Cloning and characterization of microRNAs from wheat (Triticum aestivum L.).
Cloning and characterization of microRNAs from wheat (Triticum aestivum L.).
复制标题
小麦 (Triticum aestivum L.) 的 microRNA 的克隆和表征
DOI：
10.1186/gb-2007-8-6-r96
复制
发表时间：
2007
期刊：
影响因子：
12.3
通讯作者：
Sun Q
中科院分区：
文献类型：
--
作者：
Yao Y;Guo G;Ni Z;Sunkar R;Du J;Zhu JK;Sun Q
背景
MicroRNA (miRNA) 是一类小型非编码调节 RNA，通过指导靶标 mRNA 裂解或翻译抑制来调节基因表达。迄今为止，miRNA的鉴定仅限于少数模式植物物种，如拟南芥、水稻和杨树，其基因组已被测序。小麦是世界上最重要的谷类作物之一。迄今为止，仅预测了小麦中的少数保守 miRNA，并且小麦 miRNA 的计算鉴定需要基因组序列，而该序列是未知的。
结果
为了鉴定小麦 (Triticum aestivum L.) 中新颖且保守的 miRNA，我们构建了一个小型 RNA 文库。该文库的高通量测序和随后的分析揭示了 58 个 miRNA 的鉴定，其中包括 43 个 miRNA 家族。其中，35 个 miRNA 属于 20 个保守的 miRNA 家族。其余 23 个 miRNA 是新的，在小麦中形成 23 个 miRNA 家族；更重要的是，其中 4 个新 miRNA（miR506、miR510、miR514 和 miR516）似乎是单子叶植物特异性的。 Northern blot分析表明，一些新的miRNA优先在某些组织中表达。基于序列同源性，我们预测了 46 个潜在靶点。因此，我们鉴定了大量单子叶植物特异性和小麦特异性 miRNA。这些结果表明，保守的和小麦特异性的miRNA在小麦生长发育、胁迫反应和其他生理过程中发挥着重要作用。
结论
这项研究发现了 58 个小麦 miRNA，包括 43 个 miRNA 家族；其中 20 个家族在小麦中是保守的，23 个是新的。它首次对小麦 miRNA 及其预测靶标进行了大规模克隆和表征。
Background
MicroRNAs (miRNAs) are a class of small, non-coding regulatory RNAs that regulate gene expression by guiding target mRNA cleavage or translational inhibition. So far, identification of miRNAs has been limited to a few model plant species, such as Arabidopsis, rice and Populus, whose genomes have been sequenced. Wheat is one of the most important cereal crops worldwide. To date, only a few conserved miRNAs have been predicted in wheat and the computational identification of wheat miRNAs requires the genome sequence, which is unknown.
Results
To identify novel as well as conserved miRNAs in wheat (Triticum aestivum L.), we constructed a small RNA library. High throughput sequencing of the library and subsequent analysis revealed the identification of 58 miRNAs, comprising 43 miRNA families. Of these, 35 miRNAs belong to 20 conserved miRNA families. The remaining 23 miRNAs are novel and form 23 miRNA families in wheat; more importantly, 4 of these new miRNAs (miR506, miR510, miR514 and miR516) appear to be monocot-specific. Northern blot analysis indicated that some of the new miRNAs are preferentially expressed in certain tissues. Based on sequence homology, we predicted 46 potential targets. Thus, we have identified a large number of monocot-specific and wheat-specific miRNAs. These results indicate that both conserved and wheat-specific miRNAs play important roles in wheat growth and development, stress responses and other physiological processes.
Conclusion
This study led to the discovery of 58 wheat miRNAs comprising 43 miRNA families; 20 of these families are conserved and 23 are novel in wheat. It provides a first large scale cloning and characterization of wheat miRNAs and their predicted targets.
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