scholarly journals Vegetative Compatibility Groups in Cercospora kikuchii, the Causal Agent of Cercospora Leaf Blight and Purple Seed Stain in Soybean

2005 ◽  
Vol 95 (3) ◽  
pp. 257-261 ◽  
Author(s):  
G. Cai ◽  
R. W. Schneider

Nitrogen nonutilizing (Nit) mutants were used to assess vegetative compatibility of 58 isolates of Cercospora kikuchii, 55 of which were isolated from soybean plants in Louisiana. Two isolates were vegetatively self-incompatible. Of 56 self-compatible isolates, 16 were assigned to six multimember vegetative compatibility groups (VCGs), 01 to 06, with 2 or 3 isolates in each VCG. The other 40 isolates each belonged to a distinct VCG. All six multimember VCGs contained isolates from different soy bean cultivars, and three included isolates from different locations. Only one of six multimember VCGs included isolates both from soybean leaves and seed, while the other five included isolates from only leaves or seed. The likelihood of tissue specificity or preference was discussed. All isolates and tested Nit mutants produced cercosporin on potato dextrose agar under light. Significantly different amounts of cercosporin were produced among wild-type isolates, and two Nit mutants produced significantly more cercosporin than their wild-type counterparts. All isolates produced typical Cercospora leaf blight symptoms on soybean plants in greenhouse pathogenicity tests.

1999 ◽  
Vol 89 (2) ◽  
pp. 182-188 ◽  
Author(s):  
Robert L. Bowden ◽  
John F. Leslie

We developed a method for inducing sexual outcrosses in the homothallic Ascomycete fungus Gibberella zeae (anamorph: Fusarium graminearum). Strains were marked with different nitrate nonutilizing (nit) mutations, and vegetative compatibility groups served as additional markers in some crosses. Strains with complementary nit mutations were cocultured on carrot agar plates. Ascospores from individual perithecia were plated on a minimal medium (MM) containing nitrate as the sole nitrogen source. Crosses between different nit mutants segregated in expected ratios (3:1 nit-:nit+) from heterozygous perithecia. Analysis of vegetative compatibility groups of progeny of two crosses indicated two and three vegetative incompatibility (vic) genes segregating, respectively. For rapid testing of sexual recombination between nit mutants, perithecia were inverted over MM to deposit actively discharged ascospores. Development of proto-trophic wild-type colonies was taken as evidence of sexual recombination. Strains of G. zeae group 2 from Japan, Nepal, and South Africa, and from Indiana, Kansas, and Ohio in the United States were sexually interfertile. Four group 1 strains were not interfertile among themselves or with seven group 2 strains. Attempts to cross G. zeae with representatives of F. acuminatum, F. avenaceum, F. culmorum, F. crookwellense, F. oxysporum, and three mating populations of G. fujikuroi were not successful.


2002 ◽  
Vol 92 (8) ◽  
pp. 827-832 ◽  
Author(s):  
N. Nitzan ◽  
M. Hazanovsky ◽  
M. Tal ◽  
L. Tsror(Lahkim)

Black dot of potato, caused by Colletotrichum coccodes, is a disease of growing economic importance, but the degree of genetic diversity and pathogenic differentiation among isolates is unknown. Using nitrate auxotrophic (Nit) mutants, we characterized vegetative compatibility groups (VCG) diversity for C. coccodes for 110 isolates originating from Israel, The Netherlands, and France. We recovered frequencies of nit1 and NitM mutant classes at 38.5 and 7.2%, respectively, and selected 12 isolates as tester isolates. Using these testers, we defined four multimember VCGs at 7.3, 35.5, 20.0, and 10.0% frequency in this sample. Thirty isolates (27.3% of all tested isolates) could not be assigned to any of the major groups, and showed only self-compatibility. The frequency of recovery of Nit mutant sectors was highest in isolates from VCG4, with 50.9 and 13.6% recovery for nit1 and NitM, respectively. However, we did not detect differences in the frequency of mutant classes among the three countries of origin. In pathogenicity tests, isolates from VCG3 were the most aggressive to potato, as expressed by high stem colonization levels and sclerotia density on root and crown. These results suggest that there is significant VCG diversity in this species and that this VCG diversity may be correlated with pathogenic characteristics or specialization.


