Mature Pollen Research Articles

A Rapid Alkalinization Factor-like Peptide EaF82 Impairs Tapetum Degeneration during Pollen Development through Induced ATP Deficiency.

In plants, the timely degeneration of tapetal cells is essential for providing nutrients and other substances to support pollen development. Rapid alkalinization factors (RALFs) are small, cysteine-rich peptides known to be involved in various aspects of plant development and growth, as well as defense against biotic and abiotic stresses. However, the functions of most of them remain unknown, while no RALF has been reported to involve tapetum degeneration. In this study, we demonstrated that a novel cysteine-rich peptide, EaF82, isolated from shy-flowering 'Golden Pothos' (Epipremnum aureum) plants, is a RALF-like peptide and displays alkalinizing activity. Its heterologous expression in Arabidopsis delayed tapetum degeneration and reduced pollen production and seed yields. RNAseq, RT-qPCR, and biochemical analyses showed that overexpression of EaF82 downregulated a group of genes involved in pH changes, cell wall modifications, tapetum degeneration, and pollen maturation, as well as seven endogenous Arabidopsis RALF genes, and decreased proteasome activity and ATP levels. Yeast two-hybrid screening identified AKIN10, a subunit of energy-sensing SnRK1 kinase, as its interacting partner. Our study reveals a possible regulatory role for RALF peptide in tapetum degeneration and suggests that EaF82 action may be mediated through AKIN10 leading to the alteration of transcriptome and energy metabolism, thereby causing ATP deficiency and impairing pollen development.

Assembly and Comparative Analysis of the Complete Mitochondrial Genome of Two Species of Calla Lilies (Zantedeschia, Araceae).

In this study, the mitochondrial genomes of two calla species, Zantedeschia aethiopica Spreng. and Zantedeschia odorata Perry., were assembled and compared for the first time. The Z. aethiopica mt genome was assembled into a single circular chromosome, measuring 675,575 bp in length with a 45.85% GC content. In contrast, the Z. odorata mt genome consisted of bicyclic chromosomes (chromosomes 1 and 2), measuring 719,764 bp and exhibiting a 45.79% GC content. Both mitogenomes harbored similar gene compositions, with 56 and 58 genes identified in Z. aethiopica and Z. odorata, respectively. Analyses of codon usage, sequence repeats, gene migration from chloroplast to mitochondrial, and RNA editing were conducted for both Z. aethiopica and Z. odorata mt genomes. Phylogenetic examination based on the mt genomes of these two species and 30 other taxa provided insights into their evolutionary relationships. Additionally, the core genes in the gynoecium, stamens, and mature pollen grains of the Z. aethiopica mt genome were investigated, which revealed maternal mitochondrial inheritance in this species. In summary, this study offers valuable genomic resources for future research on mitogenome evolution and the molecular breeding of calla lily.

Arabidopsis pollen-specific glycerophosphodiester phosphodiesterase-like genes are essential for pollen tube tip growth.

In angiosperms, pollen tube growth is critical for double fertilization and seed formation. Many of the factors involved in pollen tube tip growth are unknown. Here, we report the roles of pollen-specific GLYCEROPHOSPHODIESTER PHOSPHODIESTERASE-LIKE (GDPD-LIKE) genes in pollen tube tip growth. Arabidopsis thaliana GDPD-LIKE6 (AtGDPDL6) and AtGDPDL7 were specifically expressed in mature pollen grains and pollen tubes and GFP-AtGDPDL6 and GFP-AtGDPDL7 fusion proteins were enriched at the plasma membrane at the apex of forming pollen tubes. Atgdpdl6 Atgdpdl7 double mutants displayed severe sterility that was rescued by genetic complementation with AtGDPDL6 or AtGDPDL7. This sterility was associated with defective male gametophytic transmission. Atgdpdl6 Atgdpdl7 pollen tubes burst immediately after initiation of pollen germination in vitro and in vivo, consistent with the thin and fragile walls in their tips. Cellulose deposition was greatly reduced along the mutant pollen tube tip walls, and the localization of pollen-specific CELLULOSE SYNTHASE-LIKE D1 (CSLD1) and CSLD4 was impaired to the apex of mutant pollen tubes. A rice pollen-specific GDPD-LIKE protein also contributed to pollen tube tip growth, suggesting that members of this family have conserved functions in angiosperms. Thus, pollen-specific GDPD-LIKEs mediate pollen tube tip growth, possibly by modulating cellulose deposition in pollen tube walls. This article is protected by copyright. All rights reserved.

