Pathogenicity,vegetative compatibility and amplified fragment length polymorphism (AFLP) analysis of <Emphasis Type="Italic">Fusarium oxysporum</Emphasis> f. sp. <Emphasis Type="Italic">radicis-cucumerinum</Emphasis> isolates from Turkish greenhouses

The objective of the current study was to characterize Fusarium oxysporum f. sp. radicis-cucumerinum isolates from cucumbers in Turkey in terms of pathogenicity, vegetative compatibility and amplified fragment length polymorphism (AFLP) variation. In the 2007 and 2008 greenhouse cucumber-growing seasons, surveys were conducted in Adana, Antalya, Hatay and Mersin provinces of the Mediterranean region of Turkey. Forty-seven fungal isolates of F. oxysporum were recovered from diseased cucumber plants. The pathogenicity of each isolate was tested on cucumber seedlings at the one-true-leaf stage. Forty of the 47 isolates of F. oxysporum were virulent on cucumber seedlings. Based on disease symptoms, the differential effect of temperatures of 17°C and 29°C on disease development, and the virulence on cucumber seedlings, these 40 isolates were identified as F. oxysporum f. sp. radicis-cucumerinum. Nitrate non-utilizing mutants were generated on minimal medium containing 1.5% KClO₃ and their phenotypes were determined. Mutants in different phenotypic classes were paired on minimal medium; of 40 F. oxysporum f. sp. radicis-cucumerinum isolates, thirty-eight were placed into VCG 0260. Remaining two strains were assigned to VCG 0261. The AFLP primers produced a total of 180 fragments between 200 and 500 bp in length for the 30 isolates tested. At a genetic similarity of 0.71, the UPGMA analysis separated the isolates into two distinct clusters. The first cluster, AFLP I, included 28 isolates, of which all belonged to VCG 0260. Two strains in the second AFLP cluster both belonged to VCG 0261.     

Physiological races and vegetative compatibility groups within Fusarium oxysporum f.sp. gladioli

The pathogenicity and vegetative compatibility of mainly Dutch isolates ofFusarium oxysporum collected from diseased gladioli and other Iridaceae were investigated. Based on their pathogenicity to two differential gladiolus cultivars, the isolates could tentatively be divided into two races. All self-compatible isolates ofFusarium oxysporum f.sp.gladioli belonged to one of three distinct vegetative compatibility groups, VCG 0340, 0341 or 0342, and were incompatible with isolates that were not pathogenic to gladiolus. Isolates of one of the two races were restricted to one VCG while isolates of the other race were present in all three VCGs.     

Characterisation ofFusarium oxysporum isolated from carnation in Australia based on pathogenicity,vegetative compatibility and random amplified polymorphic DNA (RAPD) assay

Isolates ofF. oxysporum collected from symptomless carnation cuttings from Australian carnation growers properties, together with isolates from national collections, were screened for pathogenicity and grouped according to vegetative compatibility and random amplified polymorphic DNA (RAPD) patterns. The collection of 82 Australian isolates sorted into 23 different vegetative compatibility groups (VCGs). Of 69 isolates tested for pathogenicity, 24 were pathogenic to carnations, while the remaining 45 were non-pathogenic. All pathogenic isolates were within two VCGs, one of which was also compatible with an isolate obtained from an international culture collection, and which is known to represent VCG 0021 and race 2. Race status of the two pathogenic VCGs remains unknown. The RAPD assay revealed distinct DNA banding patterns which could distinguish pathogenic from non-pathogenic isolates as well as differentiate between isolates from the two pathogenic VCGs.     

Physiological Races and Vegetative Compatibility Groups of Fusarium oxysporum f. sp. lactucae Isolated from Crisphead Lettuce in Japan