2016 ◽  
Vol 56 (2) ◽  
pp. 163-166 ◽  
Author(s):  
Maria Rataj-Guranowska

Abstract The way compatible pairs of nitrate non-utilizing mutants (nit) are selected is usually not explained and remains unclear whether these pairs are representative for an isolate or strain. In addition, tester strains of Verticillium dahliae vegetative compatibility groups (VCGs) cross-react with at least one pair of tester strains of another VCG, and although it is a common knowledge of scientists working with the fungus that reversion of nit mutants to wild type occurs far too often, this fact is rarely mentioned in papers. To overcome the above problems, a protocol was developed for the generation of large number of nit mutants from any given isolate and to ensure that compatible pairs of mutants are indeed stable and the most frequent within the putative tester mutants produced from each isolate. Thus, we provide a reproducible and objective way of selecting V. dahliae tester strains for each isolate and VCG. Although VC grouping is based on the formation of stable heterokaryons, we demonstrate in this work that cross-reactions cannot be eliminated and that strict genetic barriers between two main VCG groups are absent in V. dahliae.


1985 ◽  
Vol 63 (2) ◽  
pp. 179-183 ◽  
Author(s):  
John E. Puhalla

Twenty-one strains of Fusarium oxysporum were classified on the basis of vegetative compatibility or the ability to form hetcrokaryons. Heterokaryon formation was demonstrated by pairing mutants that were unable to reduce nitrate. These "nit mutants" could be recovered without mutagen treatment from selective media containing KClO3. On Czapek's minimal medium the nit mutants had a radial growth rate like that of wild type, but their colonies were very thin. Two genetically different nit mutants were recovered in each of the 21 strains and paired in all combinations on minimal medium. Heterokaryon formation was indicated by dense growth where the two mutant colonies touched. As a result, 16 vegetative compatibility groups (VCGs) were defined such that only strains in the same VCG were vegetatively compatible. In no case was a strain assignable to more than one VCG. There was some evidence for a correlation between VCG and forma specialis. An evolutionary model to explain this correlation is proposed. Vegetative compatibility may be a fast and easy way to distinguish pathotypes of F. oxysporum.


2008 ◽  
Vol 98 (11) ◽  
pp. 1205-1211 ◽  
Author(s):  
R. B. Martins ◽  
L. A. Maffia ◽  
E. S. G. Mizubuti

Cercospora leaf spot is a destructive fungal disease that has become a threat to the coffee industry in Brazil. Nevertheless, little is known about populations of its causal agent, Cercospora coffeicola. We evaluated the potential of using nitrogen-nonutilizing (nit) mutants and vegetative compatibility groups (VCGs) to characterize the genetic variability of the C. coffeicola population associated with coffee plantings in Minas Gerais state (MG), Brazil. A total of 90 monosporic isolates were obtained from samples collected according to a hierarchical sampling scheme: (i) state geographical regions (Sul, Mata, and Triângulo), and (ii) production systems (conventional and organic). Nit mutants were obtained and 28 VCGs were identified. The 10 largest VCGs included 72.31% of all isolates, whereas each of the remaining 18 VCGs included 1.54% of the isolates. Isolates of the largest VCGs were found in the three regions sampled. Based on the frequencies of VCGs at each sampled level, we estimated the Shannon diversity index, as well as its richness and evenness components. Genetic variability was high at all hierarchical levels, and a high number of VCGs was found in populations of C. coffeicola associated with both conventional and organic coffee plantings.