THE INFLUENCE OF CLIMATE CHANGES ON THE CONDITION AND REPRODUCTIVE PROCESSES OF PINE OF THE COMMON FINNISH ORIGIN IN THE CONDITIONS OF VINNYTSIA REGION

According to research results, it was established that all clones were characterized by intensive formation of microstrobils and pollination. Tree analysis showed that clones of Finnish origin entered the pollen maturation phase earlier compared to local origin. Externally, the anthers of the local population were much larger. The analysis of photographs showed that the clones were characterized by different intensity of pollen grain germination and also had a different proportion of pollen grains with abnormalities. A general review of the images revealed that most of the clones of Finnish origin and the local population are characterized by fairly high viability. It was determined that the average number of germinated pollen grains for the studied clones was 89, of which about half (44) had anomalies. According to the obtained research results, we determined that the largest number of germinated grains is 150 pcs. was noted in clone No. 13 (E 618). The minimum number was recorded in clone No. 21 (E1881) – 13 pcs. Accordingly, the same clones recorded the maximum and minimum number of ungerminated pollen grains. In particular, clone No. 10 had 218 units, and clone No. 21 had 223 units. The most pollen grains with anomalies were found in clone No. 10 – 95 pcs. It was found that the highest percentage of pollen without germination was characteristic of clone No. 21 and was about 94.2%. At the same time, the lowest share of such pollen was recorded in clone No. 13 – 40.5%. As a result of regular records of the preservation of clones on the plantation, a high level of preservation was noted, which is 94.8%. Out of the initial number of 413 clones, 22 died. The total loss of clones in the section of representative offices was 1-4 trees. The highest dropout was characteristic of clones: No. E2312 (33.3%), E2257 (20.0%) K801 (20.0%). According to the correlation analysis, a weak correlation was found between preservation and state of clones and preservation and fruiting (r = - 0.21 and 0.19, respectively). The analysis of correlation dependences indicates that the deterioration of the condition of clones is not the main reason for their death. It was established that the average intensity of the formation of cones during the last years fluctuated within the range of 1.3-2.0 points. The best level of seed bearing was noted in 2015 and 2018. Along with the local control, the formation of cones was more intense in clones E 80 and E 709 and averaged more than 2.0 points. As a result of conducting field research, it was determined that the average condition of clones has slightly deteriorated in recent years. In particular, there is an increase in the average status category from 1.3 to 1.7. The best condition, along with local control, is characterized by clones E 80 and E 2125 (condition category 1,2). Clones E 1591, E 2257, K 684, K 801, K 818 are in much worse condition.

Stages of Microspore Development in Eggplant (Solanum melongena L.)

Background: Microspores are small haploid spores that develop into the male gametophyte. Microsporocytes undergo meiotic division to form microspores. Microspores can be found in seedless and seed plants. The microspores in each flowering plant are different. This study aims to observe microspores on eggplant flowers. Method: Microspore observations were carried out on different flower bud sizes until the flower buds bloomed. Result: The results showed microspores in eggplant had different stages of development for each flower bud size. The stages of microspore development observed were Early uninucleate (Young microspore), Mid-uninucleate, Late uninucleate (Vacuolate microspore), early binucleate (Young bicellular pollen), mid-binucleate, and mature pollen. Conclusion: In eggplant microspore culture, anther length is a strong parameter to predict the stage of microspore development contained there in.

