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利用高通量测序技术鉴定和分析花生(Arachis hypogaea L.)中的 microRNAs。

Identification and characterization of microRNAs from peanut (Arachis hypogaea L.) by high-throughput sequencing.

机构信息

Shandong Peanut Research Institute, Qingdao, People's Republic of China.

出版信息

PLoS One. 2011;6(11):e27530. doi: 10.1371/journal.pone.0027530. Epub 2011 Nov 16.

DOI:10.1371/journal.pone.0027530
PMID:22110666
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC3217988/
Abstract

BACKGROUND

MicroRNAs (miRNAs) are noncoding RNAs of approximately 21 nt that regulate gene expression in plants post-transcriptionally by endonucleolytic cleavage or translational inhibition. miRNAs play essential roles in numerous developmental and physiological processes and many of them are conserved across species. Extensive studies of miRNAs have been done in a few model plants; however, less is known about the diversity of these regulatory RNAs in peanut (Arachis hypogaea L.), one of the most important oilseed crops cultivated worldwide.

RESULTS

A library of small RNA from peanut was constructed for deep sequencing. In addition to 126 known miRNAs from 33 families, 25 novel peanut miRNAs were identified. The miRNA* sequences of four novel miRNAs were discovered, providing additional evidence for the existence of miRNAs. Twenty of the novel miRNAs were considered to be species-specific because no homolog has been found for other plant species. qRT-PCR was used to analyze the expression of seven miRNAs in different tissues and in seed at different developmental stages and some showed tissue- and/or growth stage-specific expression. Furthermore, potential targets of these putative miRNAs were predicted on the basis of the sequence homology search.

CONCLUSIONS

We have identified large numbers of miRNAs and their related target genes through deep sequencing of a small RNA library. This study of the identification and characterization of miRNAs in peanut can initiate further study on peanut miRNA regulation mechanisms, and help toward a greater understanding of the important roles of miRNAs in peanut.

摘要

背景

MicroRNAs(miRNAs)是大约 21 个核苷酸的非编码 RNA,通过核酸内切酶切割或翻译抑制在植物中转录后调控基因表达。miRNAs 在许多发育和生理过程中发挥着重要作用,其中许多在物种间是保守的。已经在少数模式植物中对 miRNAs 进行了广泛的研究;然而,对于花生(Arachis hypogaea L.)中这些调控 RNA 的多样性知之甚少,花生是全球种植的最重要的油料作物之一。

结果

构建了花生小 RNA 文库进行深度测序。除了来自 33 个家族的 126 个已知 miRNA 外,还鉴定了 25 个新的花生 miRNA。发现了四个新 miRNA 的 miRNA*序列,为 miRNA 的存在提供了额外的证据。其中 20 个新 miRNA 被认为是种特异性的,因为在其他植物物种中没有发现同源物。qRT-PCR 用于分析不同组织和种子不同发育阶段的 7 个 miRNA 的表达,其中一些表现出组织和/或生长阶段特异性表达。此外,根据序列同源性搜索预测了这些假定 miRNA 的潜在靶标。

结论

我们通过对小 RNA 文库进行深度测序,鉴定了大量的 miRNAs 及其相关靶基因。对花生中 miRNA 的鉴定和特征研究可以启动对花生 miRNA 调控机制的进一步研究,并有助于更好地理解 miRNA 在花生中的重要作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0fb5/3217988/4325995a0ba6/pone.0027530.g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0fb5/3217988/3daefffc98ce/pone.0027530.g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0fb5/3217988/e3ff689a6d65/pone.0027530.g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0fb5/3217988/4325995a0ba6/pone.0027530.g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0fb5/3217988/3daefffc98ce/pone.0027530.g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0fb5/3217988/e3ff689a6d65/pone.0027530.g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0fb5/3217988/4325995a0ba6/pone.0027530.g003.jpg

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