花椰菜查尔酮合酶基因的克隆及功能分析  

Cloning and Functional Analysis of Sclerotinia sclerotiorum Resistance Related Gene CHS in Cauliflower

作　　者：赵风治 马钰婕 韦韬[1] 江汉民[2] 王春国[1] 宋文芹[1] 孙德岭[2] 陈成彬[1] 

机构地区：[1]南开大学生命科学学院,天津300071 [2]天津科润蔬菜研究所,天津300384

出　　处：《南开大学学报（自然科学版）》2016年第4期61-66,共6页Acta Scientiarum Naturalium Universitatis Nankaiensis

基　　金：天津市自然科学基金重点项目(13JCZDJC29000);天津市应用基础与前沿技术研究计划一般项目(14JCYBJC31000)

摘　　要：基于同源序列克隆技术,从花椰菜中克隆了查尔酮合酶(chalcone synthase;CHS)基因cDNA全长,将该基因成功转入花椰菜,经过Basta抗性筛选,PCR鉴定,得到7株花椰菜转基因过表达阳性植株.转基因花椰菜的核盘菌胁迫分析显示,与对照花椰菜相比,在病原胁迫的不同时期,转基因植株中BoCHS基因的表达量显著上升,且随着胁迫时间的增加,该基因表达量均增高,且在36 h时达到最大值,表明转基因花椰菜通过过量表达CHS来增强自身对菌核病的抗性,使植株对菌核病的抗性明显增强.与查尔酮合酶在油菜、向日葵、烟草等中的抗病作用一致,表明花椰菜BoCHS基因也是抗菌核病的重要基因.Based on homologous cloning, we isolated CHS gene from Brassica, over-expressed the gene in Brassica to investigate the function of the gene in resistance to S.sclerotiorum. The full-length c DNA sequence of CHS gene was isolated and cloned from Brassica primarily. We construct plant expression vector p C3301-Bo CHS successfully, and then transformated Brassica using Agrobacterium- mediated method. Following with screening with Basta and molecular identification,we obtained positive transgenic plants for next study. Analysis of expression changes of the gene were carried out by q RT-PCR. Results showed that transgenic Brassica over-expressing Bo CHS markedly enhanced resistance to S.sclerotiorum compared with wild type. These results are consistent with the resistant function of CHS in rapeseed, sunflower and tobacco, revealed that Bo CHS is also important disease resistance genes.

关 键 词：花椰菜 菌核病 查尔酮合酶 转基因 功能验证 

分 类 号：Q943.2[生物学—植物学]