Cloning and Expression Analysis by Real-Time PCR of 4-Coumarate:Coenzyme A ligase(4CL) Gene in Galega orientalis

doi:10.11733/j.issn.1007-0435.2010.04.010

›› 2010, Vol. 18 ›› Issue (4): 533-538.DOI: 10.11733/j.issn.1007-0435.2010.04.010

Cloning and Expression Analysis by Real-Time PCR of 4-Coumarate:Coenzyme A ligase(4CL) Gene in Galega orientalis

YANG Dong-mei¹, WANG Xue-min¹, GAO Hong-wen¹, Dzyubenko Nikolay², Chapurin Vladimir², HAN Yong-zeng¹, SONG Qing-xiao¹, DONG Jie¹

1. Institute of Animal Science, Chinese Academy of Agriculture Sciences, Beijing 100193, China;
2. N.I.Vavilov All-Russian Research Institute of Plant Industry, St.Petersburg 190000, Russia

Received:2009-12-24 Revised:2010-02-01 Online:2010-08-15 Published:2010-08-15

东方山羊豆4香豆酸:辅酶A连接酶(4CL)基因的克隆和荧光定量表达分析

杨冬梅¹, 王学敏¹, 高洪文¹, Dzyubenko Nikolay², Chapurin Vladimir², 韩永增¹, 宋清晓¹, 董洁¹

1. 中国农业科学院北京畜牧兽医研究所, 北京, 100193;
2. 俄罗斯瓦维洛夫全俄植物栽培研究所, 圣彼得堡, 190000

通讯作者: 高洪文,E-mail:gaohongwen@263.net
作者简介:杨冬梅(1985- ),女,回族,内蒙古锡林浩特市人,硕士研究生,研究方向为牧草种质资源,ydm9986@126.com.
基金资助:
现代农业产业技术体系建设专项资金;“十一五”科技支撑项目(2008BADB3B01)资助

Abstract

Abstract: Galega orientalis L.was a pasture species which was introduced to China at the end of the twentieth century.The 4CL gene was cloned in Galega orientalis to explore its gene resources.The influence of 4CL gene on the stress-resistance in Galega orientalis was well studied.All of the research formed the basis for the implantation and application of Galega orientalis in China in future.With RACE and RT-PCR methods,the full length cDNA,which encode 4-coumarate: coenzyme A ligase,was cloned from Galega orientalis.The 1,916-bp full length Go4CL had a 1,653-bp ORF,which encodes 510 amino acids residues.The results from Real-Time PCR indicated that the expression of 4CL gene was the strongest in the root of Galega orientalis,and the least in leaves.Many abiotic stress treatments,such as PEG,ABA,MeJA and SA,all up regulated the expression of Go4CL gene,which indicated that 4CL may play a role in the biotic and abiotic stress-resistance regulation pathway in plants.

Key words: Galega orientalis, 4CL, Gene clone, Real-Time PCR

摘要： 东方山羊豆(Galega orientalis L.)是20世纪末引种到我国的牧草品种,为发掘其基因资源,本实验克隆出东方山羊豆4 CL基因,并深入研究4 CL基因对东方山羊豆抗逆性的影响,为今后东方山羊豆在我国的种植和应用奠定基础。结果表明:采用RACE技术结合RT-PCR方法从东方山羊豆中获得一个编码4 CL基因的cDNA全长序列,命名为Go4CL。该序列全长1916 bp,开放阅读框1653 bp,编码510个氨基酸。利用Real-Time PCR方法检测发现,Go4CL基因在东方山羊豆的根中表达量最高,叶中表达量最少。在PEG、ABA、MeJA、SA胁迫处理下Go4CL基因的表达量均有不同程度的上调,说明此基因在植物抵御干旱、病菌侵袭和机械损伤等胁迫过程中能够起到调控作用。

关键词: 东方山羊豆, 4香豆:酸辅酸A连接酶, 基因克隆, 荧光定量PCR

CLC Number:

Q786
S541.9

YANG Dong-mei, WANG Xue-min, GAO Hong-wen, Dzyubenko Nikolay, Chapurin Vladimir, HAN Yong-zeng, SONG Qing-xiao, DONG Jie. Cloning and Expression Analysis by Real-Time PCR of 4-Coumarate:Coenzyme A ligase(4CL) Gene in Galega orientalis[J]. , 2010, 18(4): 533-538.

杨冬梅, 王学敏, 高洪文, Dzyubenko Nikolay, Chapurin Vladimir, 韩永增, 宋清晓, 董洁. 东方山羊豆4香豆酸:辅酶A连接酶(4CL)基因的克隆和荧光定量表达分析[J]. , 2010, 18(4): 533-538.

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Cloning and Expression Analysis by Real-Time PCR of 4-Coumarate:Coenzyme A ligase(4CL) Gene in Galega orientalis

东方山羊豆4香豆酸:辅酶A连接酶(4CL)基因的克隆和荧光定量表达分析

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