盐穗木miR397a增强转基因拟南芥的低温耐性  

Halostachys caspica miR397a Enhances Cold Tolerance of Transgenic Arabidopsis thaliana

作　　者：魏冉 张慧珍 陈方圆 赵丽娟 李文辉 曾幼玲 WEI Ran;ZHANG Huizhen;CHEN Fangyuan;ZHAO Lijuan;LI Wenhui;ZENG Youling(Xinjiang Key Laboratory of Biological Resources and Genetic Engineering,School of Life Science and Technology,Xinjiang University,Urumqi Xinjiang 830017,China)

机构地区：[1]新疆大学生命科学与技术学院新疆生物资源基因工程重点实验室,新疆乌鲁木齐830017

出　　处：《新疆大学学报（自然科学版）（中英文）》2023年第1期87-94,共8页Journal of Xinjiang University(Natural Science Edition in Chinese and English)

基　　金：国家自然科学基金“荒漠盐生植物盐穗木两个microRNA与靶标基因的耐盐抗旱功能研究”(31760071).

摘　　要：盐穗木(Halostachys caspica)是耐盐性极强的苋科藜属盐生植物.miRNA是长度为21∼24 nt的内源性非编码小RNA,通常在转录后水平负调控靶基因的表达介导多种生物学过程.从盐穗木高盐胁迫下根的小RNA文库和转录组数据中,克隆盐穗木miR397a(HcmiR397a)前体序列和漆酶HcLACs(Laccase,LAC)为预测的靶基因.通过实验的方法鉴定HcmiR397a与HcLACs的靶向关系及HcmiR397a在多种非生物胁迫下的表达和HcmiR397a的转基因拟南芥在低温胁迫下的功能.研究结果表明:(1)烟草瞬时表达体系和5’RLM-RACE技术确定HcmiR397a与HcLAC17之间存在靶向切割,具体的切割位点位于HcmiR397a的第10∼11碱基处;(2)qRT-PCR检测的结果显示盐、干旱和低温胁迫下,盐穗木同化枝HcmiR397a受到显著诱导;(3)异源表达HcmiR397a的转基因拟南芥表现出较强的低温耐性.Halostachys caspica is a halophyte belonging to Chenopodium of Amaranthaceae with extreme salt tolerance.miRNA is an endogenous small non-coding RNA with a length of 21»24 nt and participates in a variety of biological processes by negative regulating the expression of target genes at the post transcriptional level.The miR397a precursor were screened,cloned and LAC genes(Laccase,LAC)were predicted from the H.caspica root small RNA libraries under high salinity and this species’s transcriptome data.This paper aims to identify the targeting relationship between HcmiR397a and HcLACs by experimental methods,the expression of HcmiR397a in the H.caspica branches under various abiotic stresses and the function of HcmiR397a transgenic Arabidopsis under low temperature stress.The results were as follows:(1)Tobacco transient expression system and 5’RLM-RACE technology confirmed that there was a targeted cleavage between HcmiR397a and HcLAC17,and the specific cleavage site was located at the 10th»11th base of HcmiR397a;(2)The results of qRT-PCR showed that HcmiR397a was significantly induced under salt,drought and low temperature stress;(3)Transgenic Arabidopsis heterologously expressing HcmiR397a improved cold tolerance,compared to the wild type.

关 键 词：盐穗木miR397a(HcmiR397a) 转基因拟南芥 低温耐性 

分 类 号：Q946-33[生物学—植物学]