Beijing Engineering Research Center for Hybrid Wheat, Beijing Academy of Agriculture and Forestry Sciences, Beijing 100097, China.
The Municipal Key Laboratory of Molecular Genetic of Hybrid Wheat, Beijing 10097, China.
J Exp Bot. 2022 Apr 5;73(7):2157-2174. doi: 10.1093/jxb/erab516.
Thermosensitive genic male sterile (TGMS) wheat lines are the core of two-line hybrid systems. Understanding the mechanism that regulates male sterility in TGMS wheat lines is helpful for promoting wheat breeding. Several studies have obtained information regarding the mechanisms associated with male sterility at the transcriptional level, but it is not clear how the post-transcriptional process of alternative splicing might contribute to controlling male sterility. In this study, we performed genome-wide analyses of alternative splicing during the meiosis stage in TGMS line BS366 using PacBio and RNA-Seq hybrid sequencing. Cytological observations indicated that cytoskeleton assembly in pollen cells, calcium deposition in pollen and tapetal cells, and vesicle transport in tapetal cells were deficient in BS366. According to our cytological findings, 49 differentially spliced genes were isolated. Moreover, 25 long non-coding RNA targets and three bHLH transcription factors were identified. Weighted gene co-expression network analysis detected four candidate differentially spliced genes that had strong co-relation with the seed setting percentage, which is the direct representation of male sterility in BS366. In this study, we obtained comprehensive data regarding the alternative splicing-mediated regulation of male sterility in TGMS wheat. The candidates identified may provide the molecular basis for an improved understanding of male sterility.
温敏雄性不育(TGMS)小麦品系是两系杂交系统的核心。了解 TGMS 小麦品系中雄性不育的调控机制有助于促进小麦育种。已有几项研究从转录水平获得了与雄性不育相关的机制信息,但尚不清楚可变剪接的转录后过程如何有助于控制雄性不育。本研究采用 PacBio 和 RNA-Seq 混合测序技术,对 TGMS 系 BS366 的减数分裂阶段的可变剪接进行了全基因组分析。细胞学观察表明,BS366 花粉细胞中的细胞骨架组装、花粉和绒毡层细胞中的钙沉积以及绒毡层细胞中的囊泡运输均存在缺陷。根据我们的细胞学发现,分离出 49 个差异剪接基因。此外,还鉴定出 25 个长非编码 RNA 靶标和 3 个 bHLH 转录因子。加权基因共表达网络分析检测到四个与结实率(BS366 雄性不育的直接表现)具有强相关性的候选差异剪接基因。本研究获得了 TGMS 小麦中可变剪接介导的雄性不育调控的全面数据。鉴定出的候选基因可能为深入了解雄性不育提供分子基础。