How do fertile stamens and staminodes go differently in Kingdonia uniflora? Evidence from stamen numbers, ontogeny, and structure

Kingdonia uniflora is in Circaeasteraceae and has fertile stamens and staminodes which are both variable in number; however, the differentiating process of these two types of stamens and the nectariferous structure of staminodes are still unclear. Variation in number and ontogeny (including the degeneration stage of staminodes) of stamens in K. uniflora are investigated under SEM, TEM and LM. The results indicate that there are 63 floral formulas in 80 flowers counted, and the most frequent flower floral formula (4 flowers) is P5A°11A5G6. The correlation between the stamen number and the total floral organ number is high (0.61), while the highest correlation with the total number of floral organs is the number of carpels, and there is a weak correlation between the two types of stamens (-0.23). The stamen development period is divided into five stages, with differentiation of stamens occurring at stage 2. The anther wall formation of fertile stamens is of the basic type, and the simultaneous meiosis form tetrahedral tetrads, while mature pollens are 2-celled and tricolpate. Sporogenous tissue of staminodes fails to develop further at an earlier stage, without the formation of secondary sporogenous cells. The head of mature staminodes has nectary tissue consisting of the secretory epidermis (two types of secretory cells), nectary parenchyma (about 15 layers) and vascular tissue with vessel and sieve tube elements. There are well-developed microchannels in epidermal cell walls, and the cells related to secreting nectar are rich in mitochondria, endoplasmic reticula and small vesicles. In addition, microsporogenesis, male gametophyte development, the secretory function of staminodes, and other characteristics in K. uniflora are similar to those in Ranunculaceae. This study clarifies the differentiating stage of fertile stamens and staminodes and explores the new structure (nectary) that staminodes formed, and also, it enriched the secretion modes in Ranunculales.

Coordination of floral and fiber development in cotton (Gossypium) by hormone- and flavonoid-signalling associated regulatory miRNAs.

Various plant development activities and stress responses are tightly regulated by various microRNAs (miRNA) and their target genes, or transcription factors in a spatiotemporal manner. Here, to exemplify how flowering-associated regulatory miRNAs synchronize their expression dynamics during floral and fiber development in cotton, constitutive expression diminution transgenic lines of auxin-signaling regulatory Gh-miR167 (35S-MIM167) were developed through target mimicry approach. 'Moderate' (58% to 80%)- and 'high' (> 80%)-Gh-miR167 diminution mimic lines showed dosage-dependent developmental deformities in anther development, pollen maturation, and fruit (= boll) formation. Cross pollination of 'moderate'35S-MIM167 mimiclines with wild type (WT) plant partially restored boll formation and emergence of fiber initials on the ovule surface. Gh-miR167 diminution favored organ-specific transcription biases in miR159, miR166 as well as miR160, miR164, and miR172 alongwith their target genes during anther and petal development, respectively. Similarly, accumulative effect of percent Gh-miR167 diminution, cross regulation of its target ARF6/8 genes, and temporal mis-expression of hormone signaling- and flavonoid biosynthesis-associated regulatory miRNAs at early fiber initiation stage caused irregular fiber formation. Spatial and temporal transcription proportions of regulatory miRNAs were also found crucial for the execution of hormone- and flavonoid-dependent progression of floral and fiber development. These observations discover how assorted regulatory genetic circuits get organized in response to Gh-miR167 diminution and converge upon ensuing episodes of floral and fiber development in cotton.

ANTHER STRUCTURE IN <i>CRASSULA ERICOIDES</i>, <i>C. INTERMEDIA</i> AND <i>C. MULTICAVA</i> (CRASSULACEAE)

