Diversity and spatial distribution of vegetative compatibility types and mating types of <Emphasis Type="BoldItalic">Cryphonectria parasitica</Emphasis> in the Aydın Mountains,Turkey

In this study, the population structure of the chestnut blight fungus Cryphonectria parasitica in the Aydın Mountains was investigated to make inferences about fungal reproduction and population diversity. A total of 213 C. parasitica isolates from eight subpopulations were used to determine vegetative compatibility (vc) and mating types of the population. Furthermore geostatistical analysis was performed to define the spatial structure of the population. The results showed that the isolates were vegetatively compatible with the European vc types of either EU-1 or EU-12. Both vc types were found in almost all subpopulations, but their frequencies varied depending on location. The results of a PCR assay showed that both mating types of C. parasitica (MAT-1 and MAT-2) exist in the population. MAT-1 comprised 65% of the total isolates, and the ratio of mating types was significantly skewed from 1:1. Genotyping based on combined vc and mating type data revealed four genotypes: EU-1/MAT-1 (28.6%), EU-1/MAT-2 (34.7%), EU-12/MAT-1 (36.2%) and EU-12/MAT-2 (0.5%). Geostatistatical analysis indicated that vc types, mating types and vc/mating genotypes were spatially autocorrelated and clustered in their distributions. Results suggested that C. parasitica could have a clonal population structure that is generated by asexual reproduction. Low vc-type diversity suggests that the C. parasitica population in the Aydın Mountains may be highly suitable to hypovirus invasion, thereby providing a high potential for successful biological control. However, co-occurrence of sexually compatible strains of EU-1 and EU-12 at the same locations in close proximity creates a high risk of increase in vc-type diversity.     

New detections of chestnut blight in Great Britain during 2019–2020 reveal high Cryphonectria parasitica diversity and limited spread of the disease

Cryphonectria parasitica was detected for the first time in the United Kingdom in 2011. A 2017–2018 survey detected the disease at different sites in Berkshire, Derbyshire, Devon, Dorset and London, while the present study comprises the results of the 2019–2020 survey with new findings and additional sites in the United Kingdom (Berkshire, Buckinghamshire, Cornwall, Derbyshire, Devon, London, West Sussex) and in Jersey, reflecting the progressive detection of more infected trees. A total of 189 samples were collected from 52 sites, and 123 samples tested positive both by quantitative real-time PCR and/or isolation from 43 sites. A total of 115 isolates were tested for mating type, vegetative compatibility group (VCG) and Cryphonectria hypovirus 1 (CHV-1). Twelve VCGs were identified, with four of them being first records in United Kingdom. The highest diversity of VCGs was detected in Devon followed by West Sussex while London and Derbyshire presented the lowest. Both mating types were detected (41% MAT-1 and 59% MAT-2), and no heterokaryons were detected. Perithecia of C. parasitica were not observed at any site during this survey. CHV-1 was detected in three isolates in very low concentration from three different locations in London and was always the unmutated subtype I haplotype E-5. A greater diversity of VCGs at outbreak sites compared with previous surveys, combined with their scattered distribution and the slow spread of the pathogen, supports the hypothesis that this disease has been introduced through imports over time from Europe.     

Vegetative Compatibility and Mycotoxin Chemotypes among Fusarium graminearum (Gibberella zeae) Isolates from Wheat in Argentina

Gibberella zeae (anamorph Fusarium graminearum) is the main pathogen causing Fusarium head blight of wheat in Argentina. The objective of this study was to determine the vegetative compatibility groups (VCGs) and mycotoxin production (deoxynivalenol, nivalenol and 3-acetyl deoxynivalenol) by F. graminearum populations isolated from wheat in Argentina. VCGs were determined among 70 strains of F. graminearum isolated from three localities in Argentina, using nitrate non-utilizing (nit) mutants. Out of 367 nit mutants generated, 41% utilized both nitrite and hypoxanthine (nit1), 45% utilized hypoxanthine but not nitrite (nit3), 9% utilized nitrite but not hypoxanthine (NitM) and 5% utilized all the nitrogen sources (crn). The complementations were done by pairing the mutants on nitrate medium. Fifty-five different VCGs were identified and the overall VCG diversity (number of VCGs/number of isolates) averaged over the three locations was 0.78. Forty-eight strains were incompatible with all others, thus each of these strains constituted a unique VCG. Twenty-two strains were compatible with other isolates and were grouped in seven multimembers VCGs. Considering each population separately, the VCG diversity was 0.84, 0.81 and 1.0 for San Antonio de Areco, Alberti and Marcos Juarez, respectively. Toxin analysis revealed that of the 70 strains of F. graminearum tested, only 90% produced deoxynivalenol, 10% were able to produce deoxynivalenol and very low amounts of 3-acetyldeoxynivalenol. No isolate produced nivalenol. The results indicate a high degree of VCG diversity in the F. graminearum populations from wheat in Argentina. This diversity should be considered when screening wheat germplasm for Fusarium head blight resistance.     