One hundred and sixteen isolates of Fusarium oxysporum f. sp. lactucae obtained from 85 fields in three crisphead lettuce-producing areas in Nagano Prefecture, Japan were typed for races using differential cultivars Patriot, Banchu Red Fire and Costa Rica No. 4. They were also grouped into vegetative compatibility groups (VCGs) using complementation tests with nitrate non-utilizing (nit) mutants. Two California strains reported as F. oxysporum f. sp. lactucum, a type culture of F. oxysporum f. sp. lactucae, and 28 avirulent isolates of F. oxysporum obtained from crisphead lettuce were included for comparison. Among Nagano isolates, 66 isolates were identified as race 1, and 50 as race 2. Race 1 strains derived from Shiojiri and Komoro cities and race 2 from Kawakami village and Komoro city. All isolates of race 2 were biotin auxotrophs, and the race could be distinguished based on its requirement for biotin on minimal nitrate agar medium (MM). Pathogenic isolates were classified into two VCGs and three heterokaryon self-incompatible isolates. Strong correlations were found between race and VCG. All the race 1 strains were assigned to VCG 1 except self-incompatible isolates, and all the race 2 strains to VCG 2. The 28 avirulent isolates of F. oxysporum were incompatible with VCG 1 and VCG 2. California strains was vegetatively compatible with VCG 1, and they were assigned to race 1. Based on vegetative compatibility, these two races of F. oxysporum f. sp. lactucae may be genetically distinct, and F. oxysporum f. sp. lactucae race 1 is identical to F. oxysporum f. sp. lactucum. Received 7 May 2002/ Accepted in revised form 6 September 2002     

Vegetative Compatibility of Fusarium graminearum Isolates and Genetic Study on Their Carbendazim-Resistance Recombination in China

ABSTRACT Monoconidial isolates of 33 carbendazim-sensitive isolates and 31 carbendazim-resistant isolates of Fusarium graminearum were selected from three regions of China for vegetative compatibility group (VCG) analysis. A total of 213 and 224 nit mutants were recovered from the 33 sensitive and the 31 resistant isolates, respectively. Of all the nit mutants, the frequency of the different phenotypes was 44.6, 46.5, 5.7, and 3.2% for nit1, nit3, nitM, and nitA, respectively. VCG analysis identified 30 different VCGs among the 33 sensitive- and the 31 carbendazim-resistant isolates, with VCG diversity 0.91 and 0.97, respectively. Both, a carbendazim-sensitive and a -resistant isolate from the same field belonged to the same VCG. In all then, a total of 59 VCGs were identified among the 64 isolates with an overall VCG diversity 0.92. Direct hyphal fusion was observed in six pairs of vegetatively compatible complements, which is evidence of heterokaryon formation. It was hypothesized that carbendazim resistance could not be transferred by hyphal fusion or there is a small chance to be transferred between two compatible isolates. Three stable sexual recombinants of F. graminearum were randomly chosen from each of the three genetic crosses to study their biological properties. There were no significant differences in mycelial linear growth and pathogenicity between recombinants and their parents, but they differ in sporulation ability and capacity to produce perithecia. We concluded that sexual recombination presumably played a role in the development of carbendazim resistance under field conditions.     

Vegetative compatibility and heterokaryon stability in Fusarium oxysporum f.sp. radicis-lycopersici from Italy

Fusarium crown and root rot, caused by Fusarium oxysporum f.sp. radicis-lycopersici ( Forl ), is one of the most destructive soilborne diseases of tomato in Italy. Chlorate-resistant, nitrate-nonutilizing ( nit ) mutants were used to determine vegetative compatibility among 191 isolates of Forl collected in five geographic regions (Calabria, Emilia-Romagna, Liguria, Sardinia, Sicily) in Italy. The isolates were assigned to five vegetative compatibility groups (VCGs): 65 isolates to VCG 0090; 99 to VCG 0091; 23 to VCG 0092; two to VCG 0093; and two to VCG 0096. The population structure of Forl in Italy is similar to that reported for Israel, and differs from that found in North Atlantic European countries, where VCG 0094 is predominant. The stability of prototrophic heterokaryons originating from hyphal anastomosis between compatible complementary nit mutants was assessed through conidial analysis and mycelial mass transfer. Most monoconidial cultures (84%) recovered from 117 prototrophic heterokaryons were nit mutants, indicating that heterokaryons generally do not proliferate well through conidiation; most of the 177 prototrophic heterokaryons examined were unstable, and only 9% sustained prototrophic growth through the tenth mycelial transfer upon subculturing. The prototrophic growth is proposed to be maintained through restoration of the heterokaryotic state by continual anastomosis between adjacent homokaryotic hyphae. Since heterokaryosis is a prerequisite for parasexual recombination, we speculate that this mechanism is unlikely to play a major role in generating the VCG diversity found among Forl or other strains of F. oxysporum.     

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.     

