Transcriptional regulation of male-sterility in 7B-1 male-sterile tomato mutant.
Transcriptional regulation of male-sterility in 7B-1 male-sterile tomato mutant.
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DOI：
10.1371/journal.pone.0170715
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发表时间：
2017
期刊：
影响因子：
3.7
通讯作者：
Fellner M
中科院分区：
文献类型：
--
作者：
Omidvar V;Mohorianu I;Dalmay T;Zheng Y;Fei Z;Pucci A;Mazzucato A;Večeřová V;Sedlářova M;Fellner M
7B-1番茄（Solanum lycopersicum L. cv Rutgers）是一种雄性不育突变体，具有较强的抗非生物胁迫能力，是一种潜在的杂交种子育种和胁迫工程的候选品种。为了揭示7B-1雄性不育的分子调控机制，利用mRNA测序（RNA-Seq）技术研究了7B-1雄性不育和野生型（WT）花药的转录组学特征。共鉴定出768个差异表达基因（deg），包括132个上调转录本和636个下调转录本。DEGs的基因本体（GO）富集分析表明，7B-1突变对代谢过程（如蛋白质水解和碳水化合物分解过程）有普遍影响。利用qRT-PCR和原位杂交技术对16个在花药发育调控中起关键作用的候选基因进行进一步分析。细胞学研究表明，7B-1突变体花药发育存在一些缺陷，包括花药成熟不同步、减数分裂功能障碍、小孢子受阻、胼胝质降解缺陷和绒毡层发育异常。TUNEL实验显示7B-1花药毡层细胞程序性死亡（PCD）存在缺陷。目前的研究提供了对7B-1突变体转录组的见解。我们在7B-1中发现了几个表达水平改变的基因（包括β -1,3葡聚糖酶、GA2oxs、胱抑素、半胱氨酸蛋白酶、果胶酯酶、TA29和肌动蛋白），这些基因可能调节花药发育过程，如减数分裂、绒毡层发育和细胞壁形成/降解。
The 7B-1 tomato (Solanum lycopersicum L. cv Rutgers) is a male-sterile mutant with enhanced tolerance to abiotic stress, which makes it a potential candidate for hybrid seed breeding and stress engineering. To underline the molecular mechanism regulating the male-sterility in 7B-1, transcriptomic profiles of the 7B-1 male-sterile and wild type (WT) anthers were studied using mRNA sequencing (RNA-Seq). In total, 768 differentially expressed genes (DEGs) were identified, including 132 up-regulated and 636 down-regulated transcripts. Gene ontology (GO) enrichment analysis of DEGs suggested a general impact of the 7B-1 mutation on metabolic processes, such as proteolysis and carbohydrate catabolic process. Sixteen candidates with key roles in regulation of anther development were subjected to further analysis using qRT-PCR and in situ hybridization. Cytological studies showed several defects associated with anther development in the 7B-1 mutant, including unsynchronized anther maturation, dysfunctional meiosis, arrested microspores, defect in callose degradation and abnormal tapetum development. TUNEL assay showed a defect in programmed cell death (PCD) of tapetal cells in 7B-1 anthers. The present study provides insights into the transcriptome of the 7B-1 mutant. We identified several genes with altered expression level in 7B-1 (including beta-1,3 glucanase, GA2oxs, cystatin, cysteine protease, pectinesterase, TA29, and actin) that could potentially regulate anther developmental processes, such as meiosis, tapetum development, and cell-wall formation/degradation.