Pollen Formation Research Articles

Full-length transcriptome analysis provides new insights into the early bolting occurrence in medicinal Angelica sinensis

Angelica sinensis (Oliv.) Diels root part is an integral component of traditional Chinese medicine, widely prescribed to improve blood circulation and blood stasis. However, early bolting of A. sinensis compromises the quality of the roots and hence is a major limitation for yield of medicinal materials. To date, little information about the molecular mechanisms underlying bolting is available for this important medicinal plant. To identify genes putatively involved in early bolting, we have conducted the transcriptome analysis of the shoot tips of the early-bolting plants and non-bolting (normal) plants of A. sinensis, respectively, using a combination of third-generation sequencing and next-generation sequencing. A total of 43,438 non-redundant transcripts were collected and 475 unique differentially expressed genes (DEGs) were identified. Gene annotation and functional analyses revealed that DEGs were highly involved in plant hormone signaling and biosynthesis pathways, three main flowering pathways, pollen formation, and very-long-chain fatty acids biosynthesis pathways. The levels of endogenous hormones were also changed significantly in the early bolting stage of A. sinensis. This study provided new insights into the transcriptomic control of early bolting in A. sinensis, which could be further applied to enhance the yield of medicinally important raw materials.

New genomic resources and comparative analyses reveal differences in floral gene expression in selfing and outcrossing Collinsia sister species.

The evolutionary transition from outcross- to self-fertilization is one of the most common in angiosperms and is often associated with a parallel shift in floral morphological and developmental traits, such as reduced flower size and pollen to ovule ratios, known as the “selfing syndrome.” How these convergent phenotypes arise, the extent to which they are shaped by selection, and the nature of their underlying genetic basis are unsettled questions in evolutionary biology. The genus Collinsia (Plantaginaceae) includes seven independent transitions from outcrossing or mixed mating to high selfing rates accompanied by selfing syndrome traits. Accordingly, Collinsia represents an ideal system for investigating this parallelism, but requires genomic resource development. We present a high quality de novo genome assembly for the highly selfing species Collinsia rattanii. To begin addressing the basis of selfing syndrome developmental shifts, we evaluate and contrast patterns of gene expression from floral transcriptomes across three stages of bud development for C. rattanii and its outcrossing sister species Collinsia linearis. Relative to C. linearis, total gene expression is less variable among individuals and bud stages in C. rattanii. In addition, there is a common pattern among differentially expressed genes: lower expression levels that are more constant across bud development in C. rattanii relative to C. linearis. Transcriptional regulation of enzymes involved in pollen formation specifically in early bud development may influence floral traits that distinguish selfing and outcrossing Collinsia species through pleiotropic functions. Future work will include additional Collinsia outcrossing-selfing species pairs to identify genomic signatures of parallel evolution.

Ubiquitin-dependent Argonauteprotein MEL1 degradation is essential for rice sporogenesis and phasiRNA target regulation

MEIOSIS ARRESTED AT LEPTOTENE1 (MEL1), a rice (Oryza sativa) Argonaute (AGO) protein, has been reported to function specifically at premeiotic and meiotic stages of germ cell development and is associated with a novel class of germ cell-specific small noncoding RNAs called phased small RNAs (phasiRNAs). MEL1 accumulation is temporally and spatially regulated and is eliminated after meiosis. However, the metabolism and turnover (i.e. the homeostasis) of MEL1 during germ cell development remains unknown. Here, we show that MEL1 is ubiquitinated and subsequently degraded via the proteasome pathway in vivo during late sporogenesis. Abnormal accumulation of MEL1 after meiosis leads to a semi-sterile phenotype. We identified a monocot-specific E3 ligase, XBOS36, a CULLIN RING-box protein, that is responsible for the degradation of MEL1. Ubiquitination at four K residues at the N terminus of MEL1 by XBOS36 induces its degradation. Importantly, inhibition of MEL1 degradation either by XBOS36 knockdown or by MEL1 overexpression prevents the formation of pollen at the microspore stage. Further mechanistic analysis showed that disrupting MEL1 homeostasis in germ cells leads to off-target cleavage of phasiRNA target genes. Our findings thus provide insight into the communication between a monocot-specific E3 ligase and an AGO protein during plant reproductive development.

Floral anatomy, embryology, seed, and fruit development in Cephalanthus (Naucleeae-Rubiaceae), with emphasis on C. glabratus.

