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1.
ABSTRACT A degree of genetic diversity may exist among Verticillium dahliae isolates within vegetative compatibility groups (VCGs) that bears phytopathological significance and is worth investigating using molecular tools of a higher resolution than VCG characterization. The molecular variability within and among V. dahliae VCGs was studied using 53 artichoke isolates from eastern-central Spain, 96 isolates from cotton, 7 from cotton soil, and 45 from olive trees in countries of the Mediterranean Basin. Isolates were selected to represent the widest available diversity in cotton- and olive-defoliating (D) and -nondefoliating (ND) pathotypes, as well as for VCG. The VCG of 96 cotton and olive isolates was determined in this present study. Molecular variability among V. dahliae isolates was assessed by fluorescent amplified fragment length polymorphism (AFLP) analysis and by polymerase chain reaction (PCR) assays for DNA fragments associated with the D (462 bp) and ND (824 bp) pathotypes, as well as a 334-bp amplicon associated with D pathotype isolates but also present in some VCG2B isolates. Isolates from cotton were in VCG1A, VCG1B, VCG2A, VCG2B, and VCG4B and those from olive trees were in VCG1A, VCG2A, and VCG4B. Artichoke isolates included representatives of VCG1A, VCG2A, VCG2B (including a newly identified VCG2Ba), and VCG4B. AFLP data were used to generate matrixes of genetic distance among isolates for cluster analysis using the neighbor-joining method and for analysis of molecular variance. Results demonstrated that V. dahliae isolates within a VCG subgroup are molecularly similar, to the extent that clustering of isolates correlated with VCG subgroups regardless of the host source and geographic origin. VCGs differed in molecular variability, with the variability being highest in VCG2B and VCG2A. For some AFLP/VCG subgroup clusterings, V. dahliae isolates from artichoke grouped in subclusters clearly distinct from those comprising isolates from cotton and olive trees. In addition, VCG2B isolates from artichoke formed two distinct clusters that correlated with PCR markers of 334 bp (VCG2B(334)) or 824 bp (VCG2B(824)). Artichoke isolates in the VCG2B(334)/2beta(334) cluster were molecularly similar to isolates of VCG1A. The molecular difference found among artichoke isolates in VCG2B correlates with virulence of isolates to artichoke and cotton cultivars demonstrated in a previous study.  相似文献   

2.
A multiplex-nested-PCR procedure was developed for in planta detection of Verticillium dahliae isolates infecting artichoke and assessment of their vegetative compatibility groups (VCGs). PCR markers were identified and assigned to V. dahliae VCGs, including: i) a 334 bp marker amplified from VCG1A or VCG2B334 isolates; ii) a 688 bp marker amplified from VCG2A or VCG4B isolates; and iii) a 688 bp and a 964 bp PCR marker amplified from VCG2B824 isolates. The infecting V. dahliae VCGs were identified in artichoke tissues according to specific patterns of amplified markers after two rounds of PCR. The PCR-based 'molecular tool box' was first optimized using DNA extracted from artichoke plants artificially inoculated with isolates representative of known VCGs. Thereafter, the efficiency of the molecular procedure was tested using DNA extracted from naturally-infected artichoke plants showing a range of symptom severity as well as from symptomless plants. The novel multiplex-nested-PCR assay was clearly superior in detecting the pathogen compared to conventional isolation procedures, and in addition was informative about the VCGs. Moreover, the PCR method allowed the detection and VCG identification of V. dahliae infections in symptomless but infected plants, which had yielded false negatives when checked by microbiological isolation procedures. This 'molecular tool box' has uncovered the presence of several V. dahliae VCGs infecting the same artichoke plants in the Comunidad Valenciana Region. In addition, it is useful for genetic and pathogenicity diversity studies of V. dahliae populations infecting artichoke, and may help in predicting the severity of verticillium wilt epidemics.  相似文献   

