Genetic diversity of Fusarium oxysporum f.sp. niveum responsible of watermelon Fusarium wilt in Tunisia and Spain

Naima Boughalleb-M'Hamdi; Ibtissem Ben Salem; Najwa BenFradj; Paloma Abad-Campus

Naima Boughalleb-M'Hamdi Department of Biological Sciences and Plant Protection, University of Sousse, UR13AGR3, Higher Agronomic Institute of Chott Meriem, 4042, Sousse, Tunisia
Ibtissem Ben Salem Department of Biological Sciences and Plant Protection, University of Sousse, UR13AGR3, Higher Agronomic Institute of Chott Meriem, 4042, Sousse, Tunisia
Najwa BenFradj Department of Biological Sciences and Plant Protection, University of Sousse, UR13AGR3, Higher Agronomic Institute of Chott Meriem, 4042, Sousse, Tunisia
Paloma Abad-Campus Mediterranean Agroforestry Institute, Polytechnic University of Valencia, Camino de Vera s / n, 46022, Valencia- Spain

Abstract

Fusarium wilt is a serious disease, caused by Fusarium oxysporum f. sp. niveum (FON) and responsible of significant yield losses in watermelon crop in Tunisia. Thirty-nine FON isolates were collected from different infected watermelon fields in Tunisia and were identified on the basis of morphological features and by using one pair of species-specific primers Fn-1/Fn-2. Genetic diversity of the twenty-six FON isolates, originated from Spain and Tunisia, was studied by applying Inter simple sequence repeat genotyping. PCR amplification appears to be efficient to identify FON isolates amplifying only a single PCR band of approximately 800 bp. The RAMS study using two, bi and four trinucleotide microsatellites primers, showed a different genetic similarity degree among FON isolates. Seventy-one bands were amplified by four ISSR primer combinations. Diversity in the banding patterns obtained by DNA fingerprinting was always >50% and allowed us to distinguish all the isolates tested, according to number and size of the fragments, which ranged from 300 to 2800 bp. The genetic similarity values are comprised between 8 and 97%. UPGMA grouped the 45 genotypes into six main clusters at a similarity index value above 0.5, showing a relative genetic homogeneity and no correlation has been found among FON isolates and their origins. The most abundant Cluster VI comprising thirty-nine FON isolates.

References

Appel DJ, Gordon TR, 1995. Intraspecific variation within populations of Fusarium oxysporum based on RFLP analysis of the intergenic spacer region of the rDNA. Experimental Mycology 19,120-128.

Barve MP, Haware MP, Sainani MN, Ranjekar PK, Gupta VS, 2001. Potential of microsatellites to distinguish four races of Fusarium oxysporum f.sp. ciceri prevalent in India. Theoretical and Applied Genetics 102,138-147.

Belabid L, Baum M, Fortas Z, Bouznad Z, Eujayl I, 2004. Pathogenic and genetic characterization of Algerian isolates of Fusarium oxysporum f. sp. lentis by RAPD and AFLP analysis. African Journal of Biotechnology 3 (1), 25-31.

Boughalleb N, El Mahjoub M, 2006. Détection des races 0, 1 et 2 de Fusarium oxysporum f. sp. niveum et leur distribution dans les régions de production de la pastèque en Tunisie. OEPP/EPPO, Bulletin OEPP/EPPO Bulletin 35, 253–2.

Callaghan SE, Puno VI, Williams AP, Weir BS, Balmas V, Sengsoulichan K., Phantavong, S, Keovorlajak T, Phitsanoukane P, Xomphouthilath P, Phapmixay KS, Vilavong S, Liew ECY, Duckitt GS, Burgess LW, 2016. First report of Fusarium oxysporum f.sp. niveum in the Lao PDR. Australasian Plant Disease Notes 11,9.

Everts KL, Himmelstein JC, 2015. Fusarium wilt of watermelon: Towards sustainable management of a re- emerging plant disease. Crop Protection 73, 93-99.

FAOSTAT: Food and Agriculture Organization of the United Nations), 2016. http://faostat3.fao.org/browse/Q/*/E

Hantula J, Muller MM, 1997. Variations within Gremmeniella abietina in Finland and other countries as determined by random amplified microsatellites (RAMS). Mycological Research 101(2), 169–175.

Hirano Y, Arie T, 2009. Variation and phylogeny of Fusarium oxysporum isolates based on nucleotide sequences of polygalacturonase genes. Microbes and Environments 24, 113–120.

