辣椒查尔酮合成酶基因CaCHS02的克隆及功能分析  

Cloning and Expression Analysis of the Chalcone Synthase Gene CaCHS02 in Pepper

作　　者：王小迪 李宁[1] 高升华 尹延旭[1] 徐凯 詹晓慧 姚明华[2,3] 王飞 WANG Xiaodi;LI Ning;GAO Shenghua;YIN Yanxu;XU Kai;ZHAN Xiaohui;YAO Minghua;WANG Fei(Hubei Provincial Key Laboratory of Vegetable Germplasm Innovation and Genetic Improvement,Economic Crops Research Institute of Hubei Academy of Agricultural Sciences,Wuhan 430064,China;Hubei Academy of Agricultural Sciences,Wuhan 430064,China;Hubei Hongshan Laboratory,Wuhan 430070,China)

机构地区：[1]湖北省农业科学院经济作物研究所,蔬菜种质创新与遗传改良湖北省重点实验室,武汉430064 [2]湖北省农业科学院,武汉430064 [3]湖北洪山实验室,武汉430070

出　　处：《辣椒杂志》2024年第3期1-10,共10页Journal of China Capsicum

基　　金：湖北省重点研发项目(2023BBB044);湖北省支持种业高质量发展资金项目(HBZY2023B004-4);武汉市产学研项目(2023110201030679);湖北省现代农业产业技术专项资金资助(2023HBSTX4-06);湖北省农业科学院青年科学基金项目(2024NKYJJO6);湖北省自然科学基金青年项目(2024AFB379);湖北省博士后创新实践岗位项目;湖北省重点研发项目(2022BBA0060)。

摘　　要：查尔酮合成酶(Chalcone synthase,CHS)基因在植物黄酮类物质代谢过程中发挥重要作用。为研究查尔酮合成酶基因CaCHS02在辣椒(Capsicum annuum L.)中辣椒类黄酮代谢过程中的功能,本研究分析了CaCHS02基因的基因特征、蛋白特点和基因表达模式,并构建了CaCHS02过表达载体,通过农杆菌介导法获得瞬时过表达CaCHS02(35S:CaCHS02)的辣椒植株。结果发现,瞬时过表达CaCHS02的辣椒植株叶片中CaCHS02发生显著超量表达,其中编码类黄酮代谢途径中其他关键酶基因(CaCHS02、CaPAL、CaC4H、Ca4CL、CaCHI、CaFLS和CaF3H)的表达水平同步显著上调;超表达CaCHS02促进辣椒叶片中查尔酮合成酶酶活性、总黄酮含量和辣椒叶片的α-葡糖糖苷酶抑制活性的提升。研究表明,CaCHS02在辣椒类黄酮代谢过程中发挥正向调控功能,过表达CaCHS02提高了辣椒叶片的α-葡糖糖苷酶抑制活性。本研究为解析辣椒α-葡萄糖苷酶抑制剂生物合成机制奠定了基础,为选育高α-葡糖糖苷酶抑制活性的功能辣椒品种提供了理论支持。Chalcone synthase genes play an important role in the metabolism of flavonoids in plants.In order to investigate the function of the Chalcone synthase gene CaCHS02 in the flavonoid metabolism process of pepper(Capsicum annuum L.),its gene characteristics,protein characteristics,and expression patterns were analyzed in this study.Further construction of CaCHS02 overexpression vector was carried out,and pepper plants with overexpression of CaCHS02(35S:CaCHS02)were obtained through agrobacterium-mediated transgenic method.RT-qPCR analysis revealed that CaCHS02 was significantly overexpressed in the leaves of transgenic plants and the expression levels of genes encoding other key enzymes in flavonoid metabolism pathways(CaCHS02,CaPAL,CaC4H,Ca4CL,CaCHI,CaFLS,and CaF3H)were also significantly upregulated.Meanwhile,assessments on chalcone synthase activity,total flavonoid content andα-glucosidase inhibitory activity(AGI activity)showed that overexpression of CaCHS02 significantly increased chalcone synthase activity and total flavonoid content in pepper leaves,further enhancing the AGI activity in pepper leaves.In summary,CaCHS02 plays a positive regulatory role in the metabolism of flavonoids in pepper and overexpression of CaCHS02 enhances the AGI activity in pepper leaves.This study laid the foundation for analyzing the biosynthesis mechanism ofα-glucosidase inhibitor in peppers and provided the theoretical support for the breeding of functional pepper with highα-glucosidase inhibition activity.

关 键 词：辣椒 查尔酮合成酶 CaCHS02 类黄酮 α-葡糖糖苷酶抑制活性 

分 类 号：S641.3[农业科学—蔬菜学]