Information on the reproductive anatomy in genera of the tribe Naucleeae, particularly Cephalanthus, is scarce and fragmented. Of the six species in the genus, only the mature megagamethophyte of Cephalanthus occidentalis has been described. This study aims to provide information on embryological aspects in flowers of C. glabratus and to analyze the morphology and anatomy of the flowers, fruit, and seed in the six species of the genus. Cephalanthus glabratus have imperfect flowers: pistillate (PF) and staminate (SF). In the PF, the ovules are functional, while in the SF, they atrophy during the formation of the embryo sac. The mature ovule has a single integument, corresponds to the Phyllis type and the embryo sac is a Polygonum type, forming only in the PF. The presence of pollenkitt and secondary presentation of pollen were observed in the SF, as well as in the pollen formation previously described, whereas in the PF, they are absent, due to the collapse of the pollen grains inside the indehiscent anthers. The analysis of the ontogeny of the ovular excrescence in C. glabratus determined its funicular origin, calling it an aril. Its development is a pre-anthesis event, initiated during megasporogenesis. In seeds, the aril is a fleshy, white appendage which almost completely envelops the seeds of Cephalanthus, except for Cephalanthus natalensis where it is noticeably more reduced. Studies of the fruit in Cephalanthus species indicate that the infructescence is a dry schizocarp which separates into uni-seminated mericarps, except in C. natalensis that has fleshy indehiscent fruit.

A MADS-Box Gene CiMADS43 Is Involved in Citrus Flowering and Leaf Development through Interaction with CiAGL9.

MADS-box genes are involved in various developmental processes including vegetative development, flower architecture, flowering, pollen formation, seed and fruit development. However, the function of most MADS-box genes and their regulation mechanism are still unclear in woody plants compared with model plants. In this study, a MADS-box gene (CiMADS43) was identified in citrus. Phylogenetic and sequence analysis showed that CiMADS43 is a GOA-like Bsister MADS-box gene. It was localized in the nucleus and as a transcriptional activator. Overexpression of CiMADS43 promoted early flowering and leaves curling in transgenic Arabidopsis. Besides, overexpression or knockout of CiMADS43 also showed leaf curl phenotype in citrus similar to that of CiMADS43 overexpressed in Arabidopsis. Protein–protein interaction found that a SEPALLATA (SEP)-like protein (CiAGL9) interacted with CiMADS43 protein. Interestingly, CiAGL9 also can bind to the CiMADS43 promoter and promote its transcription. Expression analysis also showed that these two genes were closely related to seasonal flowering and the development of the leaf in citrus. Our findings revealed the multifunctional roles of CiMADS43 in the vegetative and reproductive development of citrus. These results will facilitate our understanding of the evolution and molecular mechanisms of MADS-box genes in citrus.

Cytological and morphology characteristics of natural microsporogenesis within Camellia oleifera.

The online version contains supplementary material available at 10.1007/s12298-021-01002-5.

Comprehensive Genome-Wide Exploration of C2H2 Zinc Finger Family in Grapevine (Vitis vinifera L.): Insights into the Roles in the Pollen Development Regulation.

Some C2H2 zinc-finger proteins (ZFP) transcription factors are involved in the development of pollen in plants. In grapevine (Vitis vinifera L.), it has been suggested that abnormalities in pollen development lead to the phenomenon called parthenocarpy that occurs in some varieties of this cultivar. At present, a network involving several transcription factors types has been revealed and key roles have been assigned to members of the C2H2 zinc-finger proteins (ZFP) family in model plants. However, particularities of the regulatory mechanisms controlling pollen formation in grapevine remain unknown. In order to gain insight into the participation of ZFPs in grapevine gametophyte development, we performed a genome-wide identification and characterization of genes encoding ZFP (VviZFP family). A total of 98 genes were identified and renamed based on the gene distribution into grapevine genome. The analysis performed indicate significant changes throughout VviZFP genes evolution explained by high heterogeneity in sequence, length, number of ZF and presence of another conserved domains. Moreover, segmental duplication participated in the gene family expansion in grapevine. The VviZFPs were classified based on domain and phylogenetic analysis into three sets and different groups. Heat-map demonstrated differential and tissue-specific expression patterns of these genes and k-means clustering allowed to identify a group of putative orthologs to some ZFPs related to pollen development. In transgenic plants carrying the promVviZFP13::GUS and promVviZFP68::GUS constructs, GUS signals were detectable in the anther and mature pollen grains. Expression profiling of selected VviZFP genes showed differential expression pattern during flower development and provides a basis for deepening in the understanding of VviZFPs role on grapevine reproductive development.

