受条斑病菌侵染的水稻cDNA文库构建  

Construction of a rice cDNA library induced by infection of Xanthomonas oryzae pv.oryzicola

作　　者：赵梅勤[1] 李玉蓉[1] 邹丽芳[1] 陈功友[1] 

机构地区：[1]南京农业大学植物保护学院/农业部病虫监测与治理重点开放实验室,江苏南京210095

出　　处：《南京农业大学学报》2009年第2期51-55,共5页Journal of Nanjing Agricultural University

基　　金：国家863计划项目(2006AA02Z180;2006AA10A210);国家自然科学基金项目(30671354)

摘　　要：提取经条斑病菌（Xanthomonas oryzae pv．oryzicola）侵染诱导后的水稻mRNA，利用SMART^TM技术于载体pGADT7-SfiAB上构建了水稻cDNA文库。检测发现，文库中含有水稻病程相关蛋白基因OsPR—1a、OsPR—1b和PAL。随机提取cDNA文库克隆的质粒，酶切分析发现，该cDNA文库的插入片段分布在500～2000bp之间，平均大小约为1kb，重组率达95％。上述结果表明，水稻受条斑病菌侵染的cDNA文库质量较好，这为研究条斑病菌致病性因子与水稻互作机制奠定了工作基础。Constructing a rice cDNA library is a fundamental basis for understanding molecular interactions of rice with Xanthomonas oryzae pv. oryzicola, the critical pathogen of bacterial leaf streak in rice. The total RNA were extracted from rice leaves previously infected for 10 hours by the pathogen. The cDNA were reversely transcripted from the mRNA using SMARTTM technique and then ligated into a vector pGADT7-SfiAB at the sites of SfiA and SfiB. Collections of the ligated plasmid DNA were the cDNA library of rice which had interactions with the pathogen. The PCR amplification of OsPR-1a, OsPR-1b and PAL as pathogenesis-related protein genes indicated that the cDNA library contained those genes. Digestion of randomly selected clones of the library with the enzyme Sfi Ⅰ demonstrated that the sizes of inserted fragments in the vector were between 500 -2 000 bp, that the average size was about 1 kb, and that 95% of the library clones were recombinants. The above suggests that the constructed cDNA library of rice is of good quality and of use for isolation of interacting proteins with pathogenicity determinants of the bacterium by a yeast twohybrid system.

关 键 词：水稻 水稻条斑病菌 CDNA文库 SMARTTM技术 病程相关蛋白 

分 类 号：Q945.8[农业科学—植物病理学]