The development and structure of the anther wall in Crassula intermedia, C. multicava and C. ericoides was studied for the first time. In the studied species, the presence of basic features of the Crassulaceae family associated with reproduction processes was confirmed. These include: tetrasporangiate anther, parietal tapetum, simultaneous microsporogenesis, predominantly tetrahedral tetrads of microspores, 2-celled and 3-colporate mature pollen grains. The microsporangium wall is 4-layered (epidermis, endothecium, middle layer and tapetum) and its development occurs within a typical variation of the centrifugal type. Tannins accumulate in epidermal cells: in C. intermedia, along the periphery of the cell protoplast, and in C. multicava and C. ericoides, the cell protoplasts are completely filled with tannins. The tapetum develops without reorganization (typical parietal tapetum). The study has shown that Crassula species are characterized by a number of unique features. They form an eccentric connective displaced from the central position towards the abaxial microsporangia, while the connective tissue does not develop between the adaxial microsporangia. Fibrous thickenings were found not only in the endothecium cells, but also in the cells around the vascular bundle of the connective tissue. The number of cell layers with fibrous thickenings and their location differ in the studied species. In C. ericoides, they are arranged in one layer in the form of a ring in the anther part completely fused with the filament. In C. intermedia and C. multicava, 2–3 layers are formed there. As regards the anther part free from the filament, in C. ericoides cells with fibrous thickenings are formed only in the endothecium, while in other two species not only in the endothecium, but also in 2–3 underlying layers of the connective tissue. In the structure of the stamen and anther of Crassula species, there are features common with Sedum and Aeonium species (the presence of cells with fibrous thickenings in the anther part free from the filament) and Kalanchoe (the stamen is wide at the base and narrow at the junction with the anther). Our data indicate that additional studies are needed, and the position of the Crassula clade continues to be controversial on the phylogenetic tree of the Crassulaceae family.

RHO OF PLANT proteins are essential for pollen germination in Arabidopsis.

Pollen germination is a process of polarity establishment, through which a single and unique growth axis is established. Although most of the intracellular activities associated with pollen germination are controlled by RHO OF PLANTs (ROPs) and increased ROP activation accompanies pollen germination, a critical role of ROPs in this process has not yet been demonstrated. Here, by genomic editing of all four Arabidopsis (Arabidopsis thaliana) ROPs that are preferentially expressed in pollen, we showed that ROPs are essential for polarity establishment during pollen germination. We further identified and characterized two ROP effectors in pollen germination (REGs) through genome-wide interactor screening, Boundary of ROP domain (BDR) members BDR8 and BDR9, whose functional loss also resulted in no pollen germination. BDR8 and BDR9 were distributed in the cytosol and the vegetative nucleus of mature pollen grains but re-distributed to the plasma membrane (PM) of the germination site and to the apical PM of growing pollen tubes. We demonstrated that the PM re-distribution of BDR8 and BDR9 during pollen germination relies on ROPs but not vice versa. Furthermore, enhanced expression of BDR8 partially restored germination of rop1 pollen but had no effects on that of the quadruple rop pollen, supporting their genetic epistasis. Results presented here demonstrate an ROP signaling route essential for pollen germination, which supports evolutionarily conserved roles of Rho GTPases in polarity establishment.

The cotton pectin methyl esterase gene GhPME21 functions in microspore development and fertility in Gossypium hirsutum L.

Pectin widely exists in higher plants' cell walls and intercellular space of higher plants and plays an indispensable role in plant growth and development. We identified 55 differentially expressed genes related to pectin degradation by transcriptomic analysis in the male sterile mutant, ms1. A gene encoding pectin methylesterase (GhPME21) was found to be predominantly expressed in the developing stamens of cotton but was significantly down-regulated in ms1 stamens. The tapetal layer of GhPME21 interfered lines (GhPME21i) was significantly thickened compared to that of WT at the early stage; anther compartment morphology of GhPME21i lines was abnormal, and the microspore wall was broken at the middle stage; Alexander staining showed that the pollen grains of GhPME21i lines differed greatly in volume at the late stage. The mature pollen surfaces of GhPME21i lines were deposited with discontinuous and broken sheets and prickles viewed under SEM. Fewer pollen tubes were observed to germinate in vitro in GhPME21i lines, while tiny of those in vivo were found to elongate to the ovary. The seeds harvested from GhPME21i lines as pollination donors were dry and hollow. The changes of phenotypes in GhPME21i lines at various stages illustrated that the GhPME21 gene played a vital role in the development of cotton stamens and controlled plant fertility by affecting stamen development, pollen germination, and pollen tube elongation. The findings of this study laid the groundwork for further research into the molecular mechanisms of PMEs involved in microspore formation and the creation of cotton male sterility materials.