3.
ABSTRACT Severe Verticillium dahliae attacks have occurred in artichoke crops in the Comunidad Valenciana region of eastern-central Spain since the late 1990s. Knowledge of genetic and virulence diversity in the pathogen population is a key factor for the management of the disease through disease risk assessment as well as development and use of resistant cultivars. V. dahliae isolates from artichoke (109 isolates) and cotton (three isolates) in that region were characterized by vegetative compatibility grouping (VCG), and specific polymerase chain reaction assays using three sets of primer pairs that differentiate the cotton-defoliating (D) and -nondefoliating (ND) V. dahliae pathotypes. In all, 35 and 39 V. dahliae isolates representative of the identified VCGs and geographic origins were tested for virulence to artichoke cvs. Nun 6374 and Nun 9444, and cotton cv. Acala SJ-2, respectively. Four VCGs were identified among 107 artichoke isolates, and 2 isolates were heterokaryon self-incompatible: VCG1A (one isolate), VCG2A (31 isolates), VCG2B (72 isolates), and VCG4B (three isolates). The three cotton isolates were VCG1A. Isolates in VCG2B were distributed across the region and were the most prevalent isolates in the northern part. Conversely, 83.9% of isolates in VCG2A were recovered from the southern part of the region. Two subgroups of isolates were identified in VCG2B based on heterokaryon compatibility with either international or local tester isolates, which further showed diversity in the amplification of 334- and 824-bp DNA fragments which are markers of the D and ND pathotypes, respectively. Virulence of isolates to artichoke and cotton correlated with VCG but the pattern of correlation varied with the host. VCG1A isolates from artichoke and cotton induced defoliation in cotton but not in artichoke. Collectively, isolates of VCG2B and VCG4B were the most virulent and isolates of VCG1A or HSI were the least virulent to artichoke; but isolates of VCG1A were more virulent to cotton than those of any other VCG. Also, molecular subgrouping in VCG2B determined by amplification of the 334- and 824-bp markers correlated with virulence of isolates to the two hosts tested.  相似文献   

4.
Severity of Verticillium wilt in olive trees in Andalusia, southern Spain is associated with the spread of a highly virulent, defoliating (D) Verticillium dahliae pathotype of vegetative compatibility group 1A (VCG1A) but the extent of this spread and the diversity of the pathogen population have never been documented. VCG typing of 637 V. dahliae isolates from 433 trees in 65 orchards from five olive-growing provinces in Andalusia indicated that 78.1% were of VCG1A, 19.8% of VCG2A, 0.6% of VCG2B, 1.4% of VCG4B, and one isolate was heterokaryon self-incompatible. A single VCG prevailed among isolates within most orchards but two and three VCGs were identified in 12 and 3 orchards, respectively, with VCG1A+VCG2A occurring in 10 orchards. VCG1A was the predominant VCG in the three most important olive-growing provinces, and was almost as prevalent as VCG2A in another one. Molecular pathotyping of the 637 isolates using specific polymerase chain reaction assays indicated that VCG1A isolates were of the D pathotype whereas isolates of VCG2A, -2B, and -4B were of the less virulent nondefoliating (ND) pathotype. The pathotype of isolates correlated with the disease syndrome affecting sampled trees. Only three (seq1, seq2, and seq4) of the seven known sequences of the V. dahliae-specific 539- or 523-bp amplicon were identified among the 637 isolates. Distribution and prevalence of VCGs and seq sequences among orchards indicated that genetic diversity within olive V. dahliae in Andalusia is higher in provinces where VCG1A is not prevalent. Log-linear analysis revealed that irrigation management, source of irrigation water, source of planting stock, and cropping history of soil were significantly associated with the prevalence of VCG1A compared with that of VCG2A. Multivariate analyses using a selected set of agricultural factors as variables allowed development of a discriminant model for predicting the occurrence of D and ND pathotypes in the area of the study. Blind tests using this model correctly indentified the V. dahliae pathotype occurring in an orchard. The widespread occurrence and high prevalence of VCG1A/D pathotype in Andalusia have strong implications for the management of the disease.  相似文献   

