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At least 19 recordsLinked to original sources

PubMed2026

Virome metatranscriptomic profiling of birch pollen reveals a diverse viral community.

INTRODUCTION: Viruses are increasingly recognized as integral components of plant-associated biological systems. However, their occurrence and diversity in the reproductive tissues of woody plants remain poorly understood. Birch (Betula spp.) produces large quantities of wind-dispersed pollen that can travel over long distances and may harbour viruses or virus-derived nucleic acids originating from the host plant and/or its associated microbiota. METHODS: We investigated the virome of birch pollen collected from trees growing in central and suburban Berlin, Germany. Metatranscriptomic analyses were performed on pooled pollen samples collected in 2020. These analyses were complemented by RT-PCR screening of individually processed pollen samples collected in 2025 from resampled trees. Primer walking was additionally used to recover an extended genome sequence of a pollen-associated birch toti-like virus. RESULTS: Multiple virus-associated contigs were identified in both pooled metatranscriptomic datasets. These included sequences corresponding to the cherry leaf roll virus (CLRV), the birch idaeovirus (BIV) and the birch toti-like virus (BTLV), as well as additional virus-like contigs provisionally assigned to lineages related to the Orthototiviridae, Botourmiaviridae, Endornaviridae, and Chrysoviridae families. RT-PCR analysis of individually processed pollen samples confirmed the continued detection of CLRV, BIV, and BTLV within the Berlin sampling framework. A near-complete genome sequence was recovered from a pollen-derived BTLV isolate from Berlin, showing high amino acid sequence identity to a recently described leaf-derived BTLV isolate from the United States. DISCUSSION: These findings demonstrate that birch pollen harbours a diverse assemblage of plant- and/or microbiome-associated viruses and expand the known tissue distribution and geographic range of BTLV. More broadly, they establish pollen as an underexplored ecological niche for virome research and provide a foundation for future studies on the ecology, transmission dynamics, and epidemiological significance of pollen-associated and pollen-transmitted viruses.

Betula↗

PubMed2026

Spatial inheritance patterns across maize ears are associated with alleles that reduce pollen fitness.

Often, more pollen grains land on recipient flowers than there are ovules to fertilize. Consequently, the haploid male gametophyte engages in post-pollination competition, one way that pollen genotype can influence inheritance. The maize (Zea mays subsp. mays L.) inflorescence (ear), with its elongated stigma and style structures (silks), has a conspicuous spatial heterogeneity, with longer silks at the base of the ear than at the apex. To evaluate the hypothesis that alleles with reduced pollen fitness influence the spatial distribution of progeny genotypes along the ear, we developed an updated phenotyping platform that maps fluorescently marked mutant (Ds-GFP) kernel phenotypes on the ear via an implementation of the Faster R-CNN machine vision model (EarVision.v2) and a statistical pipeline that evaluates the relationship between kernel position and transmission ratio (EarScape). Our dataset (1384 ears) represents 58 Ds-GFP insertion alleles. None of the 48 alleles with Mendelian inheritance showed any significant spatial trend. In contrast, 50% of alleles with a pollen-specific transmission defect (5/10) exhibited significant spatial effects. An insertional mutant of the gene encoding a putative actin-binding protein, base-to-apex gradient1* (bag1*), is associated with decreased mutant transmission at the ear base relative to the apex. Surprisingly, a mutant allele of another pollen-expressed gene (Zm00001eb236740) generates the opposite trend, decreased mutant transmission toward the ear apex; and two mutant alleles of the sperm cell attachment factor gamete expressed2 (gex2) can produce ears with transmission highest at both base and apex. We conclude that pollen fitness mutants cause unexpectedly diverse spatial patterns of progeny genotypes.

Zea mays↗

PubMed2026

Cytological Assessment of Maize Pollen Viability Using a Simplified Staining Protocol.

Anomalies of meiosis frequently result in abnormal chromosome segregation, which leads to defects in pollen formation in maize. Thus, assessing pollen viability is an important measure for examining the overall success of male sexual reproduction. Pollen viability tests are used to characterize mutants defective in meiosis and microsporogenesis, and to determine the effects of genome instability and environmental conditions on reproduction. This protocol describes a rapid method for assessing pollen viability in maize, using a simplified cytological staining approach. Traditional pollen staining methods, such as Alexander staining, often require hazardous chemicals that are increasingly restricted due to safety concerns. The method presented here uses easily accessible reagents and avoids highly toxic substances.

