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7B-1雄性不育番茄突变体雄性不育的转录调控

Transcriptional regulation of male-sterility in 7B-1 male-sterile tomato mutant.

作者信息

Omidvar Vahid, Mohorianu Irina, Dalmay Tamas, Zheng Yi, Fei Zhangjun, Pucci Anna, Mazzucato Andrea, Večeřová Vendula, Sedlářova Michaela, Fellner Martin

机构信息

Laboratory of Growth Regulators, Centre of the Region Haná for Biotechnological and Agricultural Research, Palacký University and Institute of Experimental Botany AS CR, Šlechtitelů 27, Olomouc-Holice, Czech Republic.

School of Computing Sciences, University of East Anglia, Norwich, United Kingdom.

出版信息

PLoS One. 2017 Feb 8;12(2):e0170715. doi: 10.1371/journal.pone.0170715. eCollection 2017.

Abstract

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.

摘要

7B-1番茄(Solanum lycopersicum L. cv Rutgers)是一种对非生物胁迫耐受性增强的雄性不育突变体,这使其成为杂交种子育种和胁迫工程的潜在候选材料。为了深入了解调控7B-1雄性不育的分子机制,利用mRNA测序(RNA-Seq)研究了7B-1雄性不育和野生型(WT)花药的转录组图谱。总共鉴定出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。对DEG进行的基因本体(GO)富集分析表明,7B-1突变对代谢过程有普遍影响,如蛋白水解和碳水化合物分解代谢过程。对16个在花药发育调控中起关键作用的候选基因进行了qRT-PCR和原位杂交进一步分析。细胞学研究显示7B-1突变体花药发育存在几个缺陷,包括花药成熟不同步、减数分裂功能障碍、小孢子停滞、胼胝质降解缺陷和绒毡层发育异常。TUNEL分析显示7B-1花药绒毡层细胞程序性细胞死亡(PCD)存在缺陷。本研究为7B-1突变体的转录组提供了见解。我们鉴定出7B-1中几个表达水平改变的基因(包括β-1,3葡聚糖酶、GA2氧化酶、半胱氨酸蛋白酶抑制剂、半胱氨酸蛋白酶、果胶酯酶、TA29和肌动蛋白),它们可能潜在地调控花药发育过程,如减数分裂、绒毡层发育和细胞壁形成/降解。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2e58/5298235/c8ccd974fe69/pone.0170715.g001.jpg

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