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     

Rice (Oryza sativa L.) cultivars tolerant to high temperature at flowering: anther characteristics

We examined the relationship between morphological characteristics of anthers and fertility in japonica rice cultivars subjected to high temperature (37.5(26 degrees C day/night) at flowering. Percentage fertility was negatively correlated with the number of cell layers that separated the anther locule from the lacuna that formed between the septum and the stomium. The cell layers consisted of the remaining septum and degraded tapetum, and serve to keep the adjacent two locules closed. Anther dehiscence therefore requires the rupture of the cell layers. We conclude that the tight closure of the locules by the cell layers delayed locule opening, and decreased fertility at high temperatures.     

Rapid Swelling of Pollen Grains in the Dehiscing Anther of Two-rowed Barley (Hordeum distichum L. emend. LAM .)

The role of rapid swelling of pollen grains in anther dehiscencein Hordeum distichum L. emend. L AM. and the mechanism of thisswelling were examined. Artificial opening of the floret inducedrapid swelling of pollen grains and thecae dehiscence. The thecadehisced as pollen grains became swollen and dehisced anthershad larger pollen grains than indehisced anthers. Septa in theanther segments dehisced as a result of water-induced pollenpressure. These results strongly support the theory that therapid swelling of pollen grains is a driving force for antherdehiscence. On the other hand, potassium was detected in pollengrains from dehisced anthers, but not in pollen grains in indehiscedanthers. This suggests that potassium ions function as a turgorregulator in the rapid swelling of pollen grains. The mechanismof anther dehiscence is discussed in relation to the swellingof pollen grains, as is the possible mechanism of this swelling.Copyright 2000 Annals of Botany Company Anther dehiscence, Hordeum distichum L. emend. L AM., pollen swelling, potassium ion, two-rowed barley     

Anther structure and pollen development in Melicoccus lepidopetalus (Sapindaceae): An evolutionary approach to dioecy in the family

Anther and pollen development in staminate and pistillate flowers of dioecious Melicoccus lepidopetalus (Sapindaceae) were examined by light and electron microscopy. Young anthers are similar in both types of flowers; they consist of epidermis, endothecium, two to four middle layers and a secretory tapetum. The microspore tetrads are tetrahedral. The mature anther in staminate flowers presents compressed epidermal cells and endothecium cells with fibrillar thickenings. A single locule is formed in the theca by dissolution of the septum and pollen grains are shed at two-celled stage. The mature anthers of pistillate flowers differ anatomically from those of staminate flowers. The epidermis is not compressed, the endothecium does not develop fibrillar thickenings, middle layers and tapetum are generally persisting, and the stomium is nonfunctional. Microspore degeneration begins after meiosis of microspore mother cells. At anthesis, uninucleate microspores and pollen grains with vegetative and generative nuclei with no cytokinesis are observed. Some pollen walls display an abnormal exine deposition, whereas others show a well formed exine, although both are devoid of intine. These results suggest that in the evolution towards unisexuality, the developmental differences of anther wall tissues and pollen grains between pistillate and staminate flowers might become more pronounced in a derived condition, such as dioecy.     

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 arabidopsis DELAYED DEHISCENCE1 gene encodes an enzyme in the jasmonic acid synthesis pathway

delayed dehiscence1 is an Arabidopsis T-DNA mutant in which anthers release pollen grains too late for pollination to occur. The delayed dehiscence1 defect is caused by a delay in the stomium degeneration program. The gene disrupted in delayed dehiscence1 encodes 12-oxophytodienoate reductase, an enzyme in the jasmonic acid biosynthesis pathway. We rescued the mutant phenotype by exogenous application of jasmonic acid and obtained seed set from previously male-sterile plants. In situ hybridization studies showed that during the early stages of floral development, DELAYED DEHISCENCE1 mRNA accumulated within all floral organs. Later, DELAYED DEHISCENCE1 mRNA accumulated specifically within the pistil, petals, and stamen filaments. DELAYED DEHISCENCE1 mRNA was not detected in the stomium and septum cells of the anther that are involved in pollen release. The T-DNA insertion in delayed dehiscence1 eliminated both DELAYED DEHISCENCE1 mRNA accumulation and 12-oxophytodienoate reductase activity. These experiments suggest that jasmonic acid signaling plays a role in controlling the time of anther dehiscence within the flower.     