Phylogenetic relationships of Fusarium oxysporum f. sp. melonis in Iran

Fusarium wilt of melon caused by Fusarium oxysporum f. sp. melonis is a destructive fungal disease in melon growing regions. Isolates of F. oxysporum obtained from six major melon producing provinces in Iran, from melons and other hosts, were characterized based on pathogenicity to melon, vegetative compatibility groups (VCGs) and nuclear ribosomal DNA intergenic spacer (IGS) sequencing. Thirty-four of 41 isolates from Iran in this study were identified as race 1,2 which belonged to either VCG 0134 or an unassigned VCG, which based on IGS sequencing grouped with the VCG 0135 tester isolate. The seven remaining isolates were identified as nonpathogenic to melon belonging to two undescribed VCGs. Based on sequence analyses of the IGS region of Iranian and foreign isolates, nine lineages were identified, each including one VCG. The separation of VCGs into distinct lineages based on IGS sequences is mostly consistent with Repetitive extragenic palindromic PCR (Rep-PCR) results. Exceptions are VCGs 0130 and 0131, which could be differentiated with IGS sequences, but not with Rep-PCR. Different races from the USA, France and Iran associated with VCG 0134 grouped into one IGS lineage but could be differentiated with Rep-PCR, suggesting that this VCG is more diverse than previously thought. Given the long history of melon cultivation in Iran and the Rep-PCR diversity of isolates belonging to this VCG, it could be speculated that VCG 0134 perhaps evolved in Iran.     

Genetic diversity in the mango malformation pathogen and development of a PCR assay

Malformation is a destructive disease of mango, Mangifera indica . Its causal agent possesses the morphological features of Fusarium subglutinans , a species whose taxonomy and nomenclature has recently been in a state of flux. Genetic diversity was examined among 74 F. subglutinans -like isolates from malformed mango in Brazil, Egypt, Florida (USA), India, Israel and South Africa. With nitrate-nonutilizing ( nit ) auxotrophic mutants, seven vegetative compatibility groups (VCGs) were identified. Three of the VCGs were found in a single country, and VCG diversity was greatest in Egypt and the USA where, respectively, four and three different VCGs were found. RAPD profiles generated with arbitrary decamer primers were variable among isolates in different VCGs, but were generally uniform for isolates within a VCG. In PCR assays, a 20-mer primer pair that was developed previously to identify F. subglutinans from maize (mating population [MP]-E of the Gibberella fujikuroi complex) also amplified a specific 448 bp fragment for isolates of F. sacchari from sugarcane (MP-B) and what was probably F. circinatum (pine, MP-H). With the exception of three isolates from Brazil, it did not amplify the fragment from F. subglutinans -like isolates from mango. A second pair of 20-mer primers was developed from a unique fragment in the RAPD assays. It amplified a specific 608 bp fragment for 51 of 54 isolates from mango (all but the three Brazilian isolates). It also amplified a smaller, 550 bp fragment from isolates of F. nygamai (MP-G), but did not amplify DNA of isolates of any other taxon of Fusarium that was tested.     

