草莓叶片中5种新microRNA的RT-PCR鉴定及靶基因分析  

Identification of Five Novel Micro RNAs by RT-PCR and Analysis of Their Target Genes in Strawberry Leaves

作　　者：张馨宇[1,2] 戚华彩 董向向 李贺[2] 张志宏[2] 

机构地区：[1]辽宁省农业科学院蔬菜研究所,沈阳110161 [2]沈阳农业大学园艺学院/设施园艺省部共建教育部重点实验室,沈阳110161

出　　处：《沈阳农业大学学报》2016年第2期141-147,共7页Journal of Shenyang Agricultural University

基　　金：辽宁省高等学校杰出青年学者成长计划项目(LJQ2014069);国家自然科学基金项目(31101524)

摘　　要：micro RNA(miRNA)是来自真核生物自身基因组的非编码小分子RNA,在转录后水平上负调控基因表达,在植物的生长发育和胁迫应答等方面起重要作用。以栽培草莓艳丽为试材,利用RT-PCR技术克隆草莓叶片中5种新miRNA的前体及靶基因,分析靶基因的结构特征与功能,结果表明:草莓mi R-6,mi R-12,mi R-13,mi R-14,mi R-29的前体长度分别为513 bp,309 bp,464bp,334 bp和343 bp,均能形成典型的茎环结构,其中mi R-12、mi R-13的成熟miRNA序列在茎环3'端臂上,mi R-6、mi R-14、mi R-29成熟miRNA序列在茎环的5'端臂上。mi R-14与mi R-29的靶基因分别为854bp和690bp,与森林草莓基因组对应序列一致性为98.72%和99.42%,其中mi R-29靶基因是GRP基因,编码143个氨基酸,推测mi R-29靶基因在草莓植株的胁迫响应中有重要调控作用。本研究丰富了草莓miRNA的研究,为揭示草莓miRNA的生物学功能奠定基础。microRNAs are small, non-coding RNAs from eukaryotic genomes that act to negatively regulate the expression of theirtarget genes at the post-transcriptional levels, and play important roles in the plant growing development and their response toenvironmental stresses. The precursors of five novel miRNAs and their target genes were cloned by using RT-PCR methods fromleaves of cultivated strawberry, Yanli, and characteristic and function of the targets genes were analyzed. The results showed that theprecursors lengths of miR-6, miR-12, miR-13, miR-14 and miR-29 were 513 bp, 309 bp, 464 bp, 334 bp and 343 bp,respectively. They could form typical stem-loop structure. Among them, the mature miRNAs of miR12 and miR13 located in the 3＇end of stem-loop, and the mature miRNAs of miR-6, miR-14 and miR-29 were at the 5＇ end of stem-loop. The target genes ofmiR-14 and miR-29 were 854bp and 690bp in length, and their identities of sequences were 98.72% and 99.42% comparing withcorresponding sequence in strawberry genome. The target gene of miR-29 was GRP, encoded a predicted protein of 143 amino acidresidues. So we deduce it has the important regulated function in their response to environmental stresses of strawberries. Theresults will enrich the research on strawberry miRNA and lay the foundation of the biological function of strawberry miRNA.

关 键 词：草莓 MIRNA 前体 靶基因 功能分析 

分 类 号：S663.9[农业科学—果树学]