5.
Korolev N  Katan J  Katan T 《Phytopathology》2000,90(5):529-536
A collection of 565 isolates of Verticillium dahliae, recovered between 1992 and 1997 from 13 host plant species and soil at 47 sites in Israel, was tested for vegetative compatibility using nitrate-nonutilizing (nit) mutants. Three vegetative compatibility groups (VCGs) were found and identified as VCG2A (28 isolates), VCG2B (158 isolates), and VCG4B (378 isolates) by using international reference strains. One isolate was heterokaryon self-incompatible. Of the VCG2B isolates, 92% were recovered from the northern part of Israel and 90% of VCG4B isolates were recovered from the south, with some overlap in the central region. Isolates of the minor group VCG2A were geographically scattered among the two major VCGs. Isolates of the same VCG resembled one another more than isolates from different VCGs based on colony and microsclerotial morphology, temperature responses, and, partially, pathogenicity. Different pathotypes were defined among 60 isolates tested, using cotton (cv. Acala SJ-2) and eggplant (cv. Black Beauty) as differentials. All isolates in VCG2A and 86% of the isolates in VCG4B, irrespective of their origin, induced weak to moderate symptoms on cotton and moderate to severe symptoms on eggplant and were similar to the previously described cotton nondefoliating patho-type. In contrast, all cotton isolates in VCG2B caused severe foliar symptoms, stunting, and often death, but little or no defoliation of inoculated cotton plants. These were defined as a cotton defoliating-like pathotype and induced only weak to moderate symptoms on eggplant. We concluded that vegetative compatibility grouping of V. dahliae in Israel is closely associated with specific pathogenicity and other phenotypic traits.  相似文献   

6.
Aggressiveness of Verticillium dahliae isolates from three vegetative compatibility groups (VCGs) was tested on potato and tomato. VCG4B was the most aggressive to potato and VCG2A was the most aggressive to tomato; VCG2B was the least aggressive to both potato and tomato. In potato, disease incidence, symptom severity and colonization index of stem segments were significantly higher in plants inoculated with VCG4B isolates than in those inoculated with VCG2B and VCG2A isolates. Inoculation with VCG4B and VCG2A decreased plant height and fresh weight more than inoculation with VCG2B. In tomato, VCG2A caused significantly more severe symptoms than either VCG4B or VCG2B. The colonization index in tomato plants inoculated with VCG2A was also significantly higher than in those inoculated with VCG4B and VCG2B. Similar patterns of relative aggressiveness were observed in potato and tomato when the pathogenicity of isolates of various VCGs, each originating from a specific host (cotton, potato or eggplant), was compared.  相似文献   

7.
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.  相似文献   

8.
Vegetative compatibility groups of a collection of 71 Greek Verticillium dahliae isolates obtained from cotton plants were tested. Nit mutants were generated from single spore wild strains by selecting chlorate-resistant sectors on minimal medium amended with potassium chlorate, 25g/l. These mutants were tested against tester strains from the USA and Greece of the previously described VCGs 1, 2, 3 and 4. Forty-six of 71 isolates belonged to VCG2, because they were able to anastomose with the testers of this group, two isolates belonged to VCG4 and one to VCG1, while the 22 remaining strains could not be assigned to any of the identified VCGs. Our data demonstrated that wilt of cotton is caused only by V. dahliae in Greece, and VCG2 is the most commonly detected VCG. Some strains were found to be more virulent to cotton than other strains from the same VCG. This is the first report of VCG1 of Verticillium in Greece.  相似文献   

