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Gibberella Fujikuroi Mating Population a and Fusarium Subglutinans from Teosinte Species and Maize from Mexico and Central America

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Gibberella Fujikuroi Mating Population a and Fusarium Subglutinans from Teosinte Species and Maize from Mexico and Central America 8378 Mycol. Res. 104 (7): 565-872 auly 2000). Printed in the United Kingdom. 865 Gibberella fujikuroi mating population A and Fusarium subglutinans from teosinte species and maize from Mexico and Central America A. E. DESJARDINS,t", R. D. PLAITNER1 and T. R. GORDON% 1 Mycotoxin Research Unit, National Center for Agricultural Utilization Research, US Department of Agriculture, Agricultural Research Service, 1815 N. University Street, Peoria, nIinois 61604, USA. 2 Department of Plant Pathology, University of California, Davis, California 95616, USA. E-mail: [email protected] Received 8 October 1999: accepted 29 Ociober 1999. Seed samples of maize (lea mays ssp. mays) from Mexico and of teosintes (lea spp.), the nearest wild relatives of maize, from Mexico, Guatemala, and Nicaragua were assessed for infection with Fusarium species. Strains similar in morphology to Fusarium moniliforme and F. subglutinans were the most frequent isolates from maize and from teosinte species including Z. diploperennis, Z. lu.:rurians, Z. mays ssp. me:ricana, and Z. mays ssp. parviglumis. Analysis of fertility, vegetative compatibility and mycotoxin production identified 63 % of the 70 F. moniliforme strains from teosinte as genetically diverse members of Gibberella fujikuroi mating population A, a common pathogen of maize. The F. subglutinans strains from maize and teosinte were similarly genetically diverse, but were not fertile with standard testers of G. fujikuroi mating populations B and E, common pathogens of Poaceae, or of mating population H, which causes pitch canker disease of pine. Fifty-four percent of the 80 F. subglutinans strains were fertile when crossed with female tester strains from teosinte and maize collected in a field at Netzahualcoyotyl in the state of Mexico. These strains from Mexico and Central America may comprise a new and distinct G. fujikuroi mating population, but a strain from the Netzahualcoyotyl field site was fertile with a strain of G. fujikuroi mating pQpulation H from California. Thus, F. subglutinans from teosinte and maize may have a close relationship to mating population H from pine. INTRODUCTION distinguish from domesticated maize (Bird 1995, Vibrans &: Estrada Flores 1998). The ears and seeds of teosintes and The wild teosintes (Zea spp.) of the prairies, open woodlands, maize, however, are profoundly different. Whereas the maize and roadsides of Mexico, Guatemala and Nicaragua are the ear bears hundreds of naked seeds that remain attached to the closest relatives and probable ancestors of domesticated maize ear at maturity, teosinte species bear about a dozen seeds in (Zea nmys ssp. nmys) <Doebley 1994). Nineteenth-century an ear that shatters at maturity. Each teosinte seed is botanists recognised the affinity between teosintes and maize, completely covered by a very hard and lustrous triangular and introduced Guatemalan teosinte to Europe, the United fruitcase, giving it the general appearance of a small grey or States and elsewhere as a fodder crop. In the 1920s, maize brown pebble. Despite the dramatic differences in ear and seed breeders initiated exploration and collection of teosinte morphology between teosinte and maize, all species of throughout Mexico and Central America. Since the 1960s, teosinte can form hybrids with maize under natural conditions. Wilkes, litis, Doebley and others have monitored, collected Crosses of maize with the annual teosintes Z. nmys ssp. and characterised wild populations of teosinte, and defined a mexicana and parvigIumis are highly fertile, and progeny number of taxa: Z. dipIoperennis, Z. perennis, Z. Iuxurians, Z. demonstrate a range of morpholOgical traits intermediate nmys spp. huehuetenangensis, Z. mays ssp. parvigIumis, and Z. between the parents. Doebley and others <Doebley &: Stec nmys ssp. mexicana (Wilkes 1967, Doebley et aI. 1994, Taba 1991, Dorweiler et aI. 1993, Paterson et aI. 1995) have used 1995). interspecific crosses to show that relatively few genes with Zea mays ssp. mexicana is a troublesome weed of maize large effects are involved in transformation of the teosinte ear fields where, until flowering, the plant can be difficult to into the maize ear. Wild relatives of crop plants are important sources of genes • Corresponding author. for improving disease resistance and other complex traits of Disclaimer: Names are necessary to report factually on available data. but agricultural crops (Lenne &: Wood 1991). Various teosinte the USDA neither guarantees nor warrants the standard of the products. and species have been used or proposed as germplasm sources for the use of the name by USDA implies no approval of the product to the exclusion of others that may also be suitable. breeding maize with traits such as higher protein, seed 866 Gibberella fujikuroi and Fusarium subglutinans from Zea MaHng populaHon identification hardness, perennial growth, drought tolerance