Clonal population structure and introductions of the chestnut blight fungus, <Emphasis Type="Italic">Cryphonectria parasitica</Emphasis>, in Asturias,northern Spain

To understand the history of introductions of the chestnut blight fungus, Cryphonectria parasitica, in the Principality of Asturias in northern Spain, we conducted an extensive survey of chestnut blight and collected C. parasitica from 216 sites. All 778 isolates were assayed for vegetative compatibility (vc) type, whereas a subsample of 301 isolates was assayed for mating type, and 189 isolates were genotyped at 16 microsatellite markers. We found low diversity for all markers. Nearly all isolates (95%) were compatible with vc type EU-1 and had the same microsatellite multilocus haplotype, or differed from the most common type by mutation at one locus. Approximately 5% of the isolates were vegetatively compatible with EU-13 and only two isolates (< 1%) were compatible with EU-3; five different microsatellite haplotypes were found among isolates in these latter two vc types. The overall mating-type ratio was 218 MAT-1: 81 MAT-2, with both mating types represented in each of the three vc types. Microsatellite haplotypes based on ten markers used in France showed that most isolates in Asturias were either identical to or only one marker different from one of the seven most common genotypes in France, RE103. Based on these ten markers alone, the population of C. parasitica in Asturias, would appear to have been founded by a single genotype from the C1 lineage (to which RE103 belongs) found in eastern France and northern Italy. However, additional genotyping by vc types suggests the introduction of multiple genotypes, with different vc types. The exact source for introduction into Asturias cannot be determined without additional genotyping of isolates from other locations. Regardless of their origin, the low diversity of vc types makes this population ideal for deploying hypovirulence because there will be few barriers for virus transmission between individuals.     

Characterization of Pyricularia grisea in the United States Using Independent Genetic and Molecular Markers

ABSTRACT A total of 540 isolates of Pyricularia grisea from rice in the United States were examined for vegetative compatibility, MGR586 DNA fingerprint diversity, and mating type based on hybridization with the mat1-1 and mat1-2 sexual mating type alleles. The collections contained both archived and contemporary field isolates representative of the known MGR586 lineages and races that occur throughout the United States. Complementary nitrate nonutilizing (nit) or sulfate nonutilizing (sul) mutants were used to assess vegetative compatibility in P. grisea. There was a complete correspondence between vegetative compatibility groups (VCGs), MGR586 lineage, and mating type among 527 contemporary isolates (collected between 1991 and 1997) from Arkansas, Louisiana, Missouri, Mississippi, and Texas; all isolates in MGR586 lineages A, B, C, and D belonged to VCGs US-01, US-02, US-03, and US-04, respectively. In addition, all isolates tested in VCGs US-01 and US-04 had the mat1-1 mating type allele whereas those in VCGs US-02 and US-03 had the mat1-2 allele. The strict association of independent markers during this sample period was consistent with a strictly asexual mode of reproduction. However, examination of archived isolates collected in the 1970s and 1980s and contemporary isolates revealed an incongruent relationship between the independent markers. MGR586 C and E isolates were vegetatively compatible which indicated that multiple robust MGR586 delineated lineages could be nested within certain VCGs. Although isolates in lineages C and E were vegetatively compatible, they were of opposite mating type. Several hypotheses, including recombination, could account for the incongruence between the various markers. Among the eight MGR586 lineages (A through H) that occur in the United States, all isolates in lineages A, D, E, G, and H had the mat1-1 allele, whereas isolates in lineages B, C, and F had the mat1-2 allele. Nit mutants can be recovered relatively easy from P. grisea and should allow large numbers of individuals within a population to be assessed for vegetative compatibility. VCGs may prove to be an effective multilocus marker in P. grisea. Thus, VCGs should be a useful means for characterizing genetic structure in populations of the rice blast fungus worldwide, provide a useful genetic framework to assist in interpreting molecular population data, and may provide insight into potential sexual or asexual recombination events.     

Population structure of Fusarium oxysporum f. sp. radicis-lycopersici in Florida inferred from vegetative compatibility groups and microsatellites

Fusarium crown and root rot caused by Fusarium oxysporum f. sp. radicis-lycopersici (FORL) is a serious soilborne disease reducing tomato yields in Florida, a leading state in fresh market tomato production in the United States. One hundred and twenty five isolates of FORL were collected from the three main tomato-growing counties in Florida between 2006 and 2008. Vegetative compatibility groups (VCGs) and 10 microsatellite loci were used to infer the population structure of FORL. Up to 69.8 % of the isolates could be assigned to one of three VCGs, 0094, 0098 or 0099, with frequencies of 38.6, 24.4, and 6.8 % respectively. A medium level of population differentiation (Φst?=?0.159) was detected among the three counties, and three optimal clusters (populations) were supported by discriminant analysis of principal components. In addition, each population had some individuals that likely migrated from other populations. Migration probably played an important role in shaping the population structure of FORL since repeated VCGs and multilocus genotypes were observed in the three counties. Considerable migrants (> 1.33 migrants per generation) were also detected between the three counties, resulting in an increase in the effective population size and genetic diversity of F. oxysporum f. sp. radicis-lycopersici. Although migration is an important evolutionary force, mutation and parasexual recombination could not be completely ruled out as contributing causes to the genetic diversity of FORL. Management strategies that limit the movement of F. oxysporum f. sp. radicis-lycopersici are necessary to reduce the evolutionary potential of the pathogen.     

