甘蓝型油菜BnMYC2的克隆及对植物抗虫胁迫的研究
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四川农业大学

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四川省育种攻关项目“突破性油料育种材料和方法创新及新品种选育”(2021YFYZ0018)


Study of BnMYC2 Cloning and Resistance to Insect StressJIANG Qian1#, YANG Yao1#, GOU Qixian1, ZHI Shouchuan1, LENG Junjiao1, LI Yilin1, LI Zhuang1,2*
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    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之间的关系,从甘蓝型油菜恢复系R18中克隆获得BnMYC2基因的cDNA序列,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、开花后27 d的果荚和开花后35 d的果荚中均有表达,且开花后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。

    Abstract:

    In order to study the relationship between myelocytosis protein (MYC2) gene and insect resistance of Brassica napus, the cDNA sequence of BnMYC2 gene was cloned from Brassica napus restorer line R18, and the gene expression and insect resistance of transformation model plant Arabidopsis thaliana were analyzed. 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 Brassica 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 flowering and pods 35 days after flowering, and the expression level was the highest in pods 27 days after flowering. 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 Brassica napus may participate in the response to insect stress as an important regulator.

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  • 收稿日期:2021-09-24
  • 最后修改日期:2022-01-04
  • 录用日期:2022-01-05
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