Features related to anther opening in Solanum species (Solanaceae)

The mode of anther opening and the morphological and histological variability of the stomium are described in 30 Solanum species. Poricidal, poricidal‐longitudinally dehiscing and longitudinally dehiscing anthers are observed. In the three types, the stomium may be diverse with regard to shape and histological characteristics before opening, but is always composed of small epidermal cells as the sole anther wall layer; the stomial cells may be differentiated only in part of the anther length. Particular crescent‐shaped structures in the epidermis, called ‘ridges’, are observed to line the stomium in most species. These ridges may be related to the stomium opening, working together with the cells with thickened walls of the anther. Cells with thickened walls are developed in the endothecium, middle layers and/or connective tissue at the apical end of the anther, surrounding the pore; only in the longitudinally dehiscing anthers of S. nitidum does an endothecium with thickened cell walls develop along its entire length. At least two histological features (the differentiation of small stomial epidermal cells as a unique layer, and the distribution of cells with thickened walls) seem to constrain the form of the open stomium. © 2008 The Linnean Society of London, Botanical Journal of the Linnean Society, 2008, 158 , 344–354.     

Comparative anatomy and morphology of nectar-producing Melastomataceae

Background and Aims

Most neotropical Melastomataceae have bee-pollinated flowers with poricidal anthers. However, nectar rewards are known to be produced in about 80 species in eight genera from four different tribes. These nectar-producing species are pollinated by both vertebrates and invertebrates.

Methods

The floral morphology and anatomy of 14 species was studied in six genera of nectar-producing Melastomataceae (Blakea, Brachyotum, Charianthus, Huilaea, Meriania and Miconia). Anatomical methods included scanning electron microscopy, and serial sections of paraffin-embedded flowers.

Key Results

All vertebrate-pollinated melastome flowers have petals that do not open completely at anthesis, thus forming a pseudo-tubular corolla, while closely related species that are bee pollinated have rotate or reflexed corollas. In most species, nectar secretion is related to stomatal or epidermal nectaries and not filament slits as previously reported. Moreover, the nectar is probably supplied by large vascular bundles near the release area. Blakea and Huilaea have nectary stomata located upon the dorsal anther connective appendages. Brachyotum also has nectary stomata on the anther connectives, but these are distributed lengthwise along most of the connective. Meriania may release nectar through the anther connective, but has additional nectary stomata on the inner walls of the hypanthium. Miconia has nectary stomata on the ovary apex. Charianthus nectaries were not found, but there is circumstantial evidence that nectar release occurs through the epidermis at the apex of the ovary and the lower portions of the inner wall of the hypanthium.

Conclusions

Nectar release in Melastomataceae is apparently related to nectary stomata and not filament slits. The presence of nectary stomata on stamens and on ovary apices in different lineages suggests that the acquisition of nectaries is a derived condition. Nectary location also supports a derived condition, because location is strongly consistent within each genus, but differs between genera.Key words: Blakea, Brachyotum, Charianthus, Huilaea, Meriania, Melastomataceae, Miconia, nectaries, nectary stomata, pollination     

Molecular phylogenetics of Melastomataceae and Memecylaceae: implications for character evolution