9.
Host Range Specificity in Verticillium dahliae   总被引：1，自引：0，他引：1  
Bhat RG  Subbarao KV 《Phytopathology》1999,89(12):1218-1225
ABSTRACT Verticillium dahliae isolates from artichoke, bell pepper, cabbage, cauliflower, chili pepper, cotton, eggplant, lettuce, mint, potato, strawberry, tomato, and watermelon and V. albo-atrum from alfalfa were evaluated for their pathogenicity on all 14 hosts. One-month-old seedlings were inoculated with a spore suspension of about 10(7) conidia per ml using a root-dip technique and incubated in the greenhouse. Disease incidence and severity, plant height, and root and shoot dry weights were recorded 6 weeks after inoculation. Bell pepper, cabbage, cauliflower, cotton, eggplant, and mint isolates exhibited host specificity and differential pathogenicity on other hosts, whereas isolates from artichoke, lettuce, potato, strawberry, tomato, and watermelon did not. Bell pepper was resistant to all Verticillium isolates except isolates from bell pepper and eggplant. Thus, host specificity exists in some isolates of V. dahliae. The same isolates were characterized for vegetative compatibility groups (VCGs) through complementation of nitrate nonutilizing (nit) mutants. Cabbage and cauliflower isolates did not produce nit mutants. The isolate from cotton belonged to VCG 1; isolates from bell pepper, eggplant, potato, and tomato, to VCG 4; and the remaining isolates, to VCG 2. These isolates were also analyzed using the random amplified polymorphic DNA (RAPD) method. Forty random primers were screened, and eighteen of them amplified DNA from Verticillium. Based on RAPD banding patterns, cabbage and cauliflower isolates formed a unique group, distinct from other V. dahliae and V. albo-atrum groups. Minor genetic variations were observed among V. dahliae isolates from other hosts, regardless of whether they were host specific or not. There was no correlation among pathogenicity, VCGs, and RAPD banding patterns. Even though the isolates belonged to different VCGs, they shared similar RAPD profiles. These results suggest that management of Verticillium wilt in some crops through crop rotation is a distinct possibility.  相似文献   

10.
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.  相似文献   

11.
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.  相似文献   

12.
One hundred and fourteen isolates of Verticillium dahliae obtained from cotton and eggplant in mainland China were successfully assigned to two vegetative compatibility groups (VCGs) except for one self-incompatible isolate. Eleven isolates were strongly compatible with T9, the tester strain of the cotton defoliating pathotype, forming a linear growth of wild type with abundant microsclerotia and dense mycelia between compatible nitrate-nonutilizing mutants. The remaining 102 isolates were grouped into the non-defoliating VCG2, although the strength of the reaction varied; some isolates were strongly compatible with the tester strain while others were only slightly compatible. All VCG1 isolates including T9 showed the same defoliating symptom in greenhouse inoculation tests. This study confirmed the presence of the defoliating pathotype (VCG1) of V. dahliae in mainland China.  相似文献   

13.
A collection of 30 strains of Verticillium dahliae, recovered during 2004–2006 from 12 cultivars of chrysanthemum (Chrysanthemum morifolium) in five districts of İzmir province in Turkey, was assigned to vegetative compatibility groups (VCGs) based on pairings of complementary nitrate-nonutilizing (nit) mutants induced on a chlorate-containing medium. Of these strains, nine were assigned to VCG1, seven to VCG2A, 11 toVCG2B and one to VCG4B. The remaining two strains could not be tested for vegetative compatibility because of their inability to yield nit mutants. Pathogenicity tests conducted by the root-dip method, demonstrated that wilt of chrysanthemum in Turkey is caused by V. dahliae, and most strains in VCG1 were significantly more aggressive to chrysanthemum than those in VCGs 2 and 4B. This is the first known study in the world of the VCGs of V. dahliae isolates from chrysanthemum.  相似文献   

14.
Vegetative compatibility tests and random amplification of polymorphic DNA (RAPD) were used to assess genetic relationships amongst 54 strains of Fusarium poae obtained from various geographical regions. Twenty-seven strains were assigned to eight multiple member vegetative compatibility groups (VCGs), while the other 27 isolates were found to form single-member VCGs. There was a partial correlation between VCG and geographical origin, but the relationship was not always clear. However, no correlation was observed between the VCG and the host plant of origin. RAPD patterns were closely associated with VCGs in all cases. Members of VCGs that were interconnected by bridging isolates formed common branches in the phenogram constructed on the basis of the RAPD patterns, while strains that belonged to single-member VCGs were scattered throughout the phenogram. These data demonstrate that the combination of traditional and molecular methodologies allows reliable intraspecific subdivisions in an asexual fungus, which is a secondary invader of a wide range of host plants, and so has never been subject to the intense selection pressure of a single host species and lacks pathogenic subgroups.  相似文献   

