马铃薯miR397的克隆及靶基因筛选

miR397广泛参与植物逆境胁迫响应。本研究从马铃薯叶片中分离出miR397的前体序列,克隆得到StmiR397基因,对潜在靶基因进行预测,采用实时荧光定量PCR（qRT-PCR）方法对低温不同表现型材料进行表达模式分析。结果表明：马铃薯miR397前体序列为81 bp,能形成稳定的茎环结构,成熟序列位于5′端,上游调控区含有温度响应等多种逆境应答作用元件。低温处理下,StmiR397在耐低温材料DR-2叶中上调表达,根中表达无差异,而在低温敏感材料费乌瑞它叶中表达无差异,根中上调表达。通过对预测的10个靶基因进行表达模式分析,初步筛选出反向抑制靶基因氧化还原酶结构域蛋白、锌指家族蛋白和正向激活靶基因磷酸果糖激酶家族蛋白、生长素诱导的β-葡萄糖苷酶,为进一步研究StmiR397与靶基因的调控网络提供了一定参考。

Cloning and target gene screening of miR397 in potato

miR397 is widely involved in plant stress response. In this study, the precursor sequence of miR397 was isolated from potato leaves, the StmiR397 was cloned and its potential target genes were predicted. The qRT-PCR was used to analyze the expression patterns of different low-temperature phenotypic materials, the results showed: the length of precursor sequence of potato miR397 was 81 bp, which could form stable stem-loop structure, and the mature sequence was located at the 5' end. The upstream regulatory region contained various stress response elements such as temperature response. Under low temperature treatment, the expression of StmiR397 was up-regulated in the leaves of the low-temperature resistant material DR-2, and the expression showed no difference in the roots, while the expression of StmiR397 showed no difference in the leaves of the low-temperature sensitive material Favorite, and the expression was up-regulated in the roots. According to the expression patterns analyses of the 10 predicted target genes, oxidoreductase-like domain-containing protein gene, zinc knuckle (CCHC-type) family protein gene of the reverse inhibitory target genes and the positive activation target genes 6-phosphate 1-phosphotransferase subunit alpha gene and auxin-induced beta-glucosidase gene were initially screened. This study provides a certain reference for further research on the regulatory network of StmiR397 and target genes.