Melastomataceae are among the most abundant and diversified groups of plants throughout the tropics, but their intrafamily relationships and morphological evolution are poorly understood. Here we report the results of parsimony and maximum likelihood (ML) analyses of cpDNA sequences from the rbcL and ndhF genes and the rpl16 intron, generated for eight outgroups (Crypteroniaceae, Alzateaceae, Rhynchocalycaceae, Oliniaceae, Penaeaceae, Myrtaceae, and Onagraceae) and 54 species of melastomes. The sample represents 42 of the family's currently recognized ～150 genera, the 13 traditional tribes, and the three subfamilies, Astronioideae, Melastomatoideae, and Memecyloideae (= Memecylaceae DC.). Parsimony and ML yield congruent topologies that place Memecylaceae as sister to Melastomataceae. Pternandra, a Southeast Asian genus of 15 species of which five were sampled, is the first- branching Melastomataceae. This placement has low bootstrap support (72%), but agrees with morphological treatments that placed Pternandra in Melastomatacaeae because of its acrodromal leaf venation, usually ranked as a tribe or subfamily. The interxylary phloem islands found in Memecylaceae and Pternandra, but not most other Melastomataceae, likely evolved in parallel because Pternandra resembles Melastomataceae in its other wood characters. A newly discovered plesiomorphic character in Pternandra, also present in Memecylaceae, is a fibrous anther endothecium. Higher Melastomataceae lack an endothecium as do the closest relatives of Melastomataceae and Memecylaceae. The next deepest split is between Astronieae, with anthers opening by slits, and all remaining Melastomataceae, which have anthers opening by pores. Within the latter, several generic groups, corresponding to traditional tribes, receive solid statistical support, but relationships among them, with one exception, are different from anything predicted on the basis of morphological data. Thus, Miconieae and Merianieae are sister groups, and both are sister to a trichotomy of Bertolonieae, Microlicieae + Melastomeae, and Dissochaeteae + Blakeeae. Sonerileae/Oxysporeae are nested within Dissochaeteae, Rhexieae within Melastomeae, and African and Asian Melastomeae within neotropical Melastomeae. These findings have profound implications for our understanding of melastome morphological evolution (and biogeography), implying, for example, that berries evolved from capsules minimally four times, stamen connectives went from dorsally enlarged to basal/ventrally enlarged, and loss of an endothecium preceded poricidal dehiscence.     

The taxonomic value of floral characters in Rapateaceae (Poales-Monocotyledons)

The floral anatomy of Cephalostemon, Monotrema, Rapatea, Spathanthus, and Stegolepis was studied for taxonomic purposes. All species studied share colleters between the floral parts; sepals, petals, anthers, and style covered by an ornamented cuticle; short epidermal cells with sinuous walls on the abaxial surface of the petals; tetrasporangiate anthers with phenolic idioblasts in the epidermis; endothecium with spiral thickenings; incompletely septate ovary; and anatropous, bitegmic ovules. The floral anatomy is useful not only for characterizing the family, but also for delimiting the subfamilies and genera. Sepals with silica bodies in the epidermal cells; mature anther wall composed of epidermis, endothecium, and middle layer; absence of phenolic idioblasts in the sepals, filaments, and ovary; and stylar epidermal cells with thickened external periclinal wall support Rapateoideae. Cephalostemon and Rapatea show a great number of similarities, corroborating their close relationship indicated in the phylogenetic analyses of the family. Monotrema shares few characters with the genera of Rapateoideae, corroborating its placement in Monotremoideae. Stegolepis shows several distinctive characters, probably related to the greater diversity found in this genus.     

Some histochemical features of anther wall of <Emphasis Type="Italic">Leucojum aestivum</Emphasis> (Amaryllidaceae) during pollen development

In this study, polysaccharide and RNA contents of anthers were investigated on different phases of sporogenesis by using light microscopy techniques from histological and cytological point of view in Leucojum aestivum. Paraffin and semi-thin sections of anthers were stained with toluidine blue and PAS. Anthers were tetrasporangiate. The wall of the anther consists of an epidermis, endothecium, middle layer and glandular tapetum. During one nucleated microspore and mature pollen phase microspores and tapetum cells began to degenerate and they were become very rich of RNA in L. aestivum. And also RNA content was increased in endothecium and middle layer cells except the epidermis cells of anther wall. An increase in RNA content indicates cell activation. Polysaccharides were not seen in young anther wall but they were seen in older ones. They were generally condensed in the cell walls and especially in the cell walls of vascular bundles of connective tissue. This could be thought that insoluble polysaccharides were used in metabolic events in early developmental stages. Appearance of polysaccharides in late phases was indicated that polysaccharides were used in the formation of cuticule and differentiation of endothelium cell walls.     

