百合炭疽病病原菌PCR检测技术  

Study on the Technique of PCR Detection of Colletotrichum lilii

作　　者：戴富明[1] 曾蓉[1] 戚龙君 陆金萍[1] 

机构地区：[1]上海市农业科学院植物保护研究所上海市设施园艺重点实验室,上海201403 [2]上海市出入境检验检疫局,上海200135

出　　处：《上海交通大学学报（农业科学版）》2011年第4期55-60,共6页Journal of Shanghai Jiaotong University(Agricultural Science)

基　　金：国家质检总局科研项目(2009IK252)

摘　　要：百合炭疽病是百合(Lilium longiflorum)种球上重要病害之一,其病原为百合炭疽病菌(Colletotrichum lilii Plakidas)。通过百合炭疽病菌与常见胶孢炭疽病菌(C.gloeosporioides)等4种不同炭疽病菌种类的核糖体DNA ITS序列比对设计特异性引物,通过特异性检验,确定BT73/BT-R510和BT-F98/BT-R510。2对引物均可以分别扩增出438bp和413bp的百合炭疽病菌目标片段,而对常见4种炭疽病菌及百合球茎上2种常见的病原菌百合枯萎病菌及灰霉病菌均不能扩增出相同的目标片段。用引物对F73/BT-R510和BT-F98/BT-R510特异性PCR扩增百合炭疽病菌的灵敏度分别都在50pg以上。用外圈引物对F73/BT-R510和内圈引物对BT-F98/BT-R510进行巢式PCR扩增,对百合炭疽病检测的灵敏度提高不明显。Lily anthracnose is one of the most common diseases infecting lily bulbs caused by Colletotrichum lilii. Specific primer pairs were designed by comparing rDNA ITS sequences of C. lilii, C. g｜oeosporioides and other three Colletotrichurn species. By the specific assessment of PCR detection, two pairs of primer pairs, BT73/BT-R510 and BT-F98/BT-R510 were selected to amplify the target DNA sequence of C. lilii,which produced 438 bp and 413 bp specific fragments of C. lilii, respectively, and no any DNA fragments of other four Colletotrichum species and two species,Botrytis cinerea and Fusarium oxysporum f. sp. lilii from the lily bulb were amplified. PCR with the above two primer pairs could detect 50pg DNA of C. lilii at least,but the nested PCR with BT73/BT-R510 as the outer primers and BT-F98/ BT-R510 as the inner primers didn＇t improved the sensitivity of detection.

关 键 词：百合 炭疽病 PCR 检测 

分 类 号：S436.3[农业科学—农业昆虫与害虫防治]