木薯MeLHCB4基因的克隆及表达分析  

Cloning and Expression Analysis of MeLHCB4 from Cassava

作　　者：丁凯旋 郑婉茹 李琳琳 潘月云 张银东[1,2] 耿梦婷 陈银华[1,2] DING Kaixuan;ZHENG Wanru;LI Linlin;PAN Yueyun;ZHANG Yindong;GENG Mengting;CHEN Yinhua(College of Tropical Crops,Hainan University,Haikou,Hainan 570228,China;Hainan Key Laboratory for Sustainable Utiliza-tion of Tropical Bioresources,Haikou,Hainan 570228,China)

机构地区：[1]海南大学热带作物学院,海南海口570228 [2]海南省热带生物资源可持续利用国家重点实验室培育基地,海南海口570228

出　　处：《热带作物学报》2021年第10期2813-2818,共6页Chinese Journal of Tropical Crops

基　　金：海南省重点研发计划项目(No.ZDYF2019063);海南省研究生创新科研课题(No.Hys2019-133)。

摘　　要：本研究采用RT-PCR技术克隆了木薯叶绿素a/b结合蛋白基因MeLHCB4编码区序列。通过生物信息学对其基因结构、基因编码蛋白的理化性质等进行分析,并对不同物种的LHCB4氨基酸序列进行比对和构建进化树。结果表明,木薯MeLHCB4基因的CDs序列全长858 bp,编码285个氨基酸,蛋白理论相对分子质量约为30.9 kDa,理论等电点为5.47,该蛋白属于稳定的亲水性蛋白,预测该蛋白可能定位于细胞核或细胞质中。实时荧光定量PCR检测该基因的表达模式发现,MeLHCB4基因主要在木薯叶片和茎中表达,受到茉莉酸甲酯(JA)、水杨酸(SA)和乙烯前体(ACC)等激素的诱导表达,推测其可能参与了JA、SA、ACC信号途径。细菌性枯萎病病原菌侵染木薯叶片12 h后,MeLHCB4的表达量显著提高,表明MeLHCB4参与了木薯对病原菌的响应过程。In this study,the MeLHCB4 coding region of cassava chlorophyll a-binding b binding protein gene was cloned by the RT-PCR technique.The structure of the gene and the physical and chemical properties of the protein encoded by the gene were analyzed by bioinformatics,and the LHCB4 amino acid sequences of different species were compared and the evolutionary tree was constructed.The results showed that the CDs sequence of cassava MeLHCB4 was 858 BP,encoding 285 amino acids,the theoretical molecular weight of the protein was about 30.9 kDa,and the theoretical isoelectric point was 5.47.The protein was a stable hydrophilic protein,and it was predicted that the protein might be located in the nucleus and cytoplasm.The expression pattern of MeLHCB4 was detected by real-time fluorescence quantitative PCR.It was found that MeLHCB4 was mainly expressed in the leaves and stems of cassava,and was induced by hormones such as JA,SA and ACC,which was speculated to be involved in JA,SA,ACC signal pathway.After 12 hours of Xanthomonas axonopodis pv.manihotis infecting cassava leaves,the expression of MeLHCB4 increased significantly,indicating that MeLHCB4 was involved in the response process of cassava to pathogens.

关 键 词：木薯 叶绿素结合蛋白 MeLHCB4 基因克隆 基因表达 

分 类 号：S533[农业科学—作物学]