Slowing Development Facilitates Arabidopsis mgt Mutants to Accumulate Enough Magnesium for Pollen Formation and Fertility Restoration.

Magnesium (Mg) is an abundant and important cation in cells. Plants rely on Mg transporters to take up Mg from the soil, and then Mg is transported to anthers and other organs. Here, we showed that MGT6+/− plants display reduced fertility, while mgt6 plants are fertile. MGT6 is expressed in the anther at the early stages. Pollen mitosis and intine formation are impaired in aborted pollen grains (PGs) of MGT6+/− plants, which is similar to the defective pollen observed in mgt5 and mgt9 mutants. These results suggest that Mg deficiency leads to pollen abortion in MGT6+/− plants. Our data showed that mgt6 organs including buds develop significantly slower and mgt6 stamens accumulate a higher level of Mg, compared with wild-type (WT) and MGT6+/− plants. These results indicate that slower bud development allows mgt6 to accumulate sufficient amounts of Mg in the pollen, explaining why mgt6 is fertile. Furthermore, we found that mgt6 can restore fertility of mgt5, which has been reported to be male sterile due to defects in Mg transport from the tapetum to microspores and that an additional Mg supply can restore its fertility. Interestingly, mgt5 fertility is recovered when grown under short photoperiod conditions, which is a well-known factor regulating plant fertility. Taken together, these results demonstrate that slow development is a general mechanism to restore mgts fertility, which allows other redundant magnesium transporter (MGT) members to transport sufficient Mg for pollen formation.

Genome-Wide Identification of Copper Stress-Regulated and Novel MicroRNAs in Mulberry Leaf.

Copper (Cu) is an essential trace element for plant growth and development. It is widely involved in respiration, photosynthesis, pollen formation, and other biological processes. Therefore, low or excessive copper causes damage to plants. Mulberry is an essential perennial economic tree. At present, research on the abiotic stress responses in mulberry is mainly focused on the identification of resistant germplasm resources and cloning of resistant genes. In contrast, studies on the resistance function of microRNAs and the regulatory gene responses to stress are rare. In this study, small RNA libraries (control and copper stressed) were constructed from mulberry leaf RNA. High-throughput sequencing and screening were employed, a total of 65 known miRNAs and 78 predicted novel mature miRNAs were identified, among which 40 miRNAs were differentially expressed under copper stress. Subsequently, expression patterns were verified for 14 miRNAs by real-time fluorescence quantitative PCR (qPCR). The target genes of miRNAs were validated by 5' RLM-RACE. Our results provide the bases for further study on the molecular mechanism of copper stress regulation in mulberry.

Flower size dimorphism, perianth shape and sex ratio in a gynodioecious Prunella vulgaris (Lamiaceae)

Prunella vulgaris is a gynodioecious species that forms two types of flowers, perfect and female, on different plants. These two sexual types of flowers have significant differences in some characters of calyx, corolla, androecium, and gynoecium. The distinctiveness of gynodioecy in P. vulgaris consists in insignificant size differences in anthers, the formation of sterile pollen in female flowers, and the shape of a corolla in two sexual types of flowers. Hermaphrodite plants are dominated in P. vulgaris (59.8 to 87 percent). In flood plain meadows, the proportion of females is significantly higher (from 25.9 to 40.2 percent) than in dry and forest meadows (from 13 to 19.1 percent).

SPECTRUM OF CAUSATIVELY RELEVANT AEROALLERGENS OF SOUTHERN UKRAINE WHICH PROVOKE ALLERGIC REACTION IN PATIENTS WITH POLLINOSIS ON THE EXAMPLE OF KHERSON REGION

