Anther structure and pollen development in Jurinea kilaea Azn. (Asteraceae)

In this study, Gagea villosa (Bieb.) Duby was investigated by using light microscopy methods in cytological and cytoembryological respects. Anthers were tetrasporangiate. Anther wall was formed with an epidermis, endothecium, middle layer and tapetum. Tapetum was glandular type and it began to degenerate when microspores released from tetrads. Tapetum cells generally have one or two nuclei. Mitosis seen in tapetum cells was generally normal but micronuclei were found in some of them. Fibrous thickenings were determined in endothecium. Microsporogenesis and pollen mitosis were generally regular. Cytokinesis was successive type. Meiosis in pollen mother cells was asynchronous in one anther locus. Mature pollen grains were 2-celled. Pollen sterility was found to be 24%. Some of the fertile pollen grains, smaller than the normal were seen at the end of the pollen mitosis. Microgametophyte development was examined in vivo and in vitro. Germination ratio of pollen grains in vitro was 4%. Generally swollen pollen tube tips and weak development of some curled pollen tubes were seen. Callose plug formation was seen only in vivo pollen tube growth.

Long and short particles of the Brazilian tobacco rattle virus (BTRV) were visualized in cells of the anther tissues of infected tomato plants. The BTRV appeared mostly as aggregates of parallel particles associated with mitochondria, though isolated aggregates and individual particles were seen in microspores and mature pollen grains. The BTRV particles could be seen in the cytoplasm of infected anther cells during the various stages of microsporogenesis from the stage of premeiotic cells to mature pollen grains. Virus distribution through meristems is the main mechanism that permits the BTRV to reach the pollen. Virus passage from somatic cells to the microspore-precursor cells while plasmodesmata are still functional may be complementary in case of early infection or the only way of access in late-infected plants. Both short and long particles could be seen in the cytoplasm of the vegetative and generative cells of the pollen. No virus particle was ever seen in the nuclei of infected cells, associated with the cellulose or callose walls, or on the outside wall of pollen grains. There was a small number of virus particles present in the microspore mother cells and microspores, and a large number of short- and long-particle aggregates in the mature pollen grains. These results indicate that virus increase occurs during pollen grain maturation and is not only the result of distribution. Microsporogenesis is comparable between infected and noninfected tissues. However, pollen grains from infected plants showed a lower germination than normal pollen.

The formation of mature and fertile pollen grains, taking place inside the anther, depends on supply of assimilates, in the form of sucrose, provided mainly by the leaves. Data is limited, however, with respect to the understanding of sucrose metabolism in microspores and the supporting tissues. The aims of the present work were to 1) follow the changes in total and relative concentrations of sucrose, glucose, fructose and starch in the stamen parts and microspores up until anthesis, 2) follow the activities of sucrose-metabolism-related enzymes, in the anther walls fraction and microspores of the crop plant tomato. Sucrose was found to be partially cleaved in the filament, decreasing by more than twofold in the anther wall layers and the locular fluid, and to accumulate in the mature pollen grains, constituting 80% of total soluble sugars. Thus, sucrose was both the starting sugar, supporting microspore development, and the main carbohydrate accumulated at the end of the pollen-development program. The major invertase found to be active in both the anther wall layers and in maturing microspores was cell-wall-bound invertase. High fructokinase 2 and sucrose phosphate synthase activities during pollen maturation coincided with sucrose accumulation. The potential importance of sucrose accumulation during pollen dehydration phase and germination is discussed.

The study aimed to investigate the microsporogenesis, tapetum and pollen development in Petunia hybrida Juss. under control and air pollution condition. The connective shows a well-developed placentoid, giving the sporogenous tissue a crescent-shape. The sporogenous tissue of each of the four anther locules is surrounded by dimorphic tapetum. The outer (towards the epidermis) and inner (towards the connective) tapetal layers differ in shape, staining intensity and degree of vacuolization. During microsporogenesis and pollen maturation, the tapetum undergoes several changes and ultimately degenerates after pollen mitosis. The tapetum is the secretory type. Microsporogenesis with simultaneous cytokinesis forms tetrahedral tetrads of microspores. Mature pollen grains are prolate, tricolporate/tricolporoidate with furrows disposed along the polar axis and three lowly developed pores with striate-reticulate sculpture. Under air pollution, microsporogenesis was normal till tetrad stage. After this stage, some tetrahedral tetrads were still present and some pollen grains were irregular shaped, shrunk and fragile. Some tapetal cells were smaller and the number of their nuclei was less compared to those of the control. Cellular material release was higher in polluted pollen. SDS-PAGE pattern in polluted pollen did not show significant difference compared to the control.

