大豆GmPR10基因启动子的克隆及序列分析

Cloning and Sequence Analysis of the GmPR10 Gene Promoter from Soybean( Glycine max L.)

GmPR10基因是病程相关蛋白PR10(pathogenesis-related proteins 10)在大豆中的同源基因。为探明大豆GmPR10基因的表达调控规律,应用PCR技术从大豆抗疫霉根腐病品种绥农10号中克隆了GmPR10基因上游2 235 bp的启动子序列pGmPR10,定向替换pBI121载体的CaMV35S组成型启动子,构建植物表达载体pBI121/pGmPR10/GUS,并转化农杆菌侵染烟草叶盘。GUS染色结果表明,pGmPR10受聚乙二醇(PEG)、低温(4℃)、水杨酸(SA)、茉莉酸(JA)和脱落酸(ABA)诱导表达,因此推测GmPR10基因可能参与植物激素调节植物生长发育的过程,以及生物胁迫和非生物胁迫条件下植物对环境响应的过程。此外,利用PLACE和Plant CARE在线启动子预测工具分析pGmPR10,结果表明:pGmPR10含有启动子的一般结构TATA-box和CAAT-box,光应答元件,生长素和细胞分裂素响应元件,热激元件,低温应答元件,干旱应答元件以及ABA、SA、JA应答元件等。

GmPRIO was homology with the PRIO （pathogenesis-related proteins 10） gene in other plants. To investigate the expression and regulation of the GmPRIO gene in soybean, the 2 235 bp promoter region of 5＇-flanking upstream of GmPRIO gene was isolated from the genomic DNA of soybean cuhivars Suinong 10（ with high resistance to Phytophthora sojae in Heilongjiang, China） by PCR, defined as pGmPRI0. It was directionally replaced CaMV35S promoter of the expression vector pBI121. The plant expression vector of pBII21/pGmPRIO/GUS was used to drive expression of the GUS reporter gene in the pBI121, and the construct was transformed into tobacco leaves by Agrobacterium-mediated transformation. GUS staining results showed that, pGmPR10 activity was highly induced by drought （PEG）, low temperature （4%）, salicylic acid （SA）, jas- monic acid （JA） and abscisic acid （ABA）. Therefore, it was speculated that GmPRIO gene may be involved in the process of plant growth and development with the regulation of plant hormones, as well as the process under abiotic and abiotic stresses responses to the environment. In addition, sequence analysis by PLACE and PlantCARE revealed that pGmPR10 contained promoter general structure TATA-box and CAAT-box, light response element, auxin and cytokinin response element, heat shock element, cold response element, drought responsive element and ABA, SA and JA response elements.