Mitochondrial DNA Variability in Fusarium Proliferatum (Gibberella Intermedia)

Restriction fragment length polymorphisms (RFLP) were used to assess genetic diversity of mitochondrial DNA (mtDNA) among 184 isolates of Fusarium proliferatum recovered from maize, asparagus, palms and reed. All strains were cross-fertile with standard mating type tester strains of Gibberella intermedia. Sixteen mitochondrial haplotypes were identified following digestion of DNAs with HaeIII, with seven, seven, five and six different haplotypes from maize, asparagus, palms and reed, respectively. Four haplotypes (I, III, IV and VII) were found on more than one host. Of these four, haplotype I was dominant on maize, representing 71% of the isolates. The banding patterns for haplotypes III and IV were >90% similar to the banding pattern of haplotype I. Haplotypes I, III and IV accounted for 87% of the isolates from maize, but were less common on the other hosts, accounting for 70%, 52% and 33% of the isolates from asparagus, palms and reed, respectively. Thirteen of the 16 haplotypes were recovered from only a single host plant species. When comparing the banding patterns and frequencies of these haplotypes, at least five were recovered at a higher frequency from one host relative to the others. Our results suggest that mtDNA RFLP analysis is a useful indicator of genetic divergence in Fusarium proliferatum.     

Genetic diversity among isolates of Fusariumoxysporum f.sp. canariensis

Fusarium oxysporum f.sp. canariensis causes vascular wilt disease of Phoenix canariensis , the Canary Island date palm. Seventy-two isolates of this fungus were obtained from diverse geographic locations including France, Japan, Italy, the Canary Islands, and California, Florida and Nevada, USA. The isolates were tested for vegetative compatibility and for similarities based on mitochondrial DNA (mtDNA), single-copy sequences and repetitive DNA (pEY10) polymorphisms. Seventy-one percent of the isolates belonged to a single vegetative compatibility group (VCG 0240), and four closely related mitochondrial RFLP patterns were found. A subset of the isolates was further tested for single-copy RFLPs and repetitive DNA fingerprints. Only four single-copy RFLP haplotypes were found among 25 representative isolates of F. oxysporum f.sp. canariensis tested, using nine polymorphic single-locus probe/enzyme combinations. Finally, 32 different pEY10 DNA fingerprints were found out of 57 isolates examined. Overall the results indicate that F. oxysporum f.sp. canariensis is a single lineage with a low to moderate level of genetic diversity.     

Ribotyping of EntomopathogenicVerticittium lecanii in Japan

To estimate the genetic diversity in 30 isolates ofVerticillium lecanii from aphids, whiteflies, mite and black pine in Japan, including two commercialized strains (Mycotal and Vertalec), DNA polymorphisms in ribosomal DNA of those isolates were analyzed using polymerase chain reaction (PCR). The internal transcribed spacer (ITS) and intergenic spacer (IGS) regions of the nuclear ribosomal RNA gene of each isolate were analyzed by PCR-RFLP (restriction fragment length polymorphism). The size of the PCR product from the ITS region was ~ 580 bp in 27 of the isolates. A 600 bp ITS product was detected in Mycotal and Vertalec. One Japanese isolate produced both the 580 bp and 600 bp products. Enzymatic digestion of the ITS region with Sau3A I,Msp I,Hae III andRsa I revealed RFLPs that consisted of eight haplotypes. Mycotal and Vertalec were specific haplotypes that differed from other isolates. The Japanese isolates had a complex relationship with the original host, but we identified several specific haplotypes common to an aphid origin. Ten distinct IGS haplotypes were detected in the IGS region, some of which were associated with aphid and whitefly origins. These results suggest that the haplotype of rDNA RFLP analysis can be used for studying genetic diversity inV. lecanii.     

Fusarium oxysporum f. sp. cubense Consists of a Small Number of Divergent and Globally Distributed Clonal Lineages

