玉蜀黍赤霉有性重组体的诱导及其生物学特性

 本研究以硝酸盐营养缺陷型突变体(nit)和多菌灵抗性为遗传标记,在5个所选菌株中设计了3个玉蜀黍赤霉病菌Gibberella zeae的杂交组合,使各菌株之间进行杂交,从而诱导有性重组体。从各杂交组合的后代中任意挑选出3个有性重组体,比较了这些有性重组体与其亲本在无性和有性阶段的主要生物学性状。结果表明,玉蜀黍赤霉中的nit基因及对杀菌剂多菌灵的抗药性基因可以通过有性杂交的方式重组,即发生了有性重组。有性重组体与其亲本在菌落生长、培养性状和致病性方面没有显著差异;但某些有性重组体中,产孢量和产子囊壳能力方面存在一定差异。总体看来,有性重组体仍然保持了较高的适合度。因此,可以认为有性重组在玉蜀黍赤霉群体对多菌灵抗药性发生发展以及群体遗传进化中可能起着重要的作用。     

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.     

灰葡萄孢菌nit突变体的诱导及其在营养体亲和性测定中的应用初探

 从山西运城、临汾、长治、晋中、大同等地保护地黄瓜灰霉病病株上采集、分离的分属于3个不同菌丝融合群的8个灰葡萄孢菌单孢菌株,经氯酸盐诱导处理,共获得了抗氯酸盐的硝酸盐利用缺陷突变体(nit突变体)59株,其中nit1型38株,nit3型10株,nitM型11株。所有nit突变株分别在PDA斜面转管培养3次(21 d)后,除6株恢复成野生菌株外,其余多数nit突变菌株表现稳定。来源于同一野生菌株的不同类型nit突变体间或同一菌丝融合群不同野生菌株的nit突变体间可产生互补反应而形成异核体,其中以nitM型突变株互补性最好,在利用nit突变体测定灰葡萄孢菌营养体亲和性时应作为标准菌株。来源于不同菌丝融合群的nit突变体间不能产生互补反应。     

福建省香蕉枯萎病菌硝酸盐营养突变体的营养体亲和性测定

 Fourteen strains of Fusarium oxysporum f. sp. cubense were induced to produce 146 nitrate-nonutilizing(nit) mutants on a chlorate-containing medium. Among them, there were 117 nit1 mutants(80.14%), 17 nit3 mutants(11.64%) and 12 nitM mutants(8.22%). These strains were divided into two vegetative compatibility groups(VCGs) by the vegetative compatibility tests. Twelve strains of F. oxysporum f. sp. cubense from Musa AAA belonged to VCG1, two trains from Musa ABB belonged to VCG2.     

Use of nitrate non-utilising (Nit) mutants to determine vegetative compatibility in Botryotinia fuckeliana (Botrytis cinerea)

Nitrate non-utilising (Nit) mutants of six field strains and three single-ascospore strains of Botryotinia fuckeliana were selected by plating mycelial plugs onto a nitrate-containing minimal medium amended with chlorate. Mutants were characterised by growth responses on minimal medium amended with various sole nitrogen sources. For each parental strain two mutants were produced: nit1, defective in nitrate reductase apoenzyme; and NitM, defective in the molybdenum cofactor pathway. Complementation on nitrate minimal medium was observed between nit1 and NitM mutants from the same parent in all instances. However, complementation was not observed between any such combinations when the mutants were derived from different parents. It is concluded that a vegetative incompatibility system operates in B. fuckeliana resulting in multiple vegetative compatibility groups. One of the single-ascospore strains was derived from a cross between two of the field strains, indicating that new compatibility groups can be generated during sexual reproduction. Mycelial interaction zones were formed between all parental strains when they were paired on NaCl-amended medium, indicating congruence between mycelial incompatibility and vegetative incompatibility.     