Journal Article↗

PubMed2026

PAT: An Image Analysis Tool for Automated Scoring of Pollen in Alexander-Stained Anthers.

Quantitative pollen viability analysis is a critical but labor-intensive step in plant reproductive biology. Existing deep-learning Segment Anything Models (SAM) fail to reliably segment viable pollen in Alexander-stained anthers. To address this, we fine-tuned an existing Cellpose-SAM model for pollen segmentation. We integrated it into PAT (Pollen Analysis Tool), a cross-platform desktop application. PAT features instance segmentation with interactive quality control, an in-app model retraining module, and publication-ready statistical outputs. We deployed PAT in an EMS suppressor screen of semi-sterile Arabidopsis smg7-6 mutants, enabling efficient candidate prioritization for whole-genome sequencing and mapping of the candidate mutation. This screen led to the identification of a point mutation in CAP-D2 (capd2-2), a Condensin I subunit, that rescues the smg7-6 meiotic phenotype. Notably, mutation in a Condensin II subunits (CAP-D3 and CAP-H2) does not confer rescue. Further characterization suggests the capd2-2 allele is hypomorphic, showing no defects in vegetative growth, chromocenter compaction, or transposable element silencing. Collectively, we demonstrate that accessible AI tools have the potential to bridge gaps in plant phenotyping and accelerate the pace of biological discovery.

Alexander staining↗

PubMed2026

Pan-Genomic Dissection of GH1 β-Glucosidases in Brassica rapa Identifies BrBGLU10 as an Important Regulator of Pollen Development.

Glycoside hydrolase family 1 (GH1) β-glucosidases (BGLUs) play diverse roles in plant development and stress responses. However, a comprehensive pan-genomic characterization of this gene family across diverse Brassica rapa accessions is still lacking. Here, we conducted a pan-genome-wide analysis of BGLU genes across 21 B. rapa accessions. A total of 1840 BGLU genes were identified and clustered into 57 orthologous gene groups (OGGs), comprising 22 core, 19 dispensable, and 16 private groups. Phylogenetic reconstruction assigned these OGGs to five subgroups, and duplication analysis revealed whole-genome duplication as the predominant driver of family expansion, accounting for 47.51% of duplicated genes. Expression profiling identified two core genes, BrBGLU10 and BrBGLU56, as specifically expressed in fertile floral buds and differentially regulated between fertile and sterile lines. CRISPR/Cas9-mediated knockout of BrBGLU10 resulted in approximately 36% pollen abortion and drastically reduced seed set upon self-pollination, supporting its important role in pollen development. Collectively, these findings establish BrBGLU10 as an important regulator of pollen development and a potential target for fertility-related applications via gene editing in B. rapa and related Brassica crops.

BrBGLU10↗

PubMed2026

Circadian- and light-regulated oscillatory expression of CSA in rice leaves is required for pollen fertility.

The oscillatory expression of CSA in rice leaves is regulated by the circadian clock and red/far-red light signals, mediated through DOF5 and PIL11, and is required for normal pollen fertility. Photoperiod-sensitive male-sterile lines represent a pivotal innovation in the development of hybrid rice. However, the underlying mechanisms governing photoperiod-sensitive male reproductive development remain poorly understood. Our previous studies demonstrated that the carbon starved anther (csa) mutant exhibits male sterility under short-day (SD) conditions but partial fertility under long-day (LD) conditions. In this study, we report that CSA expression follows an oscillatory rhythm in rice leaves under both SD and LD conditions, a pattern regulated by both circadian clock and light signals. Tissue-specific RNA interference knockdown of CSA in leaves was associated with reduced pollen viability, suggesting that CSA expression in leaves contributes to normal male fertility. Promoter truncation assay results indicate that distinct regions of the CSA promoter contribute differentially to the regulation of CSA expression in leaves versus anthers, and that both the CSA expression level in anthers and the rhythmic expression pattern of CSA in leaves are associated with the restoration of male fertility. Using dual-luciferase, yeast one-hybrid, and electrophoretic mobility shift assays, we identified two proteins, PIL11 and DOF5, which directly bind to specific motifs (an E-box and T/AAAAG motif) within the CSA promoter truncation, thereby regulating its transcription. These findings elucidate novel mechanisms linking light sensing to the expression of circadian-controlled genes, thus connecting photoperiod with male reproductive development in rice.

