the primitive nature of the species.     

Seed-coat anatomy of japanese species ofCorydalis andDicentra (Papaveraceae; Fumarioideae)

Development and structure of seed-coat were examined in 16 species ofCorydalis and two species ofDicentra. Neither the tegmen (developed ii) nor the testa (developed oi) is multiplicative during seed development. Mature seed-coat is consistent in all the species examined in having mechanical structure in the exo- or endotesta. Differences are found in (1) which of the integumentary layers develops into the main mechanical layer (exotesta or endotesta), (2) whether a mesotesta is differentiated or not, and (3) whether the endotegmen is persistent as a layer of thick-walled cells, is persistent only partially, or is thoroughly degenerated. Theses seed-coat characters distinguish six groups of species (i.e., four groups inCorydalis and two inDicentra), which represent well infrageneric taxa proposed on the other characters. Evidence from seed-coat anatomy further suggests thatDicentra spectabilis (subg.Hedycapnos) retains the primitive (endotestal) seed-coat of Fumarioideae.     

Pollen morphology ofPyrola and its taxonomic significance

Pollen from 152 collections representing 16Pyrola species was examined with LM, SEM and TEM. The genusPyrola is stenopalynous in having tetrads and 3-colporoidate grains. However, a continuous and serial variation in the exine sculpture and the tetrad diameter (D) was revealed within the genus. Křísa's system ofPyrola (1971) was re-examined in the light of these palynological characters which may have a taxonomic significance, as well as the external morphology. P. minor (only member of the subgenusAmelia), which is characterized by having small (D=30–35 μm) pollen tetrads with verrucate sculpture must best be placed near sectionPyrola of the subgenusPyrola. P. faurieana must be segregated from sectionChlorantha and together withP. media placed intermediate between the subgenusAmelia and sectionPyrola of the subgenusPyrola based on their larger (D=ca. 40 μm) pollen tetrads with verrucate sculpture. Two series of sectionChlorantha excludingP. faurieana are differentiated in the exine sculpture. Therefore, they are regarded as the distinct taxonomic groups. SectionScotophylla is characterized by having psilate sculptured pollen tetrads, which indicates that its sectional rank is sufficiently asserted. The evolutionary trend in the exine sculpture from verrucate through rugulate to psilate is suggested withinPyrola.     

Critical studies on the sexuality of southern Argentinian Ericaceae

Summary The family Ericaceae is considered to be mostly hermaphroditic and only in a few cases unisexual or dioecious. Five Argentinian species (Gaultheria antarctica Hook, f., G. caespitosa P. et E., G. phillyreaefolia (Pers.) Sleumer, G. tenuifolia (Phil.) Sleumer and Pernettya insana (Molina) Gunckel), previously described as hermaphroditic, were found by us to have two types of flowers: female and male. Female flowers have both a rudimentary androecium with stamens, in which the anthers are small, collapsed and without pollen grains or are reduced to filaments only, and a perfect gynoecium with well-developed ovules filling the carpel locules. The stigma is expanded and of the wet, papillate type, with copious to slight secretion. The stigma protrudes above the anthers. Male flowers have a well-developed androecium included within the corolla and a reduced gynoecium. Anthers are as long as the filaments, except in G. caespitosa. Pollen is shed as tetrads which are 100% viable. Ovule development ranges from total absence, passing through aborted ovules of various sizes to apparently normal ones. Stigmata are neither expanded nor papillate, except in Pernettya insana. Only in two species do the style and stigma extend beyond the level of the anthers. Heterostyly does not occur in any of the five species studied. Functional dioecy thus characterizes the five species considered and is reported for the first time in the genus Gaultheria.     