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。     

THE ASSOCIATION OF DRUSE CRYSTALS WITH THE DEVELOPING STOMIUM OF CAPSICUM ANNUUM (SOLANACEAE) ANTHERS

Each of the two stomiums in the anther of Capsicum annuum (sweet pepper) consists of a single layer of cells immediately below the epidermis between two adjacent locules. Each stomium extends the entire length of the anther and splits open at pollen maturity. Many calcium oxalate druse crystals form within the vacuoles of the stomium cells in association with membrane complexes and paracrystalline bodies. These latter structures are reported here for the first time and each is considered to be a nucleation site for druse crystal formation. Prior to the appearance of membrane complexes and crystals within the vacuoles, plasmalemmasomes are visible next to the stomium cell walls and contain vesicles and fibrous material. We propose that these bodies carry wall materials, including calcium ions and possibly oxalate ions, into the vacuoles. Their presence coincides with crystal formation. Two other types of crystals occur in the connective tissue between stomiums and the single vascular strand. These crystals, along with those in the two stomiums, form at precise times during anther development. Contrary to the more numerous suggestions that crystals protect against predators or are metabolic waste products, we believe their formation aids in degradation and weakening of the cell walls between the locules and, thus, contributes to the release mechanism for the pollen.     

Mechanism of Septum Opening in Anthers of Two-rowed Barley (Hordeum vulgare L.)

To test the hypothesis that the rapid swelling of pollen grainsdriven by potassium movement opens the septum in anthers ofpoaceous plants, we studied (1) the behaviour of pollen grainsduring unfolding of the locule and (2) the distribution of potassiumin the locule in two-rowed barley. In the first experiment,the unfolding of decapitated anthers was observed. The pollengrains paved the inner wall of the locule during the unfoldingprocess, suggesting that the pollen grains bend the locule walloutward when they adhere to the wall. In the second experiment,the distribution of potassium in transverse sections of loculesin dehisced and indehisced anthers was observed. In indehiscedanthers, potassium was detected outside the pollen grains. Incontrast, in dehisced anthers, potassium was detected insidepollen grains. This suggested potassium ions moved from theinter-pollen space (locular fluid) into the pollen grains inthe locule at the time of pollen-grain swelling. Copyright 2000Annals of Botany Company Hordeum vulgare L., locule unfolding, pollen grain swelling, potassium ion, two-rowed barley     

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.     

The diversity of anther structures and dehiscence patterns among Hamamelididae

HUFFORD, L. D. & ENDRKSS, P. K., 1989. The diversity of anther structures and dehiscence patterns among Hamamelididae. This survey of anther structures and dehiscence patterns focuses on the range of diversity among extant Hamamelididae. The definition and structure of the anther stomium are considered in detail to provide a basis for characterizing dehiscence patterns. We are concerned particularly with the structural basis and distribution of so-called valvate dehiscence, which we define here as occurring only in those anthers that possess stomial bifurcations or markedly eccentric stomia. Valvate dehiscence is restricted to Trochodendrales and Hamamelidales among Hamamelididae, although some Hamamelidaceae possess only linear, not markedly eccentric stomia that lead to longitudinal dehiscence patterns. Anther forms are somewhat variable and do not appear to be highly correlated with stomial patterns, although stomial bifurcations occur most frequently in anthers with broad, thick connectives that extend for the full length (or nearly so) of the thecae. Valvate dehiscence usually occurs in anthers in which the pollen sacs are embedded in bulky superficial tissues. An evolutionarily secondary extension of the stomium around the thecal shoulders seems to have occurred in taxa with a nonextensive connective and may facilitate a broader anther opening in cases of longitudinal dehiscence. An endothecial-like connective hypodermis is a notable characteristic among examined 'Lower Hamamelididae' (except Disanthus) and is also present in Daphnipfiyllum and Eucommia. We hypothesize that this specialized connective hypodermis facilitates a broader opening of the anther.     

Secretory tapetum of Brassica oleracea L.: polarity and ultrastructural features

Summary The ultrastructure of the secretory, binucleate tapetum of Brassica oleracea in the micro spore mother cell (MMC) stage through to the mature pollen stage is reported. The tapetal cells differentiate as highly specialized cells whose development is involved in lipid accumulation in their final stage. They start breaking down just before anther dehiscence. Nuclei with dispersed chromatin, large nucleoli and many ribosomes in the cytoplasm characterize the tapetal cells. The wall-bearing tapetum phase ends at the tetrade stage. The dissolution of tapetal walls begins from the inner tangential wall oriented towards the loculus and proceeds gradually along the radial walls to the outer tangential one. The plasmodesmata transversing the radial walls between tapetal cells persist until the mature microspore, long after loss of the inner tangential wall. After wall dissolution, the tapetal protoplasts retain their integrity and position within the anther locule. The tapetal cell membrane is in direct contact with the exine of the microspores/pollen grains and forms tubular evaginations that increase its surface area and appear to be involved in the translocation of solutes from the tapetal cells to the microspores/ pollen grains. The tapetal cells exhibit a polarity expressed by spatial differentiation in the radial direction.     

