Identification and characterization of 27 conserved microRNAs in citrus.
Identification and characterization of 27 conserved microRNAs in citrus.
复制标题
柑橘中 27 个保守 microRNA 的鉴定和表征。
DOI：
10.1007/s00425-009-0971-x
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发表时间：
2009-09
期刊：
影响因子：
4.3
通讯作者：
Chao, C. Thomas
中科院分区：
文献类型：
--
作者：
Song, Changnian;Fang, Jinggui;Li, Xiaoying;Liu, Hong;Chao, C. Thomas
MicroRNA (miRNA) 是一类非蛋白质编码的小RNA。考虑到许多 miRNA 基因在不同植物基因组中的保守性，从非模式生物中鉴定 miRNA 既实用又有助于解决 miRNA 引导的基因调控问题。柑橘是重要的主食果树，公开的柑橘表达序列标签（EST）数据库正在不断增加。然而，到目前为止，人们对柑橘中的 miRNA 知之甚少。在本研究中，在柑橘EST数据库中搜索了来自拟南芥的27个已知miRNA的miRNA前体，其中搜索到的13个前体序列可以形成与拟南芥相似的折叠结构。通过Northern blotting可以在柑橘叶、幼芽、花、果实和根中检测到这13种柑橘microRNA和其他13种潜在柑橘miRNA的普遍表达，其中一些在不同组织中表现出差异表达。基于 miRNA 与其靶序列表现出完美或近乎完美的互补性，15 个柑橘 miRNA 共鉴定出 41 个潜在靶点。大多数靶标是在柑橘发育（包括叶、芽和根发育）中发挥重要作用的转录因子。此外，一些其他靶基因似乎在不同的生理过程中发挥作用。通过检测枳壳中 miRNA 介导的 mRNA 裂解，已通过实验验证了四个靶基因。总体而言，本研究对柑橘中miRNA的鉴定和表征可以启动柑橘miRNA调控机制的进一步研究，并有助于我们更多地了解柑橘中miRNA的重要作用。
MicroRNAs (miRNAs) are a class of non-protein-coding small RNAs. Considering the conservation of many miRNA genes in different plant genomes, the identification of miRNAs from non-model organisms is both practicable and instrumental in addressing miRNA-guided gene regulation. Citrus is an important staple fruit tree, and publicly available expressed sequence tag (EST) database for citrus are increasing. However, until now, little has been known about miRNA in citrus. In this study, 27 known miRNAs from Arabidopsis were searched against citrus EST databases for miRNA precursors, of which 13 searched precursor sequences could form fold-back structures similar with those of Arabidopsis. The ubiquitous expression of those 13 citrus microRNAs and other 13 potential citrus miRNAs could be detected in citrus leaf, young shoot, flower, fruit and root by northern blotting, and some of them showed differential expression in different tissues. Based on the fact that miRNAs exhibit perfect or nearly perfect complementarity with their target sequences, a total of 41 potential targets were identified for 15 citrus miRNAs. The majority of the targets are transcription factors that play important roles in citrus development, including leaf, shoot, and root development. Additionally, some other target genes appear to play roles in diverse physiological processes. Four target genes have been experimentally verified by detection of the miRNA-mediated mRNA cleavage in Poncirus trifoliate. Overall, this study in the identification and characterization of miRNAs in citrus can initiate further study on citrus miRNA regulation mechanisms, and it can help us to know more about the important roles of miRNAs in citrus.
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