ABSTRACT A worldwide collection of Fusarium oxysporum f. sp. cubense was analyzed using anonymous, single-copy, restriction fragment length polymorphism (RFLP) loci. Several lines of evidence indicated that this pathogen has a clonal population structure. Of the 165 isolates examined, only 72 RFLP haplotypes were identified, and nearly half the isolates were represented by the five most common haplotypes. Individuals with identical haplotypes were geographically dispersed, and clone-corrected tests of gametic disequilibrium indicated significant nonrandom association among pairs of alleles for 34 of 36 loci tested. Parsimony analysis divided haplotypes into two major branches (bootstrap value = 99%) that together contained eight clades supported by significant bootstrap values. With the exception of two isolates, all isolates within a vegetative compatibility group were in the same clade and clonal lineage. Clonal lineages were defined by isolates having coefficients of similarity between 0.94 and 1.00. Ten clonal lineages were identified, and the two largest lineages had pantropical distribution. Minor lineages were found only in limited geographical regions. Isolates composing one lineage (FOC VII) may represent either an ancient genetic exchange between individuals in the two largest lineages or an ancestral group. The two largest lineages (FOC I and FOC II) and a lineage from East Africa (FOC V) are genetically distinct; each may have acquired the ability to be pathogenic on banana independently.     

Characterization of Phytophthora infestans populations of southern Brazil in 2004 and 2005

The populations of Phytophthora infestans (Pi) in southern Brazil in 2004 and 2005 are characterized herein. The isolates were collected from potato and tomato plants in the states of Paraná (PR), Santa Catarina (SC), and Rio Grande do Sul (RS). The mating type of 131 potato and 32 tomato isolates was determined. Forty-nine isolates from potatoes and 11 from tomatoes were analyzed for their Gpi phenotype. A subset of 35 isolates was evaluated for mitochondrial (mtDNA) polymorphisms. A sample of 146 isolates was tested for sensitivity to the fungicide metalaxyl, and most isolates (64%) were moderately sensitive. Fifty-nine isolates were classified as A1 mating type and 103 as A2. One isolate behaved as both A1 and A2 mating type. All tomato isolates were A1 mating type and presented the 86/100 pattern for the enzyme GPI and mtDNA Ib, indicating that these isolates belong to the US-1 clonal lineage. Of the 131 potato isolates, 103 were A2, 27 were A1 and one was A1/A2 mating type. Among the potato isolates 27 exhibited the Gpi phenotype 100/100, the same as BR-1, and 20 were 86/100, the same as US-1. Potato isolates presented the mitochondrial haplotypes Ia (74%) and IIa (26%). The data suggest the presence of only the BR-1 clonal lineage on potatoes in the states of PR and SC. However, in the state of RS, more than one clonal lineage was observed infecting potatoes, and there may be sexual reproduction between the lineages.     

Differentiation of Rhizoctonia AG-D Isolates from Turfgrass into Subgroups I and II Based on rDNA and RAPD Analyses

Binucleate Rhizoctonia anastomosis group (AG) D is the cause of rhizoctonia-patch and elephant-footprint diseases of zoysiagrass, and winter-patch disease of bentgrass. Rhizoctonia AG-D is also known as the causal pathogen of other diseases such as sharp-eye-spot of cereals, foot-rot of cereals and winter-stem-rot of mat rush. Isolates of AG-D have been divided into the two subgroups AG-D (I) and AG-D (II), based on the results of cultural characteristics and pathogenicity tests. Isolates obtained from zoysiagrass exhibiting symptoms of rhizoctonia-patch disease, from bentgrass with winter-patch disease, from wheat with foot-rot disease, and from mat rush with winter-stem-rot disease were reported to belong to subgroup AG-D (I). On the other hand, isolates obtained from zoysiagrass with elephant-footprint disease were assigned to subgroup AG-D (II). To confirm the existence of these two subgroups in AG-D, the genetic structure of AG-D isolates from turfgrass and other crops was compared. RFLP analysis of the ITS region from rDNA after digestion with the restriction enzymes EcoRI, HaeIII, HhaI, HinfI, and MboI separated AG-D isolates into two groups corresponding to AG-D (I) and AG-D (II). Furthermore, other AGs except AG-Q (AGs-A, Ba, Bb, C, E, F, G, I, K, L, O, P, and R. solani AG1-IC) did not have the same patterns that were seen for the two AG-D subgroups. AG-Q isolates from bentgrass showed the same patterns as AG-D (I). The results of the RAPD analysis also revealed the existence of two groups that corresponded to AG-D (I) and AG-D (II). These analyses revealed that Rhizoctonia AG-D isolates from turfgrass could be divided into two subgroups consistent with those based on cultural characteristics and pathogenicity. In addition, isolates of foot-rot disease of wheat and isolates of winter-stem-rot disease of mat rush whose cultural characteristics were the same as those of AG-D (I) also showed similar RFLP and RAPD patterns to those of AG-D (I) isolates from turfgrass.     