and resistance to viral diseases (Wilkes 1967, Taba 1995). Although teosintes All strains identified as F. moniliforme (syn. F. verticillioides) are reported to be susceptible to some fungi that are were crossed twice as males with tester strains of G. fufikuroi pathogenic to maize, including rusts, smut, downy mildew and mating population A (MP-A) and once with G. fujikuroi leaf blights (Shurtleff 1980, McGee 1988), there has been little MP-F (syn. G. thapsina) (Klittich et al. 1997). Strains of F. interest in occurrence of fungal pathogens in wild teosinte moniliforme that were not fertile with MP-A or MP-F were populations. Furthermore, we have found no published crossed twice as males with tester strains of G. fu.iikuroi MP­ information on susceptibility of teosinte to Fusarium species D (anamorph F. proliferatum) and MP-G (syn. G. nygamai) that are common pathogens of maize, or on the occurrence of (Klaasen & Nelson 1996), and with one set of tester strains Fusarium species in wild teosinte populations (Shurtleff 1980, of MP-H (syn. G. circinata) (Nirenberg & O'Donnell 1998) McGee 1988, Farr et al. 1989). Thus, the potential of teosinte from California and one set from South Africa. All strains species as sources of genes for improving maize resistance to identified as F. subglutinans were crossed twice as males with Fusarium is unknown. tester strains of G. fujikuroi MP-B (anamorph F. sacchari), MP­ The major goal of this study was to identify and E (anamorph F. subglutinans), and MP-H, and selected strains characterise Fusarium species from the closest wild relatives were crossed twice as males with MP-A, MP-D and MP-G. of maize. Our specific objectives were: (1) to survey Fusarium Crosses were made on carrot agar as described (Klittich & species infecting seeds of wild teosinte populations in Mexico, Leslie 1988, Klaasen & Nelson 1996), but male parents were Guatemala and Nicaragua; and (2) to determine whether F. cultured on V-8 juice agar (Stevens 1981) rather than subglutinans strains from wild teosinte and maize are members complete medium. Incubation conditions were either constant of a known or new mating population of Gibberella fujikuroi. light at 21°C, or a 12 h, 25° light/21° dark alternating cycle. Crosses were scored as positive when ascospores were MATERIALS AND METHODS observed upon microscopic examination of the contents of enlarged perithecia. Standard tester strains of seven mating Seed sample collection populations were obtained from the Fusarium Research Center Teosinte samples consisted primarily of seed collected during and other sources. Strain numbers were (MATA-2) M-3120, the 1970s, 1980s, and 1990s from wild populations in open (MATA-I) M-3125, (MATB-2) M-6865, (MATB-1) M-6866, woodlands, prairies, and roadsides, but some seed samples had (MATC-2) M-6883, (MATC-1) M-6884, (MAID-2) M-6992, been increased by cultivation (Doebley et al. 1984). Teosinte (MAID-1) M-6993, (MATE-2) M-3693, (MATE-I) M-3696, and maize seeds also were collected in November 1996 from (MATF-2) M-6563, (MATF-1) M-6564, (MATG-2) M-7491, plants growing together in a field at Netzahualcoyotyl near and (MATG-1) M-7492 (Kerenyi et al. 1999). Tester strains of Texcoco, Mexico (Bird 1995). Approximately 500 mature mating population H were (matH+) Fsp52 and (matH-) Fsp90 teosinte seeds were randomly collected from several dozen from California, and (matH+) MRC 6213 and (matH-) MRC plants of Z. mays ssp. mexicana. Approximately 300 symptom­ 7488 from South Africa (Britz et al. 1999). less, undamaged, maize seeds were randomly collected from To identify testers for a new mating population, selected 14 ears of blue and white maize. strains of F. subglutinans from teosinte and maize were crossed pairwise on carrot agar. Two strains, Fst51 and Fst55, from Netzahualcoyotyl teosinte and maize, respectively, were IsolaHon of Fusarium strains selected as testers because they had the best female fertility of all strains tested. All strains were tested twice as males, and Teosinte fruitcases were split, and the kernels were removed. strains were scored as fertile if perithecia containing mature Kernels of teosinte and maize were surface-disinfected by ascospores were obtained in one or more tests. For viability placing them in 0.5 % sodium hypochlorite for 1 min, and and mating type tests, single ascospores were picked at were rinsed twice in sterile water. Surface-disinfected teosinte random from crushed perithecia, transferred individually to V­ kernels were placed on the surface of a plate of Fusarium 8 juice agar and assessed for germination and for fertility as selective agar medium containing 0.1 % pentachloronitro­ males in crosses to strains Fst51 and Fst55. These two strains benzene (Nelson et al. 1983). Maize kernels were softened in were also tested in reciprocal crosses with tester strains of sterile water for approximately 1 h, then, split open with a mating populations A-H. sterile knife, and placed, cut surface down, on the same selective medium. Kernels were incubated for 5-7 d, then one VegetaHve compaHbility analysis colony per kernel was reisolated from a single spore, and identiifed to species using morphological criteria (Nelson et al.
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