Genetic diversity of the entomopathogen<Emphasis Type="Italic">Verticillium lecanii</Emphasis> on the basis of vegetative compatibility

Forty-three isolates ofVerticillium lecanii from insects, phytopathogenic fungi and other substrates were tested for vegetative compatibility by observing heterokaryon formation among complementary nitrate-nonutilizing (nit) mutants.nit mutants were isolated from 42/43 strains examined. Twenty-one isolates were self-incompatible, and the remaining 21 isolates were divided into 14 vegetative compatibility groups (VCGs): ten containing only a single strain each, and the remaining four containing two to four isolates each. Members of isolates in each of these VCGs all shared the same IGS haplotype. Further, the isolates within a VCG were correlated with one another in part by fragment patterns of mt-LrDNA, -SrDNA, Bt-2 and H4 region, by PCR-RFLP and -SSCP, but not by dsRNA. Two isolates belonging to VL-J2 have high virulence to aphids, whereas strains from VL-J1 lack this character. These findings indicate that two VCGs (VL-J1 and -J2) may originate from two distinct clonal lineages. Alternatively, high VCG diversity and HSI frequency ofV. lecanii might be associated with an array of distinct lineages. These data not only suggest relationships among DNA polymorphisms, virulence, and VCG, but also demonstrate genetic heterogeneity ofV. lecanii. http://www.phytoparasitica.org posting Sept. 30, 2003.     

High vegetative compatibility diversity of Cryphonectria parasitica infecting sweet chestnut (Castanea sativa) in Britain indicates multiple pathogen introductions

Chestnut blight, caused by Cryphonectria parasitica, was identified in Devon, UK, in December 2016. Intensive surveys detected the disease at further sites in Devon (seven), Berkshire (one), Dorset (one), Derbyshire (four) and a cluster of eight sites in southeast London. Over 570 survey samples were tested, and 227 were positive for C. parasitica by isolation and real-time PCR. A total of 227 isolates were tested for mating type, and 197 screened for vegetative compatibility group (VCG) and compared with VCGs known from mainland Europe. The same isolates were also screened for the presence of Cryphonectria hypovirus 1 (CHV-1). Eleven VCGs were identified within the UK population. Five corresponded to already known European VCGs but six were unique. The European VCGs mainly came from the Devon, Dorset, Berkshire and Derbyshire disease outbreaks, whilst unique VCGs were almost exclusively from the southeast London cluster. Both mating types were detected, but only one mating type was present at each site, with the exception of a single Devon site. Perithecia of C. parasitica were never observed at any site. CHV-1 was found in seven isolates from three different locations and was always subtype-I, which has limited hypovirulence. Therefore, although CHV-1 is associated with C. parasitica at some outbreaks, it probably has limited impact on virulence. The diversity of VCGs and their distribution at outbreak sites, together with findings of CHV-1, suggests C. parasitica has been introduced to the UK multiple times over at least two decades through international plant trade.     

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

Fusarium culmorum is a major pathogen able to cause foot and root rot and the incitant of Fusarium head blight in wheat in Tunisia. The aims of the present study were to evaluate by PCR the type of mycotoxins produced, to determine the mating type and to analyse the genetic diversity by microsatellite markers of 82?F. culmorum isolates recovered from two separated Tunisian fields. Specific sequences in the Tri6-Tri5 intergenic region, Tri7 and Tri13 were used to identify 3-AcDON- or 15-AcDON-. All studied F. culmorum isolates, were of the 3-AcDON- type. No 15-AcDON- and NIV types were detected in this research. Both mating types MAT1-1 and MAT1-2 were recovered from the two fields in approximately equal proportions. Five polymorphic microsatellite markers were applied to F. culmorum isolates, to determine the genetic variation in and among populations. Sixty-four haplotypes were identified; the analysis of the population structure did not reveal a strong variation between fields. Total gene diversity (H _T ?=?0.505; H _S ?=?0.497) and analysis of molecular variance confirmed that most of the genetic variability was within populations (Φ _ST ?=?0.033; P?<?0.0039). Gene flow (N _m ?=?31.05) indicated little differentiation among populations. Based on these results, the F. culmorum isolates collected from different fields might be part of one large panmictic population and in addition the low linkage disequilibrium values with high genetic variation within populations suggest that the population is recombining sexually.     