Evidence for Natural Selection in the Mitochondrial Genome of Mycosphaerella graminicola

ABSTRACT Pathogenicity assays were combined with restriction fragment length polymorphism (RFLP) markers in the mitochondrial and nuclear genomes to compare Mycosphaerella graminicola populations adapted to bread wheat (Triticum aestivum) and durum wheat (T. turgidum) in the Mediterranean Basin. The majority of isolates had unique nuclear DNA fingerprints and multilocus haplotypes. Only six mitochondrial DNA (mtDNA) haplotypes were identified among 108 isolates assayed. There were minor differences in frequencies of alleles at nuclear RFLP loci between the two host-adapted populations, but differences in the frequencies of mtDNA haplotypes were highly significant (P < 0.0001). mtDNA haplotype 1 dominated on the isolates adapted to bread wheat, and its frequency was twice as high as for the isolates adapted to durum wheat. mtDNA haplotype 4, which contained a unique approximately 3-kb insertion, was detected only in isolates showing specificity toward durum wheat and was the dominant haplotype on this species. We propose that the low mitochondrial diversity in this pathogenic fungus is due to a selective sweep and that differences in the frequencies of mtDNA haplotypes between the two host-adapted populations were due to natural selection according to host species.     

Phenotypic and genotypic structure of Phytophthora infestans populations on tomato and potato in the North of Thailand in 2000–2002

A total of 80 single–lesion isolates of Phytophthora infestans were collected from tomatoes and potatoes in several locations in Chiang Mai and Tak provinces in 2000–2002. These isolates were analyzed for mating type, metalaxyl sensitivity, mitochondrial DNA (mtDNA) haplotype RFLP pattern as determined by probe RG57, and for microsatellite markers. All isolates were A1 mating type. Isolates from tomato were usually sensitive to metalaxyl, but isolates from potato were usually resistant to metalaxyl. With one exception, all tomato isolates were related to the US-1 clonal lineage. With two exceptions, all potato isolates were related to two European lineages. In these two provinces, the populations of P. infestans on tomatoes are clearly different from those on potatoes.     

Pathogenic, Genetic and Molecular Characterisation of Fusarium oxysporum f.sp. lilii

Isolates of Fusarium oxysporum from lily were screened for pathogenicity, vegetative compatibility and DNA restriction fragment length polymorphisms, and compared to reference isolates of F. oxysporum f.sp. gladioli and F. oxysporum f.sp. tulipae to justify the distinction of F. oxysporum f.sp. lilii. Twenty-four isolates from different locations in The Netherlands (18 isolates), Italy (4 isolates), Poland and the United States (1 isolate each) shared unique RFLP patterns with probes D4 and pFOM7, while hybridization did not occur with a third probe (F9). Except for a self-incompatible isolate, these 24 isolates all belonged to a single vegetative compatibility group (VCG 0190). Isolates belonging to VCG 0190 were highly pathogenic to lily, but not to gladiolus or tulip, except for a single nonpathogenic isolate. Six saprophytic isolates of F. oxysporum from lily were nonpathogenic or only slightly aggressive to lily, gladiolus and tulip, belonged to unique VCGs and had distinct RFLP patterns. Three pathogenic isolates previously considered to belong to F. oxysporum f.sp. lilii were identified as F. proliferatum var. minus; all three belonged to the same VCG and shared unique RFLP patterns. These three isolates were moderately pathogenic to lily and nonpathogenic to gladiolus and tulip. The reference isolates of F. oxysporum f.sp. tulipae were pathogenic to tulip, but not to lily and gladiolus; they shared a distinct RFLP pattern, different from those encountered among pathogenic and saprophytic isolates from lily, and formed a separate new VCG (VCG 0230). Reference isolates of F. oxysporum f.sp. gladioli belonging to VCG 0340 proved pathogenic to both gladiolus and lily, but not to tulip. These isolates, as well as isolates belonging to VCGs 0341, 0342 and 0343 of F. oxysporum f.sp. gladioli, had RFLP patterns different from those encountered among the isolates from lily or tulip. These findings identify F. oxysporum f.sp. lilii as a single clonal lineage, distinct from F. oxysporum f.sp. gladioli and f.sp. tulipae.     

