摘要
以高抗菌核病的油菜品种湘油15和高感菌核病的油菜品种98C40为材料,采用RT–PCR方法克隆BnPMEI1基因的编码区全长,利用生物信息学软件和实时荧光定量PCR分析BnPMEI1的结构和表达特性,并构建该基因超表达载体,分析该基因的功能。序列分析结果表明,该基因编码区全长615bp,编码204个氨基酸,具有1个信号肽和1个PMEI结构域,属于PMEI家族基因,命名为BnPMEI1(GenBank登录号为ON254917)。表达分析结果表明,BnPMEI1在根、茎、花、叶、角果皮和籽粒中均有表达,在叶中的表达量最高,根中的表达量最低;湘油15的茎、叶、花中的表达量均显著高于98C40的。在核盘菌、茉莉酸甲酯(Me JA)和水杨酸(SA)处理下,BnPMEI1在湘油15中的表达上调,在98C40中的表达则受到抑制;乙烯处理下,BnPMEI1在2个品种中的表达均下调,在湘油15中下调的幅度更大;离体叶片接种结果显示,超表达Bn PMEI1的油菜突变体叶片病斑明显小于野生型叶片的。综合分析,BnPMEI1基因可能介导了SA和茉莉酸途径的调控,诱导油菜菌核病抗性的提高。
To understand the expression characters of BnPMEI1 gene in Brassica napus we cloned the gene from Xianyou 15 with the high resistance to Sclerotinia sclerotiorum and 98C40 with high sensitivity to S. sclerotiorum by PCR. Then the sequences were subjected to bioinformatics anlysis and expression in bacterial host. Sequence analysis showed that the coding region of the gene was full-length 615 bp, encoding 204 amino acids, with a signal peptide and a PMEI domain, belonging to the PMEI family gene, named BnPMEI1(GenBank accession number is ON254917).Expression analysis showed that BnPMEI1 was expressed in root, stem, flower, leaf, pod skin and grain, but the expression was the highest in leaf and the lowest in root, and the expression in stem, leaf and flower of Xiangyou15 were significantly higher than those of 98C40. Under the treatment of S. sclerotiorum, methyl jasmonate(MeJA) and salicylic acid(SA), the expression of BnPMEI1 in Xiangyou15 was up-regulated, while that in 98C40 was inhibited;under ethylene treatment, the expression of BnPMEI1 in both varieties were down-regulated, and the down-regulation was more significant in Xiangyou15. The results of inoculation in vitro showed that the disease spot of rape mutants overexpressing BnPMEI1 was significantly smaller than that of wild-type leaves. Comprehensive analysis showed that BnPMEI1 gene might be regulated by SA and jasmonic acid pathway, which induced the resistance of B. napus to S. sclerotiorum.
作者
王峰
刘木兰
马肖
张秋平
WANG Feng;LIU Mulan;MA Xiao;ZHANG Qiuping(College of Agronomy,Hunan Agricultural University,Changsha,Hunan 410128,China)
出处
《湖南农业大学学报（自然科学版）》
CAS
CSCD
北大核心
2023年第1期25-34,共10页
Journal of Hunan Agricultural University(Natural Sciences)
基金
国家自然科学基金项目(31601339)
湖南省自然科学基金面上项目(2021JJ30352)
湖南省教育厅项目(16C0780)。