Oryza↗

PubMed2026

The DELAYED ABAXIAL TRICHOMES Helitron has dual functions in vegetative and pollen development in Arabidopsis thaliana.

Transposons drive genetic diversity and evolution by altering the genomic landscape over time. Here, we describe DELAYED ABAXIAL TRICHOMES (DAB), a Helitron/RC transposable element in Arabidopsis thaliana that has a role in vegetative phase change and gametogenesis. A genome-wide association study (GWAS) for the timing of abaxial trichome development (an adult leaf trait) in A. thaliana revealed a conserved haplotype of polymorphisms within DAB that delays abaxial trichome production. CRISPR-Cas9-induced deletions of DAB are gametophytic pollen-lethal, indicating that this locus is also required for pollen production. DAB produces 24-nucleotide siRNAs with sequence complementarity to genes involved in embryogenesis, gametogenesis, and seed development. DAB also impacts the expression of ARGONAUTE genes, genes involved in RNA-directed DNA methylation (RdDM), as well as genes in several key genetic pathways. This global effect on gene expression suggests that DAB may have functions beyond those identified in this study.

Arabidopsis↗

PubMed2025

Genetic interactions and natural variation underlying S-RNase-independent unilateral incompatibility in Solanum.

Pistils of self-incompatible (SI) species/populations typically reject pollen of related self-compatible (SC) species/populations, but not vice versa, a pattern known as unilateral incompatibility (UI). UI is complex and includes both S-RNase-dependent and S-RNase-independent mechanisms. Pistils of Solanum pennellii LA0716 (SC, no S-RNase) reject pollen of cultivated tomato, Solanum lycopersicum (SC); UI in this system involves the expression of ornithine decarboxylase2 (ODC2) and HT-A/-B genes in the pistil, and farnesyl pyrophosphate synthase2 (FPS2), ui6.2, and ui12.2 in pollen. We show that IL12-3 (HT-A/-B) × IL3-3 (ODC2) double introgression lines reject S. lycopersicum pollen, while odc2 or ht-a mutants do not, demonstrating that ODC2 and HT-A are required for UI. Transmission ratio distortion in favor of pennellii alleles was observed in interspecific F2 S. lycopersicum × S. pennellii near ui6.2 and ui12.2, and in F2 IL12-3 × IL3-3 near ui12.2. Equivalent populations made with odc2 mutants segregate in Mendelian ratios, while ht-a mutants have little effect, indicating ui6.2 and ui12.2 interact primarily with ODC2. Pollen from fps2 mutants in S. pennellii LA0716 are incompatible on pistils of all tested S. pennellii and some Solanum habrochaites accessions, but compatible with all other tomato clade species, suggesting ODC2-dependent UI evolved in a common ancestor to S. pennellii and S. habrochaites. Within S. habrochaites, fps2 pollen rejection was observed mainly in SI or mixed mating populations, suggesting an association with outcrossing. Triple mutants of S. pennellii and S. habrochaites lacking functional ODC2, HT-A/-B, and S-RNase are cross-compatible as female parents with S. lycopersicum, allowing transfer of their cytoplasmic genomes into cultivated tomato.

Solanum↗

PubMed2026

Magnetic nanoparticle-mediated genetic transformation and gene editing system in loquat (Eriobotrya japonica).

Loquat (Eriobotrya japonica Lindl.) is a valuable subtropical fruit tree whose genetic improvement has been significantly constrained by the absence of an efficient genetic transformation system. Although Agrobacterium-mediated transformation is the most widely used method, it proves ineffective in loquat due to the species' recalcitrance to in vitro regeneration. Pollen-based transformation offers a promising alternative by bypassing the need for tissue culture. However, the pollen wall poses a major physical barrier to the uptake of exogenous DNA. In this study, we investigated magnetic nanoparticle (MNP)-mediated transformation as a novel strategy for loquat. We confirmed that loquat pollen contains tricolporate apertures with diameters ranging from 3.0 to 5.0 μm, which are structurally suitable for the entry of MNPs-DNA. Based on this finding, we developed and optimized a transformation protocol using polyethyleneimine-coated Fe3O4 nanoparticles to deliver genetic material into loquat pollen grains. Using this approach, we successfully generated stable transgenic loquat lines, including both overexpression and gene-edited mutants. To our knowledge, this is the first report of successful MNP-mediated pollen transformation in a woody plant species. This work establishes a robust and efficient genetic transformation platform for loquat, providing a valuable tool for functional genomics and molecular breeding, as well as a potentially applicable strategy for other recalcitrant woody plants.

