Genetic male sterility (GMS) facilitates hybrid seed production in crops including cotton (Gossypium hirsutum). However, the genetic and molecular mechanisms specifically involved in this developmental process are poorly understood. In this study, small RNA sequencing, degradome sequencing, and transcriptome sequencing were performed to analyze miRNAs and their target genes during anther development in a GMS mutant ('Dong A') and its fertile wildtype (WT). A total of 80 known and 220 novel miRNAs were identified, 71 of which showed differential expressions during anther development. A further degradome sequencing revealed a total of 117 candidate target genes cleaved by 16 known and 36 novel miRNAs. Based on RNA-seq, 24, 11, and 21 predicted target genes showed expression correlations with the corresponding miRNAs at the meiosis, tetrad and uninucleate stages, respectively. In addition, a large number of differentially expressed genes were identified, most of which were involved in sucrose and starch metabolism, carbohydrate metabolism, and plant hormone signal transduction based on Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis. The results of our study provide valuable information for further functional investigations of the important miRNAs and target genes involved in genetic male sterility and advance our understanding of miRNA regulatory functions during cotton anther development.

中文翻译：

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通过高通量RNA测序，在雄性不育突变体与其野生型棉花之间的花药发育过程中鉴定和分析microRNA和差异表达的基因。

遗传雄性不育（GMS）促进包括棉花（陆地棉）在内的农作物的杂交种子生产。但是，人们对这种发育过程特别涉及的遗传和分子机制了解甚少。在这项研究中，在GMS突变体（'Dong A'）及其可育野生型（WT）的花药发育过程中，进行了小RNA测序，降解组测序和转录组测序来分析miRNA及其靶基因。总共鉴定出80种已知和220种新颖的miRNA，其中71种在花药发育过程中显示差异表达。进一步的降解组测序显示，共有117个候选靶基因被16个已知miRNA和36个新miRNA切割。根据RNA-seq，预测的24、11和21个靶基因在减数分裂时与相应的miRNA表达相关，四核和单核阶段。此外，根据基因本体论（GO）和《京都基因与基因组百科全书》（KEGG）分析，鉴定出大量差异表达基因，其中大多数涉及蔗糖和淀粉代谢，碳水化合物代谢以及植物激素信号转导。 。我们的研究结果为重要的miRNA和参与遗传雄性不育的靶基因的进一步功能研究提供了有价值的信息，并加深了我们对棉花花药发育过程中miRNA调控功能的了解。基于基因本体论（GO）和《京都基因与基因组百科全书》（KEGG）分析的植物激素信号转导。我们的研究结果为重要的miRNA和参与遗传雄性不育的靶基因的进一步功能研究提供了有价值的信息，并加深了我们对棉花花药发育过程中miRNA调控功能的了解。基于基因本体论（GO）和《京都基因与基因组百科全书》（KEGG）分析的植物激素信号转导。我们的研究结果为重要的miRNA和参与遗传雄性不育的靶基因的进一步功能研究提供了有价值的信息，并加深了我们对棉花花药发育过程中miRNA调控功能的了解。