低温诱导泡核桃中查尔酮合成酶基因克隆及功能分析  

Cloning and Functional Analysis of Chalcone Synthase Gene in Cold-induced Juglans sigillata

作　　者：王毅[1,2] 肖良俊[1] 马婷[1] 宁德鲁 

机构地区：[1]云南省林业科学院经济林研究所,云南省木本油料工程技术研究中心,昆明650201 [2]云南省林业科学院,云南省森林植物培育与开发利用重点实验室,国家林业局云南珍稀濒特森林植物繁育和保护重点实验室,昆明650201

出　　处：《分子植物育种》2018年第2期386-391,共6页Molecular Plant Breeding

基　　金：云南省林业科技创新项目(2014CX01); 国家科技支撑课题(2011BAD46B01)共同资助

摘　　要：查尔酮合成酶是植物中黄酮类物质生物合成的第一关键酶,在植物抗性生理中扮演重要作用。本研究利用RT-PCR技术从泡核桃中成功克隆获得一个受冷害诱导的查尔酮合成酶基因(Js CHS1),其基因全长1 490 bp,包含1个内含子,开放阅读框全长为1 170 bp,编码389个氨基酸,登录号为:KX657834。JSCHS1与核桃查尔酮合成酶蛋白序列同源性高达98%,而泡核桃和核桃查尔酮合成酶基因内含子DNA同源性为95%,核桃CHS内含子与泡核桃相比有8个碱基的缺失。系统进化树分析显示其与核桃形成一个独立分支。半定量PCR显示:泡核桃在常温及低温(4℃)处理后都有微弱表达,而在低温处理6 h后强烈表达,这说明Js CHS1是典型的受冷害诱导的查尔酮合成酶基因。本研究为揭示泡核桃抗寒机理以及查尔酮合成酶在冷害胁迫下的作用提供研究基础,并为利用基因工程手段培育抗寒新品种提供理论依据。Chalcone synthase is the first key enzyme of flavonoid biosynthesis in plant,and plays an important role in the physiology of plant stress resistance.In this study,we cloned a cold-induced chalcone synthase gene（JsCHS1） by RT-PCR technology from Juglans sigillata.The gene full length was 1 490 bp including one intron,which consisted of 1 170 bp open reading frame（ORF） and encoded 389 amino acids,and its accession number in Gen Bank was KX657834.The deduced JsCHS1 protein shared 98% similarity with chalcone synthase of Juglans regia.The chalcone synthase gene intron DNA homology of J.sigillata and J.regia was 95%.There were 8 nucleotide de JsCHS1 and CHS from J.regia had the same branch.Semi-quantitative PCR results showed that JsCHS1 was weakly expressed in leaf of J.sigillata after normal and low temperature（4℃） treatments,and strongly expressed under low temperature treatment after 6 h.This result showed that JsCHS1 was a typical cold-induced chalcone synthase gene.This study would provide research basis for revealing the cold resistance mechanism of J.sigillata,as well as the role of chalone synthase in cold water stress,and also provide theoretical basis for breeding new varieties which would be resistant to low temperature by genetic engineering technology.

关 键 词：泡核桃 查尔酮合成酶 冷害 基因表达 基因克隆 

分 类 号：Q943.2[生物学—植物学] S664.1[农业科学—果树学]