Development of the molecular methods for rapid detection and differentiation of Fusarium oxysporum and F. oxysporum f. sp. niveum in Taiwan.
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.
Lin
Ying-Hong
YH
Department of Plant Pathology, National Chung Hsing University, Taichung City 402, Taiwan, ROC.
Chen
Kan-Shu
KS
Chang
Jing-Yi
JY
Wan
Yu-Ling
YL
Hsu
Ching-Chi
CC
Huang
Jenn-Wen
JW
Chang
Pi-Fang Linda
PF
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
D017931:DNA Primers; D005819:Genetic Markers