Genetic and pathogenic characterization of <Emphasis Type="Italic">Verticillium dahliae</Emphasis> isolates from eggplant in Turkey

During 2005 to 2007, eggplant fields in 19 provinces from three different regions (western, southern and southeastern Anatolia regions) of Turkey were surveyed for Verticillium wilt. Sixty-seven isolates of Verticillium dahliae from wilted eggplants were collected and used for vegetative compatibility analysis using nitrate non-utilizing mutants and reference tester strains of vegetative compatibility groups (VCGs) 1A, 2A, 2B, 3, 4A and 4B. Among all isolates, 33 (12 from western, 15 from southern and six from southeastern Anatolia) were assigned to VCG2B, 23 (four from western, eight from southern and 11 from southeastern Anatolia) to VCG2A, six (four from southern, one from western, and one from southeastern Anatolia) to VCG4B and five (one from western, one from southern and three from southeastern Anatolia) to VCG1A, whereas VCG3 and VCG4A were not defined among isolates. In order to test if there is a correlation between VCG and pathogenicity in V. dahliae, pathogenicity of 30 isolates, representing the four multimember VCGs, were tested on Solanum melongena cvs. ‘Kemer’ and ‘Aydın Siyahı’ in an unheated greenhouse. All isolates were found to be pathogenic on both cultivars and there was no difference in susceptibility between the two cultivars. VCG4B isolates collectively led to higher vascular discoloration index (VDI) on both cultivars and higher disease severity index (DSI) on ‘Kemer’ compared with other VCGs. Similarly, VCG1A caused lower VDI on both cultivars and lower DSI on ‘Kemer’. Isolates within each of VCGs 1A, 2A and 4B caused similar VDI on both cultivars. Isolates of VCG2B were found to vary in their VDI values on both cultivars. To the best of our knowledge, the present study is the first report of natural infections of eggplant by VCG1A.     

Vegetative compatibility grouping in Botrytis cinerea using sulphate non-utilizing mutants

Twenty-one strains of Botrytis cinerea isolated from six plant species on ten sites throughout Israel, as well as a strain from France, were tested for vegetative and mycelial incompatibility, pathogenicity, resistance to the fungicides carbendazim and iprodione, and colony morphology. Selenate-resistant mutants were isolated from the strains as spontaneous, fast-growing sectors arising from restricted colonies on medium amended with sodium selenate with a mean frequency of 0.04 sectors/colony; 81% of the sectors were sulphate non-utilizing (sul) mutants. One hundred and four sul mutants were divided into two complementary groups: resistant (66 mutants) and sensitive to chromate. Based on compatibility reactions between chromate-resistant and chromate-sensitive sul mutants, 12 strains were compatible only with themselves and were each classified as belonging to different vegetative compatibility groups (VCGs). Nine strains were each compatible with one to three other strains, and were assembled into three multi-member VCGs. Mycelial incompatibility between wild-type strains (barrage), in the form of a zone of dark pigmentation or sparse mycelium with or without dark pigmentation of the agar along the line of confrontation, was observed for 70% of the inter-strain pairings. There was no correspondence in compatibility between strains revealed by two approaches: strains in different VCGs did not necessarily produce a barrage. However, self-compatibility was observed both as heterokaryon formation between complementary sul mutants and as an absence of barrages between mycelia of wild-type strains; wild-type strains belonging to the same VCG did not exhibit strong barrages, although weak antagonistic reactions were observed. Strains in two multi-member VCGs showed the same patterns of resistance to carbendazim and iprodione; the third multi-member VCG contained isolates with different patterns of resistance. Four morphological types were revealed among wild-type strains: conidial (five strains), sclerotial (six strains), intermediate (ten strains), and mycelial (one strain). On bean leaves, conidial strains were more aggressive than sclerotial strains.     

Vegetative compatibility and pathogenicity of<Emphasis Type="Italic">Verticillium dahliae</Emphasis> isolates from the aegean region of Turkey

A total of 101Verticillium dahliae isolates were recovered from cotton plants at 57 sites in the Aegean region of Turkey between 2003 and 2004. Isolates were tested for vegetative compatibility by observing heterokaryon formation among complementary nitrate-nonutilizing (nit) mutants. Forty-six isolates were assigned to VCG 1, 12 to VCG 2A, 33 to VCG 2B and four to VCG 4B. The remaining six isolates could not be tested for vegetative compatibility because of their inability to yieldnit mutants. All isolates recovered were tested for pathogenicity on cotton cultivars Acala SJ-1 and Deltapine 15-21 by the stem-injection method. The isolates of VCG 2 and 4B, irrespective of their origin, induced weak to severe symptoms on cotton and were similar to the previously described cotton non-defoliating pathotype. In contrast, all cotton isolates of VCG1 caused severe foliar symptoms, stunting, defoliation and often death. This is the first report on VCG 1 ofV. dahliae in Turkey. http://www.phytoparasitica.org posting May 4, 2007.     

