College of Agronomy and Biotechnology, China Agricultural University, Beijing, China.
PLoS One. 2012;7(4):e33696. doi: 10.1371/journal.pone.0033696. Epub 2012 Apr 6.
To date, no miRNAs have been identified in the important diploid cotton species although there are several reports on miRNAs in upland cotton. In this study, we identified 73 miRNAs, belonging to 49 families, from Asiatic cotton using a well-developed comparative genome-based homologue search. Several of the predicted miRNAs were validated using quantitative real time PCR (qRT-PCR). The length of miRNAs varied from 18 to 22 nt with an average of 20 nt. The length of miRNA precursors also varied from 46 to 684 nt with an average of 138 ±120 nt. For a majority of Asiatic cotton miRNAs, there is only one member per family; however, multiple members were identified for miRNA 156, 414, 837, 838, 1044, 1533, 2902, 2868, 5021 and 5142 families. Nucleotides A and U were dominant, accounted for 62.95%, in the Asiatic cotton pre-miRNAs. The Asiatic cotton pre-miRNAs had high negative minimal folding free energy (MFE) and adjusted MFE (AMFE) and high MFE index (MFEI). Many miRNAs identified in Asiatic cotton suggest that miRNAs also play a similar regulatory mechanism in diploid cotton.
迄今为止,尽管在陆地棉中有几个关于 miRNA 的报道,但在重要的二倍体棉花物种中尚未发现 miRNA。在这项研究中,我们使用成熟的基于比较基因组同源搜索的方法,从亚洲棉中鉴定出 73 个 miRNA,属于 49 个家族。使用定量实时 PCR(qRT-PCR)验证了几个预测的 miRNA。miRNA 的长度从 18 到 22 个核苷酸不等,平均为 20 个核苷酸。miRNA 前体的长度也从 46 到 684 个核苷酸不等,平均为 138±120 个核苷酸。对于大多数亚洲棉 miRNA,每个家族只有一个成员;然而,miRNA 156、414、837、838、1044、1533、2902、2868、5021 和 5142 家族都鉴定出了多个成员。核苷酸 A 和 U 占优势,占亚洲棉前体 miRNA 的 62.95%。亚洲棉前体 miRNA 具有高负最小折叠自由能(MFE)和调整的 MFE(AMFE)和高 MFE 指数(MFEI)。在亚洲棉中鉴定出的许多 miRNA 表明,miRNA 也在二倍体棉中发挥类似的调节机制。