冬小麦miR172响应抗寒的特征分析

东农冬麦1号（Dn1）是我国首个能在黑龙江省高寒地区安全越冬的强抗寒冬小麦品种。为了解tae-miR172在Dn1抗寒中的调控机制,本研究克隆了Dn1中tae-miR172的前体pre-tae-miR172,根据生物信息学分析预测其靶基因为AP2dn1(TraesCS5D02G486600)。通过农杆菌介导的烟草瞬时表达试验,进一步证明tae-miR172靶向AP2dn1;利用qRT-PCR技术对大田越冬期Dn1分蘖节中pre-tae-miR172和Ap2dn1的相对表达量进行分析。结果表明,pre-tae-miR172的表达量随温度降低呈先升后降的变化趋势,而靶基因Ap2dn1的表达量变化趋势则相反,说明tae-miR172负调控Ap2dn1的表达。

Analysis on the Characteristics of miR172 Responding to Cold Resistance in Winter Wheat

Dongnongdongmai 1(Dn1) is the first winter wheat variety in China that can survive the winter safely in the high and cold regions of Heilongjiang Province. In order to understand the regulatory mechanism of tae-miR172 in Dn1 cold resistance, the pre-tae-miR172 of tae-miR172 precursor in Dn1 was cloned, and AP2dn1(TraesCS5D02G486600) was predicted as the target gene according to bioinformatics analysis, The negative regulation of AP2dn1 was verified on transcription level by tae-miR172 through Agrobacterium mediated tobacco transient expression experiment. qRT-PCR was used to analyze the relative expression of pre-tae-miR172 and AP2dn1 in the tillering nodes of Dn1 during the overwintering period in the field. The results showed that the expression level of pre-tae-miR172 increased first and then decreased with decreasing temperature, while the expression level of Ap2dn1 showed the opposite trend, indicating that tae-miR172 negatively regulated the expression of Ap2dn1.