Eriobotrya↗

PubMed2026

Maize Gametophytic factor loci Ga3 through Ga11 modify reproductive barriers.

Gametophytic factor (Ga) barriers are maize (Zea mays ssp. mays) reproductive barriers controlled by molecular incompatibilities between pollen and silks. Twelve distinct Ga loci have been identified in maize populations since the first genetic evidence of a Ga barrier was reported in 1901. Of the twelve, however, only three have been validated by modern molecular, functional and genomic studies: Ga1, Ga2, and Tcb1. The remaining "higher" Ga loci, spanning Ga3 to Ga11, were reported in the historical literature, but their associated phenotypes segregated in unexpected ways or disappeared over subsequent generations. Here we introduce and explore the hypothesis that the higher Ga loci represent modifiers of Ga1, Ga2, and Tcb1 barrier functions. By revisiting the historical literature, we found that barrier phenotypes fall into two phenotypic and functional categories. Phenotypically, the two categories represented healthy pollen with a silk-length effect and unhealthy pollen without a silk-length effect. These phenotypic categories were supported by genomic data; we identified candidate genes in each higher Ga locus by comparing historical linkage mapping data to the corresponding genomic sequence of maize reference line B73. We discovered candidate genes related to two broad pathways: pollen tube growth and RNA-directed DNA methylation. We conclude that the past century of evidence aligns with our hypothesis that maize loci Ga3 through Ga11 modify rather than directly control Ga barriers. This brief investigation provides a starting point for geneticists and evolutionary biologists to explore how strong reproductive barriers are shaped by epistatic interactions.

Epistasis↗

PubMed2026

Toward simple, rapid, and deep plant proteome analysis with an in-cell proteomics strategy.

While liquid chromatography-mass spectrometry (LCMS) has revolutionized plant proteomics over the past decade, plant sample preparation remains a major challenge due to rigid cell walls, abundant secondary metabolites, and wide dynamic range of protein abundance. These hurdles demand laborious tissue disruption, complex precipitation, and extensive cleanup prior to LCMS analysis, limiting the widespread adoption of proteomic technologies within the plant biology community. To overcome these barriers, we introduced an "in-cell proteomics" strategy that bypasses cell lysis and protein extraction by performing digestion directly inside methanol-fixed cells. We systematically benchmarked this strategy against conventional lysate-based workflows across 4 model plants (Arabidopsis thaliana, Nicotiana benthamiana, Zea mays, and Sorghum bicolor) and 3 tissue types (leaves, pollen, and seeds). Combined with minimal input material and single-shot LCMS, the in-cell approach consistently identified 9,000 to 12,000 proteins from leaves, 7,000 to 9,000 from pollen grains, and approximately 8,000 from seeds. Our comprehensive dataset demonstrates that this in-cell digestion approach substantially simplifies plant sample preparation while delivering proteomic performance equivalent to established workflows. Finally, to demonstrate the biological utility of this approach, we characterized the proteomes of N. benthamiana leaves infected with 2 fungal strains that exhibit different host specificities. Our in-depth proteomic data revealed distinct host response signatures differentiating the host-adapted Colletotrichum destructivum from the nonhost-adapted Colletotrichum sublineola strain. Overall, this study provides a simple, unbiased alternative for plant proteomic analysis that can be readily applied to tackle complex agricultural and physiological challenges in plant biology.

Proteomics↗

PubMed2025

Alternative splice acceptor site in MSH4 gene is responsible for male sterility conferred by ms5 in soybean.