Pathogenicity, vegetative compatibility and RAPD analysis of Fusarium oxysporum isolates from tobacco fields in Extremadura

Fusarium wilt of tobacco could be caused by Fusarium oxysporum f. sp. batatas or f. sp. vasinfectum since f. sp. nicotianae was rejected because there was no evidence of isolates specific to tobacco. Forty isolates of F. oxysporum from soil and plants from tobacco fields in Extremadura (south-western Spain) were characterized by pathogenicity on burley and flue-cured tobacco, for vegetative compatibility group (VCG), and by random amplified polymorphic DNA (RAPD). Isolates from burley were identified as race 1 of F. oxysporum f. sp. batatas based on pathogenicity on tobacco, sweet potato and cotton, and those from flue-cured as race 2. Most isolates from soil were heterokaryon self-incompatible (HSI) and the remaining isolates from soil and tobacco were grouped into four VCGs: VCG 1 (5 isolates from burley), VCG 2 (17 isolates from flue-cured and 4 from soil), VCG 3 (2 isolates from flue-cured) and VCG 4 (2 isolates from soil). This is the first report of the two races and VCGs of F. oxysporum f. sp. batatas in Spain. Analysis of RAPD revealed two clusters (C-I and C-II) related to race and VCGs. C-I included race 1 (VCG 1) isolates from burley and nonpathogenic (VCG 4 or HSI) isolates from soils. C-II included nonpathogenic (VCG 2) and race 2 (VCG 2 or VCG 3) isolates from flue-cured. VCG and RAPD markers were effective in distinguishing race 2 from race 1, suggesting that there are two genetically differentiated groups of F. oxysporum f. sp. batatas on tobacco in Extremadura.     

Phytopathogenic, morphological, genetic and molecular characterization of a Verticillium dahliae population from Crete, Greece

A population of 84?V. dahliae isolates mainly originating from Crete, Greece, was characterized in terms of pathogenicity and virulence on different hosts, in parallel with morphological/physiological characterization, vegetative compatibility grouping and mating type determination. Tomato race 2 was found to have supplanted race 1 and was more virulent on a tomato-susceptible cultivar than race 1. Using a differential host classification system which tests pathogenicity to tomato, eggplant, sweet pepper and turnip, 59 isolates were assigned to tomato, 19 to eggplant, one to sweet pepper and five to tomato-sweet pepper pathogenicity groups. All isolates from Crete fell into VCG subgroups 2A, 2B and 4B, while a remarkably high incidence of bridging isolates (compatible with two or more VCGs) was recorded. The tomato-sweet pepper pathogenicity group was morphologically quite distinct from the others, while conidial length and pigment intensity were discriminatory parameters among VCGs 2A, 2B and 4B. PCR-based molecular marker Tr1/Tr2 was reliable in race prediction among tomato-pathogenic isolates, except for members of VCG 4B, while the application of markers Tm5/Tm7 and 35-1/35-2 was highly successful for tomato-pathogenic isolates. E10 marker was related to VCG 2B, rather than to pathogenicity groups. A single nucleotide polymorphism in the ITS2 region, and two novel molecular markers, M1 and M2, proved useful for the fast and accurate determination of major VCGs 2A, 2B and 4B, and can be used for high-throughput population analyses in future studies. The mating type was unrelated to VCG classification and probably does not control heterokaryon incompatibility in V. dahliae.     