Genetic Relationships Between Cercospora kikuchii Populations from South America and Japan

ABSTRACT A collection 160 isolates of Cercospora kikuchii was made from South America and 245 from Japan. DNA fingerprint patterns were analyzed based on amplified fragment length polymorphism among the sample isolates, dividing the isolates into seven lineages (I to VII). Partial nucleotide sequence analyses of the beta-tubulin gene supported this division into seven lineages. Lineages I and III commonly existed in South America and Japan. In all, 136 of the 160 isolates from South America and 223 of the 245 isolates from Japan belonged to lineage I, indicating that lineage I was the major lineage in each area; 5 isolates from South America and 8 isolates from Japan belonged to lineage III. Lineages II (12 isolates) and IV (2 isolates) were specific to Japan and lineages V (3 isolates), VI (1 isolate), and VII (15 isolates) specifically existed in South America. These results suggest that the population genetic structure of C. kikuchii was different between South America and Japan, but the dominance of lineage I was common between the two areas.     

Detection of the source of infection of apple trees by Cylindrocarpon heteronema using DNA polymorphisms

Restriction fragment length polymorphisms (RFLPs) of the ribosomal DNA (rDNA) and mitochondrial DNA (mtDNA) of isolates of Cylindrocarpon heteronema were analysed using rDNA from Saccharomyces carlsbergensis and mtDNA from C. heteronema as probes. These analyses revealed intraspecific heterogeneity; four rDNA and six mtDNA restriction pattern categories were observed among the isolates tested. The two main rDNA RFLP categories, both of which subdivided into two mtDNA categories, detected within the UK isolates could not be associated with the localities from which they were obtained, but the majority of isolates originating from trees produced in the same nursery, irrespective of where they were finally planted, belonged to the same category.     

Phylogenetic and Pathotypic Analysis of Rice Bacterial Blight Race 3

Eight Philippine races of Xanthomonas oryzae pv. oryzae have been identified based on virulence phenotypes observed on a set of five differential varieties. One of these, Race 3, was found to consist of two phylogenetically distinct lineages based on DNA fingerprinting analysis. To determine, if the two lineages could be differentiated based on host-specificity, 186 strains of Race 3 were analyzed with additional fingerprints and 76 selected isolates with additional differential rice varieties. The strains were separated into 36 haplotypes clustering in three groups (IS1113-B, -C, and -G) at the 75% similarity level. Isolates varied in their reaction to a rice line carrying the resistance gene Xa7, however, the variability was not consistent within lineage. Aggressiveness of isolates belonging to lineage IS1113-B and -G was significantly greater when tested during the dry season than when tested during the wet season. However, no such differences were evident for isolates from lineage IS1113-C, indicating that environmental effects presumably light and temperature are genotype-specific.     

江西省稻区稻瘟病菌遗传宗谱与致病型的关系

为了探寻稻瘟病菌无性世代DNA水平的变异,明确江西省稻区稻瘟病菌遗传宗谱与致病型之间的对应关系,利用rep-PCR(repetitive element-based PCR)分子指纹分析技术,对稻区稻瘟病菌的群体结构和遗传多样性进行分析,并用41株代表性菌株对35个水稻品种进行了致病性测定。结果表明,以相似度75%为界,可以将不同稻区采集的99个菌株划分为14个遗传宗谱,其中,宗谱4、1和10为优势宗谱,分别包含37、18和12个菌株,占总数的37.37%、18.18%和12.12%;稻瘟病菌遗传宗谱与致病型间存在复杂的关系,同一宗谱的菌株对应多个致病型,而同一致病型的菌株,分属于不同的遗传宗谱,两者之间不存在简单的对应关系。     

Potential of sexual reproduction among host-adapted populations of Phytophthora infestans sensu lato in Ecuador

To determine the potential of sexual reproduction among host-adapted populations of Phytophthora infestans sensu lato in Ecuador, 13 A1 isolates belonging to clonal lineages US-1, EC-1 and EC-3, and 11 A2 isolates belonging to the clonal lineage EC-2, were paired on agar plates to induce crossing. In the first experiment, six A1 isolates (three US-1, two EC-1 and one EC-3) were each crossed with three A2 isolates (total = 18 crosses). Matings involving isolates of the EC-1 lineage produced more oospores of healthy appearance than did matings with isolates of US-1 or EC-3. In the second experiment, the oospores of 35 crosses (21 EC-1 × EC-2; 10 US-1 × EC-2; four EC-3 × EC-2) were dispersed on water agar to assess oospore germination. Overall, germination percentages were low. Only one cross produced enough progeny for evaluation. Twenty-three single-oospore offspring were isolated and evaluated for mating type; electrophoretic patterns of glucose-6-phosphate isomerase ( Gpi ) and peptidase ( Pep ) alloenzyme loci; mitochondrial DNA haplotype; and genomic DNA fingerprint. Multilocus genotype data indicated that all 23 isolates resulted from meiotic recombination. Four progeny with homothallic phenotype appeared to be unstable heterokaryons. Markers at several loci segregated according to simple Mendelian expectations for a diploid organism, but the ratios of three RFLP loci and the Pep locus were not consistent with Mendelian expectations. All progeny were nonpathogenic on hosts of the parental genotypes. Reduced mating success and reduced pathogenic fitness of progeny appear to be postmating mechanisms of reproductive isolation in populations of P. infestans sensu lato in Ecuador.     