大百合小孢子发生和雄配子体发育研究

用常规石蜡切片技术和压片法对大百合小孢子发生和雄配子体发育进行观察。结果表明：花药4室,花药壁由表皮、药室内壁、中层和腺质绒毡层组成,花药壁发育方式为单子叶型,药室内壁部分细胞发育后期发生纤维状加厚。小孢子母细胞减数分裂过程的胞质分裂为连续型,四分体多数为左右对称型,偶有四面体型。成熟花粉为2细胞型,具1个萌发沟。经TTC法检验,成熟花粉生活力为86.3%。从小孢子的发生及雄配子体发育的整个过程看,未见异常现象,能形成大量正常的成熟花粉。     

THE PROCESSES OF ANTHER DEHISCENCE AND POLLEN DISPERSAL III. THE DEHYDRATION OF THE FILAMENT TIP AND THE ANTHER IN THREE MONOCOTYLEDONOUS SPECIES

The relationship between anther dehiscence and dehydration of the filament tip and the connective tissue were followed in Gasteria verrucosa (Mill.) H. Duval, Allium cepa L. and Lilium hybrida cv.'Enchantment'using various microscopical techniques, including direct observations of living tissues in the SEM.
In G. verrucosa anther dehiscence immediately follows anthesis, with concomitant dehydration and shrivelling of the filament tip. In L. hybrida the filament tip dehydrates and shrivels before anthesis. The anthers start to dehisce immediately after anthesis. In A. cepa the filament tip dehydrates and shrivels slowly after anthesis and dehiscence, which may take up to several days, correlates with the rate of extension of the filament.
The attenuated period of dehiscence in A. cepa may be related to the absence of stomata on the anther. Experiments on xylem conductivity and transpiration reveal that the presence of continuously open stomata on the anthers sets the time of dehiscence after anthesis in G. verrucosa and L. hybrida.
The shrinking filament tip in all three species functions as a hinge which suspends the dangling anther after dehiscence. The uniformly thickened epidermal cell walls on the anthers opposite the filament prevent the outwardly bending locule walls from embracing the filament.     

毛钩藤的小孢子发生和雄配子体发育

在光学显微镜和透射电镜下观察了毛钩藤(Uncaria hirsuta Havil.)的小孢子发生和雄配子体发育过程.结果表明,毛钩藤花两性,具5枚雄蕊,花药4室,花药壁由表皮、药室内壁、中层和绒毡层组成,花药开裂时,药室内壁高度纤维化带状加厚.花药壁的发育方式属于双子叶型,小孢子母细胞减数分裂的胞质分裂为同时型.小孢子在四分体时期开始沉积花粉外壁,小孢子大液泡化时期开始沉积花粉内壁.成熟花粉为2-细胞型.毛钩藤的花粉发育特征和茜草科植物基本一致.毛钩藤绒毡层属于分泌型,双重起源,分别起源于次生周缘层和药隔细胞.小孢子发育早期绒毡层开始降解并分泌形成大量乌氏体,花药开裂时绒毡层完全消失,剩下少量乌氏体.小孢子早期内壁加厚突出形成,小孢子细胞核分裂以后内壁加厚开始脱落,花药开裂时,只剩下少量的内壁加厚突出.初步推测,内壁加厚突出与乌氏体共同作用为雄配子体的发育提供营养物质.     

Glyphosate-induced anther indehiscence in cotton is partially temperature dependent and involves cytoskeleton and secondary wall modifications and auxin accumulation

Yield reduction caused by late application of glyphosate to glyphosate-resistant cotton (Gossypium hirsutum; GRC) expressing CP4 5-enol-pyruvylshikmate-3-P synthase under the cauliflower mosaic virus-35S promoter has been attributed to male sterility. This study was aimed to elucidate the factors and mechanisms involved in this phenomenon. Western and tissue-print blots demonstrated a reduced expression of the transgene in anthers of GRC compared to ovules of the same plants. Glyphosate application to GRC grown at a high temperature regime after the initiation of flower buds caused a complete loss of pollen viability and inhibition of anther dehiscence, while at a moderate temperature regime only 50% of the pollen grains were disrupted and anther dehiscence was normal. Glyphosate-damaged anthers exhibited a change in the deposition of the secondary cell wall thickenings (SWT) in the endothecium cells, from the normal longitudinal orientation to a transverse orientation, and hindered septum disintegration. These changes occurred only at the high temperature regime. The reorientation of SWT in GRC was accompanied by a similar change in microtubule orientation. A similar reorientation of microtubules was also observed in Arabidopsis (Arabidopsis thaliana) seedlings expressing green fluorescent protein tubulin (tubulin alpha 6) following glyphosate treatment. Glyphosate treatment induced the accumulation of high levels of indole-3-acetic acid in GRC anthers. Cotton plants treated with 2,4-dichlorophenoxyacetic acid had male sterile flowers, with SWT abnormalities in the endothecium layer similar to those observed in glyphosate-treated plants. Our data demonstrate that glyphosate inhibits anther dehiscence by inducing changes in the microtubule and cell wall organization in the endothecium cells, which are mediated by auxin.     