瓜类尖孢镰刀菌的营养体亲和群研究

 应用营养亲和性方法研究了尖孢镰刀菌菌株抗氯酸盐突变体和nit突变体的诱发规律及分布特性,以及菌株营养体亲和群(VCG)的划分。研究表明,不同寄主(黄瓜、甜瓜和西瓜)分离的尖孢镰刀菌菌株形成的抗氯酸盐突变体数目差异不显著,平均为每个接种点产生0.89~0.98个;但寄主不同部位(根部、茎基部和茎中部)分离的菌株间差异显著,形成的数目分别为1.27、0.75及0.76个。菌株产生的nit1突变体比例(75.40%)显著高于nitM突变体比例(13.17%);nit1突变体数目会因菌株的寄主及菌株寄主部位的不同而有差异,寄主为黄瓜、甜瓜和西瓜的菌株产生的比例依次为67.73%、83.71%和77.50%,根部、茎基部及茎中部分离菌株产生的比例依次为81.82%、78.48%和68.64%,而在致病菌株与非致病菌株间无显著差异,分别为74.43%和79.63%;nitM突变体数目受菌株寄主影响较小,所占比例在11.17%~13.92%之间;而在寄主不同部位分离的菌株及致病菌株与非致病菌株间差异显著,分离自茎基部的菌株所占比例最高为15.97%,茎中部菌株所占比例最低为9.87%,致病菌株与非致病菌株所占比例分别为14.08%和9.26%。供试菌株分为7个VCGs,其特点为来源于不同寄主的尖孢镰刀菌菌株互不亲和,同一寄主的致病菌株与非致病菌株均不亲和,同一寄主不同部位分离的菌株可亲和。     

尖孢镰孢(Fusarium oxysporum)硝酸盐营养突变株及其营养体亲和性

从荸荠、棉花、西瓜、黄瓜、胡瓜、大豆、花生等12种寄主作物上获得尖孢镰孢(F—usarium oxysporum)菌株20个,已知15个菌株分属于6个专化型。所有20个菌株在含有氯酸盐的培养基上产生抗氯酸盐的突变体,其突变体在以硝酸盐作为氮源的培养基上表现为无气生菌丝生长,即不能利用硝酸盐的突变株(nit),共获得nit突变株181个。同一专化型的nit突变株之间在以硝酸盐为氮源的培养基上配对时,在菌丝接触处可产生互补作用,即营养体亲和性,表现为旺盛气生菌丝生长。不同专化型的nit突变株之间则不能互补产生异核体而表现为非亲和性仍为突变型生长。每个菌株均获得能产生互补作用的nit突变株。根据营养体亲和性反应,20个菌株归入13个营养体亲和群(VCG),并显示出VCGs与菌株专化型之间的相关性。nit突变株在一般条件下是稳定的,而且某些nit株的亲和能力特强。     

Vegetative compatibility of Fusarium oxysporum f.sp. dianthi from carnation in Israel

Auxotrophic mutants were used to determine vegetative relatedness among isolates of Fusarium oxysporum f.sp. dianthi (F.o.d.) , the vascular wilt pathogen of carnation. At the first stage, different nitrate-non-utilizing (nit) mutants were produced from 11 isolates of F.o.d. collected in Israel. Complementation (heterokaryon) tests showed that all the isolates belonged to a single vegetative compatibility group (VCG), and two mutants were chosen as its testers. Additional isolates of Fusarium from carnation, collected during 1986-88, were analysed for pathogenicity and vegetative compatibility with the testers. A total of 170 Fusarium isolates, obtained from 42 cultivars at 40 sites, were tested. All the nit mutants of all the 132 pathogenic isolates formed heterokaryons with the testers, indicating that they belonged to the same VCG. None of the 38 non-pathogenic isolates was vegetatively compatible with the testers. The nit mutants retained pathogenicity to carnation. The F.o.d. testers were not compatible with testers of five other formae speciales of F. oxysporum. Thus, F.o.d. appears to constitute a distinct genetic population within the F. oxysporum complex.     

Mating Behaviour and Vegetative Compatibility in Spanish Populations of Botryotinia fuckeliana

Mating behaviour and vegetative compatibility were studied in Spanish populations of Botryotinia fuckeliana. Fifty-seven isolates out of the 61 tested were sexually fertile with one or more of the reference strains of known mating type (MAT1-1 or MAT1-2). Thirty-nine isolates were heterothallic, giving fertile crosses when mated with the MAT1-1 (24 isolates) or the MAT1-2 (15 isolates) reference strain. Eighteen isolates crossed successfully with both MAT1-1 and MAT1-2 reference strains, and were referred to as homothallic or MAT1-1/2. Both mating types were widespread, being represented in isolates from two regions, from the same and different greenhouses, from different hosts, and from different years of isolation. Isolates were paired on Malt Extract Agar + NaCl to evaluate vegetative compatibility. Most of the paired isolates were unable to fuse and showed a different reaction of incompatibility. Nitrate-non-utilising (nit) mutants were selected by growth on a medium amended with 30–50gl^–1 potassium chlorate. Over 600 chlorate-resistant sectors were recovered from 40 isolates at a mean frequency of 0.15–2.39 sectors per colony, but only 11% were identified as nit mutants by their thin growth with no aerial mycelium on minimal medium. However, most of these nit mutants reverted to wild type during six months of storage on chlorate-amended medium. Genetic complementation between nit mutants occurred only in two cases between mutants from the same isolate.     

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.     

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.     

