Long non-coding RNAs (lncRNAs) play a crucial role in regulating gene expression vital for the growth and development of plants. Despite this, the role of lncRNAs in Chinese cabbage (Brassica rapa L. ssp. pekinensis) pollen development and male fertility remains poorly understood. In this study, we characterized a recessive genic male sterile mutant (366–2 S), where the delayed degradation of tapetum and the failure of tetrad separation primarily led to the inability to form single microspores, resulting in male sterility. To analyze the role of lncRNAs in pollen development, we conducted a comparative lncRNA sequencing using anthers from the male sterile mutant line (366–2 S) and the wild-type male fertile line (366–2 F). We identified 385 differentially expressed lncRNAs between the 366–2 F and 366–2 S lines, with 172 of them potentially associated with target genes. To further understand the alterations in mRNA expression and explore potential lncRNA-target genes (mRNAs), we performed comparative mRNA transcriptome analysis in the anthers of 366–2 S and 366–2 F at two stages. We identified 1,176 differentially expressed mRNAs. Remarkably, GO analysis revealed significant enrichment in five GO terms, most notably involving mRNAs annotated as pectinesterase and polygalacturonase, which play roles in cell wall degradation. The considerable downregulation of these genes might contribute to the delayed degradation of tapetum in 366–2 S. Furthermore, we identified 15 lncRNA-mRNA modules through Venn diagram analysis. Among them, MSTRG.9997-BraA04g004630.3 C (β-1,3-glucanase) is associated with callose degradation and tetrad separation. Additionally, MSTRG.5212-BraA02g040020.3 C (pectinesterase) and MSTRG.13,532-BraA05g030320.3 C (pectinesterase) are associated with cell wall degradation of the tapetum, indicating that these three candidate lncRNA-mRNA modules potentially regulate pollen development. This study lays the foundation for understanding the roles of lncRNAs in pollen development and for elucidating their molecular mechanisms in regulating male sterility in Chinese cabbage.

中文翻译：

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转录组分析揭示了参与大白菜（Brassica rapa L. ssp. pekinensis）遗传雄性不育和育性的潜在lncRNA-mRNA模块

长链非编码RNA (lncRNA) 在调节对植物生长和发育至关重要的基因表达方面发挥着至关重要的作用。尽管如此，lncRNA 在大白菜（Brassica rapa L. ssp. pekinensis）花粉发育和雄性生育力中的作用仍然知之甚少。在这项研究中，我们鉴定了一种隐性基因雄性不育突变体（366-2S），其中绒毡层降解延迟和四分体分离失败主要导致无法形成单个小孢子，从而导致雄性不育。为了分析lncRNA在花粉发育中的作用，我们使用雄性不育突变系（366-2 S）和野生型雄性可育系（366-2 F）的花药进行了比较lncRNA测序。我们在 366-2 F 和 366-2 S 系之间鉴定了 385 个差异表达的 lncRNA，其中 172 个可能与靶基因相关。为了进一步了解 mRNA 表达的变化并探索潜在的 lncRNA 靶基因 (mRNA)，我们在两个阶段对 366–2 S 和 366–2 F 的花药进行了比较 mRNA 转录组分析。我们鉴定了 1,176 个差异表达的 mRNA。值得注意的是，GO 分析揭示了 5 个 GO 术语的显着富集，最显着的是涉及注释为果胶酯酶和多聚半乳糖醛酸酶的 mRNA，它们在细胞壁降解中发挥作用。这些基因的显着下调可能导致 366-2 S 中绒毡层的降解延迟。此外，我们通过维恩图分析鉴定了 15 个 lncRNA-mRNA 模块。其中，MSTRG.9997-BraA04g004630.3 C（β-1,3-葡聚糖酶）与胼胝质降解和四分体分离有关。此外，MSTRG.5212-BraA02g040020.3 C（果胶酯酶）和 MSTRG.13,532-BraA05g030320.3 C（果胶酯酶）与绒毡层细胞壁降解相关，表明这三个候选 lncRNA-mRNA 模块可能调节花粉发育。该研究为了解lncRNA在花粉发育中的作用以及阐明其调节大白菜雄性不育的分子机制奠定了基础。