Caryedon serratus and its parasitoids in the Savanna around Lamto,Ivory Coast

In the savanna around Lamto,Caryedon serratus develops, in the beginning of the dry season, on maturing seeds ofPiliostigma thonningii (Caesalpinioideae). A 2^nd generation, and sometimes a 3^nd one, develops in dry seeds. There is no pupal diapause during the rainy season and adults seems to spend this period in a semi-lethargic state. The parasitoid complex ofC. serratus is a chalcid-dominated one. It includes 5 species: one oophagous (Uscana caryedoni) and 4 larvo-nymphal parasitoids. Three species are regular (Anisopteromalus caryedophagus, Bracon sp. andU. caryedoni), the others are sporadic species (Proconura serratocida, Eurytoma caryedocida). Biological data are given for each species.      

Formation of the arthropod labrum by fusion of paired and rotated limb-bud-like primordia

The nature and origin of the arthropod labrum is a matter much under dispute. We show here that in Tribolium castaneum (Herbst, 1797) the labrum develops from two individual primordia, termed labral buds. Expression of the genes decapentaplegic (dpp) and wingless (wg) in these buds is identical to the buds of the metameric appendages (e.g. thoracic legs), except that the patterns are reversed. We propose that this reversal is the result of the rotation of the labral buds through an angle of approximately 180°. We also for the first time study dpp and wg expression in the fully developed labrum of older embryonic stages. Here, gene expression patterns show that the labrum proper is formed by fusion of the labral buds along their dorsal sides, while their ventral sides are facing outward forming the lateral sides of the fused labrum. Furthermore, we show that there are very similar patterns in another arthropod species, the spider Cupiennius salei (Keyserling, 1877), although in this species the labrum develops as a single structure and not from two separate primordia. However, in C. salei the expression of engrailed is also reversed in addition to the reversal of dpp and wg expression: engrailed is expressed in the anterior half of the labrum, and not in the posterior half like in the remaining appendages. Our results suggest that the arthropod labrum is derived evolutionarily from paired limb-bud-like primordia by rotation and fusion, and that this process is recapitulated ontogenetically to a different extent in different arthropod species.     

Paraveinal mesophyll: Review and survey of the subtribeErythrininae (Phaseoleae,Papilionoideae, Leguminosae)

Paraveinal mesophyll (PVM) is a distinctive anatomical feature of the leaf mesophyll of some plant taxa that may represent a specialized physiological compartment. A comprehensive review of the 42 published references that mention PVM or similar cell layers and a survey of 121 of the 272 species of all nine genera of thePhaseoleae subtribeErythrininae demonstrate that PVM is nearly exclusively found inLeguminosae. InLeguminosae, PVM is either rare or absent in subfamilyCaesalpinioideae, uncommon inMimosoideae, and extensively distributed amongPapilionoideae. In subtribeErythrininae, PVM is ubiquitous inErythrina, and occurs in four other genera. ThreeErythrininae genera (Apios, Mucuna, andCochlianthus) lack PVM. Unique chloroplast-poor, enlarged conical cells (pellucid palisade idioblasts) occur in 80 species ofErythrina but not in any other genus ofErythrininae.     

Embryological Studies on Narcissus tazetta var. chinensis

Chinese narcissus (Narcissus tazetta var. chinensis Roem) blooms but has no seeds. Embryological studies on the species were conducted to discover the causes of its sterility. Its anther wall is composed of four layers of cells, and its tapetum is of the secretory type. The cytokinesis of microspore mother cells is of the successive type, and the tetrad is tetrahedral. During meiosis of microspore mother cells, some chromosomes lagged, and several micronuclei were found in tetrads. Only 27.7% of the pollen grains contained full cytoplasm, and 1.3% of them germinated in culture medium. No pollen grain, however, could germinate on the stigma. The ovary is trilocular with axile placenta, and the ovules are bitegmic, tenuinucellate, and anatropous. Its embryo sac is of the polygonum type. Most embryo sacs degenerated, and only about 4.5% of the ovules contained a normal embryo sac with an egg cell, two synergids, three antipodal, and a central cell containing two polar nuclei. One reason for the sterility of Chinese narcissus is the abnormality of microsporogenesis and megasporogenesis, in which only a few functional pollen grains and embryo sacs are produced. The other reason is that the pollen grains cannot germinate on the stigma. This paper was translated from Journal of Xiamen University (Natural Science), 2005, 44(1) (in Chinese)     