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     

Genotypic diversity in a South African population of the pitch canker fungus Fusarium subglutinans f.sp. pini

The genotypic diversity in a South African population of Fusarium subglutinans f.sp. pini ( F.s. pini ) was determined, based on the number of vegetative compatibility groups (VCGs). Isolates of F.s. pini from South Africa (69), California (five) and Florida (19) were included in the study. The nit1 (or nit3 ) and NitM mutants were selected as chlorate resistant sectors and paired on minimal medium. The South African isolates of F.s. pini were assigned to 23 different VCGs. No heterokaryons formed between isolates from South Africa, California and Florida. The high degree of genotypic diversity in the South African population of F.s. pini is probably due to some level of sexual reproduction in the population.     

Genetic characteristics of Fusarium verticillioides isolates from maize in Costa Rica

Thirty-nine isolates of Fusarium verticillioides from maize seeds from three regions of Costa Rica were classified on fertility, fumonisin production, vegetative compatibility and pathogenicity. The identity of the isolates was verified by sexual crosses with standard tester strains and by isozyme analysis. Twenty-three isolates (59%) were mating type A⁻ and 16 (41%) were A⁺; 29 (74%) were female fertile. The isolates produced high amounts of fumonisin B₁ when grown on sterilized maize grits, 32 isolates producing more than 1000 μg g⁻¹, as determined by TLC, and 7 less than 1000 μg g⁻¹. Vegetative compatibility tests by pairing nit mutants identified 34 vegetative compatibility groups (VCGs), of which 29 had one member and 5 had two members. Isolates belonging to the same VCG were obtained from the same seed sample. Two pathogenicity tests with different inoculation methods were performed: on toothpick inoculation of 7-week-old maize stalks, 71% of the isolates were pathogenic according to the length of the necrosis formed in the stalk, and on sand inoculation of maize seedlings all the isolates were pathogenic, according to shoot length and dry weight production. Differences in aggressiveness between some of the isolates were recorded. It is concluded that natural populations of F. verticillioides in Costa Rica consist of genetically diverse, highly fertile and pathogenic isolates that represent a potential risk for disease development and fumonisin accumulation in maize crops.     

Monogenic resistance to a new fungicide, JS399-19, in Gibberella zeae

The fungicide JS399-19 is a novel cyanoacrylate fungicide active against Gibberella zeae , and has been marketed in China for control of fusarium head blight (FHB) on wheat. Forty-three isolates sensitive to fungicide JS399-19 were collected from three commercial wheat fields in China. Forty-five isolates resistant to JS399-19, obtained from five sensitive isolates by selection for resistance to JS399-19, were selected. Three sensitivity levels were identified: sensitive (S), moderately resistant (MR) and highly resistant (HR) to JS399-19, based on a previous study. Eight isolates representing the three sensitivity-level phenotypes were randomly selected for a study on the inheritance of JS399-19 resistance by analysing the sensitivity of hybrid F₁ progeny. A nitrate-non-utilizing mutant ( nit ) was used as a genetic marker to confirm that individual perithecia were the result of outcrossing. Five crosses were assessed: S × S, S × HR, MR × HR, HR × HR and MR × S. In crosses between parents with different sensitivity levels, such as S × HR, MR × HR and MR × S, the progeny fitted a 1:1 segregation ratio of the two parental phenotypes. No segregation was observed in the crosses S × S and HR × HR. It was concluded that the MR and HR phenotypes in G. zeae were conferred by different allelic mutations within the same locus. In these isolates, resistance to JS399-19 was not affected by modifying genes or cytoplasmic components.     

尖孢镰孢(Fusarium oxysporum)硝酸盐营养突变株及其营养体亲和性

 从荸荠、棉花、西瓜、黄瓜、胡瓜、大豆、花生等12种寄主作物上获得尖孢镰孢(F-usarium oxysporum)菌株20个,已知15个菌株分属于6个专化型。所有20个菌株在含有氯酸盐的培养基上产生抗氯酸盐的突变体,其突变体在以硝酸盐作为氮源的培养基上表现为无气生菌丝生长,即不能利用硝酸盐的突变株(nit),共获得nit突变株181个。同一专化型的nit突变株之间在以硝酸盐为氮源的培养基上配对时,在菌丝接触处可产生互补作用,即营养体亲和性,表现为旺盛气生菌丝生长。不同专化型的nit突变株之间则不能互补产生异核体而表现为非亲和性仍为突变型生长。每个菌株均获得能产生互补作用的nit突变株。根据营养体亲和性反应,20个菌株归入13个营养体亲和群(VCG),并显示出VCGs与菌株专化型之间的相关性。nit突变株在一般条件下是稳定的,而且某些nit株的亲和能力特强。     