The normal development of anthers and the formation of functional pollen are the prerequisites for successful pollination and fertilization. In this study, we observed dynamic changes in inflorescence and anther development in the chinquapin (Castanea henryi) using stereomicroscopy, light microscopy, and transmission electron microscopy. We found that cytokinesis during meiosis in microsporocytes was of the simultaneous type, and that the tetrads were mainly tetrahedral. Mature pollen grains contained two cells with three germ pores. The anther wall was of the basic type and composed of epidermis, endothecium, middle layers, and tapetum. Mature anthers had no middle layer and tapetum. The tapetum was of the glandular type. At the early microspore stage, a large number of starch granules appeared in the endothecium, which was deformed at the late microspore stage. Lipid droplets appeared in tapetum during the early microspore stage, and a few lipid droplets were still found during tapetum degeneration. The mature pollen accumulated a large amount of starch and lipids. These findings demonstrated that the anther wall provides nutrients and protection for pollen development. There is relatively stable correspondence between the external morphological characteristics of male flowers and internal structure of anther development.

The primary objective of this study is to describe the anther surface morphology and pollen characteristics (pollen count, size, shape, aperture, exine ornamentation and viability) of Momordica cymbalaria Hook. F to assist with taxonomic classification. Mature anthers and pollen grains were processed using traditional techniques and examined through light microscopy (LM), scanning electron microscopy (SEM), and fluorescence microscopy (FM). The androecium is a (3) + (2) structure with bell (Ո)-shaped and crescent (C)-shaped anthers, respectively. SEM observations reveal that both anther types are monothecal and bisporangiate, displaying similar morphologies with irregular, polygonal cells. Prominent ridges are observed between microsporangia, along with multicellular and glandular trichomes at the connective base and between the folds of the thecae. Bell (Ո)-shaped and crescent (C)-shaped anthers produced an average of 904 and 911 pollen grains per anther, respectively. The mature pollen grain is monad, large, oblate, and three-colporate, and varies from circular in polar view to elliptical in equatorial view, featuring coarsely reticulate exine ornamentation. The average diameters of pollen grains are 42–43 μm and 46–47 μm for polar (P) and equatorial (E) views, respectively, with a P/E ratio of 0.92 for both bell (Ո)-shaped and crescent (C)-shaped anthers. The study found a 95 ± 0.87% pollen viability, confirmed by the fluorescein diacetate test. The present study is the first investigation into the anther surface morphology and pollen characteristics of the genus, which play a significant role in the taxonomy of Momordica, particularly M. cymbalaria. Pollen count and viability studies are crucial for future breeding programmes and commercial fruit production.

Rapid diffusion of allergenic proteins in isotonic media has been demonstrated for different pollen grains. Upon contact with stigmatic secretion or with the mucosa of sensitive individuals, pollen grains absorb water and release soluble low-molecular-weight proteins, these proteins enter in the secretory pathway in order to arrive at the cell surface. In this study we located allergenic proteins in mature and hydrated-activated pollen grains of Parietaria judaica L. (Urticaceae) and studied the diffusion of these proteins during the first 20 min of the hydration and activation processes. A combination of transmission electron microscopy and immunocytochemical methods was used to locate these proteins in mature pollen and in pollen grains after different periods of hydration and activation processes. Activated proteins reacting with antibodies in human serum from allergic patients were found in the cytoplasm, wall, and exudates from the pollen grains. The allergenic component of these pollen grains changes according to the pollen state; the presence of these proteins in the exine, the cytoplasm, and especially in the intine and in the material exuded from the pollen grains, is significant in the hydrated-activated studied times, whereas this presence is not significant in mature pollen grains. The rapid activation and release of allergenic proteins of P. judaica pollen appears to be the main cause of the allergenic activity of these pollen grains.