Development of polarity in the ovules of the F2-progeny of theOenothera hybridalbicans · hhookeri

In the F₂-progeny of hybrids from crosses betweenOenothera biennis orsuaveolens andOe. hookeri with theRenner-complexesalbicans and^hhookeri, the development of callose pattern in meiocytes and megaspore tetrads is the same as in the F₁ and the parentOe. hookeri. During the development of the megaspore tetrads and the embryo sacs primary and secondary heteropolarity as well as homopolarity is observed. Estimates for the initial frequency of homo- and heteropolar tetrads at the end of the degeneration of megaspores in the tetrads immediately before the start of embryo sac development could be calculated. The F₂-plants can be arranged in three groups, distinguished by the frequency of the two polarity types. One of these groups behaves similar to the parentOe. hookeri, the two others have more homopolar tetrads. The segregation can be interpreted as recombination of genes, which influence the development of the polarity in the ovules. This is possible by crossing-over of genes between the twoRenner-complexes of the hybrid.     

Significance of palynology for phylogeny ofAnnonaceae: Experiments with removal of pollen characters

Phylogenetic analyses based on morphology placeAnaxagorea and other taxa with granular monosulcate pollen, as in otherMagnoliales, at the base ofAnnonaceae. Taxa with columellar tetrads, granular tetrads, and inaperturate monads form a derived clade. To test the systematic importance of palynology, we analyzed the data set with pollen characters removed. The result was lower resolution and a different rooting of the family, betweenUvariopsis and other groups with columellar tetrads.Anaxagorea and other monosulcates are higher in the tree, implying that granular monosulcate pollen, laminar stamens, and irregular endosperm ruminations are reversals. This rooting is highly unparsimonious when pollen characters are included, and only weakly supported over theAnaxagorea rooting when pollen is excluded. Together with preliminary molecular analyses, these experiments confirm the special value of palynology in systematics ofAnnonaceae. This paper is dedicated to emer. Univ.-Prof. DrFriedrich Ehrendorfer on the occasion of his 70th birthday     

珍稀濒危植物大花万代兰的花粉团发育及其分类学意义

万代兰属的属间界限划定及其亲缘关系重建是兰科分类系统中的难解之谜。该研究采用常规石蜡切片技术观察了珍稀濒危植物大花万代兰的一对深裂花粉团的形成机制、花药壁发育模式、小孢子发生及雄配子体发育等的胚胎学特征。结果表明：(1)大花万代兰早期的花药原基分化出一对侧生药室,每个药室的小孢子囊中央分化出一条在花药成熟时会降解的不育隔膜组织,形成两个不等深裂的花粉团。(2)发育完整的花药壁有5~9层,包括2~6层药室内壁,符合多层型花药壁发育类型;绒毡层细胞为单核,腺质型,在花药成熟时,表皮、中层和绒毡层皆降解,仅留下2~6层纤维性加厚的药室内壁。(3)小孢子母细胞经过连续型胞质分裂形成正四面体和左右对称的小孢子四分体,小孢子四分体继续保持在同一个胼胝质内,完成有丝分裂形成了2 细胞型的四合花粉;四合花粉两两紧密排列,且由于隔膜组织的降解,最终发育为一对深裂的花粉团。根据现有兰花花药发育资料,分析了大花万代兰花粉团发育的胚胎学特征的分类学意义,为万代兰属错综复杂的系统分类提供了新资料。     

