热带作物学报2024，Vol.45Issue(4)：653-662,10.DOI:10.3969/j.issn.1000-2561.2024.04.001
巴西橡胶树HbPSKR2基因克隆及互作蛋白鉴定
Cloning and Interacted Protein Identification of HbPSKR2 Gene from Rubber Tree
摘要
Abstract
Phytosulfokine(PSK)is a small peptide hormone unique to higher plants,widely involved in biological proc-esses such as plant growth and development,division and differentiation,biotic and abiotic stress.PSK receptor(PSKR)is a direct receptor for PSK and is crucial for PSK signal transduction.In the present study,HbPSKR2 was cloned from rubber tree by RT-PCR(reverse transcription-polymerase chain reaction),and its bioinformatics,gene expression pattern,interacted proteins screening and identification were also analyzed.Results showed that the open reading frame of reading frame of HbPSKR2 had a total length of 3159 bp and encoded 1052 amino acids with molecular weight 114.84 kDa and theoretical isoelectric point 6.34.Domain analysis showed that HbPSKR2 was a typical trans-membrane protein.The first 640 amino acids were the antenna structures composed of leucine rich repeat(LRR),the 692 to 714 amino acids were transmembrane domains,and the 765 to 1052 amino acids were kinase domains.Multiple comparisons were conducted on the membrane kinase domain of HbPSKR2 and the membrane kinase domain of PSKR homologous proteins in Arabidopsis and rice.The results showed the existence of conserved sites such as ATP binding site,CaM binding site,Activation segment,GC Centre.Expression pattern analysis showed that HbPSKR2 was highly expressed in the cambium region of rubber tree.The expression level of HbPSKR2 was significantly increased at the early stage of coronatine(COR)treatment.Twelve candidate proteins interacted with HbPSKR2 were screened by yeast two hybrid technology,and the interaction between two protein kinases(HbPBL8 and HbPIX13)and HbPSKR2 was further verified by luciferase complementary imaging.The strong fluorescence signal was observed by co-transformation HbPSKR2-nLUC/HbPBL8-cLUC and HbPSKR2-nLUC/HbPIX13-cLUC into tobacco,suggesting the interaction be-tween HbPSKR2-HbPBL8 and HbPSKR2-HbPIX13 in vivo.The cloning of HbPSKR2 and identification of HbPSKR2 interacted protein would provide new insights into the molecular mechanism of laticifer differentiation in rubber tree.关键词
巴西橡胶树/HbPSKR2/酵母双杂交/荧光素酶互补成像/互作蛋白Key words
Hevea brasiliensis/HbPSKR2/yeast two-hybrid/luciferase complementary imaging/interaction protein分类
农业科技引用本文复制引用
杜晓愚,赵一杰,张世鑫,田维敏,晁金泉..巴西橡胶树HbPSKR2基因克隆及互作蛋白鉴定[J].热带作物学报,2024,45(4):653-662,10.基金项目
热带作物生物育种全国重点实验室科研项目(No.NKLTCB202309) （No.NKLTCB202309）
现代农业产业技术体系建设专项资金(No.CARS-33-YZ1) （No.CARS-33-YZ1）
海南省自然科学基金高层次人才项目(No.322RC781). （No.322RC781）
