PCR-based analysis of disease in tomato singly or mixed inoculated with <em>Fusarium oxysporum</em> f. sp. <em>lycopersici</em> races 1 and 2

Abstract

The pathogenic response of two tomato cultivars to races of Fusarium oxysporum f. sp.. lycopersici (cv. Momotaro, insensitive to race 1 of the pathogen, and cv. Ponderosa sensitive to race 1), was studied in greenhouse and laboratory experiments by inoculating the cultivars singly with race 1 or race 2, and in mixed inoculation with the two races of the pathogen. A pre-symptom PCR assay two weeks after inoculation showed that a fragment of the intergenic spacer region (IGS) of ribosomal DNA was amplifi ed by DNA templates from leaf samples of cv. Momotaro tomato plants inoculated with only race 2, or with race 1+2, but in the cv. Ponderosa the fragment was amplifi ed only in plants inoculated with race 1+2. Race-specifi c analysis using the sp13 and sp23 primers confi rmed that the amplifi ed fragment was from race 2 in cv. Momotaro and from races 1+2 in cv. Ponderosa. Later wilt symptoms mirrored the pre-symptom and post-symptom molecular analytical results: cv. Momotaro plants inoculated with only race 1 remained symptomless, while the ‘Momotaro’ plants inoculated with both races (1+2) did not manifest more severe wilt symptoms than plants inoculated with race 2 alone; cv. Ponderosa plants that were mixed-inoculated with race 1+2 manifested more severe symptoms, and at an earlier date than plants inoculated with only race 2. Growth parameters such as number of leaves and plant height showed the race 1+2 infected cv. Ponderosa were significantly retarded in growth, suggesting that significant synergism between the fungal races in tomato pathosystem can occur only when the host cultivar is sensitive to both races. An additional important finding is that pre-symptom leaf sampling of apparently healthy plants is useful in PCR diagnostic analysis to predict impending fusarial wilt outbreaks in tomato especially in infested soil.