The aim: the analysis of spectrum of causatively relevant aeroallergens of southern Ukraine which provoke allergic reaction in patients with pollinosis. Materials and methods: ambulatory medical records data and skin prick-testing results of 477 patients afflicted with pollinosis in Kherson (250 women - 52,41 % and 227 men - 47,59 %) aged from 19 to 66 years old (38,05±0,45) have been analyzed retrospectively. Results: all patients revealed clinical implications mainly during the September-October period. Clinically 151 patients (31,66%) showed severe and 326 (68,34%) moderate to severe clinical progression. Composite family plants (composites) proved to be the most widespread plants (69,76 %) among all pollen aeroallergens of Kherson region. Weeds and sunflower showed the highest percentage among composites: Ambrosia topped to 79,87 %, Helianthus (sunflower)- 75,68 %, Artemisia - 42,98 %, and Cyclachaena (sump weed) - 36,48 %. Wild and cultivated grains' aeroallergens toped to 28,22 %. Among them the sensibility to Lolium perenne (19,28 %), Festuca pratensis (17,82 %), Dactylis glomerata (17,82 %) has been noticed more often. Tree aeroallergens topped to 2,03 %. Conclusions: The third wave of pollen formation (September-October period) appeared to be the most significant and clinically hard for Kherson region. According to the specific allergologic research data, allergens of the composite family plants, among which pollen of Ambrosia, Helianthus, Artemisia and Cyclachaena cause more frequent sensibility, turned to be the most widespread causatively relevant aeroallergens in Kherson region which provoke allergic reaction in patients with pollinosis. The second place was taken by wild and cultivated grains' aeroallergens, among which sensibility to Lolium perenne, Festuca pratensis, Dactylis glomerata proved to be the most common.

The Arabidopsis AGC kinases NDR2/4/5 interact with MOB1A/1B and play important roles in pollen development and germination.

Pollen formation and pollen tube growth are essential for the delivery of male gametes into the female embryo sac for double fertilization. Little is known about the mechanisms that regulate the late developmental process of pollen formation and pollen germination. In this study, we characterized a group of Arabidopsis AGC kinase proteins, NDR2/4/5, involved in pollen development and pollen germination. The NDR2/4/5 genes are mainly expressed in pollen grains at the late developmental stages and in pollen tubes. They function redundantly in pollen formation and pollen germination. At the tricellular stages, the ndr2 ndr4 ndr5 mutant pollen grains exhibit an abnormal accumulation of callose, precocious germination and burst in anthers, leading to a drastic reduction in fertilization and a reduced seed set. NDR2/4/5 proteins can interact with another group of proteins (MOB1A/1B) homologous to the MOB proteins from the Hippo signaling pathway in yeast and animals. The Arabidopsis mob1a mob1b mutant pollen grains also have a phenotype similar to that of ndr2 ndr4 ndr5 pollen grains. These results provide new evidence demonstrating that the Hippo signaling components are conserved in plants and play important roles in sexual plant reproduction.

A Functional Kinase Is Necessary for Cyclin-Dependent Kinase G1 (CDKG1) to Maintain Fertility at High Ambient Temperature in Arabidopsis.

Maintaining fertility in a fluctuating environment is key to the reproductive success of flowering plants. Meiosis and pollen formation are particularly sensitive to changes in growing conditions, especially temperature. We have previously identified cyclin-dependent kinase G1 (CDKG1) as a master regulator of temperature-dependent meiosis and this may involve the regulation of alternative splicing (AS), including of its own transcript. CDKG1 mRNA can undergo several AS events, potentially producing two protein variants: CDKG1L and CDKG1S, differing in their N-terminal domain which may be involved in co-factor interaction. In leaves, both isoforms have distinct temperature-dependent functions on target mRNA processing, but their role in pollen development is unknown. In the present study, we characterize the role of CDKG1L and CDKG1S in maintaining Arabidopsis fertility. We show that the long (L) form is necessary and sufficient to rescue the fertility defects of the cdkg1-1 mutant, while the short (S) form is unable to rescue fertility. On the other hand, an extra copy of CDKG1L reduces fertility. In addition, mutation of the ATP binding pocket of the kinase indicates that kinase activity is necessary for the function of CDKG1. Kinase mutants of CDKG1L and CDKG1S correctly localize to the cell nucleus and nucleus and cytoplasm, respectively, but are unable to rescue either the fertility or the splicing defects of the cdkg1-1 mutant. Furthermore, we show that there is partial functional overlap between CDKG1 and its paralog CDKG2 that could in part be explained by overlapping gene expression.