Anther ontogeny in <Emphasis Type="Italic">Brachypodium distachyon</Emphasis>

Brachypodium distachyon has emerged as a model plant for the improvement of grain crops such as wheat, barley and oats and for understanding basic biological processes to facilitate the development of grasses as superior energy crops. Brachypodium is also the first species of the grass subfamily Pooideae with a sequenced genome. For obtaining a better understanding of the mechanisms controlling male gametophyte development in B. distachyon, here we report the cellular changes during the stages of anther development, with special reference to the development of the anther wall. Brachypodium anthers are tetrasporangiate and follow the typical monocotyledonous-type anther wall formation pattern. Anther differentiation starts with the appearance of archesporial cells, which divide to generate primary parietal and primary sporogenous cells. The primary parietal cells form two secondary parietal layers. Later, the outer secondary parietal layer directly develops into the endothecium and the inner secondary parietal layer forms an outer middle layer and inner tapetum by periclinal division. The anther wall comprises an epidermis, endothecium, middle layer and the secretory-type tapetum. Major documented events of anther development include the degradation of a secretory-type tapetum and middle layer during the course of development and the rapid formation of U-shaped endothecial thickenings in the mature pollen grain stage. The tapetum undergoes degeneration at the tetrad stage and disintegrates completely at the bicellular stage of pollen development. The distribution of insoluble polysaccharides in the anther layers and connective tissue through progressive developmental stages suggests their role in the development of male gametophytes. Until sporogenous cell stage, the amount of insoluble polysaccharides in the anther wall was negligible. However, abundant levels of insoluble polysaccharides were observed during microspore mother cell and tetrad stages and gradually declined during the free microspore and vacuolated microspore stages to undetectable level at the mature stage. Thus, the cellular features in the development of anthers in B. distachyon share similarities with anther and pollen development of other members of Poaceae.     

Shifts from specialised to generalised pollination systems in Miconieae (Melastomataceae) and their relation with anther morphology and seed number

Most species in Melastomataceae have poricidal anthers related to specialised bee buzz‐pollination, while some have anthers with large openings associated to non‐bee pollination systems. We tracked the evolution of anther morphology and seed number on the Miconieae phylogenetic tree to understand the evolutionary shifts in such pollination systems. Anther morphometric data and seed number were recorded for 54 taxa. Pollinators (bees, flies, wasps) were recorded for 20 available species. Ancestral state reconstruction was made using Maximum Likelihood from nrITS sequences. We used phylogenetic eigenvector regressions to estimate phylogenetic signal and the adaptive component for these traits. Species pollinated by bees or bees and wasps tend to have smaller pores and fruits with more seeds. Species pollinated by flies or flies and bees and/or wasps tend to have larger pores and fruits with less seeds. Independent evolution occurred three times for anthers with large pores and twice for fruits with few seeds. We detected a phylogenetic signal in both traits, and negative correlated evolution between them. In actinomorphic small‐flowered Miconieae, changes in anther morphology can be related to generalisation in the pollination system incorporating flies and wasps as pollinators and lessening the importance of buzzing bees in such process. Differences in pollen removal and deposition may explain differences in anther morphology and seed number in Miconieae.     

