甘蓝型油菜BnMYC2基因克隆及对植物抗虫胁迫的研究  

Study of BnMYC2 Cloning and Resistance to Insect Stress

作　　者：姜茜 杨瑶 郭世星[1,2] 吴永成 李壮[1,2] JIANG Xi;YANG Yao;GUO Shixing;WU Yongcheng;LI Zhuang(College of Agronomy, Sichuan Agricultural University, Chengdu 611130, China;2Rapeseed Research Center, Sichuan Agricultural University, Chengdu 611130, China)

机构地区：[1]四川农业大学农学院,成都611130 [2]四川农业大学油菜研究中心,成都611130

出　　处：《西北植物学报》2022年第2期201-209,共9页Acta Botanica Boreali-Occidentalia Sinica

基　　金：四川农业大学科研兴趣项目(2020050)。

摘　　要：为了研究髓细胞组织增生蛋白(Myelocytomatosis protein,MYC2)基因与甘蓝型油菜抗虫性的关系,该研究从甘蓝型油菜恢复系R18中克隆获得BnMYC2基因的cDNA序列,并进行了基因表达及转化模式植物拟南芥的抗虫分析。生物信息学分析表明,BnMYC2基因gDNA不具有内含子结构,cDNA完整开放阅读框(ORF)为1833 bp,编码610个氨基酸,推测BnMYC2基因位于甘蓝型油菜C6染色体上,具有bHLH型转录因子的保守结构域HLH。基因同源性分析结果表明,BnMYC2与拟南芥、甘蓝、萝卜的亲缘关系最近,氨基酸的相似度都在98%以上。组织特异性表达分析显示,BnMYC2在甘蓝型油菜的根、茎、叶、花、授粉后27 d的果荚和授粉后35 d的果荚中均有表达,且授粉后27 d的果荚中的表达水平最高。抗虫胁迫实验的结果显示,过表达BnMYC2拟南芥的抗虫基因VSP2的表达水平显著高于对照,推测甘蓝型油菜BnMYC2基因可能作为重要的调节因子参与虫害胁迫响应;MeJA诱导6 h后,VSP2基因在不同基因型拟南芥的表达量都明显增加,与诱导前相比差异显著,表明BnMYC2正向调控VSP2基因的表达,推测BnMYC2通过调控包括VSP2在内的多个标记基因的表达使植物响应虫害胁迫。In order to study the relationship between myelocytosis protein(MYC2)gene and insect resistance of Brassica napus,we cloned the cDNA sequence of BnMYC2 gene from B.napus restorer line R18,and analyzed the gene expression and insect resistance of transformation model plant Arabidopsis thaliana.Bioinformatics analysis showed that BnMYC2 gene gDNA lack of intron structure.The cDNA complete open reading frame(ORF)was 1833 bp,encoding 610 amino acids.The putative gene was located on the C6 chromosome of B.napus and had the conserved domain HLH of bHLH transcription factor.The results of gene homology analysis showed that BnMYC2 had the closest genetic relationship with Arabidopsis,cabbage and radish,and the similarity of amino acids was more than 98%.Tissue specific expression analysis showed that BnMYC2 was expressed in all tested tissues including roots,stems,leaves,flowers,pods 27 days after pollination and pods 35 days after pollination,and the expression level was the highest in pods 27 days after pollination.The results of insect resistance experiment showed that the expression level of insect resistance gene VSP2 in overexpressing BnMYC2 Arabidopsis was significantly higher than that in the control.It is speculated that BnMYC2 gene in B.napus may participate in the response to insect stress as an important regulator.After 6 hours of MeJA induction,the expression of VSP2 gene in different genotypes of Arabidopsis was significantly higher than that before induction,indicating that BnMYC2 positively regulates the expression of VSP2 gene.It is s-peculated that BnMYC2 can make plants respond to insect stress by regulating the expression of multiple marker genes including VSP2.

关 键 词：甘蓝型油菜 克隆 茉莉酸 表达分析 抗虫 

分 类 号：Q785[生物学—分子生物学] Q786