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中国农学通报

中国农学通报 ›› 2012, Vol. 28 ›› Issue (30): 184-190.doi: 10.11924/j.issn.1000-6850.2012-0815

所属专题： 马铃薯

• 生物技术科学 • 上一篇    下一篇

利用solexa测序技术发掘木薯microRNAs

张何芳 陈新 卢诚 章伟雄 王文泉   

  • 收稿日期:2012-03-09 修回日期:2012-04-24 出版日期:2012-10-25 发布日期:2012-10-25

Identification of miRNAs in Cassava by Solexa Sequencing

  • Received:2012-03-09 Revised:2012-04-24 Online:2012-10-25 Published:2012-10-25

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摘要：

为了发掘木薯中的microRNAs (miRNAs)，了解其在木薯中的调控机制，本研究利用Solexa测序技术在木薯中发现了大量的miRNAs，并利用一种新的miRNA定量检测技术（Multiplexed RT法）对其进行实验验证。通过对构建的3个小RNA文库进行solexa测序与生物信息学分析，最终获得19个家族93个保守的miRNA和74个新的miRNA，实验验证了测序获得的83个miRNA。在Arg7块根中特异表达的有6个miRNA，分别为miR172d、miR396c、miR398a、miR398b、miR399e、miR399f，叶片中特异表达的有13个，须根中特异表达的有3个，这些miRNAs将为深入了解木薯块根发育和淀粉累积机理提供了一个新的视角。

关键词: 聚类分析, 聚类分析

Abstract:

In order to identify miRNA (microRNA) in Cassava, to understand its mechanism of accumulation in cassava. In this study, lots of miRNAs were identificated in Cassava by Solexa sequencing。By using a new PCR based assay platform (Multiplexed RT assay), miRNAs were verified in cassava. 93 conserved miRNAs comprising 19 families and 74 novel miRNAs were verified through sequencing and bioinformatics analysing three small RNA library, 83 miRNAs were verified by Multiplexed RT assay. 6 of miRNA, miR172d, miR396c, miR398a, miR398b, miR399e and miR399f, were specifically expressed in Arg7 tuber; 13 in leaves; and 3 in fibrous roots. These miRNAs provides a new perspective to intensive study tuber development and the mechanism of starch accumulation in the cassava.