The triticale mature pollen and stigma proteomes – assembling the proteins for a productive encounter

Triticeae crops are major contributors to global food production and ensuring their capacity to reproduce and generate seeds is critical. However, despite their importance our knowledge of the proteins underlying Triticeae reproduction is severely lacking and this is not only true of pollen and stigma development, but also of their pivotal interaction. When the pollen grain and stigma are brought together they have each accumulated the proteins required for their intended meeting and accordingly studying their mature proteomes is bound to reveal proteins involved in their diverse and complex interactions. Using triticale as a Triticeae representative, gel-free shotgun proteomics was used to identify 11,533 and 2977 mature stigma and pollen proteins respectively. These datasets, by far the largest to date, provide unprecedented insights into the proteins participating in Triticeae pollen and stigma development and interactions. The study of the Triticeae stigma has been particularly neglected. To begin filling this knowledge gap, a developmental iTRAQ analysis was performed revealing 647 proteins displaying differential abundance as the stigma matures in preparation for pollination. An in-depth comparison to an equivalent Brassicaceae analysis divulged both conservation and diversification in the makeup and function of proteins involved in the pollen and stigma encounter. SignificanceSuccessful pollination brings together the mature pollen and stigma thus initiating an intricate series of molecular processes vital to crop reproduction. In the Triticeae crops (e.g. wheat, barley, rye, triticale) there persists a vast deficit in our knowledge of the proteins involved which needs to be addressed if we are to face the many upcoming challenges to crop production such as those associated with climate change. At maturity, both the pollen and stigma have acquired the protein complement necessary for their forthcoming encounter and investigating their proteomes will inevitably provide unprecedented insights into the proteins enabling their interactions. By combining the analysis of the most comprehensive Triticeae pollen and stigma global proteome datasets to date with developmental iTRAQ investigations, proteins implicated in the different phases of pollen-stigma interaction enabling pollen adhesion, recognition, hydration, germination and tube growth, as well as those underlying stigma development were revealed. Extensive comparisons between equivalent Triticeae and Brassiceae datasets highlighted both the conservation of biological processes in line with the shared goal of activating the pollen grain and promoting pollen tube invasion of the pistil to effect fertilization, as well as the significant distinctions in their proteomes consistent with the considerable differences in their biochemistry, physiology and morphology.

OsCCRL1 is Essential for Phenylpropanoid Metabolism in Rice Anthers

Phenylpropanoid metabolism and timely tapetal degradation are essential for anther and pollen development, but the underlying mechanisms are unclear. In the current study, to investigate this, we identified and analyzed the male-sterile mutant, osccrl1 (cinnamoyl coA reductase-like 1), which exhibited delayed tapetal programmed cell death (PCD) and defective mature pollen. Map-based cloning, genetic complementation, and gene knockout revealed that OsCCRL1 corresponds to the gene LOC_Os09g32020.2, a member of SDR (short-chain dehydrogenase/reductase) family enzyme. OsCCRL1 was preferentially expressed in the tapetal cells and microspores, and localized to the nucleus and cytoplasm in both rice protoplasts and Nicotiana benthamiana leaves. The osccrl1 mutant exhibited reduced CCRs enzyme activity, less lignin accumulation, delayed tapetum degradation, and disrupted phenylpropanoid metabolism. Furthermore, an R2R3 MYB transcription factor OsMYB103/OsMYB80/OsMS188/BM1, involved in tapetum and pollen development, regulates the expression of OsCCRL1. Finally, the osmyb103 osccrl1 double mutants, exhibited the same phenotype as the osmyb103 single mutant, further indicating that OsMYB103/OsMYB80/OsMS188/BM1 functions upstream of OsCCRL1. These findings help to clarify the role of phenylpropanoid metabolism in male sterility and the regulatory network underlying the tapetum degradation.

The hydroxyproline O-arabinosyltransferase FIN4 is required for tomato pollen intine development.

