title: Development of the molecular methods for rapid detection and differentiation of Fusarium oxysporum and F. oxysporum f. sp. niveum in Taiwan.
abstract: Fusarium wilt, caused by Fusarium oxysporum (Fo), is one of the most important fungal diseases worldwide. Like other plant pathogens, Fo displays specialized forms in association with its hosts. For example, F. oxysporum f. sp. niveum (Fon) is the damaging pathogen causing Fusarium wilt disease on watermelon, whereas F. oxysporum f. sp. cubense is the pathogen that infects banana. A rapid and reliable pathogen identification or disease diagnosis is essential for the integrated disease management practices in many crops. In this study, two new primer sets, Fon-1/Fon-2 and FnSc-1/FnSc-2, were developed to differentiate Fon and Fo, respectively. The PCR method using the novel primer sets has high sensitivity to detect Fon when the DNA concentration was as low as 0.01 pg or when the conidia number was as few as 5. In comparison with the published primer set, the Fon-1/Fon-2 primer set, derived from the sequence of OP-M12 random primer-amplified fragment, produced a 174 bp DNA fragment, and was more specific to Fon in Taiwan. In addition, with optimized PCR parameters, the molecular method using the Fon-1/Fon-2 primer set could directly detect Fon even when watermelon samples were collected in its early stage of disease development.
authors.affiliation: Department of Plant Pathology, National Chung Hsing University, Taichung City 402, Taiwan, ROC.
authors.forename: Ying-Hong
authors.initials: YH
authors.lastname: Lin
authors.forename: Kan-Shu
authors.initials: KS
authors.lastname: Chen
authors.forename: Jing-Yi
authors.initials: JY
authors.lastname: Chang
authors.forename: Yu-Ling
authors.initials: YL
authors.lastname: Wan
authors.forename: Ching-Chi
authors.initials: CC
authors.lastname: Hsu
authors.forename: Jenn-Wen
authors.initials: JW
authors.lastname: Huang
authors.forename: Pi-Fang Linda
authors.initials: PF
authors.lastname: Chang
mesh_terms: D001419:Bacteria / Q000302:isolation & purification; D029773:Citrullus / Q000382:microbiology; D017931:DNA Primers / Q000378:metabolism; D005670:Fusarium / Q000145:classification* / Q000302:isolation & purification*; D005819:Genetic Markers; D018546:Hypocotyl / Q000382:microbiology; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D016533:Mycological Typing Techniques / Q000379:methods*; D010935:Plant Diseases / Q000382:microbiology; D016133:Polymerase Chain Reaction; D019105:Random Amplified Polymorphic DNA Technique; D013045:Species Specificity; D013624:Taiwan
chemical_list: D017931:DNA Primers; D005819:Genetic Markers