中国农业科技导报 ›› 2020, Vol. 22 ›› Issue (12): 10-19.DOI: 10.13304/j.nykjdb.2019.0287

• 生物技术 生命科学 • 上一篇    下一篇

盐胁迫下水稻体内miRNA表达谱分析

张秀妍1,2,徐妙云2,郑红艳2,邹俊杰2,张笑宇1*,王磊2*   

  1. 1.内蒙古农业大学园艺与植物保护学院, 呼和浩特 010018;
    2.中国农业科学院生物技术研究所, 北京 100081
  • 收稿日期:2019-04-11 出版日期:2020-12-15 发布日期:2019-05-24
  • 通讯作者: 张笑宇 E-mail:zxy2000@126.com;王磊 E-mail:wanglei01@caas.cn
  • 作者简介:张秀妍 E-mail:13384756699@163.com
  • 基金资助:
    国家转基因生物新品种培育科技重大专项(2018ZX0801103B

Analysis of miRNA Expression Profiles in Rice Under Salt Stress

ZHANG Xiuyan1,2, XU Miaoyun2, ZHENG Hongyan2, ZOU Junjie2, ZHANG Xiaoyu1*, WANG Lei2*   

  1. 1.College of Horticulture and Plant Protection, Inner Mongolia Agricultural University, Huhhot 010018, China;
    2.Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, China
  • Received:2019-04-11 Online:2020-12-15 Published:2019-05-24

摘要: miRNA是生物体内起重要调控作用的一类非编码小RNA分子,在转录后水平调控基因表达,参与生长发育、抗逆等生物学过程。高通量测序法是鉴定植物中miRNA最有效的方法之一。提取盐胁迫处理不同时间的水稻根、茎和叶部的总RNA,从中分离18~30 nt小分子RNA构建文库,进行高通量测序,对miRNA表达谱特性进行了分析。结果表明,所有样品共检测到248个已知的miRNA,来源于132个不同的家族,根、茎、叶中分别有31、11、9个差异表达miRNA,分属于29个家族,靶向592个功能基因,并对其中5个差异显著且与非生物胁迫相关的重要miRNA进行了qPCR分析检测,结果显示Osa-miR166hOsa-miR166k上调表达,Osa-miR169hOsa-miR530下调表达,针对Osa-miR169h的4个靶基因进一步定量检测,其中的3个靶基因上调表达,这与Osa-miR169h的下调表达结果相一致,以上结果说明miRNA在植物抵御非生物逆境胁迫的过程中扮演重要角色。

关键词: miRNA, 表达谱, 盐胁迫, 水稻

Abstract: miRNA is a kind of non-coding small RNA which plays an important regulatory role in vivo. miRNA mainly regulates gene expression at the post-transcriptional level and involves in various biological processes such as development and stress tolerance. High-throughput sequencing is one of the most effective methods for the identification of miRNA in plants. The total RNA was extracted from roots, stems and leaves from rice treated with high salt stress. Small RNAs were isolated to construct sRNA libraries. The libraries were sequenced using high-throughput method, and the miRNA expression profiles were analyzed. The results showed that 248 known miRNA from 132 different families were detected in all samples. Totally, 31 differentially expressed miRNAs were identified in root, 11 in stem and 9 in leaf, which belonged to 29 miRNA families, targeting 592 functional genes. Five differentially expressed miRNAs, responding stress tolerance, were further analyzed. The results showed that Osa-miR166h and miR166k were up-regulated, and miR169h and miR530 were down-regulated. The miR169 target genes were investigated using qPCR, and three of four target genes were up-regulated with miR169 down-regulation. Above results indicated that miRNA played an important role in plants stress tolerance.

Key words: miRNA, expression profile, salt stress, rice