Identification of miRNAs with potential roles in regulation of anther development and male-sterility in 7B-1 male-sterile tomato mutant.
Identification of miRNAs with potential roles in regulation of anther development and male-sterility in 7B-1 male-sterile tomato mutant.
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DOI：
10.1186/s12864-015-2077-0
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发表时间：
2015-10-28
期刊：
影响因子：
4.4
通讯作者：
Fellner M
中科院分区：
文献类型：
--
作者：
Omidvar V;Mohorianu I;Dalmay T;Fellner M
7 B-1番茄品系（Solanum lycopersicum cv. Rutgers）是一个光敏感雄性不育突变体，在杂交制种中具有潜在的应用价值。番茄中的小分子RNA（sRNAs）主要与果实发育和成熟有关，但在花发育和雄性不育调控方面还没有研究。利用sRNA测序技术，我们鉴定了7 B-1突变体中可能参与花药发育和雄性不育调控的miRNAs。对来自7 B-1和野生型（WT）花药的两个sRNA文库进行测序，并且在两个文库中鉴定了32个已知的miRNAs家族和23个新的miRNAs。在7 B-1花药中，miR 390、miR 166、miR 159表达上调，miR 530、miR 167、miR 164、miR 396、miR 168、miR 393、miR 8006和两种新的miRNAs miR#W和miR#M表达下调。Ta-siRNA没有差异表达，可能与7 B-1雄性不育无关。利用5′-RACE验证了在花药发育中具有潜在作用的miRNA靶标。QPCR分析显示了7 B-1花药和茎中目标miRNA/靶标对的差异表达，表明它们可能调节这些组织中的不同生物学过程。除极少数miRNA/靶标对外，大多数miRNA/靶标对的表达水平呈负相关。原位杂交结果表明，miR 159、GAMYBL 1、PMEI和cystatin在绒毡层、四分体和小孢子中的表达占优势。总之，我们鉴定了在7 B-1的花药发育和雄性不育调控中具有潜在作用的miRNA。此外，还首次从番茄中发现了一些新的miRNAs。我们的数据可以作为一个基准，为未来的研究在其他作物雄性不育的分子机制。本文的在线版本（doi：10.1186/s12864-015-2077-0）包含补充材料，可供授权用户使用。
The 7B-1 tomato line (Solanum lycopersicum cv. Rutgers) is a photoperiod-sensitive male-sterile mutant, with potential application in hybrid seed production. Small RNAs (sRNAs) in tomato have been mainly characterized in fruit development and ripening, but none have been studied with respect to flower development and regulation of male-sterility. Using sRNA sequencing, we identified miRNAs that are potentially involved in anther development and regulation of male-sterility in 7B-1 mutant. Two sRNA libraries from 7B-1 and wild type (WT) anthers were sequenced and thirty two families of known miRNAs and 23 new miRNAs were identified in both libraries. MiR390, miR166, miR159 were up-regulated and miR530, miR167, miR164, miR396, miR168, miR393, miR8006 and two new miRNAs, miR#W and miR#M were down-regulated in 7B-1 anthers. Ta-siRNAs were not differentially expressed and likely not associated with 7B-1 male-sterility. miRNA targets with potential roles in anther development were validated using 5′-RACE. QPCR analysis showed differential expression of miRNA/target pairs of interest in anthers and stem of 7B-1, suggesting that they may regulate different biological processes in these tissues. Expression level of most miRNA/target pairs showed negative correlation, except for few. In situ hybridization showed predominant expression of miR159, GAMYBL1, PMEI and cystatin in tapetum, tetrads and microspores. Overall, we identified miRNAs with potential roles in anther development and regulation of male-sterility in 7B-1. A number of new miRNAs were also identified from tomato for the first time. Our data could be used as a benchmark for future studies of the molecular mechanisms of male-sterility in other crops. The online version of this article (doi:10.1186/s12864-015-2077-0) contains supplementary material, which is available to authorized users.