利用抑制差减杂交技术分离受水稻抗性调控的褐飞虱基因  

Application of suppression subtractive hybridization (SSH) to clone differentially expressed genes in Nilaparvata lugens (Homoptera:Delphacidae) responding to rice resistance

作　　者：杨之帆[1] 陈永勤[1] 李春华[1] 蒋思婧[1] 

机构地区：[1]湖北大学生命科学学院,武汉430062

出　　处：《昆虫学报》2009年第10期1059-1067,共9页Acta Entomologica Sinica

基　　金：国家自然科学基金项目(30500328)

摘　　要：为分离受水稻抗性调控的褐飞虱Nilaparvata lugens基因,以取食感虫水稻台中1号和高抗水稻B5的2叶1芯秧苗24h的褐飞虱4龄若虫为起始材料,采用抑制差减杂交技术构建了两个群体间的正反向差减cDNA文库。通过斑点杂交从差减文库中筛选代表受水稻抗性调控的基因的cDNA克隆,进行测序和功能分析,挑选具功能的基因进行Northern杂交验证。结果表明,通过斑点杂交筛选到的98个阳性克隆代表92个互不重复的单基因,其中25个与动物的已知蛋白基因存在较高的同源性。Northern杂交表明,这25个基因有11个表达上调,8个表达下调,提示它们可能在褐飞虱适应抗性水稻过程中发挥了重要作用。本研究结果为克隆上述新基因的全长cDNA序列及进一步研究其在褐飞虱与水稻互作中的功能奠定了基础。To isolate the differentially expressed genes in brown planthopper Nilaparvata lugens (Homoptera: Delphacidae) responding to host rice resistance,suppression subtractive hybridization (SSH) was carried out using the 4th instar larvae of brown planthopper feeding on the susceptible rice Taichung Native 1 (TN1) and the highly resistant rice B5 seedlings at the 3rd leaf stage for 24 h. A forward and a reverse subtracted cDNA libraries specific to host rice resistance were constructed,followed by differential s...

关 键 词：褐飞虱 水稻品种抗性 抑制差减杂交(SSH) 斑点杂交 差异表达基因 

分 类 号：S435.112.3[农业科学—农业昆虫与害虫防治]