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

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。

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

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.