Jimenez-Gasco MM, Milgroom MG, Jimenez-Diaz RM, 2004. Stepwise evolution of races in Fusarium oxysporum f.sp. ciceris inferred from fingerprinting with repetitive DNA sequences. Phytopathology 94, 228-235.

Leong SK, Latiffah Z, Baharuddin S, 2010. Genetic diversity of Fusarium oxysporum f. sp. cubense isolates from Malaysia. African Journal of Microbiology Research 4 (11), 1026-1037.

Lin YH, Chen KS, Chang JY, Wan YL, Hsu CC, Huang JW, Chang PFL, 2010. Development of the molecular methods for rapid detection and differentiation of Fusarium oxysporum and F. oxysporum f. sp. niveum in Taiwan. New Biotechnology 27, 409–418.

Lin YH, Chen KS, Liou TD, Huang JW, Chang PFL, 2009. Development of a molecular method for rapid differentiation of watermelon lines resistant to Fusarium oxysporum f. sp. niveum. Botanical Studies 50, 273–280.

Ma LJ, Geiser DM, Proctor RH, Rooney AP, O'Donnell K, Trail F, Gardiner DM, Manners JM, Kazan K, 2013. 'Fusarium pathogenomics'. Annual review of microbiology 67, 399-416.

Ma LJ, van der Does HC, Borkovich KA, Coleman JJ, Daboussi MJ, Di Pietro A, Dufresne M, Freitag M, Grabherr M, Henrissat B, Houterman PM, Kang S, Shim WB, Woloshuk C, Xie X, Xu JR, Antoniw J, Baker SE, Bluhm BH, Breakspear A, Brown DW, Butchko RAE, Chapman S, Coulson R, Coutinho PM, Danchin EGJ, Diener A, Gale LR, Gardiner DM, Goff S, Hammond-Kosack KE, Hilburn K, Hua-Van A, Jonkers W, Kazan K, Kodira CD, Koehrsen M, Kumar L, Lee YH, Li L, Manners JM, Miranda-Saavedra D, Mukherjee M, Park G, Park J, Park SY, Proctor RH, Regev A, Ruiz-Roldan MC, Sain D, Sakthikumar S, Sykes S, Schwartz DC, Turgeon BG, Wapinski I, Yoder O, Young S, Zeng Q, Zhou S, Galagan J, Cuomo CA, Kistler HC, Rep M, 2010. Comparative genomics reveals mobile pathogenicity chromosomes in Fusarium. Nature 464, 367-73.

Mahfooz S, Maurya DK, Srivastava AK, Kumar S, Arora DK, 2012. A comparative in silico analysis on frequency and distribution of microsatellites in coding regions of three formae speciales of Fusarium oxysporum and development of EST-SSR markers for polymorphism studies. FEMS Microbiology Letters 328: 54-60.

Mahuku GS, Henrıquez MA, Munoz J, Buruchara R, 2002. Molecular markers dispute the existence of the Afro-Andean group of the bean angular leaf spot pathogen, Phaeoisariopsis griseola. Phytopathology 92, 580–9.

Nelson AJ, Elias KS, Arévalo GE, Darlington LC, Bailey BA, 1997. Genetic characterization by RAPD analysis of isolates of Fusarium oxysporum f. sp. erythroxyli associated with an emerging epidemic in Peru. Phytopathology 87, 1220-1225.

Pina C, Texeiro P, Leite P, Villa P, Belloch C, Brito L, 2005. PCR fingerprinting and RAPD approaches for tracing the source of yeast contamination in a carbonated orange juice production chain. Journal of Applied Microbiology 98, 1107–1114.

Rebib H, Bouraoui H, Rouaissi M, Brygoo Y, Boudabbous A, Hajlaoui MR, Sadfi-Zouaoui N, 2014. Genetic diversity assessed by SSR markers and chemotyping of Fusarium culmorum causal agent of foot and root rot of wheat collected from two different fields in Tunisia. European Journal Plant Pathology 139 (3), 481–495.

Wehner T, 2008. Watermelon. Vegetables I: Asteraceae, Brassicaceae, Chenopodicaceae, and Cucurbitaceae, pp. 381-418 In: J. Prohens and F. Nuez (eds.), Springer, New York.

Zhang Z, Zhang J, Wang Y, Zheng X, 2005. Molecular detection of Fusarium oxysporum f. sp. niveum and Mycosphaerella melonis in infected plant tissues and soil. FEMS Microbiology Letters 249, 39–47.