巴西橡胶树橡胶转移酶激活因子HbRTA的克隆、表达及定位  

Cloning,Expression and Localization of HbRTA Gene Encoding Rubber Transferase Activator from H.brasiliensis

作　　者：石波明 杨礼富 康桂娟[3] 曾日中[3] Shi Boming;Yang Lifu;Kang Guijuan;Zeng Rizhong(College of Horticulture,Hainan University,Haikou,570228;Institute of Scientific and Technical Information,Chinese Academy of Tropical Agricultural Science,Haikou,570216;Rubber Research Institute,Chinese Academy of Tropical Agricultural Sciences,Haikou,571101)

机构地区：[1]海南大学园艺学院,海口570228 [2]中国热带农业科学院科技信息研究所,海口570216 [3]中国热带农业科学院橡胶研究所,海口571101

出　　处：《分子植物育种》2020年第23期7718-7725,共8页Molecular Plant Breeding

基　　金：中国热带农业科学院基本科研业务费专项资金(1630022019001;1630022020027);海南省自然科学基金项目(319MS089)共同资助。

摘　　要：为深入了解天然橡胶次生代谢生物合成过程,本研究从橡胶树胶乳中分离得到一个橡胶转移酶激活因子基因HbRTA,并对其进行了克隆、表达、定位以及生物学分析。结果表明,HbRTA的基因组序列为3632 bp,含有7个外显子和6个内含子,其开放阅读框(ORF)为774 bp,编码蛋白含257个氨基酸,分子量为29.38 kD,理论等电点为8.57。酵母实验及亚细胞定位表明,HbRTA蛋白没有转录激活活性,且定位在细胞质中。实时荧光定量PCR分析发现,HbRTA在叶片、雌花、雄花、胶乳、树皮中均表达,其中胶乳表达量高于其它组织,并且胶乳中该基因的表达受割胶、茉莉酸(JA)和乙烯利(ET)刺激的影响。综合分析结果推测:HbRTA的表达可能与巴西橡胶树橡胶生物合成及胶乳产量有关。To further understand the biosynthesis process of natural rubber polygenes,in this study,a gene encoding the rubber transferase activator,HbRTA,was isolated from the latex of Hevea brasiliensis(rubber tree).Its cloning,expression,localization and biological analysis were carried out.The results showed that the HbRTA genome sequence was 3632 bp,containing 7 exons and 6 introns.The open reading frame(ORF)of HbRTA was774 bp,encoding a protein of 257 amino acids,with molecular weight 29.38 kD and theoretical isoelectric point8.57.Yeast experiments and subcellular localization showed that HbRTA protein had no transcriptional activation activity and was located in the cytoplasm.Real-time fluorescence quantitative PCR analysis showed that HbRTA gene was expressed in leaves,female flowers,male flowers,latex and bark,and the expression level in latex was higher than that in other tissues.Moreover,the expression of HbRTA gene in latex was affected by bark tapping,jasmonic acid(JA)and ethrel(ET)stimulation.These results suggested that the expression of HbRTA might be related to the rubber biosynthesis and lattex yield of rubber tree.

关 键 词：巴西橡胶树(Hevea brasiliensis) 胶乳 橡胶转移酶激活因子(HbRTA) 基因表达 

分 类 号：S794.1[农业科学—林木遗传育种]