香蕉黑腐病菌(Botryodiplodia theobromae)的PCR检测  

Detection of Botryodiplodia theobromae causing banana black rot disease by polymerase chain reaction

作　　者：郭立佳[1] 黄俊生[1] 王国芬[1] 邓国平[1] 杨腊英[1] 

机构地区：[1]中国热带农业科学院环境与植物保护研究所海南省热带农业有害生物检测监控重点实验室,海南儋州571737

出　　处：《植物病理学报》2007年第3期248-254,共7页Acta Phytopathologica Sinica

基　　金：科技部"国家科技基础条件平台工作"(2004DKA30560);农业部南亚热作专项(LY2004-5)

摘　　要：根据香蕉黑腐病菌可可球二孢菌(Botryodiplodia theobromae)与其它香蕉病原真菌核糖体基因转录间隔区(rDNA-ITS)ITS1和ITS2间序列差异,设计了特异引物Bth-S(5′-TCTCCCACCCTTTGTGAAC-3′)和Bth-A(5′-AAAAGT-TCAGAAGGTTCGTC-3′),利用此引物对包括可可球二孢菌在内的21个菌株基因组DNA进行PCR扩增,结果只有4个可可球二孢菌菌株扩增到422bp特异带,其它17个菌株无扩增产物。灵敏度测试结果表明此特异引物能对1pg的可可球二孢菌基因组DNA进行扩增。对自然感染黑腐病的香蕉果实组织和接种可可球二孢菌或多种香蕉病原真菌混合接种的果实组织进行检测,Bth-S和Bth-A引物对不仅能够在自然感染黑腐病果实组织中特异检测到可可球二孢菌,而且能在未显症和发病的接菌香蕉果实组织中特异检测得到可可球二孢菌。这为香蕉可可球二孢菌潜伏侵染检测提供了技术支持。Based on the differences in the internal transcribed spacer （ITS1 and ITS2 ） sequences of Botryodiplodia theobromae and other banana fungal pathogens, a pair of primers specific for B. theobromae causing banana black rot disease was designed. Primer pair Bth-S and Bth-A consistently amplified a single product of 422 bp in DNA prepared from 4 isolates of B. theobromae, but not from 17 other isolates. The sensitivity of detection with primers Bth-S / Bth-A was 1 pg genomic DNA. B. theobromae could be specifically detected by PCR assay with the primer pair Bth-S and Bth-A from banana fruit tissues infected naturally, and that inoculated with the pathogen or pathogen complex including B. theobromae, Colletotrichum musae and Fusarium semitectum even before the visible symptom. These provided great help for detecting latent pathogen causing banana black rot disease in fruits.

关 键 词：香蕉黑腐病 可可球二孢菌 基因转录间隔区 PCR检测 

分 类 号：S436.67[农业科学—农业昆虫与害虫防治]