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当前位置: 首页 > 文章 > 油菜柠檬酸合酶基因的克隆及在逆境下的表达 作物学报 2009,35 (1) 33-40
Position: Home > Articles > Cloning of Citrate Synthase Gene in Rapeseed (Brassica napus L.) and Its Expression under Stresses Acta Agronomica Sinica 2009,35 (1) 33-40

油菜柠檬酸合酶基因的克隆及在逆境下的表达

作  者：
童晋;詹高淼;王新发;刘贵华;华玮;王汉中
单  位：
中国农科院油料作物研究所/农业部油料作物遗传改良重点开放实验室
关键词：
油菜;柠檬酸合酶;表达模式;胁迫
摘  要：
柠檬酸合酶是植物多种代谢途径的限速酶,及代谢变化的标志酶。为了解油菜柠檬酸合酶基因的生理功能,以甘蓝型油菜叶片cDNA为模板,设计特异性引物,获得一编码柠檬酸合酶基因的cDNA序列,全长1 659 bp,ORF区含476个氨基酸残基,序列比对显示其蛋白序列与拟南芥有较高的同源性(96.0%),氨基末端含一线粒体靶信号。聚类分析表明,油菜柠檬酸合酶基因与其他植物内该基因高度同源。对油菜幼苗进行植物生长调节物质、高温和低温、强光照和弱光照、盐、菌核病、干旱和水渍等处理,采用半定量PCR法对油菜叶片柠檬酸合酶基因的表达模式进行检测,发现在盐胁迫、暗光和强光的处理下,柠檬酸合酶基因的表达基本没有变化;在水渍、干旱、IAA和6-BA胁迫下,其表达有所升高,但出现峰值的时间不同,ABA对表达模式的影响与IAA相反;感染菌核病后其表达降低;对GA3的应答呈鞍型。对部分处理采用荧光定量PCR验证,其结果与半定量PCR结果基本一致。
译  名：
Cloning of Citrate Synthase Gene in Rapeseed (Brassica napus L.) and Its Expression under Stresses
作  者：
TONG Jin,ZHAN Gao-Miao,WANG Xin-Fa,LIU Gui-Hua,HUA Wei,and WANG Han-Zhong Oil Crops Research Institute,Chinese Academy of Agricultural Sciences,Wuhan 430062,China
关键词：
Rapeseed(Brassica napus L.);Citrate synthase;Expression pattern;Stress
摘  要：
As a key enzyme of some metabolisms,citrate synthase shows a sign for metabolism.In order to investigate the func-tion of citrate synthase gene,the cDNA encoding citrate synthase was cloned from rapeseed leaf by RT-PCR.It was 1 659 bp long and encoded a protein with 476 amino acids.The deduced protein sequence had a mitochondrial targeting signal in N-terminal,which was very similar to the citrate synthase in Arabidopsis thaliana(96.0%).Alignment analysis showed that citrate synthase gene had high homology in plants.Under different stresses,we tested the expression of citrate synthase gene in rapeseed leaf by using semi-quantitative PCR.The expression of citrate synthase gene had no obvious change in stresses of salt,dark,high illumi-nation,while was increased at different time in treatments of water logging,drought,IAA,and 6-BA.Interestingly the effect of ABA was contrary to that of IAA.In the treatment of sclerotium blight,the expression of citrate synthase gene was depressed.There was a saddle curve of citrate synthase gene expression in the treatment of gibberellin.The results from real-time PCR of several treatments were mainly in accord with that from semi-quantitative PCR.

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