摘要
应用聚合酶链式反应（ＰＣＲ）技术检测甘蔗茎白条病无症样品和细菌纯培养及ＤＮＡ，并以改良半选择培养基（ｍＸＡＭ）加以验证。结果表明，ＰＣＲ能特异性地检出白条黄单胞包括标准菌株３３９１５ＡＴＣＣ和血清型Ⅰ，Ⅱ等１９个菌株。但不能扩增其它５个分离自蔗茎的腐生黄单胞和１３个其它属种的植物细菌。能够检出少至１０ｐｇ靶细菌的总ＤＮＡ或２０个活细菌。ＰＣＲ检测灵敏度较ＥＬＩＳＡ和ＤＩＡ高１００～１０００倍。田间样品检测结果与实际发病符合度为９３．４６％。该项技术适用于国内外甘蔗种质资源交换和口岸检疫以及病害流行学研究。
The PCR (Polymerase chain reaction) amplification assay based on the degenerated primer pair XAF1/XAR1 was applied to assess the extracts of the stalk of sugarcane with or without the symptoms of leaf scald, the suspension of pure pathogen and genome DNA. The PCR technique succeeded in detecting 19 strains of Xanthomonas albilineans , including 33915 ATCC and serovars Ⅰ and Ⅱ but failed to amplify 5 other strains of Xanthomonas and other genera and species of bacteria. This PCR technique proved to be 100 to 1000 fold more sensitive than ELISA or DIA, being able to detect the genome DNA of as little as 10 pg target bacteria or of as few as 20 living cells. The agreement between the average percentage of detecting positive samples and the actual incidence of LSD in the ratoon of the same field was up to 93.46%. It is therefore recommended to employ this technique in domestic and international exchange of germplasms, in plant disease quarantine and in epidemiological research.
出处
《西南农业大学学报（自然科学版）》
CSCD
1998年第4期328-333,共6页
Journal of Southwest Agricultural University
