木薯MeTPP1基因克隆、结构变异及其表达分析

doi:10.13560/j.cnki.biotech.bull.1985.2017-0714

生物技术通报 ›› 2018, Vol. 34 ›› Issue (1): 97-103.doi: 10.13560/j.cnki.biotech.bull.1985.2017-0714

木薯MeTPP1基因克隆、结构变异及其表达分析

丁泽红, 铁韦韦, 付莉莉, 胡伟

中国热带农业科学院热带生物技术研究所,海口 571101

收稿日期:2017-08-29 出版日期:2018-01-26 发布日期:2018-01-22
作者简介:丁泽红,男,副研究员,研究方向：植物分子生物学;E-mail：dingzehong@itbb.org.cn
基金资助:
国家自然科学基金项目（31600198）,中央级公益性科研院所基本科研业务费专项资金资助项目（1630052016012）

Gene Cloning,Structure Variation,and Expression Analysis of MeTPP1 in Cassava

DING Ze-hong, TIE Wei-wei, FU Li-li, HU Wei

Institute of Tropical Bioscience and Biotechnology,Chinese Academy of Tropical Agricultural Science,Haikou 571101

Received:2017-08-29 Published:2018-01-26 Online:2018-01-22

摘要/Abstract

摘要： 旨在研究木薯MeTPP1基因在干旱、低温等非生物胁迫响应中的作用。用同源基因克隆的方法从木薯叶片中克隆MeTPP1基因,用MEGA软件构建Neighbor-joining系统进化树,用DnaSP软件分析基因结构变异,用实时荧光定量PCR技术分析MeTPP1基因在不同胁迫处理下的表达特性。结果表明,MeTPP1基因含有一个1 131 bp的开放阅读框,编码376个氨基酸,含有TPP家族保守结构域。系统进化树分析表明,MeTPP1与杨树和杞柳中同源基因的亲缘关系较近,序列相似性分别为77.8%和74.5%。基因结构变异发现,MeTPP1在木薯野生种和栽培种之间共有9个错义突变,它们可能与MeTPP1的表达有关。实时荧光定量PCR分析表明,MeTPP1表达量受到干旱、低温和ABA处理的响应。上述结果表明,MeTPP1在转录水平参与ABA介导的木薯干旱和低温胁迫,可作为候选基因进一步研究其在木薯抗逆中的功能。

关键词: 海藻糖-6-磷酸酯酶, MeTPP1, 干旱, 低温, 表达分析

Abstract: This work is to reveal the roles of MeTPP1 gene in abiotic stresses such as drought and cold in cassava. Homology-based cloning method was used to clone MeTPP1 gene from cassava leaves,MEGA software to construct its neighbor-joining phylogenetic tree,DnaSP software to analyze its structural variations,and quantitative RT-PCR（qRT-PCR）to explore its expression characteristics under different abiotic treatments. The results showed that gene,MeTPP1 had a 1 131 bp open reading frame encoding 376 amino acids,and contained a conserved domain of TPP gene family. Phylogenetic analysis revealed that MeTPP1 had close genetic relationship to its homologues from Populus trichocarpa and Salix purpurea,and the sequence similarity was 77.8% and 74.5%,respectively. Genetic structural variation showed that a total of nine mis-sense mutations,which might be related to the expression of MeTPP1,were identified between cassava wild and cultivated species. qRT-PCR analysis demonstrated that the expression of MeTPP1 significantly changed in response to drought,cold,and ABA treatments. Together,these results indicate that MeTPP1 is involved in ABA-mediated drought and cold stresses of cassava at the transcriptional level,and can be served as a candidate to further study its functions in resistance of abiotic stress in cassava.

Key words: trehalose-6-phosphate, phosphatase,MeTPP1,drought,cold,expression, analysis

丁泽红, 铁韦韦, 付莉莉, 胡伟. 木薯MeTPP1基因克隆、结构变异及其表达分析[J]. 生物技术通报, 2018, 34(1): 97-103.

DING Ze-hong, TIE Wei-wei, FU Li-li, HU Wei. Gene Cloning,Structure Variation,and Expression Analysis of MeTPP1 in Cassava[J]. Biotechnology Bulletin, 2018, 34(1): 97-103.

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木薯MeTPP1基因克隆、结构变异及其表达分析

Gene Cloning,Structure Variation,and Expression Analysis of MeTPP1 in Cassava

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