In soybean breeding, using the recessive male-sterile ms5 gene, derived from fast neutron mutagenesis, for recurrent selection is advantageous because of the d2 locus, which controls cotyledon color in mature seeds and can be used as a phenotypic selection marker for ms5 male sterility. However, occasional self-fertilization occurs because of the elimination of d2 linkage and instability of male sterility. Elucidating the mechanism and the gene responsible for ms5 male sterility may resolve these problems. Using fine mapping with 15 simple sequence repeat (SSR) markers, we narrowed down the candidate ms5 locus to a 54-kbp region. Bulked-DNA analysis using next-generation sequencing revealed a deletion as a candidate variation in the region. This 15-bp deletion and a nucleotide substitution were identified in intron 1 of MutS homolog (GmMSH4), which modulates chromosomal recombination in meiosis. The ms5 transcript contained a novel exon with a premature termination codon. This exon originated from an alternative splice acceptor site caused by the deletion and nucleotide substitution, disrupting gene function. Co-segregation of male sterility with five independent mutations in GmMSH4 was confirmed using progeny of mutant lines. Mutations in GmMSH4 led to biased DNA partitioning during meiosis, resulting in collapsed or enlarged pollen and suggesting that ms5 male sterility is caused by the failure of pollen formation during meiosis due to the loss of function of GmMSH4. These findings could help explain the mechanism of instability of ms5 male sterility and improve the efficiency of recurrent selection using DNA markers in soybean breeding.

Glycine max↗

PubMed2025

Genetic basis for broad interspecific compatibility in Solanum verrucosum.

Solanum verrucosum Schlechtendal (2x = 2n = 24) is unique among the clade 4 Solanum Sect Petota species. In addition to being one of the only fully self-compatible diploid potato species, S. verrucosum is the only clade 4 species that lacks prezygotic interspecific reproductive barriers. This allows S. verrucosum to accept pollen from a broad range of Solanum species and thereby serving as a genetic "bridge" between the cultivated or primary potato gene pool and distantly related wild relatives in the tertiary gene pool. The genetic mechanisms underlying self-compatibility in Solanum often underpin interspecific compatibility interactions, which in S. verrucosum, has been attributed to the lack of S-RNase expression. Using an interspecific F2 mapping population (n = 150), we investigated the genetic mechanisms responsible for the lack of interspecific reproductive barriers in S. verrucosum. This F2 population was evaluated for the ability to accept pollen from two clade 1, 1 EBN species (S. pinnatisectum and S. tarnii); from which two QTL for interspecific compatibility were identified on chromosomes 1 and 11, explaining 56.6% of the phenotypic variation observed. To identify the genetic basis of interspecific compatibility, we generated a chromosome-scale genome assembly of S. verrucosum MSII1813-2 and performed gene expression profiling of reproductive organs. Differential gene expression of S-RNase, located within the chromosome 1 QTL, confirmed the central role of the S-locus and specifically, S-RNase, in interspecific compatibility. Discovery of a non-S-locus QTL is consistent with previous findings that other non-S-locus factors are necessary for interspecific compatibility in S. verrucosum.

Solanum↗

PubMed2025

Translation Co-factor PABP-interacting protein 11 moonlights as a transcriptional activator to modulate callose synthesis gene expression.

The development of rice fertility is a complex process, which is precisely regulated by numerous genes. In this study, we cloned and characterized OsPAIP11, a PABP-interacting protein that functions as an auxiliary factor in translation initiation. The ospaip11 exhibited multiple defects, including impaired callose synthesis, delayed tapetum apoptosis, and abnormal pollen wall development, which are essentially consistent with the phenotype of the allelic mutant dcet1. Subcellular localization analysis revealed that OsPAIP11 is localized in both the cytoplasm and nucleus. Interestingly, further investigation demonstrated that the RRM2 domain of OsPAIP11 exhibits transcriptional activation activity. Moreover, OsPAIP11 directly binds to the promoter of the callose synthesis-related genes GLUCAN SYNTHASE-LIKE 5 (OsGSL5) and OsGAMYB, thereby regulating their transcription and influencing callose biosynthesis during pollen development. Additionally, OsPAIP11 also interacts with the translation initiation factor and auxiliary factors. These findings suggest that OsPAIP11 modulates male fertility primarily by regulating the transcription of callose synthesis-related genes and may also participate in the translation process.

Glucans↗

PubMed2026

Heat stress impact on rice reproductive processes: challenges and new approaches.

Heat stress represents one of the most severe abiotic constraints to rice (Oryza sativa L.) productivity and is expected to intensify under ongoing climate change, particularly affecting the reproductive phase and leading to substantial yield and grain quality losses. This review synthesizes current knowledge on the impacts of heat stress on rice reproduction, with a focus on both male and female reproductive structures and their interactions. Evidence from anatomical, physiological, transcriptomic, and metabolomic studies to describe how elevated temperatures disrupt key reproductive processes, including microsporogenesis, anther dehiscence, pollen viability, pollen-pistil interactions, fertilisation, and embryo sac development were integrated in this review. It further discusses the genotype-dependent differences in reproductive thermotolerance; and key genes, metabolites, and pathways associated with heat stress perception, signalling, and tolerance are highlighted. Finally, it is briefly discussed how recent advances in breeding strategies, functional genomics and genome-editing technologies, particularly CRISPR-based approaches, are providing new opportunities to enhance reproductive resilience to heat stress and how it is essential to close the existing molecular knowledge gaps in the development of heat-tolerant rice varieties capable of sustaining productivity in a warming climate.