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.     

The elegans fusaria causing wilt disease of carnation. II. Distinction of vegetative compatibility groups

The vegetative compatibility patterns among isolates ofElegans fusaria causing wilt disease of carnation were investigated. Nitrate non-utilizing mutants were generated from 16 isolates labelledF. redolens, nine of which came from carnation, and from 33 isolates labelledF. oxysporum, 19 of which came from carnation. Pairings of the mutants revealed five vegetative compatibility groups among the isolates from carnation, corresponding withF. oxysporum f.sp.dianthi race 1 (VCG1), race 2 (VCG2) and race 4 (VCG3),F. redolens f.sp.dianthi (VCG4) andF. redolens isolates from foot rot-diseased carnations (VCG5). Besides three isolates typical ofF. redolens, VCG4 comprised a now slightly deviating subculture of the type isolate ofF. redolens f.sp.dianthi of which the cultural characteristics correspond toF. oxysporum instead ofF. redolens. This observation may be taken to support previous conclusions that the distinction between both taxa is not justified. Otherwise, the compatibility patterns did not provide decisive evidence to accept or reject conspecificity of both taxa. Isolates from carnation did not form heterokaryons with other formae speciales ofF. oxysporum.Samenvatting De vegetatieve compatibiliteitspatronen bij isolaten vanElegans-fusaria die verwelkingsziekte bij anjer veroorzaken werden onderzocht. Van 16 isolaten vanF. redolens, waarvan negen afkomstig van anjers, en van 33 isolaten vanF. oxysporum, waarvan 19 afkomstig van anjers, werden mutanten gegenereerd die zonder een organische stikstofbron geen luchtmycelium meer konden vormen. Paringen tussen mutanten van isolaten afkomstig van anjers brachten een vijftal vegetatieve compatibiliteitsgroepen aan het licht, die overeenkwamen metF. oxysporum f.sp.dianthi fysio 1 (VCG 1), fysio 2 (VCG 2) en fysio 4 (VCG3),F. redolens f.sp.dianthi (VCG4) enF. redolens isolaten afkomstig van aan voetrot lijdende anjers (VCG5). Naast drie voorF. redolens karakteristieke isolaten omvatte VCG4 ook een afwijkende subculture van het type-isolaat vanF. redolens f.sp.dianthi, die in cultuureigenschappen overeen kwam metF. oxysporum in plaats vanF. redolens. Deze waarneming geeft enige steun aan eerdere conclusies dat het onderscheid tussen beide taxa niet gerechtvaardigd is. Daarbuiten gaven de compatibiliteitspatronen geen uitsluitsel over de mogelijke conspecificiteit van beide taxa. Isolaten afkomstig van anjers vormden geen heterokaryons met andere formae speciales vanF. oxysporum.     

Characterization of biotin-autotrophic isolates derived from naturally occurring auxotrophic race 2 isolates of <Emphasis Type="Italic">Fusarium oxysporum</Emphasis> f. sp. <Emphasis Type="Italic">lactucae</Emphasis>

Race 2 isolates of Fusarium oxysporum f. sp. lactucae have been recognized as biotin auxotrophs and consequently have restricted growth on Puhalla's minimal medium (MM), which contains no biotin. Biotin-autotrophic isolates were raised from race 2 isolates through cultural mutation that grew as well on MM as they did on MM supplemented with biotin. These autotrophs were identical to the parental isolates in pathogenicity on race differential cultivars of lettuce (Patriot, Banchu Red Fire, and Costa Rica No. 4), and thus were designated as race 2. A vegetative compatibility test indicated that the autotrophic isolates fell into the same vegetative compatibility group as the parents. Culture filtrates of the autotrophs allowed abundant growth of the parental auxotroph on MM, and, through a competitive enzyme-binding assay, biotin was detected in the culture filtrates. These results suggest that biotin auxotrophy in the natural race 2 isolates has no direct relation to pathogenicity, qualitatively defined as physiological race, or to vegetative compatibility.     

