梨叶片中苹果褪绿叶斑病毒的引物原位标记检测.pdfVIP

园　艺 　学 　报 　2009 , 36 ( 1) : 15 - 20 A cta Horticu lturae Sin ica 梨叶片中苹果褪绿叶斑病毒的引物原位标记检测 赵 　英 , 牛建新 (石河子大学农学院 , 新疆石河子 832003) 摘 　要 : 选用带苹果褪绿叶斑病毒的香梨叶片 , 制备成适合进行原位 R TPCR 的石蜡切片 。设计特异 引物 , 利用引物原位标记 ( Prim ed in s itu labeling, PR IN S) 技术对切片组织中的病毒进行了检测研究 , 结 果表明 , PR IN SF ITC染色法和 PR IN SRhodam ine染色法均能获得 良好的检测效果 , 有病毒部位分别显示绿 色荧光和红色荧光 , 而且荧光出现的位置与原位 R TPCR 检测的结果一致 。由此证明引物原位标记技术可 用于果树病毒的原位检测 。 关键词 : 梨 ; 引物原位标记 ; 苹果褪绿叶斑病毒 ; 病毒检测方法 ; 染色 中图分类号 : S 66 12　　文献标识码 : A 　　文章编号 : 0513353X (2009) 0 1001506 D etection of A pp le ch loro tic lea f spo t viru s in Pear by Pr im ed in situ L abeling ZHAO Ying and N IU J ianxin (A g ricu ltu ra l Colleg e, S h ihez i Un ivers ity, S h ihez i, X inj iang 832003, Ch ina) A b stract: The slice s of p ear leaf samp le s infected by App le ch lorotic leaf sp ot virus were p rep ared for in s itu R TPCR and were detected by p rim ed in s itu labeling techn ique u sing sp ecial p rim er. It wa s showed that both stain ing m ethod s of PR IN SF ITC and PR IN SRhodam ine can get good detection re su lts in wh ich the p arts have viru se s showed green and red fluore scence ligh t, re sp ectively, and the site s of show ing fluore scence showed the sam e site s a s the site s detected by R TPCR m ethod. Therefore, p rim ed in s itu labeling techn ique can be p erfectly u sed for viru s in s itu detection of fru it tree s, and it is also a rap id, simp le and reliab le in s itu detection m ethod. Key words: p ear; p rim ed in s itu labeling; App le ch lorotic leaf sp ot virus; viru s detection m ethod; stain ing 苹果褪绿叶斑病毒 (App le ch lorotic leaf sp ot virus, ACL SV ) 为纤毛病毒属 ( Trichovirus) 的代表成 员 (洪健和周雪平 , 2005) ,