Characterization of a new cultural type (LP) of Rhizoctonia solani AG2-2 isolated from warm-season turfgrasses, and its genetic differentiation from other cultural types

Isolates of Rhizoctonia solani AG2-2 obtained from turf with symptoms of large-patch disease of warm-season turfgrasses were compared with known AG2-2 isolates belonging to cultural types IIIB and IV. Some isolates that were previously identified as type IV have been separated here and named LP isolates. Comparisons among isolates were based on cultural morphology, hyphal growth rate, pathogenicity and restriction fragment length polymorphism (RFLP) analysis in the nuclear encoded ribosomal DNA (rDNA) genes. The cultural characteristics of LP isolates varied from those of types IIIB and IV. LP isolates did not show distinct sclerotial formation and zonation, and the colour of their mycelia and pigment deposition was dark brown. LP isolates had slower hyphal growth rates than types IIIB and IV, with an optimum temperature of 25°C compared with 28°C for types IIIB and IV. LP isolates were less virulent on radish but highly virulent on zoysia grass when compared with isolates of types IIIB and IV. Genomic DNA was digested separately with Eco RI, Ban III, Xba I and Sal I, and probed with cloned rDNA from Alternaria alternata in Southern hybridizations. LP isolates had one RFLP pattern, while both IIIB and IV possessed four different patterns each. Cluster analysis of RFLPs showed that R. solani AG2-2 is divided into three genetic subgroups, consisting of the IIIB, IV and LP isolates, respectively. The polymerase chain reaction (PCR) amplified rDNA internally transcribed spacer (ITS) regions of the IIIB, IV and LP isolates had the same length but produced different restriction patterns when digested with Msp I and Taq I. These results indicate that there are three cultural types in R. solani AG2-2, namely IIIB, IV and LP.     

Genetic characterization of the Golovinomyces cichoracearum complex in Australia

To characterize the evolutionary lineages of the Golovinomyces cichoracearum complex on introduced plants in Australia, the rDNA ITS regions from 47 herbarium specimens were compared by RFLPs and sequencing. Six RFLP groups were found, each corresponding to a previously reported evolutionary lineage in the complex. The largest of these groups (Group 1) contained 15 specimens infecting a range of tribes in the Asteraceae, despite a previous report that this lineage is largely restricted to the Tribe Heliantheae (Asteraceae). This group contained the only specimens with ascomata. Curved foot cells were formed by two lineages; one (Groups 5 & 6) containing specimens from a range of host families including the Asteraceae (Tribe Lactucae only), the other (Groups 2 & 3) containing members of the Asteraceae (Tribe Anthemidae only). Group 5 may represent G. orontii sensu stricto , while Group 6 is currently unnamed. Golovinomyces cichoracearum sensu stricto (specimens from Scorzonera ) formed a distinct group that did not contain any specimens from Australia. Suggestions are made for future species delimitation in the complex.     

Molecular marker analysis of European Setosphaeria turcica populations

Setosphaeria turcica is the causal agent of northern corn leaf blight, a foliar maize disease of worldwide economic importance. In Europe, its severity increases. To investigate the pathogen's population-genetic structure in central Europe, a total of 80 isolates was sampled in Germany, Switzerland, France, Austria, and Hungary and investigated with 52 random amplified polymorphic DNA (RAPD) markers. The mating type of the isolates was determined in testcrosses. Among the 73 isolates from maize there were 26 different RAPD haplotypes. All isolates with identical haplotype are considered clonemates. The haplotype shared by most members was represented by 22 isolates from Germany, Switzerland, and France, indicating high fitness and substantial migration. Only a single clone had members in both southeastern Austria and southwestern Switzerland, suggesting that the Alps constitute a major barrier for this pathogen. Several haplotypes differed by only one or two RAPD bands from the predominant haplotype and may have arisen by mutation. Few other clonal lineages were detected. The evolution of some haplotypes could not be explained by mutation alone. Sexual recombination may rarely occur. In population samples from Germany, Switzerland, and France, mating type MAT2 was predominating, while most isolates from Austria and Hungary had MAT1. Seven isolates from Johnson grass (Sorghum halepense), an alternative host of S. turcica, were clonemates and very different in RAPD haplotypes from all isolates collected from maize.     