Oryza sativa (L.)↗

PubMed2026

Molecular diagnostics and integrated management challenges of tobacco streak virus: Current status and future perspectives.

Tobacco streak virus (TSV) is an economically important viral pathogen causing severe yield and quality losses in several agricultural, horticultural and medicinal crops worldwide. Its complex epidemiology involving sap transmission, infected pollen and pollen-feeding thrips, together with symptom similarity to other necrosis-inducing pathogens, frequently results in misdiagnosis and delayed disease management. This review critically evaluates recent advances in TSV diagnostics and integrated disease management strategies. Particular emphasis is placed on the transition from conventional biological and serological assays to advanced molecular diagnostics including reverse transcription polymerase chain reaction (RT-PCR), quantitative real-time PCR, multiplex PCR and emerging isothermal amplification technologies such as recombinase polymerase amplification (RPA) and loop-mediated isothermal amplification (LAMP). The review also highlights emerging innovations including CRISPR/Cas-based diagnostics in addition, integrated management approaches involving phytosanitation, weed reservoir management, vector ecology-based, host resistance breeding, RNA interference (RNAi) and genome editing technologies are critically analysed. Major challenges including inadequate field validation, limited multiplex capability, poor assay standardization and scarcity of resistant cultivars are discussed. Future objectives to develop quick, field-adaptable and durable TSV detection and management methods are additionally discussed.

CRISPR/Cas diagnostics↗

PubMed2026

Site-Specific Measurement of Meiotic Crossing-Over Rate with Droplet Digital PCR.

Understanding the frequency and distribution of meiotic crossovers (COs) is critical for both fundamental studies on meiosis and for practical applications in plant breeding, where controlling recombination can accelerate crop improvement. Determining CO rates at specific genomic loci has traditionally relied on labor-intensive methods that require the production and genotyping of large progenies. Here, we present a high-throughput protocol for site-specific quantification of meiotic COs in maize using droplet digital PCR (ddPCR). The method is based on genotyping individual pollen nuclei from hybrid plants to detect recombinant and nonrecombinant alleles at defined chromosomal intervals. By distributing several thousands of pollen nuclei into nanoliter-sized droplets and performing PCR with allele-specific fluorescent probes, this method allows precise quantification of CO frequency with high sensitivity. The protocol provides detailed guidance for nuclei isolation, probe master mix preparation, droplet generation, and data interpretation. This method can be easily adapted for use in other plants.

Journal Article↗

PubMed2026

A single-nucleus transcriptome atlas of soybean anthers.

Anther development is crucial for plant sexual reproduction. However, a high-resolution, cell-type-specific transcriptomic atlas of this process is lacking for the legume crop soybean (Glycine max). Here, we construct a comprehensive transcriptional atlas of developing soybean anthers using single-nucleus RNA sequencing (snRNA-seq). We identify and characterize nine distinct cell types spanning both somatic and reproductive lineages. Our analysis reveals robust transcriptional continuity across anther developmental stages and dynamic reprogramming during key transitions. Notably, the shift from diploid meiocytes to haploid unicellular microspores is marked by the induction of previously inactive genes, despite an overall reduction in transcript abundance. Subsequently, within bicellular microspores, generative and vegetative cell lineages exhibit sharply divergent transcriptional programs: generative cells specialize in mRNA export and turnover, whereas vegetative cells up-regulate translational machinery. Evolutionary analysis further indicates that generative-cell-specific genes are subject to more relaxed purifying selection compared to those specific to vegetative cells. Functional validation using mutants generated by CRISPR/Cas9-mediated genome editing and EMS mutagenesis reveals the essential roles of OSD1A and PKSA in pollen development and fertility. This high-resolution atlas provides fundamental insights into the transcriptional regulation of soybean anther development and serves as a valuable resource for manipulating male fertility to advance hybrid breeding programs. The data are available at https://databases.genedenovo.com/pollen.

Glycine max↗