[1]安飞飞,崔梦佳,杨 龙,等.木薯MeGalt1基因克隆、在采后生理腐烂过程中表达分析及载体构建[J].福建农林大学学报(自然科学版),2021,50(02):244-249.[doi:10.13323/j.cnki.j.fafu(nat.sci.).2021.02.014]
　AN Feifei,CUI Mengjia,YANG Long,et al.Cloning and expression of MeGalt1 and vector construction in cassava during physiological deterioration after harvesting[J].,2021,50(02):244-249.[doi:10.13323/j.cnki.j.fafu(nat.sci.).2021.02.014]
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木薯MeGalt1基因克隆、在采后生理腐烂过程中表达分析及载体构建()
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福建农林大学学报（自然科学版）[ISSN:1671-5470/CN:35-1255/S]

卷:
50卷
期数:
2021年02期
页码:
244-249
栏目:
生命科学
出版日期:
2021-02-15

文章信息/Info

Title:
Cloning and expression of MeGalt1 and vector construction in cassava during physiological deterioration after harvesting
文章编号:
1671-5470(2021)02-0244-06
作者:
安飞飞1;  崔梦佳2;  杨 龙3;  陈松笔1
1.中国热带农业科学院热带作物品种资源研究所/农业部木薯种质资源保护与利用重点实验室, 海南 海口 571101; 2.南京农业大学农学院,江苏 南京 210000; 3.贵州省亚热带作物研究所,贵州 兴义 562400
Author(s):
AN Feifei1;  CUI Mengjia2;  YANG Long3;  CHEN Songbi1
1.Tropical Crops Genetic Resources Institute/Key Laboratory of Ministry of Agriculture for Germplasm Resources Conservation and Utilization of Cassava, Chinese Academy of Tropical Agricultural Sciences, Haikou, Hainan 571101, China; 2.College of Agriculture, Nanjing Agricultural University, Nanjing, Jaingsu 210000, China; 3.Subtropical Crops Research Institute of Guizhou Province, Xingyi, Guizhou 562400, China
关键词:
木薯;  采后生理腐烂;  MeGalt1;  克隆;  表达分析
Keywords:
Manihot esculenta;  post-harvest physiological deterioration;  MeGalt1;  cloning;  expression analysis
分类号:
S533
DOI:
10.13323/j.cnki.j.fafu(nat.sci.).2021.02.014
文献标志码:
A
摘要:
β-1,3-半乳糖基转移酶(Galt)是形成糖蛋白中Lewis a结构的关键酶,从木薯块根中克隆MeGalt1基因,分析其在木薯块根不同采后腐烂程度中的表达,旨在揭示MeGalt1基因在木薯块根采后生理腐烂中的作用.进化分析表明,木薯中存在3个MeGalt1基因并成功克隆MeGalt1-1、MeGalt1-2和MeGalt1-3,分别具有1 890、1 893和1 902 bp的CDS区,编码629、630和633个氨基酸.Western Blot显示木薯块根采后生理腐烂过程受到蛋白质N-糖基化修饰调控.定量分析表明MeGalt1-1和MeGalt1-3随着腐烂程度加深其表达量也随之升高.在此基础上,成功构建MeGalt1基因的植物过表达载体pCAMBIA1300r-MeGalt1-1和pCAMBIA1300r-MeGalt1-3,为进一步解析MeGalt1在木薯块根采后生理腐烂中的分子作用机制提供材料.
Abstract:
β-1, 3-galactosyltransferase(Galt)is a key enzyme in the formation of Lewis a structure in glycoproteins. To reveal the role of MeGalt1 gene in the post-harvest physiological deterioration(PPD)of Manihot esculenta Crantz, the MeGalt1 gene was cloned from cassava tuberous roots. Then the expression of the MeGalt1 gene was characterized under different degrees of PPD. Evolution analysis showed that 3 MeGalt1 genes were existed in cassava, named MeGalt1-1, MeGalt1-2 and MeGalt1-3, and they were further successfully cloned from cassava tuberous roots. The sizes of the coding sequence regions of the 3 genes were 1 890, 1 893 and 1 902 bp, encoding 629, 630 and 633 amino acids, respectively. Western blotting showed that PPD was regulated by N-glycosylation. RT-PCR revealed that the expressions of MeGalt1-1 and MeGalt1-3 were increased with the severity of PPD. On this basis, the plant overexpression vectors pCAMBIA1300r-MeGalt1-1 and pCAMBIA1300r-MeGalt1-3 were successfully constructed, which provides materials for the further analysis of MeGalt1 gene in the PPD of cassava tuberous roots.

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备注/Memo

备注/Memo:
收稿日期:2020-05-27 修回日期:2020-09-16
基金项目:国家重点研发计划项目(2019YFD1000500); 中央级公益性科研院所基本科研业务费专项(1630032019009); 海南省自然科学基金项目(320MS100); 贵州省科技计划项目(黔科合支撑[2019]2358号).
作者简介:安飞飞(1983-),女.研究方向:木薯蛋白质组学.Email:aff85110@163.com.通信作者陈松笔(1966-),男,研究员,博士生导师.研究方向:木薯遗传育种; Email:songbichen@catas.cn.
更新日期/Last Update: 2021-02-15