Characterization of Fusarium oxysporum f. sp. radicis-cucumerinum attacking melon under natural conditions in Greece

A severe root and stem rot disease of melon was observed during the 2001 growing season on four glasshouse crops in Heraklio, Greece. A total of 43 isolates of F. oxysporum , obtained in Crete from glasshouse-grown melon and showing fusarium wilt or root and stem rot symptoms, were characterized by pathogenicity and vegetative compatibility. The majority of these isolates was also fingerprinted via amplified fragment length polymorphic (AFLP) analysis. Of the total number of isolates, 22 were identified by pathogenicity tests as F. oxysporum f. sp. melonis , 20 as F. oxysporum f. sp. radicis-cucumerinum , while one isolate was nonpathogenic on cucumber, melon, sponge gourd and pumpkin. All 22 isolates of F. oxysporum f. sp. melonis were assigned to vegetative compatibility group (VCG) 0134, and all 20 isolates of F. oxysporum f. sp. radicis-cucumerinum to VCG 0260. Isolates of F. oxysporum f. sp. radicis-cucumerinum were incompatible with isolates of F. oxysporum f. sp. melonis. AFLP fingerprinting allowed for the clustering of the isolates of the two formae speciales of F. oxysporum along two separate phenetic groups: f. sp. melonis to AFLP major haplotype I, and f. sp. radicis-cucumerinum to AFLP major haplotype II. Overall, pathogenicity, vegetative compatibility grouping and AFLP analysis were correlated and effectively distinguished isolates of F. oxysporum from melon. This appears to be the first report of natural infection of melon by F. oxysporum f. sp. radicis-cucumerinum worldwide.     

Intraspecific variation in Diplodia seriata isolates occurring on grapevines in Spain

Variation of Diplodia seriata, a fungal species associated with botryosphaeria dieback of grapevine, was investigated with respect to its genetic, phenotypic and pathogenic characteristics. The inter‐simple sequence repeat (ISSR) technique was used to investigate the genetic diversity of 83 isolates of D. seriata. Five ISSR primers were able to provide reproducible and polymorphic DNA fingerprint patterns, thus showing a relevant genetic variability in the species. Analyses of ISSR data by different clustering methods grouped the isolates into two distinct clusters through the Bayesian and DAPC analyses. No relationships between either geographic or host origin of isolates and genetic clusters were observed. Several representative isolates from each genetic cluster were chosen for studying their conidial dimensions, in vitro mycelial growth, vegetative and mating compatibility, and pathogenicity on detached grapevine canes and potted vines. No significant differences in conidial dimensions were detected among the groups. Vegetative compatibility reactions were observed among isolates but this was not related with the genetic clustering. Production of sexual fruiting bodies in vegetative compatible crossings was not observed under the experimental conditions used in the study. All 14 isolates tested for pathogenicity were confirmed to be pathogenic according to the length of the necrotic lesions that they caused and their reisolation frequencies from the infected plant tissues. Differences in the length of necrosis were detected among isolates, thus revealing the existence of different virulence levels in the species.     

Pathogenicity and Virulence of the Two Dutch VCGs of Verticillium dahliae to Woody Ornamentals

Two experiments were performed in two consecutive years to test whether isolates of different vegetative compatibility groups (VCGs) differ in their ability to cause disease in woody ornamentals, to study the host specificity of the isolates and to get an insight into disease development in woody hosts. A range of woody ornamental plant species, including Acer campestre, Acer platanoides, Acer pseudoplatanus, Catalpa bignonioides, Cotinus coggygria, Robinia pseudoacacia, Rosa canina, Syringa vulgaris and Tilia cordata, were root-dip inoculated with six isolates of Verticillium dahliae, belonging to the two VCGs that occur in the Netherlands (VCG NL-1 and VCG NL-2). Isolates belonging to each VCG caused severe symptoms of verticillium wilt in most plant species tested. Disease progress differed between plant species, but was generally the same for the two VCGs. No overall differences in virulence were observed between the two VCGs for external wilt symptoms, number of dead plants, or shoot length. No significant VCG × plant species interactions were present for these characteristics. However, isolates of VCG NL-1 caused more vascular discolouration than did isolates of VCG NL-2. Isolates within VCGs often differed considerably in their virulence to certain hosts, as shown by highly significant isolate × plant species interactions. Isolates were more virulent on their original host. These findings imply that VCG identification does not contribute to disease prediction for a range of woody hosts.     

