摘要
为探究gma-miR1514b与靶基因GmNAC120之间的调控作用,利用生物信息学方法预测gma-miR1514b的靶基因及结合位点,构建野生型和突变型GmNAC120的瞬时转化烟草及双荧光素酶载体,将gma-miR1514b与野生型和突变型GmNAC120分别组合后共转化烟草叶片,检测GmNAC120的表达量、双荧光素酶活性及荧光强度。结果显示:成功构建野生型与突变型GmNAC120基因的表达载体;相对表达量结果表明,共转化gma-miR1514b与野生型GmNAC120烟草叶片中GmNAC120的表达量较低,而gma-miR1514b与突变型GmNAC120烟草叶片中GmNAC120基因的表达量较高;双荧光素酶活性及荧光表达结果表明,gma-miR1514b可以降低野生型GmNAC120的双荧光素酶活性和荧光强度,而对于突变型GmNAC120的双荧光素酶活性和荧光强度则没有抑制作用。研究表明gma-miR1514b能够靶向切割GmNAC120 mRNA,并负调控GmNAC120的表达。研究结果可为研究大豆gma-miR1514b调控的抗逆网络提供依据。
In order to study the regulatory effect between gma-miR1514b and target gene GmNAC120,the target gene and binding site of gma-miR1514b were predicted by bioinformatics methods.We constructed transient transformed tobacco and dual luciferase vector of wild-type and mutant GmNAC120 and transformed with Agrobacterium,combined gma-miR1514b with wild-type and mutant GmNAC120 and co-transformation tobacco leaves,and detect the expression level,dual luciferase activity and fluorescence intensity of GmNAC120.The results show that:The expression vector of wild-type and mutant GmNAC120 gene was successfully constructed,the relative expression results indicate that,lower expression of GmNAC120 in co-transformation gma-miR1514b and wild-type GmNAC120 tobacco leaves,however,the GmNAC120 gene expression was higher in gma-miR1514b and mutant GmNAC120 tobacco leaves.The dual luciferase activity and fluorescence expression results showed that,the gma-miR1514b could decrease the dual luciferase activity of and fluorescence intensity of wild-type GmNAC120,and not the inhibition for the dual luciferase activity and fluorescence intensity of the mutant GmNAC120.The results indicated that gma-miR1514b is able to target cleavage GmNAC120 mRNA and negatively regulate GmNAC120 expression.The results of this study provide a basis for studying the stress resistance network regulated by gma-miR1514b in soybean.
作者
王嘉玉
倪志勇
于月华
WANG Jiayu;NI Zhiyong;YU Yuehua(Agricultural College,Xinjiang Agricultural University,Urumqi 830052,Xinjiang,China;College of Life Science,Xinjiang Agricultural University/Xinjiang Key Laboratory for Ecological Adaptation and Evolution of Extreme Environment Biology,Urumqi 830052,Xinjiang,China)
出处
《大豆科学》
北大核心
2025年第3期61-67,共7页
Soybean Science
基金
新疆维吾尔自治区自然科学基金(2022D01E14)
天山英才培养计划(2023TSYCCX0012)
国家自然科学基金(32360454)。