The pollen grain cell wall is a highly specialized structure composed of distinct layers formed through complex developmental pathways. The production of the innermost intine layer, composed of cellulose, pectin and other polymers, is particularly poorly understood. Here we demonstrate an important and specific role for the hydroxyproline O-arabinosyltransferase (HPAT) FIN4 in tomato intine development. HPATs are plant-specific enzymes which initiate glycosylation of certain cell wall structural proteins and signaling peptides. FIN4 was expressed throughout pollen development in both the developing pollen and surrounding tapetal cells. A fin4 mutant with a partial deletion of the catalytic domain displayed significantly reduced male fertility in vivo and compromised pollen hydration and germination in vitro. However, fin4 pollen that successfully germinated formed morphologically normal pollen tubes with the same growth rate as the wild-type pollen. When we examined mature fin4 pollen, we found they were cytologically normal, and formed morphologically normal exine, but produced significantly thinner intine. During intine deposition at the late stages of pollen development we found fin4 pollen had altered polymer deposition, including reduced cellulose and increased detection of pectin, specifically homogalacturonan with both low and high degrees of methylesterification. Therefore, FIN4 plays an important role in intine formation and, in turn pollen hydration and germination and the process of intine formation involves dynamic changes in the developing pollen cell wall.

Changes in the content of pollen total lipid and TAG in Arabidopsis thaliana DGAT1 mutant as11.

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.

Mapping QTL for Phenological and Grain-Related Traits in a Mapping Population Derived from High-Zinc-Biofortified Wheat.

Genomic regions governing days to heading (DH), days to maturity (DM), plant height (PH), thousand-kernel weight (TKW), and test weight (TW) were investigated in a set of 190 RILs derived from a cross between a widely cultivated wheat-variety, Kachu (DPW-621-50), and a high-zinc variety, Zinc-Shakti. The RIL population was genotyped using 909 DArTseq markers and phenotyped in three environments. The constructed genetic map had a total genetic length of 4665 cM, with an average marker density of 5.13 cM. A total of thirty-seven novel quantitative trait loci (QTL), including twelve for PH, six for DH, five for DM, eight for TKW and six for TW were identified. A set of 20 stable QTLs associated with the expression of DH, DM, PH, TKW, and TW were identified in two or more environments. Three novel pleiotropic genomic-regions harboring co-localized QTLs governing two or more traits were also identified. In silico analysis revealed that the DArTseq markers were located on important putative candidate genes such as MLO-like protein, Phytochrome, Zinc finger and RING-type, Cytochrome P450 and pentatricopeptide repeat, involved in the regulation of pollen maturity, the photoperiodic modulation of flowering-time, abiotic-stress tolerance, grain-filling duration, thousand-kernel weight, seed morphology, and plant growth and development. The identified novel QTLs, particularly stable and co-localized QTLs, will be validated to estimate their effects in different genetic backgrounds for subsequent use in marker-assisted selection (MAS).

Duplicate mutations of GhCYP450 lead to the production of ms5m6 male sterile line in cotton.

The duplicated male sterile genes ms5m6 in cotton were map-based cloned and validated by the virus-induced gene silencing assays. Duplicate mutations of the GhCYP450 gene encoding a cytochrome P450 protein are responsible for the male sterility in cotton. The utilization of male sterility in cotton plays a vital role in improving yield and fiber quality. A complete male sterile line (ms5ms6) has been extensively used to develop hybrid cotton worldwide. Using Zhongkang-A (ZK-A) developed by transferring Bt and ms5ms6 genes into the commercial cultivar Zhongmiansuo 12, the duplicate genes were map-based cloned and confirmed via the virus-induced gene silencing (VIGS) assays. The duplicate mutations of GhCYP450 genes encoding a cytochrome P450 protein were responsible for producing male sterility in ms5ms6 in cotton. Sequence alignment showed that GhCYP450-Dt in ZK-A differed in two critical aspects from the fertile wild-type TM-1: GhCYP450-Dt has three amino acid (D98E, E168K, G198R) changes in the coding region and a 7-bp (GGAAAAA) insertion in the promoter domain; GhCYP450-At appears to be premature termination of GhCYP450 translation. Further morphological observation and cytological examination of GhCYP450-silenced plants induced by VIGS exhibited shorter filaments and no mature pollen grains. These results indicate that GhCYP450 is essential for pollen exine formation and pollen development for male fertility. Investigating the mechanisms of ms5ms6 male sterility will deepen our understanding of the development and utilization of heterosis.

