巴西橡胶树铁螯合物还原酶基因(HbFRO)的克隆及表达  

Cloning and Expression Analysis of Ferric-chelate Reductase Gene(HbFRO) in Hevea brasiliensis

作　　者：方佳俊 黎秀琼[1] 罗红丽[1] 

机构地区：[1]海南大学海南省热带生物资源可持续利用重点实验室,海南海口570228

出　　处：《热带生物学报》2015年第2期105-112,共8页Journal of Tropical Biology

基　　金：国家自然科学基金项目(31160151);2011"新世纪优秀人才支持计划"(NCET-11-0928)

摘　　要：在橡胶树转录组测序的基础上,通过RT-PCR方法扩增到橡胶树的1个铁螯合物还原酶基因,命名为Hb FRO。该基因包含1个2 217 bp的开放阅读框,可编码739个氨基酸的多肽。生物信息学分析结果表明,Hb FRO蛋白除了含有Ferric_reduct,FAD_binding和NAD_binding保守结构域外,还含有10个跨膜结构域;Hb FRO蛋白的氨基酸序列与木薯、蓖麻、杨树和拟南芥的FRO蛋白同源,同源性分别为85%,80%,73%和63%。半定量分析结果表明,Hb FRO基因在胶乳、花和叶片中的表达丰度明显高于在树皮和芽中的表达;当橡胶树受到炭疽病菌和白粉病菌侵染时,Hb FRO基因在叶片中的表达被明显抑制。以上数据暗示Hb FRO基因在不同组织中的表达存在差异,可能参与植物的抗病应答反应。A ferric-chelate reductase gene of rubber tree （Hevea brasiliensis） was cloned by RT-PCR and named as HbFRO based on the data of RNA-Seq of rubber tree. The sequencing result revealed that the ORF of HbFRO is 221 7 bp, encoding a 739 amino acids peptides. Bioinformatics analysis showed that the HbFRO protein con- tains conserved Ferric_reduct, FAD_binding and NAD_binding domains, which are similar to those of the FRO from other plants, and had ten transmembrane domains. The amino acid sequence of HbFRO is 85% , 80% , 73% and 63% similar to that of the FRO from Manihot esculenta, Ricinus communis, Populus tomentosa and Ar- abidopsis thaliana, respectively. Semi-quantitative RT-PCR analysis showed that the expression level of the Hb- FRO was significantly higher in leaf, flower and latex than in bark and buds, but was significantly decreased in leaves inoculated with Colletotrichum gloeosporoiodes and Odium heveae Steinm. These implied that the HbFRO was differently expressed in different tissues and probably involved in disease defense.

关 键 词：巴西橡胶树 铁螯合物还原酶(FRO) 表达分析 

分 类 号：S794.1[农业科学—林木遗传育种]