15.
ABSTRACT The monophyletic origin of host-specific taxa in the plant-pathogenic Fusarium oxysporum complex was tested by constructing nuclear and mitochondrial gene genealogies and amplified fragment length polymorphism (AFLP)-based phylogenies for 89 strains representing the known genetic and pathogenic diversity in 8 formae speciales associated with wilt diseases and root and bulb rot. We included strains from clonal lineages of F. oxysporum f. spp. asparagi, dianthi, gladioli, lilii, lini, opuntiarum, spinaciae, and tulipae. Putatively nonpathogenic strains from carnation and lily were included and a reference strain from each of the three main clades identified previously in the F. oxysporum complex; sequences from related species were used as outgroups. DNA sequences from the nuclear translation elongation factor 1alpha and the mitochondrial small subunit (mtSSU) ribosomal RNA genes were combined for phylogenetic analysis. Strains in vegetative compatibility groups (VCGs) shared identical sequences and AFLP profiles, supporting the monophyly of the two single-VCG formae speciales, lilii and tulipae. Identical genotypes were also found for the three VCGs in F. oxysporum f. sp. spinaciae. In contrast, multiple evolutionary origins were apparent for F. oxysporum f. spp. asparagi, dianthi, gladioli, lini, and opuntiarum, although different VCGs within each of these formae speciales often clustered close together or shared identical EF-1alpha and mtSSU rDNA haplotypes. Kishino-Hasegawa analyses of constraints forcing the monophyly of these formae speciales supported the exclusive origin of F. oxysporum f. sp. opuntiarum but not the monophyly of F. oxysporum f. spp. asparagi, dianthi, gladioli, and lini. Most of the putatively nonpathogenic strains from carnation and lily, representing unique VCGs, were unrelated to F. oxysporum f. spp. dianthi and lilii, respectively. Putatively nonpathogenic or rot-inducing strains did not form exclusive groups within the molecular phylogeny. Parsimony analyses of AFLP fingerprint data supported the gene genealogy-based phylogram; however, AFLP-based phylogenies were considerably more homoplasious than the gene genealogies. The predictive value of the forma specialis naming system within the F. oxysporum complex is questioned.  相似文献   

16.
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.  相似文献   

17.
Isolates of Verticillium dahliae were collected from affected trees (Acer spp., Tilia spp. and Robinia spp.) and soils in Belgian ornamental nurseries. Nitrate non-utilizing mutants were produced and vegetative compatibility groups (VCGs) were classified based on complementation tests with reference tester strains. Of the 30 isolates analysed, 12 were classified as VCG2B and 18 as VCG4B following the American classification. In order to distinguish VCG2B from VCG4B, specific polymerase chain reaction primers were designed based on the sequence of a VCG2B-associated Direct Amplification of Minisatellite-region DNA (DAMD) band generated with the core sequence of the phage M13 minisatellite DNA. Using this test, amplification products were generated for all the VCG2B isolates characterized in this study. In contrast, no signal was seen on ethidium–bromide agarose gel for VCG4B isolates. Pathogenicity tests were carried out in a glasshouse on maple-rooted cuttings inoculated with conidial suspensions of V. dahliae belonging to both groups (VCG2B/VCG4B). Some strains proved to be highly aggressive, while others did not. However, these different behaviours were not correlated with the VCGs.  相似文献   

