木薯查尔酮合酶MeCHS基因家族特征与表达分析  

Characteristics and Expression Analysis Of Chalcone Synthase MeCHS Family in Cassava

作　　者：安飞飞[1,2] 齐剑雄 陈松笔 罗秀芹 蔡杰[2] 薛晶晶 AN Feifei;QI Jianxiong;CHEN Songbi;LUO Xiuqin;CAI Jie;XUE Jingjing(Tropical Crops Genetic Resources Institute,Chinese Academy of Tropical Agricultural Sciences/Key Laboratory of Conservation and Utilization of Cassava Germplasm Resources,Ministry of Agriculture and Rural Affairs,Haikou,Hainan 571101,China;Sanya Research Institute,Chinese Academy of Tropical Agricultural Sciences,Sanya,Hainan 572025,China)

机构地区：[1]中国热带农业科学院热带作物品种资源研究所/农业农村部木薯种质资源保护与利用重点实验室,海南海口571101 [2]中国热带农业科学院三亚研究院,海南三亚572025

出　　处：《热带作物学报》2023年第12期2384-2391,共8页Chinese Journal of Tropical Crops

基　　金：中央级公益性科研院所基本科研业务费专项(No.1630052023016);海南省自然科学基金项目(No.320MS100)。

摘　　要：查尔酮合酶(chalcone synthase,CHS)是类黄酮合成途径的第一个限速酶,在植物花色形成、生长发育以及非生物胁迫中发挥着重要作用。为明确木薯MeCHS基因家族的特性及在木薯块根采后腐烂(PPD)中的表达,本研究利用生物信息学方法对木薯MeCHS基因家族成员进行理化性质、蛋白质结构等分析。采用实时荧光定量PCR(qRT-PCR)检测MeCHS基因在不同品种、不同组织及块根PPD过程中的表达情况,同时克隆并构建3个高表达的MeCHS基因的亚细胞定位载体并进行烟草瞬时转化以确定其定位。在木薯基因组中共鉴定出5个MeCHS基因家族成员,蛋白质二级结构以α-螺旋和不规则卷曲为主。qRT-PCR分析发现:MeCHS基因表达具有明显的组织特异性,在茎中表达水平最高;在木薯中主要表达的MeCHS基因为MeCHS1、MeCHS2和MeCHS3,且3个基因随着SC8块根贮藏时间的延长表达量逐步升高。将成功克隆得到的MeCHS1、MeCHS2和MeCHS3基因经农杆菌介导瞬时转化烟草下表皮细胞,显微观察表明3个基因均定位于细胞质,与预测结果相符。该研究结果可为进一步揭示木薯MeCHS基因家族的功能与提高木薯块根耐PPD能力提供理论依据。Chalcone synthase(CHS)is the first limiting enzyme in flavonoid synthesis pathway and plays an important role in flower color formation,growth and development,and abiotic stress.In order to clarify the characteristics of cas-sava MeCHS gene family and its expression in cassava postharvest physiological deterioration(PPD),the MeCHS gene family was analyzed using bioinformatics,including physicochemical property and subcellular location.qRT-PCR was carried out to detect the expression levels of MeCHS genes in different varieties,different tissues and PPD process.The genes with significant relative expression were selected for cloning and subcellular localization verification.A total of 5 MeCHS gene members were identified in the cassava genome.The secondary structure analysis showed that the MeCHS gene family was mainly composed ofα-helices and random coils.Tissue specific expression indicated that the expres-sion levels of MeCHS genes were higher in stems.The major expressed MeCHS genes in cassava were MeCHS1,MeCHS2 and MeCHS3;and the expression levels of these three genes were gradually increase with PPD.MeCHS1,MeCHS2 and MeCHS3 were successfully cloned;and fluorescence transient expression showed that MeCHS1,MeCHS2 and MeCHS3 were localized in the cytoplasm,which was consistent with the predicted results.This study will provide a theoretical basis for further revealing the function of MeCHS gene family and improving the PPD tolerance of cassava.

关 键 词：木薯 查尔酮合酶 QRT-PCR 克隆 表达分析 

分 类 号：S533[农业科学—作物学] S435.33