木薯MePCS1基因的克隆、表达分析及功能验证  

Cloning,Expression Analysis,and Functional Verification of the MePCS1 Gene in Cassava

作　　者：梁宝娟 葛玉建 侯静怡 张慧敏 任治欣 耿梦婷 王亚杰 郭建春[2,3] 姚远 LIANG Baojuan;GE Yujian;HOU Jingyi;ZHANG Huimin;REN Zhixin;GENG Mengting;WANG Yajie;GUO Jianchun;YAO Yuan(College of Tropical Agriculture and Forestry,Hainan University,Haikou,Hainan 570228,China;Institute of Tropical Biosci-ence and Biotechnology,Chinese Academy of Tropical Agricultural Sciences/Key Laboratory of Biology and Genetic Resources of Tropical Crops,Ministry of Agriculture and Rural Affairs/Hainan Institute for Tropical Agricultural Resources/Key Laboratory for Biology and Genetic Resources of Tropical Crops of Hainan Province,Haikou,Hainan 571101,China;Sanya Research Institute,Chinese Academy of Tropical Agricultural Sciences/National key Laboratory for Tropical Crop Breeding,Sanya,Hainan 572000,China)

机构地区：[1]海南大学热带农林学院,海南海口570228 [2]中国热带农业科学院热带生物技术研究所/农业农村部热带作物生物学与遗传资源利用重点实验室/海南热带农业资源研究院/海南省热带农业生物资源保护与利用重点实验室,海南海口571101 [3]中国热带农业科学院三亚研究院/热带作物生物育种全国重点实验室,海南三亚572000

出　　处：《热带作物学报》2024年第11期2243-2250,共8页Chinese Journal of Tropical Crops

基　　金：海南省自然科学基金项目(No.320RC709,No.324QN318);国家自然科学基金项目(No.32301740)

摘　　要：植物络合素合酶(PCS)是催化植物络合素(PC)合成的关键酶,PC可通过络合作用减轻重金属对植物的毒害。土壤重金属污染增加木薯食用安全风险,对木薯络合素合酶基因MePCS1进行功能分析,对木薯重金属减控研究具有重要意义。本研究通过PCR技术克隆了SC8木薯品种的MePCS1基因的编码区,全长为1512 bp,共编码503个氨基酸。蛋白序列分析表明:MePCS1蛋白为亲水性蛋白,含有41个磷酸化位点和2个糖基化位点,不含有信号肽。构建系统进化树发现,木薯MePCS1蛋白与橡胶树的PCS蛋白亲缘关系最近。利用qPCR技术分析表明,MePCS1基因在茎中表达量最高;在木薯块根发育过程中,MePCS1基因在块根膨大期的表达量最高;铅(Pb)胁迫诱导MePCS1基因表达。MePCS1基因能提高BY4741酵母对Pb的耐受能力。本研究结果为进一步解析MePCS1基因功能特性及其对木薯的Pb减控机制提供新的信息。Phytochelatin synthase(PCS)is a key enzyme that catalyzes the synthesis of phytochelatin(PC).PC can re-duce the toxicity of heavy metals to plants through complexation.Heavy metal pollution in soil increases the risk of cassava food safety.It is of great significance to study the reduction and control of heavy metals in cassava by analyzing the function of MePCS1 gene.In this study,the coding region of MePCS1 gene in cassava variety SC8 was cloned by PCR.The full length was 1512 bp,encoding 503 amino acids.Protein sequence analysis showed that MePCS1 protein was a hydrophilic protein,containing 41 phosphorylation sites and 2 glycosylation sites,without signal peptide.The phylogenetic tree showed that the cassava MePCS1 protein had the closest relationship with the PCS protein of the rubber tree.The qPCR analysis showed that the MePCS1 gene had the highest expression level in the stem.During the development of cassava roots,the expression level of MePCS1 gene was the highest in the root enlargement period.Lead(Pb)stress induced MePCS1 gene expression.MePCS1 gene can improve the tolerance of BY4741 yeast to Pb.The re-sults of this study provide new information for further analysis of the functional characteristics of the MePCS1 gene and its mechanism for reducing Pb in cassava tuber roots.

关 键 词：植物络合素合酶 重金属Pb胁迫 酵母表达 

分 类 号：S533[农业科学—作物学]