Department of Agricultural Biotechnology, National Academy of Agricultural Science, Rural Development Administration, 150 Suin-ro Gwonseon-gu, Suwon, 441-707, Korea.
BMC Plant Biol. 2012 Nov 19;12:218. doi: 10.1186/1471-2229-12-218.
BACKGROUND: MicroRNAs (miRNAs) are one of the functional non-coding small RNAs involved in the epigenetic control of the plant genome. Although plants contain both evolutionary conserved miRNAs and species-specific miRNAs within their genomes, computational methods often only identify evolutionary conserved miRNAs. The recent sequencing of the Brassica rapa genome enables us to identify miRNAs and their putative target genes. In this study, we sought to provide a more comprehensive prediction of B. rapa miRNAs based on high throughput small RNA deep sequencing. RESULTS: We sequenced small RNAs from five types of tissue: seedlings, roots, petioles, leaves, and flowers. By analyzing 2.75 million unique reads that mapped to the B. rapa genome, we identified 216 novel and 196 conserved miRNAs that were predicted to target approximately 20% of the genome's protein coding genes. Quantitative analysis of miRNAs from the five types of tissue revealed that novel miRNAs were expressed in diverse tissues but their expression levels were lower than those of the conserved miRNAs. Comparative analysis of the miRNAs between the B. rapa and Arabidopsis thaliana genomes demonstrated that redundant copies of conserved miRNAs in the B. rapa genome may have been deleted after whole genome triplication. Novel miRNA members seemed to have spontaneously arisen from the B. rapa and A. thaliana genomes, suggesting the species-specific expansion of miRNAs. We have made this data publicly available in a miRNA database of B. rapa called BraMRs. The database allows the user to retrieve miRNA sequences, their expression profiles, and a description of their target genes from the five tissue types investigated here. CONCLUSIONS: This is the first report to identify novel miRNAs from Brassica crops using genome-wide high throughput techniques. The combination of computational methods and small RNA deep sequencing provides robust predictions of miRNAs in the genome. The finding of numerous novel miRNAs, many with few target genes and low expression levels, suggests the rapid evolution of miRNA genes. The development of a miRNA database, BraMRs, enables us to integrate miRNA identification, target prediction, and functional annotation of target genes. BraMRs will represent a valuable public resource with which to study the epigenetic control of B. rapa and other closely related Brassica species. The database is available at the following link: http://bramrs.rna.kr [1].
背景: microRNAs (miRNAs) 是参与植物基因组表观遗传调控的功能非编码小 RNA 之一。尽管植物基因组中既包含进化保守的 miRNAs,也包含物种特异性的 miRNAs,但计算方法通常只能识别进化保守的 miRNAs。最近对油菜基因组的测序使我们能够识别 miRNAs 和它们的假定靶基因。在这项研究中,我们试图基于高通量小 RNA 深度测序,提供更全面的油菜 miRNAs 预测。
结果: 我们从幼苗、根、叶柄、叶片和花五种组织中测序了小 RNA。通过分析映射到油菜基因组的 275 万个独特的reads,我们鉴定了 216 个新的和 196 个保守的 miRNAs,它们预测靶向大约 20%的基因组蛋白质编码基因。对五种组织中的 miRNAs 进行定量分析表明,新的 miRNAs 在不同组织中表达,但表达水平低于保守 miRNAs。油菜和拟南芥基因组之间的 miRNAs 比较分析表明,油菜基因组中保守 miRNAs 的冗余拷贝可能在全基因组三倍体化后被删除。新的 miRNA 成员似乎是从油菜和拟南芥基因组中自发产生的,表明了 miRNA 的物种特异性扩张。我们已经将这些数据公开在一个名为 BraMRs 的油菜 miRNA 数据库中。该数据库允许用户从这里研究的五种组织类型中检索 miRNA 序列、它们的表达谱以及它们的靶基因描述。
结论: 这是首次使用全基因组高通量技术从芸薹属作物中鉴定新的 miRNAs。计算方法和小 RNA 深度测序的结合为基因组中 miRNAs 的预测提供了可靠的依据。大量新的 miRNAs 的发现,许多 miRNA 只有少数靶基因和低表达水平,表明 miRNA 基因的快速进化。miRNA 数据库 BraMRs 的开发使我们能够整合 miRNA 的鉴定、靶基因的预测和功能注释。BraMRs 将成为研究油菜和其他密切相关的芸薹属物种的表观遗传调控的宝贵公共资源。该数据库可在以下链接访问:http://bramrs.rna.kr[1]。
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