Anther development in Catalpa bignonioides Walter was investigated from the sporogenous cell to the mature pollen grain stages to determine whether the pollen and anther wall development follows the basic scheme in angiosperms. In order to follow pollen ontogeny through successive stages of pollen development, anthers at different developmental stages were embedded in epon according to the usual method, and semi-thin sections, taken from the epon embedded anthers, were stained with toluidine blue for general histological observations under light microscopy. The young anther wall of C. bignonioides consists of four layers; from the exterior, the epidermis, endothecium, middle layer, and a secretory tapetum. The tapetum is dual in origin and dimorphic. Ubisch bodies were observed on the inner tangential walls of the tapetal cells. The number of the anther wall layers changes depending on the developmental stage and region of the anther. In contrast to the other anther wall layers , epidermis and endothecium layers remain intact until anthesis. Endothecial cells enlarge and develop thickenings at maturity. During microspore development , meiocytes undergo meiosis and simultaneous cytokinesis leading to the formation of permanent tetrahedral, isobilateral and rarely linear tetrads. Pollen tetrads are shed from the anther as compound pollen grains. Results of the study revealed that pollen and anther wall development in C. bignonioides follows the basic scheme in angiosperms.

This study has compared mature pollen grains while still in the anther, as well as post-pollination responses, from untreated and CHATM Chemical Hybridizing Agent-treated wheat plants using light (bright-field, phase-contrast, and fluorescence), scanning, and transmission electron microscopy. The chemical, azetidine-3-carboxylic acid (A3C), was applied at three treatment levels of 0.75, 1.0 and 1.5 kg/ha for the mature pollen investigations. It was found that the major effect of A3C on mature pollen is an alteration of the wall precursor vesicles (wp-vesicles), which form a high proportion of the contents of the mature grass pollen grain. The degree of deformation of the wp-vesicles is dose dependent. There is some evidence that increased aggregations of ribosomes are formed in treated pollen cytoplasm. Pollination studies (all at a treatment level of 1.0 kg/ha) show that, in most cases, the treated pollen does not germinate, and a high percentage of the pollen grains burst (60% burst grains in treated material compared to 28% in controls). In about 20% of the cases from treated plants, a short pollen tube forms, but no tubes were seen to grow far enough to enter the stigma hairs of the pistil. Thus, A3C does not act by preventing pollen formation, but by the prevention of normal pollen tube growth. There appears to be a specific targeting of the wp-vesicles such that, even in cases where the ultra-structure of the vesicles is not altered, the normal course of events leading to the incorporation of their contents into the extending tube wall is arrested. Further studies must be undertaken to determine the significance of the effect of CHATM Chemical Hybridizing Agent on wp-vesicle composition.

In mature pollen grains, lipids are primarily stored in the form of lipid droplets that provide energy and act as a carbon source for normal pollen development and germination. Triacylglycerol (TAG) is the major form of stored plant lipids. Diacylglycerol transferase, which is encoded by DGAT1 in Arabidopsis thaliana, is an important enzyme regulating triacylglycerol synthesis. Within the seeds of the DGAT1 mutant as11, the content of TAG is significantly decreased and the fatty acid composition also differs from the wild type. Transcriptome data of mature anthers showed that the genes involved in the TAG synthesis pathway were downregulated in as11. Analysis of gene expression patterns via transcriptome data also revealed that the expression of PDAT1, which functions in a manner complementary to the DGAT1 gene, was significantly decreased in as11, whereas the amylopectin synthase genes SS1 and SS2 were upregulated in mutant as11. We also detected lower total lipid, TAG and fatty acid contents in mature as11 pollen, with palmitic acid (C16:0) and linolenic acid (C18:3) being the major fatty acids in mature pollen. The cytological results showed that the lipid droplet content was reduced in mature as11 pollen. In the binuclear pollen grain II stage, WT pollen contained lipid droplets that were primarily accumulated around the generative nucleus, whereas the pollen in the mutant as11 was rich in starch grains that were primarily distributed around the vegetative nucleus. Ultrastructural analysis indicated that during pollen development in as11, the amount of endoplasmic reticulum in tapetal cells and pollen grains decreased, whereas the Golgi body content increased, which directly or indirectly led to a decrease in the levels of lipidosomes and an increase in the starch content in as11. Changes in the lipid content and fatty acid composition of the mutant as11 differ from those in the wild type during pollen development.

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.

