毛竹miR398和miR408的克隆及其表达分析

Cloning and Expression Analysis of miR398 and miR408 in Phyllostachys edulis

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-MIR398和ped-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-miR398和ped-miR408)均位于5′端臂上。ped-miR398和ped-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-miR398与pedmiR408的表达量均上调;CuSO_4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-miR398与ped-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。

To investigate the expression patterns of miR398and miR408in Phyllostachys edulis,the precursor sequences of ped-MIR398and ped-MIR408were 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-MIR398and ped-MIR408were83bp and92bp,respectively,which both could fold into stable stem-loop structures,and the mature sequences(ped-miR398and ped-miR408)were generated at5′end of the arm in the stem-loop structures,respectively.The qPCR results indicated that both ped-miR398and ped-miR408were constitutively expressed,among which was most abundant in leaf blades.The expression of ped-miR398and ped-miR408were both up-regulated by the treatments of high light intensity,sucrose and GA3,and they were both down-regulated by the treatments of CuSO4and ABA.Under the treatments of darkness,NaCl and4℃,the expression of ped-miR398was up-regulated,and that of ped-miR408was down-regulated.Therefore,it was suggested that ped-miR398and ped-miR408might play different regulatory roles in the process of abiotic stress adaptation in bamboo.