Localization of ubiquitin in anthers and pistils of Nicotiana

Ubiquitin-conjugated compounds were localized in anthers and pistils of Nicotiana alata by immuno-cytochemistry. In young anthers, antibodies to ubiquitin bound to callose cell walls surrounding pollen mother cells and to organelles in the endothecium. At the freespore stage, antibodies bound to circular-cell cluster cells subtending the stomium and to organelles and cell walls of endothecial cells. Near anther dehiscence, locular material was labeled. In pistils, cell walls of stylar transmitting tissue were labeled in a beaded pattern. Antibodies bound to a thin layer surrounding ovules, to the lining of embryo sacs, to cytoplasm of eggs and synergids, and to starch grains in central cells. Sites of localization were tissue- and time-specific, suggesting a regulatory role for ubiquitin in development of reproductive structures in flowering plants.     

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.     

ASSOCIATION OF FOUR DIFFERENT CALCIUM CRYSTALS IN THE ANTHER CONNECTIVE TISSUE AND HYPODERMAL STOMIUM OF CAPSICUM ANNUUM (SOLANACEAE) DURING MICROSPOROGENESIS

The anther connective tissue and hypodermal stomium between adjacent locules in the anthers of Capsicum annuum L. (Solanaceae) are the sites of formation of calcium salt crystals with four different habits. The spatial and temporal associations of these crystals and the idioblastic cells in which they form indicate that crystal sand occurs earliest in anther development near the single vascular strand, followed by spherulites and prismatic crystals farther out in the connective tissue, and finally druses occur in the hypodermal stomium. Both the druses and the crystal sand crystals are encased in crystal chambers and are associated with distinct membranes, whereas the spherulites and prismatic crystals are not bounded by any apparent membranes but they are surrounded by dense material that is rich in calcium and stains positively for polysaccharides and proteins. Quite often spherulites and prismatic crystals are observed within a single cell in contact with each other. X-ray diffraction of crystal preparations containing all four crystal habits and X-ray elemental analyses of single crystals, as well as visual observations and acid treatments, suggest that all four crystal habits consist of calcium oxalate. The hypodermal stomium and adjacent connective tissue degenerate at the pollen stage causing adjacent locules to fuse. Shortly afterward, each stomium epidermis splits open along the length of the anther releasing the pollen. It is suggested that the crystal idioblasts are involved in this process, possibly by a temporally orchestrated sequestration of calcium from both the cell cytoplasm and cell wall.     

A novel cell ablation strategy blocks tobacco anther dehiscence.

We utilized a new cell ablation strategy to ablate specific anther cell types involved in the dehiscence process. The tobacco TA56 gene promoter is active within the circular cell cluster, stomium, and connective regions of the anther at different developmental stages. We introduced a cytotoxic TA56/barnase gene into tobacco plants together with three different anticytotoxic barstar genes. The anticytotoxic barstar genes were used to protect subsets of anther cell types from the cytotoxic effects of the TA56/barnase gene. The chimeric barstar genes were fused with (1) the tobacco TP12 gene promoter that is active at high levels in most anther cell types; (2) the soybean lectin gene promoter that is active earlier in the connective, and at lower levels in the circular cell cluster and stomium, than is the TA56 promoter; and (3) the tobacco TA20 gene promoter that is active at high levels in most anther cell types but has a different developmental profile than does the TP12 promoter. Normal anther development and dehiscence occurred in plants containing the TA56/barnase and TP12/barstar genes, indicating that barstar protects diverse anther cell types from the cytotoxic effects of barnase. Anthers containing the TA56/barnase and lectin/barstar genes also developed normally but failed to dehisce because of extensive ablation of the circular cell cluster, stomium, and contiguous connective regions. Anthers containing the TA56/barnase and TA20/barstar genes failed to dehisce as well. However, only the stomium region was ablated in these anthers. The connective, circular cell cluster, and adjacent wall regions were protected from ablation by the formation of barnase/barstar complexes. We conclude that anther dehiscence at flower opening depends on the presence of a functional stomium region and that chimeric barnase and barstar genes containing promoters that are active in several overlapping cell types can be used for targeted cell ablation experiments.