1. A cytoplasmic basophily, which is removed when cells are treated with a ribonuclease enzyme, has been used as a means of identifying ribose nucleic acids and of following their distribution in the cells of the anthers of Rhoeo, and to a lesser extent in the pistil of the flower. Rapid cell growth is invariably accompanied by large amounts of ribonucleic acid in the cytoplasm and heavy deposits of chromatin in nuclei. 2. In young anthers the cytoplasm of both tapetal and sporogenous tissues is extremely rich in ribonucleic acid. As the pollen mother cells prepare for the meiotic divisions, the ribonucleic acid disappears from the cytoplasm, presumably being passed into the nucleus and changed into desoxyribose nucleic acid (chromatin). Initially the cytoplasm of microspores is devoid of basophilic material, but as they grow into mature pollen grains large amounts of ribonucleic acids accumulate in the cytoplasm. 3. The growth of tapetal cells is accompanied by one or more endomitotic nuclear division cycles. The tapetal cells, which are rich in cytoplasmic nucleic acid at the time of cytolysis, are the source of the ribonucleic acid which accumulates in the maturing pollen grains. 4. There is a similarity between microspore growth at the expense of tapetal cells and growth of oocytes in animals through the agency of nurse cells. Large amounts of ribonucleic acid are stored in mature pollen grains and used in the extremely rapid synthesis of new materials which accompanies pollen tube growth.

Jujube (Ziziphus jujuba Mill.) is an important fruit tree species in China. In this study, we studied the megasporogenesis, microsporogenesis, and female and male gametophyte development of two major jujube cultivars, "Dongzao" and "Mayazao," using the squash technique, improved paraffin section technology, and optical microscopy. Our investigation revealed that both "Dongzao" and "Mayazao" have bilocular ovaries, basal placenta, and anatropous, bitegmic, crassinucellate ovules. The tetrads formed by meiosis of megaspore mother cells are arranged in a straight line or a tetrahedron. Embryo sac development is of the Polygonum type. The flower buds contain five anthers, each having four pollen sacs. The anther wall, which is of the fundamental form, is composed of epidermis, endothecium, one or two middle layers, and glandular tapetum. Mature pollen grains are two-celled and three-colporate. Both "Dongzao" and "Mayazao" can form normal mature pollen grains. Our study, which has revealed the basic phenomena and progression of megasporogenesis, microsporogenesis, and female and male gametophyte development in jujube, has generated important data for further research on jujube cytology and reproductive biology. Finally, our explorations of the cytological mechanism of male sterility in "Dongzao" also have provided a cytological basis for crossbreeding.

Using the monoclonal antibodies JIM 5 and 7, pectin was immunolocalized and quantitatively assayed in three anther compartments of Lilium hybrida during pollen development. Pectin levels in both the anther wall and the loculus increased following meiosis, were maximal during the early microspore stages and declined during the remainder of pollen ontogenesis. In the microspores/pollen grains, pectin was detectable at low levels during the microspore stages but accumulated significantly during pollen maturation. During early microspore vacuolation, esterified pectin epitopes were detected both in the tapetum cytoplasm and vacuoles. In the anther loculus, the same epitopes were located simultaneously in undulations of the plasma membrane and in the locular fluid. At the end of microspore vacuolation, esterified pectin epitopes were present within the lipids of the pollenkitt, and released in the loculus at pollen mitosis. Unesterified pectin epitopes were hardly detectable in the cytoplasm of the young microspore but were as abundant in the primexine matrix as in the loculus. During pollen maturation, both unesterified and esterified pectin labelling accumulated in the cytoplasm of the vegetative cell, concurrently with starch degradation. In the mature pollen grain, unesterified pectin epitopes were located in the proximal intine whereas esterified pectin epitopes were deposited in the distal intine. These data suggest that during early microspore development, the tapetum secretes pectin, which is transferred to the primexine matrix via the locular fluid. Further, pectin is demonstrated to constitute a significant component of the pollen carbohydrate reserves in the mature grain of Lilium.

Biotechnological and selection investigations for making of wheat high-yielding resistant varieties require a significant amount of high-quality mature pollen. It is important to assess It is important to assess the quality of pollen grains in mature anthers in planta of the wheat genotypes included in the experiments. In the course of research the analysis of the histological status of mature anthers was carried out and qualitative cytological evaluation inside them pollen grain of spring soft wheat Zhnitsa cultivar was done in in planta conditions. It was shown that the structure of the wall locule anther presented by exothecium and endothecium is typical for cereals. It was established that 3-cell mature pollen grains are fertile (87-92%) and viable (75–80%) in general. At the same time in every mature anther abnormal pollen grains were observed and the cellular, nuclear, cytoplasmic and architectonic deviations from the norm were revealed. On the base of literature date the analysis of the reasons for the formation of abnormal pollen grains in planta was done and the classification of anomalies of plant pollen grains and the terminology used in this field of research were discussed.