Viability of black currant pollen before and after cryopreservation in liquid nitrogen, and its morphological features

Background.Learning about morphological and biological features of pollen is very important to make correct estimations of plant productivity and breeding efficiency.Materials and methods. The viability of black currant pollen was analyzed in 2019 at the Laboratory for Long-Term Storage of Plant Genetic Resources, N.I. Vavilov All-Russian Institute of Plant Genetic Resources (VIR). Eleven black currant cultivars of various ecogeographic and genetic origin maintained at Pushkin and Pavlovsk Laboratories of VIR served as the material for the research. Pollen viability was tested on an artificial medium with 10% sucrose and 0.8% agar. Morphological studies were carried out using light and confocal laser scanning microscopy at the Palynology Laboratory and the Core Centrum of Cell and Molecular Technologies in Plant Science housed by the Komarov Botanical Institute (BIN).Results and conclusion. The prevailing adverse weather conditions in 2019 produced a negative impact on the process of male generative structure formation. The level of pollen viability, depending on the cultivar, ranged from 17.98 to 58.60%. After exposure to liquid nitrogen (–196°C) for 6 months, the number of germinated pollen grains increased 1.1 to 3.2 times in all the studied cultivars, except one (‘Pozdnyaya poslevoennaya’, k-7652). A palynomorphological study revealed that reduced pollen viability in some cultivars (‘Krasnoyarskaya 1018’, k-7607; ‘Tsema’, k-25900; ‘Pozdnyaya poslevoennaya’, k-7652) was associated with morphological deviations in the sporoderm structure (small pollen grains, exine thickening and formation of verrucate mesoporium surfaces). A low positive correlation was observed between the level of pollen viability and the diameters of the pore (r = 0.43) and pollen grain (r = 0.27). An insignificant negative correlation was apparent between the level of viability and the exine thickness (r = –0.33).

Callose Deposition during Pollen Development.

Callose is a cell wall component that is dynamically deposited and degraded during pollen development. Thanks to a new article investigating pollen formation in cotton ( Gossypium hirsutum ), we now know that a pollen-specific protein regulates callose deposition by inhibiting the action of a

Overexpression of MaTPD1A impairs fruit and pollen development by modulating some regulators in Musa itinerans

BackgroundPollen formation and development is important for crop fertility and is a key factor for hybrid development. Previous reports have indicated that Arabidopsis thaliana TAPETUM DETERMINANT1 (AtTPD1) and its rice (Oryza sativa) homolog, OsTPD1-like (OsTDL1A), are required for cell specialization and greatly affect pollen formation and development. Little is known about the role of the TPD1 homolog in banana pollen development.ResultsHere, we report the identification and characterization of TPD1 homologs in diploid banana (Musa itinerans) and examine their role in pollen development by overexpressing the closest homolog, MaTPD1A. MaTPD1A exhibits high expression in stamen and localizes in the plasma membrane. MaTPD1A-overexpressing plants produce no pollen grains and smaller and seedless fruit compared to wild-type plants. Transcriptome analysis showed that in plant hormone, starch and sucrose metabolism, and linolenic acid metabolism-related pathways were affected by overexpression of MaTPD1A, and the expression of several key regulators, such as PTC1 and MYB80, which are known to affect anther development, is affected in MaTPD1A-overexpressing lines.ConclusionsOur results indicate that MaTPD1A plays an important role in pollen formation and fruit development in diploid banana, possibly by affecting the expression of some key regulators of pollen development.

Macroecological patterns of the terrestrial vegetation history during the end-Triassic biotic crisis in the central European Basin: A palynological study of the Bonenburg section (NW-Germany) and its supra-regional implications

Abstract The end-Triassic mass extinction is often linked to environmental and climate change triggered by the activity of the Central Atlantic Magmatic Province ca. 201 Mya. In the German Triassic Basin, the transition from pre-extinction to the so-called extinction interval is documented from the fossil-rich Contorta Beds to the mostly barren Triletes Beds. However, despite the lack of macrofossils, plant microfossils are present and studying palynomorph diversity and assemblages still give us a detailed insight into the impact of environmental changes on the flora. Here, we present data from 64 samples taken from the new Triassic-Jurassic section ‘Bonenburgʼ, which originates from a brick quarry in North Rhine-Westphalia (Germany), and encompasses the Triassic–Jurassic transition in the Germanic Basin. Using palynofacies analysis, we document changes in the depositional environment. Based on terrestrial and marine palynological analysis, we also document quantitative changes in four assemblage zones for correlation with other European sections. Further, we discuss the vegetation history with special focus on the transition from the pre-extinction to extinction interval (Contorta to Triletes Beds). Additionally, we investigate palynofloral diversity patterns especially prior to the Triassic-Jurassic transition, to evaluate the ecological impact of environmental upheaval on the flora. Furthermore, we document intraspecific palynomorph variability, indicating a variety of aberrant spore, pollen, and tetrad formation, in the middle Rhaetian, the lowermost upper Rhaetian, and the lowest Hettangian. Our study supports existing paleogeographical reconstructions for the region and reveals, that vegetation underwent gradual changes with intermediate successional stages rather than dramatic extinction or drastic turnover as documented for animals. Diversity patterns coinciding with aberrant palynomorph occurrences, potentially associated with episodes of increased environmental stress, suggest three disturbances pulses of probably increasing severity. Although environmental stress in the context of diversity patterns seems a likely explanation for the occurrence of aberrant palynomorphs, further investigations are needed to better understand the underlying mechanisms and their evolutionary significance and temporal connection with biotic crises.