Multigene phylogenetic analysis of inter- and intraspecific relationships in <Emphasis Type="Italic">Venturia nashicola</Emphasis> and <Emphasis Type="Italic">V. pirina</Emphasis>

Isolates of Venturia species isolated from pear in Japan, China, Taiwan and Israel were used in this study to analyze their molecular phylogenetic relationship. The nucleotides of rDNA-ITS, partial β-tubulin and elongation factor 1α genes were sequenced directly after PCR. Based on these sequence data two phylogenetic groups could be distinguished. Isolates collected from Asian pears such as Japanese and Chinese pears formed a distinct evolutionary lineage from those derived from European and Syrian pears. This result corroborated the early taxonomic separation of V. nashicola from V. pirina. In addition, trees from single-locus data sets and the combined data set showed that all isolates of V. nashicola were included in a monophyletic group and representative isolates of five pathological races originating from different locations and cultivars formed a single lineage. In contrast, two distinct evolutionary lineages were revealed in V. pirina and isolates of five races were scattered in two lineages. Israeli isolates of race 2 as well as two Japanese isolates of V. pirina formed a distinct lineage from other isolates of this species, while other Israeli isolates belonging to races 1, 3, 4, and 5 were closely related to each other and formed another lineage. It was indicated that the evolution of pathological races in V. nashicola might have occurred relatively recently as compared with V. pirina.     

Three evolutionary lineages of tomato wilt pathogen, Fusarium oxysporum f. sp. lycopersici, based on sequences of IGS, MAT1, and pg1, are each composed of isolates of a single mating type and a single or closely related vegetative compatibility group

Three evolutionary lineages of the tomato wilt pathogen Fusarium oxysporum f. sp. lycopersici were found among a worldwide sample of isolates based on phylogenetic analysis of the ribosomal DNA intergenic spacer region. Each lineage consisted of isolates mainly belonging to a single or closely related vegetative compatibility group (VCG) and a single mating type (MAT). The first lineage (A1) was composed of isolates VCG 0031 and MAT1-1; the second (A2) included VCG 0030 and/or 0032 and MAT1-1; and the third (A3) included VCG 0033 and MAT1-2. Race 1 and race 2 isolates belonged to the A1 or A2 lineages, and race 3 belonged to A2 or A3 lineages, suggesting that there is no correlation between race and lineage. However, for the isolates from Japan, race 1 (with one exception), race 2, and race 3 isolates belonged to A2, A1, and A3 lineages, respectively. These results suggest that the races could have evolved independently in each lineage; and in Japan the present races were likely to have been introduced independently after they had evolved in other locations.     

Restriction fragment length polymorphisms, races and vegetative compatibility groups within a worldwide collection of Fusarium oxysporum f.sp. gladioli

Isolates of Fusarium oxysporum f.sp. gladioli were collected from widely different geographic areas. These isolates were characterized by pathogenicity to two differential gladiolus cultivars, vegetative compatibility, and total genomic DNA restriction fragment length polymorphisms (RFLPs). RFLPs were used to estimate the genetic divergence and relationship among isolates of F. oxysporum. RFLPs were detected by Southern blot hybridization of total genomic DNA with a 3-4 kb DNA probe generated from total DNA off. oxysporum f.sp. dianthi. Cluster analysis allowed the division of pathogenic strains into three main RFLP groups, each group containing strains with similarity coefficients ranging from 78 to 100%. RFLP groups correlated with vegetative compatibility groups, not with races. Two single pathogenic isolates which could not be assigned to any of the three main vegetative compatibility groups also had distinctive RFLP patterns. Little genetic polymorphism was observed within vegetative compatibility groups, whereas the majority of RFLPs occurred between vegetative compatibility groups, suggesting a common ancestry for strains within a specific vegetative compatibility group and a polyphyletic origin for the present special form gladioli.