Genetic Diversity of Fusarium oxysporum Isolates from Cucumber: Differentiation by Pathogenicity, Vegetative Compatibility, and RAPD Fingerprinting

ABSTRACT A total of 106 isolates of Fusarium oxysporum obtained from diseased cucumber plants showing typical root and stem rot or Fusarium wilt symptoms were characterized by pathogenicity, vegetative compatibility, and random amplified polymorphic DNA (RAPD). Twelve isolates of other formae speciales and races of F. oxysporum from cucurbit hosts, three avirulent isolates of F. oxysporum, and four isolates of Fusarium spp. obtained from cucumber were included for comparison. Of the 106 isolates of F. oxysporum from cucumber, 68 were identified by pathogenicity as F. oxysporum f. sp. radicis-cucumerinum, 32 as F. oxysporum f. sp. cucumerinum, and 6 were avirulent on cucumber. Isolates of F. oxysporum f. sp. radicis-cucumerinum were vegetatively incompatible with F. oxysporum f. sp. cucumerinum and the other Fusarium isolates tested. A total of 60 isolates of F. oxysporum f. sp. radicis-cucumerinum was assigned to vegetative compatibility group (VCG) 0260 and 5 to VCG 0261, while 3 were vegetatively compatible with isolates in both VCGs 0260 and 0261 (bridging isolates). All 68 isolates of F. oxysporum f. sp. radicis-cucumerinum belonged to a single RAPD group. A total of 32 isolates of F. oxysporum f. sp. cucumerinum was assigned to eight different VCGs and two different RAPD groups, while 2 isolates were vegetatively self-incompatible. Pathogenicity, vegetative compatibility, and RAPD were effective in distinguishing isolates of F. oxysporum f. sp. radicis-cucumerinum from those of F. oxysporum f. sp. cucumerinum. Parsimony and bootstrap analysis of the RAPD data placed each of the two formae speciales into a different phylogenetic branch.     

Recent progress in understanding relationships betweenVerticillium species and subspecific groups

Improved understanding of the genetic diversity within fungi in the genusVerticillium has resulted from recent studies based on vegetative compatibility analysis and several techniques of molecular biology. Although the method used to identify vegetative compatibility groups (VCGs) does affect the results, vegetative compatibility appears to be a stable characteristic among isolates. Fairly low VCG diversity has been detected withinV. dahliae andV. albo-atrum using nitrate non-utilizing mutants. VCGs do not appear to be related to pathogenicity to particular host species, with the exception ofV. albo-atrum on alfalfa. However, there is some correlation with virulence on certain hosts and with the ability ofV. dahliae to interact with root-lesion nematodes. Studies based on DNA analysis indicate thatV. dahliae andV. albo-atrum are closely related but separate species. Restriction fragment length polymorphism (RFLP) studies have identified several subspecific groups withinV. dahliae, including two non-host-adapted groups and two that are host-adapted. They also have confirmed that alfalfa strains ofV. albo-atrum are a distinct subgroup that is probably a separate population of clonal origin. Using polymerase chain reaction (PCR), a second non-host-adapted subgroup withinV. albo-atrum was identified that was previously unknown.     

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.     

Comparative Study of Genetic Diversity and Pathogenicity Among Populations of Verticillium Dahliae from Cotton in Spain and Israel

Genetic diversity and phenotypic diversity in Verticillium dahliae populations on cotton were studied among 62 isolates from Spain and 49 isolates from Israel, using vegetative compatibility grouping (VCG), virulence and molecular assays. In Spain, defoliating V. dahliae isolates (D pathotype) belong to VCG1, and non-defoliating isolates (ND) belong to VCG2A (often associated with tomato) and VCG4B (often associated with potato). The D pathotype was not identified in Israel. The ND pathotype in Israel is comprised of VCG2B and VCG4B. Isolates in VCG2B and VCG4B ranged in virulence from weakly virulent to highly virulent. The highly virulent isolates induced either partial defoliation or no defoliation. Virulence characteristics varied with inoculation method and cotton cultivar. Highly virulent isolates from Israel were as virulent as D isolates from Spain under conditions conducive to severe disease. The D pathotype is pathologically and genetically homogeneous, whereas the ND pathotype is heterogeneous with respect to virulence, VCG, and molecular markers based on single-primer RAPD and on PCR primer pairs.