A molecular diagnostic for tropical race 4 of the banana fusarium wilt pathogen

This study analysed genomic variation of the translation elongation factor 1α (TEF‐1α) and the intergenic spacer region (IGS) of the nuclear ribosomal operon of Fusarium oxysporum f. sp. cubense (Foc) isolates, from different banana production areas, representing strains within the known races, comprising 20 vegetative compatibility groups (VCG). Based on two single nucleotide polymorphisms present in the IGS region, a PCR‐based diagnostic tool was developed to specifically detect isolates from VCG 01213, also called tropical race 4 (TR4), which is currently a major concern in global banana production. Validation involved TR4 isolates, as well as Foc isolates from 19 other VCGs, other fungal plant pathogens and DNA samples from infected tissues of the Cavendish banana cultivar Grand Naine (AAA). Subsequently, a multiplex PCR was developed for fungal or plant samples that also discriminated Musa acuminata and M. balbisiana genotypes. It was concluded that this diagnostic procedure is currently the best option for the rapid and reliable detection and monitoring of TR4 to support eradication and quarantine strategies.     

Population structure of Fusarium fujikuroi from California rice and water grass

The recent observance of Fusarium fujikuroi, the causal agent of Bakanae disease of rice, in California provides a unique opportunity to assess the population diversity of an introduced pathogen in a new environment. We collected 172 isolates of this pathogen between 2000 and 2003 from California rice and two from water grass (Echinochloa spp.). Pathogenicity of F. fujikuroi was demonstrated on early water grass (E. oryzoides) and barnyard grass (E. crus-galli) indicating that weed control should be part of Bakanae management programs. Both mating types and six unique amplified fragment length polymorphism haplotypes corresponding to six identified vegetative compatibility groups were detected. The two most frequently isolated haplotypes encompassed 94% of the collected isolates, suggesting that clonal reproduction dominates. Coefficients of similarity between the unique haplotypes ranged from 0.94 to 0.98, and indicate that there is very little genotypic variation in the F. fujikuroi population in California. The near fixation of the MAT-1 idiomorph (observed ratio 170 MAT-1:4 MAT-2), is consistent with a hypothesis of predominant or exclusive asexual reproduction. The low level of introduced genotypic diversity, in conjunction with the asexual reproductive strategy of this population will slow evolutionary processes, including adaptation to the California environment.     

香荚兰尖孢镰刀菌营养体亲和性测定的初步研究

 Eight hundred twenty four nit mutants were induced from 73 strains of Fusarium oxysporum f. sp. vanillae, and classified into four phenotypes by their abilities to utilize different nitrogen sources. Among these mutants, 64.9% were characterized as nit 1, 24.3% as nit 3, 9.8% as nit M, 1.0% as nit X. Based on complementary pairing tests of different nit mutants on the medium MM, 44 isolates belonged to 8 different VCGs, 29 isolates were classified into single and different VCGs. These results indicated that there was significant VCG diversity in Fusarium oxysporum f. sp. vanillae population. VCGs might be correlated with geographic origin of strains, but no close correlation was found between VCGs and pathogenicity.     

Vegetative compatibility groups ofVerticillium dahliae from cotton in the southeastern anatolia region of Turkey

Eighty isolates ofVerticillium dahliae from the southeastern Anatolia region and 20 isolates from the east Mediterranean region from wilted cotton plants were used for vegetative compatibility analysis employing nitrate non-utilizing mutants and reference tester strains of vegetative compatibility groups (VCGs) 1A, 2A, 2B, 3, 4A and 4B. Of the 100V. dahliae isolates, 49 were assigned to VCG1A, 39 to VCG2B, nine to VCG2A and three to VCG4B. Pathogenicity assays were conducted on susceptible cotton cv. Çukurova 1518 in the greenhouse. All VCG1A isolates induced defoliation and all VCG2B isolates caused partial defoliation symptoms. Isolates of VCG2A and VCG4B caused typical symptoms of leaf chlorosis without defoliation. This is the first report on VCGs ofV. dahliae in the southeastern Anatolia region of Turkey, which demonstrates that VCG1A of the cotton-defoliating type and VCG2B of the partially defoliating type are prevalent in this region.     

