水稻OsMAPK17-1基因的分离及转基因研究  

Isolation of Rice OsMAPK17-1 Gene and Its Genetic Transformation

作　　者：肖文芳[1,2] 陈艳[3] 汪德锋[1] 符秀梅[1] 肖晓蓉[1] 黎秀琼[3] 庞金环[3] 陈银华[1] 

机构地区：[1]海南大学热带生物资源可持续利用重点实验室,海南海口570228 [2]海南大学环境与植物保护学院,海南海口570228 [3]中国科学院微生物研究所,北京100101

出　　处：《热带作物学报》2013年第8期1490-1497,共8页Chinese Journal of Tropical Crops

基　　金：转基因生物新品种培育重大专项"水稻广谱抗病基因的系统筛选及新种质创制"(No.2009ZX08009-41B);国家自然基金(No.31260345);海南省重大科技项目(No.2D2X2013023-2)

摘　　要：OsMAPK17-1作为一种MAP激酶,受多种生物和非生物胁迫的诱导,且对病原菌的侵染有一定的防卫作用。为研究水稻OsMAPK17-1基因的功能,通过RT-PCR方法分离OsMAPK17-1基因全长cDNA,分别构建过表达与RNAi植物表达载体,采用农杆菌介导法以粳稻日本晴为受体进行遗传转化,PCR检测具有潮霉素抗性基因的转基因水稻植株,结果表明:OsMAPK17-1基因及RNAi片段均已整合到水稻基因组中。利用PCR和RT-PCR的方法对转基因后代进行筛选,获得过表达和RNAi转基因纯系各1个,模拟干旱处理结果表明,RNAi株系在干旱处理下比对照更有利于根的生长,这些结果为进一步研究OsMAPK17-1基因的功能奠定了基础。OsMAPK17-1, a MAP Kinase in rice, is induced by various biotic and abiotic stress and involved in the regulation of plant defense responses. To research the functions of flee gene OsMAPK17-1, the full length eDNA of OsMAPK17-1 was isolated by RT-PCR amplification, the over-expression and RNA interference vectors were constructed with modified pCAMBIA1300 and pTCK303 as framework plasmid, respectively, with the Hygromyein phosphotransferase gene as a selectable marker. Then the vectors were respectively transferred into the rice through Agrobacterium-mediated transformation. PCR analysis confirmed that OsMAPK17-1 and its RNAi were integrated into rice genome. The homologous transgenie lines were screened with the method of PCR. The RNAi lines were more suitable for the drought stress. These works laid the foundation for further study of OsMAPK17-1.

关 键 词：水稻 OsMAPK17-1基因 遗传转化 过表达载体 RNAI载体 

分 类 号：S511[农业科学—作物学]