香蕉枯萎病镰刀菌ITS序列的PCR扩增及其分子检测  

ITS Sequence and PCR Protocol for Detection of Banana Wilt Disease Resulted from Fusarium Oxysporum f.sp. Cubense

作　　者：王国芬[1] 彭军[1] 代鹏[1] 邓国平[1] 黄俊生[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,海南儋州571737

出　　处：《华南热带农业大学学报》2007年第3期1-5,共5页Journal of South China University of Tropical Agriculture

基　　金：中国热带农业科学院科技基金项目;编号:Rky0616;科技部农业微生物资源平台项目;编号:2004DKA30560-8。

摘　　要：根据香蕉枯萎病菌(Fusarium oxysporum f.sp.cubense)ITS序列设计合成的三条引物:FusF1(5′AACCC CTGTGAACATACCACTTG3′)、FusR1(5′GAGGAACGCGAATTAACGCGAC3′)和FusR2(5′GACGATTACCAGTAGCGAGGGT3′)构成引物对A(FusF1/R1)和B(Fus F1/R2),对尖廉孢菌古巴专化型病菌进行PCR特异扩增试验,结果表明,两对引物对引起香蕉枯萎病的镰刀菌全基因组DNA分别有338bp和408bp的特异扩增产物;在镰刀菌种以上的阶元有较好的分辨力;对目标菌株的检测下限分别是A(100fg)、B(10pg);而且B对香蕉感染枯萎病的病组织有相应的扩增产物,表明该引物对可用于香蕉枯萎病的早期鉴定和病原菌的检测。Two pairs of PRC primers , A （FusF1/R1） and B （FusF1/R2） , were designed according to the ITS sequence of rRNA gene of Frsarium oxysporum f.sp Cubense. Results from assays done with these primers showed that a 338bp fragment from A and 408bp form B of rDNA gene sequence were amplified from genomic DNA of F. oxysporum. Two pairs of primers have a good specificity for the genus Frsarium, but limited to Frsarium oxysporum f. spp. Their PCR lower limit sensitivity range were from 100fg to 10fg for DNA extracted from Frsarium oxysporum f.sp cubense mycelium. And B also can be useful for molecular identification and detection of banana wilt disease.

关 键 词：香蕉镰刀性枯萎病 香蕉感病组织 特异扩增 分子检测 

分 类 号：S436.67[农业科学—农业昆虫与害虫防治]