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毛竹miR398和miR408的克隆及其表达分析
李利超,孙化雨,杨意宏,赵韩生,王思宁,高志民
国际竹藤中心,国际竹藤中心,国际竹藤中心,国际竹藤中心,国际竹藤中心,国际竹藤中心
摘要:  为了解毛竹(Phyllostachys edulis)中miR398和miR408的表达情况,从毛竹叶片中分离了二者的前体序列,并用实时定量PCR技术对其表达模式进行了研究。结果表明,毛竹中miR398和miR408前体序列ped-MIR398ped-MIR408长度分别为83 bp和92 bp,二者均能形成稳定的茎环结构,其中成熟miRNA序列(ped-miR398ped-miR408)均位于5'端臂上。ped-miR398ped-miR408均为组成型表达,在毛竹叶中表达量最高。强光、蔗糖和GA3处理后,叶片中ped-miR398ped- miR408的表达量均上调;CuSO4和ABA处理后,叶片中二者的表达量均下调;黑暗、NaCl和4℃处理后,前者表达量上调,后者表达量下调。因此,ped-miR398ped-miR408在毛竹适应逆境胁迫过程中可能发挥着不同的调控作用。
关键词:  毛竹  miRNA  非生物胁迫  基因表达
DOI:10.11926/jtsb.3697
分类号:
基金项目:“十二五”农村领域国家科技计划项目(2015BAD04B01)资助
Cloning and Expression Analysis of miR398 and miR408 in Phyllostachys edulis
LI Li-chao,SUN Hua-yu,YANG Yi-hong,ZHAO Han-sheng,WANG Si-ning and GAO Zhi-min
International Center for Bamboo and Rattan,Key Laboratory on the Science and Technology of Bamboo and Rattan,International Center for Bamboo and Rattan,Key Laboratory on the Science and Technology of Bamboo and Rattan,International Center for Bamboo and Rattan,Key Laboratory on the Science and Technology of Bamboo and Rattan,International Center for Bamboo and Rattan,Key Laboratory on the Science and Technology of Bamboo and Rattan,International Center for Bamboo and Rattan,Key Laboratory on the Science and Technology of Bamboo and Rattan,International Center for Bamboo and Rattan,Key Laboratory on the Science and Technology of Bamboo and Rattan
Abstract:  To investigate the expression patterns of miR398 and miR408 in Phyllostachys edulis, the precursor sequences of ped-MIR398 and ped-MIR408 were isolated, and the expression of miRNAs was analyzed by real-time quantitative PCR (qPCR). The results showed that the length of precursor sequences of ped-MIR398 and ped-MIR408 were 83 bp and 92 bp, respectively, which both could fold into stable stem-loop structures, and the mature sequences (ped-miR398 and ped-miR408) were generated at 5' end of the arm in the stem-loop structures, respectively. The qPCR results indicated that both ped-miR398 and ped-miR408 were constitutively expressed, among which was most abundant in leaf blades. The expression of ped-miR398 and ped-miR408 were both up-regulated by the treatments of high light intensity, sucrose and GA3, and they were both down-regulated by the treatments of CuSO4 and ABA. Under the treatments of darkness, NaCl and 4℃, the expression of ped-miR398 was up-regulated, and that of ped-miR408 was down-regulated. Therefore, it was suggested that ped-miR398 and ped-miR408 might play different regulatory roles in the process of abiotic stress adaptation in bamboo.
Key words:  Phyllostachys edulis  miRNA  Abiotic stress  Gene expression

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