Control of plastid inheritance by environmental and genetic factors

The genomes of cytoplasmic organelles (mitochondria and plastids) are maternally inherited in most eukaryotes, thus excluding organellar genomes from the benefits of sexual reproduction and recombination. The mechanisms underlying maternal inheritance are largely unknown. Here we demonstrate that two independently acting mechanisms ensure maternal inheritance of the plastid (chloroplast) genome. Conducting large-scale genetic screens for paternal plastid transmission, we discovered that mild chilling stress during male gametogenesis leads to increased entry of paternal plastids into sperm cells and strongly increased paternal plastid transmission. We further show that the inheritance of paternal plastid genomes is controlled by the activity of a genome-degrading exonuclease during pollen maturation. Our data reveal that (1) maternal inheritance breaks down under specific environmental conditions, (2) an organelle exclusion mechanism and a genome degradation mechanism act in concert to prevent paternal transmission of plastid genes and (3) plastid inheritance is determined by complex gene–environment interactions.

The role of phytohormones and their related miRNAs in sex differentiation of Xanthoceras sorbifolium Bunge

• Determined the influence of IAA, GA, ABA, ZT on sex differentiation of Xanthoceras sorbifolium . • Identified 20 significantly differently expressed miRNAs (83 miRNA/target pairs) being related to phytohormone synthesis and signal transduction during sex differentiation. Xanthoceras sorbifolium Bunge is an important woody oilseed tree in north China. However, its yield is severely restricted by a low ratio of female to male flowers, and little is known about the physiological and molecular mechanisms underlying the sex differentiation of flowers. Therefore, carrying out the research in its sex differentiation regulation mechanism is the basis of increasing seed yield. Here, we investigated the cytology of sex differentiation, the content and their ratios of indole-3-acetic acid (IAA), gibberellins (GA 3 ), zeatin (ZT), abscisic acid (ABA), the effects of plant growth regulators on flower development, and the expression profiles of phytohormone-related miRNAs. The results showed that both functional female and male flowers were bisexual at the early stage of flower bud differentiation, while the development of the pistils of male flowers and the stamens of female flowers was arrested at later stage of sex differentiation. Furthermore, IAA, ABA and ZT were shown to be beneficial to pistil development in X. sorbifolium . GA 3 was the key phytohormone in pistil abortion. High level of ABA/ZT was beneficial for microspore mother cell meiosis. ZT/IAA was probably related to pistil abortion. High level of ZT/GA 3 contributed to ovule development. Spraying 6-BA was more effective than GA 3 in increasing female-to-male ratio regardless of CK. In addition, several miRNAs played key roles in sex differentiation, especially miR393, miR396, and miR399. They participated in the regulation of pollen maturation and multiple phytohormone pathways. The present study comprehensively identified the roles of phytohormones and their related miRNAs during sex differentiation in X. sorbifolium , which will be a valuable resource for sex regulation using genetic and physiological methods in the future.

A pollen expressed PME gene at Tcb1 locus confers maize unilateral cross-incompatibility.

A pollen expressed PME gene at Tcb1 locus confers maize unilateral cross-incompatibility.

Cell-type-specific alternative splicing in the Arabidopsis germline.

During sexual reproduction in flowering plants, the two haploid sperm cells (SCs) embedded within the cytoplasm of a growing pollen tube are carried to the embryo sac for double fertilization. Pollen development in flowering plants is a dynamic process that encompasses changes at transcriptome and epigenome levels. While the transcriptome of pollen and SCs in Arabidopsis (Arabidopsis thaliana) is well documented, previous analyses have mostly been based on gene-level expression. In-depth transcriptome analysis, particularly the extent of alternative splicing (AS) at the resolution of SC and vegetative nucleus (VN), is still lacking. Therefore, we performed RNA-seq analysis to generate a spliceome map of Arabidopsis SCs and VN isolated from mature pollen grains. Based on our de novo transcriptome assembly, we identified 58,039 transcripts, including 9,681 novel transcripts, of which 2,091 were expressed in SCs and 3,600 in VN. Four hundred and sixty-eight genes were regulated both at gene and splicing levels, with many having functions in mRNA splicing, chromatin modification, and protein localization. Moreover, a comparison with egg cell RNA-seq data uncovered sex-specific regulation of transcription and splicing factors. Our study provides insights into a gamete-specific AS landscape at unprecedented resolution.