18.
Nitrate-nonutilizing (nit) mutants were used to determine vegetative compatibility among 34 isolates of Verticillium dahliae from cotton, potato, olive, eggplant, chrysanthemum and tomato from 12 sites in Israel. Based on the formation of complementary heterokaryons, 33 isolates were assigned to two vegetative- compatibility groups (VCGs): one VCG contained 15 isolates from cotton, eggplant, chrysanthemum and olive; and the other VCG contained 18 isolates from potato, olive and cotton. The status of an additional isolate from tomato, which was compatible with both VCGs, remained unclear. In a limited pathogenicity test with 10 isolates, two (from tomato and eggplant) were pathogenic on tomato, eggplant and cotton; most isolates from cotton were pathogenic on cotton and eggplant only; and one from cotton was non-pathogenic. Fewer isolates were pathogenic on tomato than on cotton or eggplant. The diversity of vegetative compatibility found in our V. dahliae collection is comparable to that found in studies of American populations.  相似文献   

19.
ABSTRACT To better understand the genetic relationships between Verticillium dahliae isolates from lettuce and other phytopathogenic Verticillium spp. isolates from various hosts and geographic locations, the complete intergenic spacer (IGS) region of the nuclear ribosomal RNA gene (rDNA) and the beta-tubulin gene were amplified and sequenced. The sequences of the complete IGS region and the beta-tubulin gene were used alone and in combination to infer genetic relationships among different isolates of Verticillium with the maximum-likelihood distance method. Phylogenetic analyses set sequences into four distinct groups comprising isolates of V. albo-atrum, V. tricorpus, and V. dahliae from cruciferous and noncruciferous hosts. Within the four Verticillium groups, isolates of V. dahliae from cruciferous hosts displayed the closest affinity to V. dahliae from noncruciferous hosts. Isolates of V. dahliae from noncruciferous hosts could be further divided into four subgroups based on sequence similarities within the IGS region. Cross-pathogenicity tests demonstrated that most Verticillium isolates were as virulent on other hosts as on their hosts of origin. A phenogram based on the cross pathogenicity of individual isolates resembled those derived from the IGS and beta-tubulin sequence comparisons. On the basis of the data presented, the potential origin of some isolates of V. dahliae pathogenic on lettuce is proposed.  相似文献   

20.
Martin FN 《Phytopathology》2010,100(11):1231-1239
The ability to monitor mitochondrial background in Verticillium dahliae may provide an additional tool for population studies and monitoring clonal populations. Published mitochondrial genome sequences of V. dahliae (DQ351941) were used to design primers for amplification of spacer regions for assessment of mitochondrial haplotype differences among isolates. Five regions were examined (5,229 bp, or 19% of the total genome size) for 30 isolates representing a range in vegetative compatibility group (VCG), host, and geographic origin. Observed differences among isolates were due to single nucleotide polymorphisms, different numbers of bases in specific homopolymeric regions, and copies of subrepeated sequences. When the differences observed for each locus were totaled there were 28 total groupings; when the results of each locus for individual isolates were combined there were 15 mitochondrial haplotypes. Some of the observed groupings correlated with VCG. For example, five VCG-1A and VCG-1B isolates from California, Spain, and Greece had identical haplotypes; however, this was not observed for VCG-2 or VCG-4 isolates. While some VCG-2 isolates also were identical and fell into a single haplotype, five haplotypes were found for this group (five other haplotypes were observed for other isolates that had not been characterized to VCG but grouped with VCG-2 isolates in the phylogenetic analysis). Likewise, five VCG-4 isolates fell into four mitochondrial haplotypes, one of which was identical to the largest VCG-2 grouping. A heterokaryon self-incompatible isolate that was reported in the literature to cluster with VCG-2 isolates by amplified fragment length polymorphism analysis was identical with VCG-1 isolates for four of the five loci, but was intermediate between VCG-1 and VCG-2 in the haplotype analysis. Phylogenetic analysis with these regions revealed the mitochondrial background of VCG-1 and VCG2-B to be monophyletic but VCG-2A and VCG-4 could not be separated. The results obtained indicate that there is variation in mitochondrial haplotypes and this type of analysis may be a useful for characterization of isolates. While data from all five regions was used for the haplotype separation in this study, depending on the VCG or the level of variability observed within a population it is possible to use fewer loci.  相似文献   

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