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

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-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ped- 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-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。

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.