倒地铃花的形态分化与花药结构研究

利用体视显微镜、半薄切片和超薄切片法对倒地铃(Cardiospermum halicacabum Linn.)雄花和假两性花开花过程及花药发育过程进行了观察和比较研究。结果显示:(1)花蕾发育早期,倒地铃雄花和假两性花的花蕾形态没有区别;花蕾发育后期,雄花雌蕊退化,假两性花雌蕊继续发育,花蕾外部形态出现差异;开花时雄花花药开裂,假两性花花药不开裂。(2)倒地铃雄花和假两性花均具四室花药,呈蝶形;花药壁细胞从外到内依次是表皮、药室内壁、中层(2层)和绒毡层;花药壁发育为基本型,绒毡层为单核分泌型,四分体为四面体型,花粉粒两核;开花时雄花和假两性花中层都有残留;小孢子液泡化时,绒毡层开始降解,两核花粉粒时,假两性花绒毡层降解较快。(3)雄花药室内壁次生加厚完全,裂口区发育,连接同侧花粉囊的连接组织降解,花药开裂;假两性花药室内壁次生加厚不完全,具唇形细胞,药隔细胞壁未降解,同侧花粉囊未连通,花药四室,不开裂;假两性花成熟花粉粒细胞质稀少,内壁不完整。本研究结果表明,倒地铃的雄花是由两性花在发育早期雌蕊停止发育形成的,假两性花则由两性花在发育晚期雄蕊功能退化造成的。     

Characterization and genetic mapping of a mutation (ms35) which prevents anther dehiscence in Arabidopsis thaliana by affecting secondary wall thickening in the endothecium

The male sterile mutant, ms35 , of Arabidopsis thaliana was produced by X-irradiation of seeds. The mutant produces fertile pollen, but is male sterile because the anthers do not dehisce. Anther development in ms35 plants occurs as in wild-type Arabidopsis until shortly after microspores are released from meiotic tetrads. Thereafter, in the wild type, bands of lignified, cellulosic secondary wall thickenings are laid down around the cells of the anther endothecium. In contrast, wall thickenings are not formed in the endothecium of the ms35 mutant. Development of other lignified tissues, for example the vascular tissue of the stamen, occurs normally in ms35 plants. In mutant anthers, as pollen maturation is completed, the stomium is cleaved but the anther wall does not retract to release pollen. The block in anther dehiscence in ms35 plants is specifically correlated with the absence of endothecial wall thickenings. The ms35 mutation represents the first genetic evidence in support of the proposed role of the endothecium in anther dehiscence. The ms35 gene was mapped to the top arm of chromosome 3 ( hy2 -(4.17±2.31 cM)- ms35 -(32.14±5.45 cM)- gl1 ).     

The Cytological Mechanism of Low Fertility in the Naked Seed Rice

The low fertility of naked seed rice (NSR) was investigated by the following observations: somatic chromosome constitute, behavior of pollen mother cells (PMCs), the germination of mature pollen grains, the development of male and female gametes and the structure of the anther opening. The results indicated that somatic chromosomal number was 2n = 24, behavior of PMCs were normal and most of pollen grains could regularly develop further to mature male gametophytes in NSR. And dehiscence chamber and thickened endothecium cell (TEC) in numerous anthers of the NSR were developed abnormally after dicaryotic phase, result in few anthers complete opening and most partly opening or failure to opening, therefore much fewer of pollen grains attach on the stigma as compared with normal variety. Furthermore most of embryo sacs possessed abnormal structure and were sterile. All of above illustrated that the failure of the anther opening and the abortion of female gametophyte were main factors controlling the low seed-setting rate of the NSR.     

The final split: the regulation of anther dehiscence

Controlling male fertility is an important goal for plant reproduction and selective breeding. Hybrid vigour results in superior growth rates and increased yields of hybrids compared with inbred lines; however, hybrid generation is costly and time consuming. A better understanding of anther development and pollen release will provide effective mechanisms for the control of male fertility and for hybrid generation. Male sterility is associated not only with the lack of viable pollen, but also with the failure of pollen release. In such instances a failure of anther dehiscence has the advantage that viable pollen is produced, which can be used for subsequent rescue of fertility. Anther dehiscence is a multistage process involving localized cellular differentiation and degeneration, combined with changes to the structure and water status of the anther to facilitate complete opening and pollen release. After microspore release the anther endothecium undergoes expansion and deposition of ligno-cellulosic secondary thickening. The septum separating the two locules is then enzymatically lysed and undergoes a programmed cell death-like breakdown. The stomium subsequently splits as a consequence of the stresses associated with pollen swelling and anther dehydration. The physical constraints imposed by the thickening in the endothecium limit expansion, placing additional stress on the anther, so as it dehydrates it opens and the pollen is released. Jasmonic acid has been shown to be a critical signal for dehiscence, although other hormones, particularly auxin, are also involved. The key regulators and physical constraints of anther dehiscence are discussed.     

