冬小麦东农冬麦1号miR5049-3p和miR1120a前体与靶基因的克隆及低温和ABA作用下的表达调控  

Cloning of Pre-miR5049-3p,Pre-miR1120a and Their Target Genes and Expression Analysis under Low Temperature and ABA in Winter Wheat Dongnongdongmai 1

作　　者：彭瞰看 田宇[1] 杨宁[1] 吕岩[1] 赵虎 宋春华 李海丹 徐庆华[1] 苍晶[1] PENG Kankan;TIAN Yu;YANG Ning;LV Yan;ZHAO Hu;SONG Chunhua;LI Haidan;XU Qinghua;CANG Jing(Laboratory of Plant Physiology and Molecular Biology,School of Life Sciences,Northeast Agricultural University,Heilongjiang 150030,China)

机构地区：[1]东北农业大学生命科学学院植物生理与分子生物学研究室,黑龙江哈尔滨150030

出　　处：《麦类作物学报》2018年第6期641-648,共8页Journal of Triticeae Crops

基　　金：国家自然科学基金项目(31471423;31601236;31701348)

摘　　要：为研究外源ABA对miRNA响应低温胁迫的影响,利用实验室已有的冬小麦品种东农冬麦1号miRNA库,通过生物信息学手段筛选出了与糖代谢相关差异表达的miR5049-3p和miR1120a,预测出其靶基因分别为6PGL(6-磷酸葡糖酸内酯酶基因)和FBA(果糖-1,6-二磷酸醛缩酶基因)。生物信息学分析表明,两种miRNA的前体序列均能形成稳定的茎环结构,但其成熟体的碱基保守性较差,它们的启动子序列包含多种响应环境因子的顺式作用元件。经qRT-PCR分析,随着温度的降低,miR5049-3p和miR1120a分别与其靶基因呈负相关调控;外源ABA处理后,随着温度的降低,miR5049-3p和miR1120a的表达量皆呈"升-降-升"的趋势,其靶基因表达则相应明显降低,表明mi5049-3p和miR1120a可能分别负向调控靶基因6PGL和FBA,进而介导糖代谢途径来响应低温胁迫。At low temperature,exogenous ABA can regulate the expression of wheat miRNAs,and then mediate the glucose metabolism pathway in response to low temperature stress.In order to confirm this speculation,we screened out the differential expression of miRNAs,miR5049-3 p and miR1120 a,related to glycometabolism with the miRNA library of Dongnongdongmai 1 by bioinformatics,then predicted 6 PGL（6-phosphogluconolactone enzyme）and FBA（fructose-1,6-bisphosphate aldolase）as their target genes.The results showed the sequences of both pre-miR5049-3 p and premiR1120 acan form stable stem ring structure.However,the mature miR5049-3 p and miR1120 asequences have low base sequence conservatism.Their promoter sequence contains kinds of cis-elements in response to environmental factors,such as light,ABA,and temperature.The results of qRT-PCR showed that with the decrease of temperature,the expression levels of miR5049-3 p and miR1120 a were negatively correlated with those of their target genes,respectively.While after treatment with ex-ogenous ABA,the expression of miR5049-3 pand miR1120 aappeared a change of＂rise-drop-rise＂with the decrease of temperature,and the expression of target genes decreased obviously.These indicated t miR5049-3 pand miR1120 a may negatively regulate their target genes,6 PGLand FBA,to mediate glycometabolism pathway in response to low temperature stress.

关 键 词：冬小麦 MIRNAS 低温 ABA 糖代谢 

分 类 号：S512.1[农业科学—作物学] S330