Sexual recombination of carbendazim resistance in Fusarium graminearum under field conditions

BACKGROUND: Carbendazim has been the major fungicide for control of Fusarium head blight (FHB) caused by Fusarium graminearum Schwade in China. However, the effectiveness of carbendazim has been threatened by the emergence of resistant pathogen populations in the field. RESULTS: Five isolates, representing three phenotypes of different carbendazim sensitivity levels (S, MR and HR), were randomly selected to study the inheritance of carbendazim resistance by three genetic crosses under field conditions. Each parent in all crosses was marked with a different class of nitrate non‐utilizing (nit) mutation. The presence of sexual recombinants indicated that outcrossing had occurred in the crosses. Over 100 putative self‐crossing or outcrossing perithecia for each cross were randomly sampled on the surface of the haulms of dead rice for each pair of parents. Results showed that 5.7–20.9% outcrossing frequency occurred in the three crosses and confirmed sexual recombination under field conditions. There were no significant differences in mycelial linear growth and pathogenicity between the selected recombinants and their parents, but they differed in sporulation ability and capacity to produce perithecia. Nevertheless, most of the sexual recombinants possessed fitness levels comparable with those of their parents. CONCLUSION: Outcrossing between carbendazim‐sensitive and ‐resistant isolates did occur under field conditions, and sexual recombination must play a role in the development of carbendazim resistance in the field. Copyright © 2009 Society of Chemical Industry     

Vegetative compatibility groups within Verticillium dahliae isolates from different hosts in Greece

Forty-four isolates of Verticillium dahliae obtained from different diseased hosts were tested by vegetative compatibility group (VCG) analysis to investigate their genetic relatedness and correlate the results with four VCGs (1, 2, 3, 4) previously described. Based on complementarity of nit mutants, only three VCGs were identified from the Greek isolates. Seventeen isolates were assigned to VCG 2 (A or B), two to VCG 3 and eight to VCG 4 (A or B). The 17 remaining isolates could not be grouped to any of the three VCGs. All isolates belonging to a distinct VCG complemented strongly with at least one of the two tester strains of that group, or with several strains of the Greek collection belonging to that VCG.     

Barrage Zone Formation Between Vegetatively Incompatible Fusarium graminearum (Gibberella zeae) Isolates

ABSTRACT Vegetative compatibility has been used to assess the population biology of many fungal plant pathogens. However, for many species, including Fusarium graminearum, this has meant making auxotrophic mutants to force heterokaryon formation. A method was developed to observe barrage zones of thick, raised mycelium at the junctions of vegetatively incompatible F. graminearum isolates. The appearance of the barrage zones was influenced by the growth medium and the light. Barrage zones on V8 agar were thicker and better defined than those on potato dextrose agar, Spezieller Nahrstoffarmer agar, and water agar. The addition of ground wheat kernels to V8 agar enhanced barrage zone formation. Incubating the cultures under constant light at 2,150 lx produced more distinct barrage zones than constant light at 3,400 lx, constant darkness, or ambient room light. Forty-three F. graminearum isolates from 34 vegetative compatibility groups, determined previously using nit auxotrophic mutants, were paired in all combinations using these optimized conditions. Isolates in different vegetative compatibility groups typically formed distinct, thick barrage zones at their junctions. Pairs of isolates in the same vegetative compatibility group had a very slight or no visible reaction, or rarely, a distinct "line gap" of sparse mycelium. Subcultures from the same isolate typically had no visible reaction at their colony junctions; however, subcultures from some isolates had thin, slight barrage zones. This method was used to identify the proportion of each of four F. graminearum isolates from infected barley spikes in the field, inoculated previously with a mixture of conidia from these four isolates. Barrage zone formation represents a rapid method to screen vegetative compatibility groups in F. graminearum and may be useful for other Fusarium species.     

Vegetative Compatibility amongVerticillium dahliae Isolates from Watermelon in Greece

Vegetative compatibility among 17 isolates ofVerticillium dahliae obtained from watermelon, originating from eight regions of Greece, was investigated using complementation tests between nitrate-nonutilizing(nit) mutants. Among 529 chlorate-resistant sectors obtained, only 107 werenit mutants. These mutants were paired with tester strains (from Greece and other countries) of previously described vegetative compatibility groups (VCGs), and also were paired in many combinations among themselves. All isolates were self-compatible. Sixteen isolates were found to belong to VCG2. Only isolate V75 could not be assigned to a VCG, because the threenit mutants obtained from it showed negative reactions with the tester strains of four VCGs and with complementary mutants from other isolates. Based on this sample, we conclude that the population ofV. dahliae from watermelon in Greece is homogeneous in respect to VCG.