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东北农业大学培育的自主知识产权的冬小麦新品种‘东农冬麦1号’(‘dn1’)是强抗寒(可耐–30℃低温)栽培品种。本研究应用BLAST、UNAFOLD等生物信息学软件从冬小麦EST库中分析和预测抗寒相关miRNA及其靶基因,并利用qRT-PCR技术验证表达丰度高、差异显著的miRNA及其靶基因的表达特性。结果显示:生物信息学软件分析和预测到16条抗寒相关miRNAs,隶属于15个家族、对应137个靶基因;其中miR1118、miR1120、miR1128、miR1133和miR1139表达丰度高、差异显著,上述5个miRNA在–10℃时表达量显著下降、在–25℃时表达量显著升高,但是其相应靶基因在–10℃时表达量显著升高,在–25℃时显著下降。可见,上述5个miRNA及其靶基因响应低温胁迫,与‘dn1’的强抗寒性相关。本研究为冬小麦响应低温胁迫下miRNA调控的分子机制奠定基础。
Dongnong Dongmai 1 (’dn1’) with independent intellectual property rights cultivated by Northeast Agricultural University is a cultivar with strong cold resistance (tolerable -30 ℃ low temperature). In this study, bioinformatics software such as BLAST and UNAFOLD was used to analyze and predict cold-adapted miRNA and its target genes from winter wheat EST library. QRT-PCR was used to verify the expression of miRNA and its target genes with high abundance and significant difference characteristic. The results showed that 16 cold-resistant miRNAs were analyzed and predicted by bioinformatics software, belonging to 15 families, corresponding to 137 target genes. Among them, the expression levels of miR1118, miR1120, miR1128, miR1133 and miR1139 were high and the difference was significant. The expression of miRNAs decreased significantly at -10 ℃, but significantly increased at -25 ℃. However, the expression levels of miRNAs at -10 ℃ were significantly increased and significantly decreased at -25 ℃. It can be seen that the above five miRNAs and their target genes are responsive to low temperature stress and are related to the strong cold resistance of ’dn1’. This study laid the foundation for the molecular mechanism of winter wheat response to miRNAs under low temperature stress.