杨树10种miRNA的低氮胁迫差异表达及靶基因的鉴定与分析  

Differential Expression of 10 miRNAs in Poplar under Low Nitrogen Stress and the Identification and Analysis of Target Genes

作　　者：包海[1] 孙丰硕[1] 徐千惠 王延伟[1] Bao Hai;Sun Fengshuo;Xu Qianhui;Wang Yanwei(National Engineering Laboratory for Tree Breeding,Key Laboratory of Genetics and Breeding in Forest Trees and Ornamental Plants,Ministry of Edu- cation,Tree and Ornamental Plant Breeding and Biotechnology Laboratory of State Forestry Administration,Beijing Forestry University,Beijing, 100083)

机构地区：[1]北京林业大学林木育种国家工程实验室林木花卉遗传育种教育部重点实验室国家林业局树木花卉育种与生物工程重点开放实验室

出　　处：《分子植物育种》2019年第3期771-779,共9页Molecular Plant Breeding

基　　金：国家自然科学基金(31670671;31470668)资助

摘　　要：microRNA (miRNA)是一类重要的非编码小分子RNA,在植物应对逆境胁迫的响应中起着重要调控作用。为了探讨miRNA在杨树受低氮营养胁迫不同时间段的差异表达模式以初步鉴定参与低氮胁迫响应的miRNA,本研究以重要乡土树种毛白杨为材料,进行不同时间段的低氮胁迫处理,分别提取长度在200 nt以下的小片段RNA,利用实时定量PCR技术研究毛白杨在低氮胁迫后10种miRNA的差异表达模式。结果显示,miRNA的表达在低氮胁迫下有显著变化,部分miRNA的表达呈现动态反应。在低氮胁迫下,miRNA的表达呈现四种模式,大部分miRNA的表达在处理1 d后受到显著抑制。借助生物信息学手段,进一步鉴定到10个miRNA的靶基因,功能注释表明靶基因共分为6大类,降解组实验进一步验证了miR159a对靶基因的切割。该研究初步表明,miRNA在杨树对低氮营养的胁迫响应中可能发挥着重要调控作用。MicroRNAs (miRNA)is a kind of important noncoding small RNA,which plays a critical role in plant response to stress.In order to investigate the differential expression of poplar under low nitrogen nutrition stress of different treatment periods and preliminary identification of miRNAs that participate in the response of poplar to low nitrogen stress,this investigation used the important native tree species Populus tomentosa as material for treatment of low nitrogen stress at different time periods,and extracted small fragment RNA with length no more than 200 nt through real-time quantitative RT-PCR (qRT-PCR)technology to elucidate differential expression patterns of 10 miRNA under low nitrogen stress.The results showed that miRNAs were significantly changed under low nitrogen stress,and some of them showed a dynamic response.The expression of most miRNAs was significantly suppressed after I d of treatment.The target genes of 10 miRNAs were further identified in silico.The function were further annotated,which showed that the target genes were divided into 6 major categories.Moreover,the cleavage of miR159a on target was verified through degradorne analysis.This investigation showed that miRNAs might act an indispensable role in regulating the response of poplar to low nitrogen stress.

关 键 词：毛白杨 低氮营养胁迫 MICRORNA 靶基因 差异表达 

分 类 号：S792.11[农业科学—林木遗传育种]