The systematic significance of the endothecium in Begoniaceae

The endothecial thickening patterns in 173 species, representing the three genera of Begoniaceae, were investigated using cleared macerated anthers. Begoniaceae contain taxa with U-shaped thickenings, perforate base plates, entire base plates, tympanate base plates, anticlinal bars, and taxa that lack endothecial thickenings. The degree of correlation between these endothecial classes and accepted taxonomic boundaries varies: some classes are confined to a single taxonomic unit (e.g. absence of thickenings, non-perforate tympanate base plates) while others are present in several taxonomic units (e.g. U-shapes, perforate base plates). This study provides an improved understanding of the diversity of endothecial patterns in a large genus and indicates that in the Begoniaceae the endothecium is of moderate systematic value within and between closely related sections in Begonia and in Symbegonia . We conclude that the endothecium exhibits high levels of homoplasy between distantly related sections of Begonia . In addition, we show that in contrast to past observations of the endothecium in dicotyledons, apical anther dehiscence in the Begoniaceae is not correlated with a loss of endothecial thickening.     

The Mechanics of the Grass Flower: Anther Dehiscence and Pollen Shedding in Maize

In this paper on the flower mechanics of the grasses, the openingmechanism of the maize anther is studied. Both the septum betweeneach two locules and the stomium of these porate-dehiscing anthersappear to be opened due to lysis of the middle lamellae of theircells. Additional mechanical force of the expanding pollen mightbe necessary to completely dissociate the parenchyma cells ofthe septum. A number of hours before anthesis the anther isstructurally able to dehisce. At anthesis the dehydrating endotheciumcells bend the locule walls bordering the pore in outward direction.Presumably evaporation is not the only cause for this dehydration. Poaceae; Zea mays ; flower; anther; dehiscence; endothecium; pollen     

扁桃花药开裂前后壁层细胞形态变化及分析

为探明扁桃花药开裂前后壁层细胞形态变化,以鹰咀扁桃鳞片开裂期、小蕾期、大蕾期和盛花期的花蕾为研究材料,运用石蜡切片法结合铁苏木精染色法、考马斯亮蓝染色法、PAS染色法对花药壁层细胞进行染色;同时用Nikon SMZ-250体视显微镜拍摄花药开裂过程,观测花粉粒长、短轴长度。结果表明:(1)从鳞片开裂期到小蕾期,花粉粒的长、短轴长度都增大,多糖颗粒数量增多,绒毡层细胞完全消失,中层细胞和药隔处细胞逐渐溶解;药室内壁细胞切向长度增加幅度大于径向长度,内、外壁长度都增大,螺旋状纤维进一步形成;表皮细胞切向长度增加幅度大于径向长度。(2)从小蕾期到大蕾期花粉粒长、短轴长度明显增大,多糖颗粒持续增多;中层细胞和药隔处细胞大部分溶解;药室内壁细胞径向、切向长度持续增大,内壁长度增大、外壁长度趋于稳定,多糖颗粒数量减少,螺旋状纤维基本形成;表皮细胞切向减小幅度大于径向。(3)从大蕾期到花药半开裂,花粉粒长、短轴长度稍微增大;中层细胞和药隔处细胞完全溶解;药室内壁细胞切向长度持续增大,径向长度趋于稳定,内壁长度持续增大,外壁长度逐渐减小,多糖颗粒数量较少;表皮细胞切向、径向长度持续减小。(4)花药半开裂后,花粉粒长、短轴长度都减小;药室内壁细胞和表皮细胞切向、径向长度都减小;药室内壁细胞内、外壁长度减小并趋于接近,内壁长度减小趋势出现晚于外壁。研究认为,扁桃花药壁层细胞形态变化是花药开裂的基础,并与花药开裂密切相关。