巴西橡胶树 HbERFB4-1 基因的克隆和表达分析

Cloning and Expression Analysis of HbERFB4-1 Gene in Hevea brasiliensi

摘要:

根据EST序列信息,利用RACE技术分离了巴西橡胶树(Hevea brasiliensis)的1个AP2/ERF类基因,命名为HbERFB4-1。该基因cDNA全长732 bp,含有完整的开放阅读框架,编码147个氨基酸,不含内含子。推导的HbERFB4-1氨基酸序列与杨树(Populus trichocarpa)、蓖麻(Ricinus communis)、拟南芥(Arabidopsis thaliana)和大豆(Glycine max)中相应蛋白氨基酸序列的一致性分别为78%、71%、62%和55%,具有典型的AP2类蛋白结构特征。聚类分析表明,HbERFB4-1属于ERF家族B4亚族。半定量RT-PCR分析结果表明,该基因在胶乳中的表达量相对较低,但乙烯利刺激后其在胶乳中的表达量迅速增加,推测该基因可能参与了胶乳中乙烯信号的转导。

Abstract:

Based on EST sequences from an ethephon-induced latex SSH cDNA library of Hevea brasiliensis, a full-length cDNA encoding AP2/ERF type gene named as HbERFB4-1, was cloned from H. brasiliensis by RACE-PCR. The HbERFB4-1 cDNA has full length of 732 bp with an open reading frame (ORF) of 444 bp, and encodes 147 amino acid residues. No introns were detected in HbERFB4-1gene. The deduced HbERFB4-1 contains an AP2 domain and three NISs, which shows high identity to AP2/ERF in Populus trichocarpa,Ricinus communis,Arabidopsis thaliana and Glycine max, counting for 78%、71%、62% and 55%, respectively. Phyogenetic analysis showed that HbERFB4-1 belonged to the B4 subfamily of ERF family. Semi-quantitative RT-PCR analysis indicated that the transcription of HbERFB4-1 in latex was low, but it was rapidly induced by ethephon, suggesting that HbERFB4-1 is involved in the ethylene signal pathway in latex of H. brasiliensis.