Development and application of new molecular markers for analysis of genetic diversity in Verticillium dahliae populations

The aim of this study was to develop new polymorphic markers for analysis of genetic diversity in the fungal soilborne plant pathogen Verticillium dahliae. Twelve polymorphic markers (five microsatellites and seven polymorphic sequences) were developed from a genomic library enriched for microsatellites. Screening of polymorphic loci was done using a collection of 25 V. dahliae isolates of diverse geographic origins, host sources and vegetative compatibility groups (VCGs). Three methods were used to score alleles: polyacrylamide gel electrophoresis (PAGE), sequencing of PCR‐amplified loci, and capillary electrophoresis. The new markers were used to assess genetic differentiation between isolates associated with different host plants. Two collections of isolates were analysed, obtained from artichoke (30 isolates) and potato (20 isolates) from crops grown in rotation located in the same area in eastern‐central Spain. The resolution of genetic differentiation between these two collections using the new markers was compared to that provided by other often‐used markers (SCARs and VCGs). Sequence analysis of the alleles proved to be the most unambiguous technique for scoring microsatellite data. The relatively high genetic differentiation observed between isolates from different crops (genetic differentiation coefficient, G_ST = 0·24) and their high genotypic diversity suggest a divergence between V. dahliae from artichoke and potato. It is hypothesized that evolution of V. dahliae from the local resident population in association with the two host crops has occurred. The new markers are useful for resolving population structure within V. dahliae and may contribute to a better understanding of the population biology of this fungus.     

Genetic structure of <Emphasis Type="Italic">Fusarium verticillioides</Emphasis> populations isolated from maize in Argentina

Fusarium verticillioides (sexual stage Gibberella moniliformis) is a common fungal pathogen of maize worldwide that also produces fumonisin mycotoxins. Populations of this fungus can be diverse with respect to neutral and selectable genetic markers. We used vegetative compatibility groups (VCGs) and amplified fragment length polymorphisms (AFLPs) to evaluate the genetic structure of three F. verticillioides populations from commercial maize fields in Argentina. Based on work with similar populations from outside South America, we expected individuals within the populations to be genetically diverse, that genotypic variation would be distributed in a manner consistent with random mating, and that populations from different locations would be genetically indistinguishable from one another. We analysed 62 AFLP loci for 133 fungal isolates. All three populations were genotypically diverse but genetically similar and potentially part of a larger, randomly mating population, with significant genetic exchange occurring between the three subpopulations. There was no evidence for linkage disequilibrium at P = 0.05. The low values of G _ST , the lack of frequent private alleles, and the lack of a systemic pattern of linkage disequilibrium all suggest that sexual reproduction is sufficiently common in F. verticillioides and that the dispersal of strains is sufficiently efficient for the population of F. verticillioides in the main maize growing region to be a single randomly mating population with no detectable genetic subdivision. Thus differences in disease and/or toxin production observed in this region are best attributed to differences other than the genetic composition of the population.     

Mycotoxin Production and Molecular Variability of European and American Isolates of Fusarium Culmorum

The main causative agents of Fusarium head blight are Fusarium graminearum and Fusarium culmorum. We examined the mycotoxin-producing abilities and molecular variability of 37 Fusarium culmorum isolates collected from the Pan-Northern Hemisphere, together with isolates representing related species. Mycotoxin-producing abilities of the isolates were tested by thin layer chromatography and by PCR using primer pairs specific for the Tri7 and Tri13 genes. Thirty isolates belonged to chemotype I (producing deoxynivalenol and 3-acetyl-deoxynivalenol), while seven represented chemotype II (producing nivalenol and/or fusarenone X). The presence of a functional Tri7 gene correlated well with nivalenol production. Isolates belonging to chemotype I were in general more pathogenic in in vitro tests than those belonging to chemotype II. Phylogenetic analysis of the random amplified polymorphic DNA profiles (RAPD) of the isolates enabled the isolates to be clustered into different groups. Most isolates from Hungary exhibited identical RAPD profiles. A similar clustering was found on the tree based on restriction analysis of the intergenic spacer region data. Sequence analysis of a putative reductase gene fragment of the isolates was also carried out. A correlation was detected between the geographic origin of the isolates and their position on the cladogram produced based on sequence data. The presence of mating type gene homologues was also tested with primer pairs specific for MAT1-1 and MAT1-2. The isolates carried either MAT1-1 or MAT1-2 homologues. No correlation was observed between clustering of the isolates based on RAPD, restriction analysis of the intergenic spacer region or sequence data and the distribution of MAT idiomorphs. Similarly, no correlation was detected between mycotoxin-producing abilities or aggressiveness and molecular characteristics of the isolates. Statistical analysis of RAPD data and lack of strict correlation between trees based on different data sets supported the view that Fusarium culmorum has a recombining population structure. The presence of mating type gene homologues in the isolates indicates that the recombining population structure is caused by ongoing or past meiotic exchanges.