传统傣药竹叶兰的花粉团发育及分类学意义

该研究利用石蜡切片技术观察并描述了兰科单型属竹叶兰属的花粉团发育过程,包括花形态解剖特征、8个花粉团的形成机制、花药壁发育模式、小孢子发生及雄配子体发育等特征,为该属复杂的系统亲缘关系提供胚胎学证据。结果表明:(1)成熟花药有两个药室,每个药室有4个一簇金色的花粉团,被白色花药帽;早期花药原基分化出的一对并列侧生药室,每个药室中央的小孢子囊在极面观方向分化出两条十字交叉的纵向不育隔膜组织,将其沿花药室纵轴方向深切为4个不等大的棒状次生孢子囊,最后发育为4个花粉团。(2)花药成熟时,靠花药开裂处的隔膜组织比近药隔膜组织的降解速度快且彻底,因此每个药室内的4个花粉团在花药开裂处粘合成一簇。(3)发育完好的花药壁共有6～7层,由外到内为表皮、3～4层药室内壁、中层和双核绒毡层,符合多层型花药壁的发育模式;花药成熟时,表皮退化,纤维性加厚发生在3～4层药室内壁,中层和绒毡层彻底降解。(4)小孢子母细胞通过同时型胞质分裂产生了正四面体型、左右对称、十字交叉型排列的小孢子四分体;小孢子四分体继续保持在同一胼胝质内完成了雄配子体发育,形成了2-细胞型的四合花粉;四合花粉两两或松散或紧密排列,构成了粉质花粉团。在前人的研究基础上,本文证实、补充并分析了竹叶兰属的花粉团发育特征,为该属的亲缘关系提供了胚胎学证据。     

Sectile pollinia and relationships in theOrchidaceae

The sectile state, in which pollinia are subdivided into units larger than tetrads, is one of several systematically important orchid pollinium characters. We recognize two types of sectile pollinia, based upon form and arrangement of massulae. One type is found primarily in members ofOrchidoideae-Spiranthoideae, while the other characterizes epidendroid genera. Additional characters that have systematic potential are massula dimorphism, caudicle composition, and degree of tetrad packing. Hollow sectile pollinia uniteArethusa andCalopogon; Stereosandra andEpipogium have dissimilar pollina, but other characters support a close relationship between them. The sectile condition is interpreted as resulting from a shift in the timing of cohesion of developing pollen cells.     

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.     

龙舌草果实和种子及其种苗发育形态学研究

采用形态观察和GMA切片相结合的方法,对龙舌草果实、种子及种苗发育的形态结构进行解剖学观察研究,探讨龙舌草个体发育中表现出的与水生环境相适应的结构特征.结果显示:(1)龙舌草由佛焰苞包被的果实通常由6心皮构成,心皮边缘不完全愈合,层片状胎座,水面形成的幼嫩果实被果柄拉入水中发育成熟;种子具毛,萌发时苗端先于根端发育,在胚根分化之前先出现单细胞的下胚轴毛,随后胚根发育为初生根,并在真叶的节处发育出不定根;果皮和叶肉内的维管束仅由少数细胞构成,且细胞分化不明显,有发达的通气道;叶表皮无角质层,细胞外凸,叶肉细胞数量少;表明龙舌草的结构特征与水生环境相适应.(2)研究还发现,龙舌草的种子无胚乳,下胚轴贮存种苗发育需要的大量淀粉粒,以及初生根和不定根均有根毛的结构,且与泽泻科的慈菇和泽泻的结构一致,证明水鳖科与泽泻科具有较近的亲缘关系.     

Production and viability of unreduced gametes in triploid interspecific blueberry hybrids

Summary Three triploid (2n=3x=36) blueberry hybrids were obtained by hand-pollinating approximately 7,000 flowers of tetraploid highbush blueberry cultivars (based on Vaccinium corymbosum L.) with pollen from the diploid species V. elliottii Chapm. Meiotic analysis of these triploids revealed trivalents, bivalents and univalents in all metaphase I cells, with lagging chromosomes evident at anaphase I. Pollen of the three triploids was mostly aborted and did not stain with acetocarmine. However, the three triploids did produce from 0.9%–1.3% giant pollen grains that stained with acetocarmine and were present as monads, dyads or triads, rather than the normal tetrads. Pollination of 10,853 flowers of hexaploid V. ashei Reade cultivars with pollen from the triploids produced 266 berries, which averaged fewer than two fully-developed seeds per berry. One triploid clone showed partial female fertility when crossed to hexaploids, self-pollinated, or intercrossed with other triploids. Ploidy levels of the resulting hybrids were determined.Florida Agricultural Experiment Station Journal Series No. 8672