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丹参中五个RNA依赖的RNA聚合酶基因的鉴定、分子克隆及表达分析

Identification, molecular cloning and expression analysis of five RNA-dependent RNA polymerase genes in Salvia miltiorrhiza.

作者信息

Shao Fenjuan, Lu Shanfa

机构信息

Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, China.

出版信息

PLoS One. 2014 Apr 14;9(4):e95117. doi: 10.1371/journal.pone.0095117. eCollection 2014.

Abstract

RNA-dependent RNA polymerases (RDRs) act as key components of the small RNA biogenesis pathways and play significant roles in post-transcriptional gene silencing (PTGS) and antiviral defense. However, there is no information about the RDR gene family in Salvia miltiorrhiza, an emerging model medicinal plant with great economic value. Through genome-wide predication and subsequent molecular cloning, five full-length S. miltiorrhiza RDR genes, termed SmRDR1-SmRDR5, were identified. The length of SmRDR cDNAs varies between 3,262 (SmRDR5) and 4,130 bp (SmRDR3). The intron number of SmRDR genes varies from 3 (SmRDR1, SmRDR3 and SmRDR4) to 17 (SmRDR5). All of the deduced SmRDR protein sequences contain the conserved RdRp domain. Moreover, SmRDR2 and SmRDR4 have an additional RRM domain. Based on the phylogenetic tree constructed with sixteen RDRs from Arabidopsis, rice and S. miltiorrhiza, plant RDRs may be divided into four groups (RDR1-RDR4). The RDR1 group contains an AtRDR and an OsRDR, while includes two SmRDRs. On the contrary, the RDR3 group contains three AtRDRs and two OsRDRs, but has only one SmRDR. SmRDRs were differentially expressed in flowers, leaves, stems and roots of S. miltiorrhiza and responsive to methyl jasmonate treatment and cucumber mosaic virus infection. The results suggest the involvement of RDRs in S. miltiorrhiza development and response to abiotic and biotic stresses. It provides a foundation for further studying the regulation and biological functions of SmRDRs and the biogenesis pathways of small RNAs in S. miltiorrhiza.

摘要

RNA依赖的RNA聚合酶(RDRs)是小RNA生物合成途径的关键组成部分,在转录后基因沉默(PTGS)和抗病毒防御中发挥重要作用。然而,对于丹参这种具有重要经济价值的新兴模式药用植物,尚无关于其RDR基因家族的信息。通过全基因组预测及后续分子克隆,鉴定出了5个丹参全长RDR基因,命名为SmRDR1 - SmRDR5。SmRDR cDNA的长度在3262 bp(SmRDR5)至4130 bp(SmRDR3)之间。SmRDR基因的内含子数量从3个(SmRDR1、SmRDR3和SmRDR4)到17个(SmRDR5)不等。所有推导的SmRDR蛋白序列均包含保守的RdRp结构域。此外,SmRDR2和SmRDR4还有一个额外的RRM结构域。基于由拟南芥、水稻和丹参的16个RDR构建的系统发育树,植物RDR可分为4组(RDR1 - RDR4)。RDR1组包含1个拟南芥RDR和1个水稻RDR,还包括2个丹参SmRDR。相反,RDR3组包含3个拟南芥RDR和2个水稻RDR,但只有1个丹参SmRDR。SmRDRs在丹参的花、叶、茎和根中差异表达,并对茉莉酸甲酯处理和黄瓜花叶病毒感染有响应。这些结果表明RDRs参与了丹参的发育以及对非生物和生物胁迫的响应。这为进一步研究SmRDRs的调控和生物学功能以及丹参中小RNA的生物合成途径奠定了基础。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6acc/3986363/25bca759357a/pone.0095117.g001.jpg

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