Arabidopsis FAX1 mediated fatty acid export is required for the transcriptional regulation of anther development and pollen wall formation.

The mutation of FAX1 (Fatty Acid Export 1) disrupts ROS homeostasis and suppresses transcription activity of DYT1-TDF1-AMS-MS188 genetic network, leading to atypical tapetum PCD and defective pollen formation in Arabidopsis. Fatty acids (FAs) have multiple important biological functions and exert diverse cellular effects through modulating Reactive Oxygen Species (ROS) homeostasis. Arabidopsis FAX1 (Fatty Acid Export 1) mediates the export of de novo synthesized FA from chloroplast and loss of function of FAX1 impairs male fertility. However, mechanisms underlying the association of FAX1-mediated FA export with male sterility remain enigmatic. In this study, by using an integrated approach that included morphological, cytological, histological, and molecular analyses, we revealed that loss of function of FAX1 breaks cellular FA/lipid homeostasis, which disrupts ROS homeostasis and suppresses transcriptional activation of the DYT1-TDF1-AMS-MS188 genetic network of anther development, impairing tapetum development and pollen wall formation, and resulting in male sterility. This study provides new insights into the regulatory network for male reproduction in plants, highlighting an important role of FA export-mediated ROS homeostasis in the process.

Hermaphroditism in Marijuana (Cannabis sativa L.) Inflorescences - Impact on Floral Morphology, Seed Formation, Progeny Sex Ratios, and Genetic Variation.

Cannabis sativa L. (hemp, marijuana) produces male and female inflorescences on different plants (dioecious) and therefore the plants are obligatory out-crossers. In commercial production, marijuana plants are all genetically female; male plants are destroyed as seed formation reduces flower quality. Spontaneously occurring hermaphroditic inflorescences, in which pistillate flowers are accompanied by formation of anthers, leads to undesired seed formation; the mechanism for this is poorly understood. We studied hermaphroditism in several marijuana strains with three objectives: (i) to compare the morphological features of this unique phenotype with normal male flowers; (ii) to assess pollen and seed viability from hermaphroditic flowers; and (iii) to assess the effect of hermaphroditism on progeny male:female (sex) ratios and on genetic variation using molecular methods. The morphological features of anthers, pollen production and germination in hermaphroditic flowers and in staminate inflorescences on male plants were compared using light and scanning electron microscopy. Seeds produced on hermaphroditic plants and seeds derived from cross-fertilization were germinated and seedlings were compared for gender ratios using a PCR-based assay as well as for the extent of genetic variation using six ISSR primers. Nei’s index of gene diversity and Shannon’s Information index were compared for these two populations. The morphology of anthers and pollen formation in hermaphroditic inflorescences was similar to that in staminate flowers. Seedlings from hermaphroditic seeds, and anther tissues, showed a female genetic composition while seedlings derived from cross-fertilized seeds showed a 1:1 male:female sex expression ratio. Uniquely, hermaphroditic inflorescences produced seeds which gave rise only to genetically female plants. In PCR assays, a 540 bp size fragment was present in male and female plants, while a 390 bp band was uniquely associated with male plants. Sequence analysis of these fragments revealed the presence of Copia-like retrotransposons within the C. sativa genome which may be associated with the expression of male or female phenotype. In ISSR analysis, the percentage of polymorphic loci ranged from 44 to 72% in hermaphroditic and cross-fertilized populations. Nei’s index of gene diversity and Shannon’s Information index were not statistically different for both populations. The extent of genetic variation after one generation of selfing in the progeny from hermaphroditic seed is similar to that in progeny from cross-fertilized seeds.

A Cytological Study of Anther and Pollen Development in Chinquapin (Castanea henryi)

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.