疣粒野生稻抗白叶枯病相关基因ME281半定量RT-PCR分析  

Expression analysis of bacterial blight resistance-related Gene ME281 in Oryza granulata Baill by semi-QRT-PCR

作　　者：吕广磊[1,2] 付坚[2] 和志娇[2,3] 白现广[2,4] 蔺忠龙[2,4] 黄兴奇[2] 杨和生 程在全[2] 

机构地区：[1]云南农业大学农学与生物技术学院,云南昆明650201 [2]云南省农业科学院生物技术与种质资源研究所,云南昆明650223 [3]云南省农业科学院高山经济植物研究所,云南丽江674100 [4]云南大学生命科学学院,云南昆明650091 [5]陆良县种子管理站,云南陆良655600

出　　处：《西南农业学报》2008年第5期1289-1293,共5页Southwest China Journal of Agricultural Sciences

基　　金：国家自然科学基金面上项目(30460019);云南省科技攻关和面上基金项目(2006NG34和2005C0063M)

摘　　要：以从疣粒野生稻受白叶枯病菌诱导所建的差减文库中筛选出的一个具有CC-NBS-LRR抗病结构域的基因(克隆号为ME281),用半定量RT-PCR法,以肌动蛋白(β-actin)基因为内参,研究疣粒野生稻抗白叶枯病相关基因ME281基因mRNA的表达水平。通过对PCR体系中循环次数及Mg2+浓度的优化,最终建立了一个稳定、方便的半定量RT-PCR体系。通过ME281基因与β-actin基因PCR产物的灰度之比,确定ME281为诱导性表达,并进一步推测其为疣粒野生稻抗白叶枯病相关基因,甚至为抗病基因。A CC-NBS-LRR domain structure of the resistance gene （ cloning,ME281 ） had been screened from the suppression subtractive hybridization（SSH） of Oryza rneyeriana induced by Xanthornonaz oryzae pv. oryzae bacteria in the laboratory preliminary work. Using semiquantitative RT-PCR method, with actin （β-actin） gene as internal control, the expression levels of resistance to bacterial blight related gene ME281 gene mRNA in Oryza meyeriana were sturdied. Through the PCR system cycles and Mg^2 ＋ concentration optimization, a stable and convenient semi-quantitative RT-PCR system was established. By the ratio of speculated as ME281 gene and β-actin gene PCR product the gray,ME281 was determined as induced expression, and furthefly it was speculated as Oryza meyeriana related gene resistance to Xanthomonas oryzae pv. oryzae, and even as the resistance gene .

关 键 词：半定量RT-PCR ME281基因 疣粒野生稻 诱导性表达 

分 类 号：S511.9[农业科学—作物学]