1991 ◽  
Vol 69 (1) ◽  
pp. 168-172 ◽  
Author(s):  
T. R. Gordon ◽  
D. Okamoto

One hundred isolates of Fusarium oxysporum collected from soil in the San Joaquin Valley in 1988 were tested for their ability to form intra- and inter-isolate heterokaryons. Five thousand and fifty isolate combinations were tested for vegetative compatibility, using complementary nitrate nonutilizing mutants. Ninety-eight combinations (2%) produced weakly positive reactions, 248 (5%) produced strong reactions, and 4704 combinations (93%) were negative for heterokaryon formation. This result suggests that anastomosis occurs infrequently among isolates in this population. Isolates were assigned to vegetative compatibility groups based on strong reactions. By this procedure 100 isolates were placed in 29 vegetative compatibility groups. At least one isolate from each of the 29 vegetative compatibility groups was tested and found to be vegetatively incompatible with the muskmelon wilt pathogen, F. oxysporum f. sp. melonis, which occurs in the same field. Each of these isolates also was avirulent to muskmelon in greenhouse pathogenicity tests. Forty isolates of F. oxysporum were collected in 1989 and 32 (80%) were compatible with isolates collected in 1988. The remaining eight isolates represented eight different vegetative compatibility groups. Key words: anastomosis, fungi, heterokaryon.


1989 ◽  
Vol 67 (8) ◽  
pp. 2420-2424 ◽  
Author(s):  
J. A. LaMondia ◽  
W. H. Elmer

Isolates of Fusarium moniliforme (Sheld.) emend. Snyd. & Hans., F. oxysporum (Schlecht) emend. Snyd. & Hans., and F. solani (Mart.) Appel & Wollenw. emend. Snyd. & Hans. were recovered from three 5-year-old field grown asparagus (Asparagus officinalis L. cv. Mary Washington) by isolating from symptomatic and asymptomatic feeder roots, storage roots, crown and basal stem segments. Fusarium moniliforme was more virulent than F. oxysporum on asparagus seedlings and F. solani was considered nonpathogenic. Isolates of F. moniliforme and F. oxysporum were placed into vegetative compatibility groups (VCGs) by demonstrating heterokaryosis with complementation tests using nitrate-nonutilizing (nit) mutants (pairing nitM and nit1 mutants). Ninety-seven of 135 isolates of F. moniliforme were placed in 13 vegetative compatibility groups. The remaining 38 isolates were not classified by vegetative compatibility because of poor nit mutant recovery. Eight of 18 isolates of F. oxysporum were unique and classed as single members of eight different VCGs. The other 10 isolates were not placed in VCGs. All isolates of F. moniliforme were virulent, but mean disease ratings differed among the isolates in different VCGs. There was no correlation between vegetative compatibility group and tissue substrate or symptom expression on the tissue substrate. It appears that virulence on asparagus is a common trait with few exceptions among genetically distinct populations of F. moniliforme and F. oxysporum colonizing asparagus.


2000 ◽  
Vol 90 (5) ◽  
pp. 529-536 ◽  
Author(s):  
Nadia Korolev ◽  
Jaacov Katan ◽  
Talma Katan

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.


1997 ◽  
Vol 87 (10) ◽  
pp. 1067-1070 ◽  
Author(s):  
Nadia Korolev ◽  
Talma Katan

Nitrate-nonutilizing (nit) mutants are commonly used to determine vegetative compatibility between isolates of Verticillium dahliae by complementation (heterokaryon) testing. These mutants emerge spontaneously as chlorate-resistant sectors growing out of partially restricted, wild-type colonies on chlorate-amended media. The commonly used chlorate media are based on minimal medium (MMC) or cornmeal agar (CMC), amended with potassium chlorate. nit mutants recovered on these media constituted 10 to 36%(on MMC) and 25 to 45%(on CMC) of the apparently resistant sectors. An improved water agar chlorate medium (WAC) is described that is more effective for selecting chlorate-resistant nit mutants. WAC medium consists of agar (2%), glucose (0.02%), and potassium chlorate (2 to 5%). On WAC, growth of most V. dahliae isolates was strongly inhibited, and 66 to 100%(average >80%) of the chlorate-resistant sectors formed were nit mutants. Most mutants were characterized as nit1, and about 6% as NitM.


2008 ◽  
Vol 98 (1) ◽  
pp. 108-114 ◽  
Author(s):  
Y.-K. Jo ◽  
S. W. Chang ◽  
J. Rees ◽  
G. Jung

Nitrate-nonutilizing (nit) mutants were recovered for the first time from 21 isolates of Sclerotinia homoeocarpa collected in the United States. Mutants were selected from shredded mycelium of each isolate when cultured on water agar medium amended with 4% (wt/vol) potassium chlorate. The mutants could be classified into three phenotypes: nit1, nit3, and NitM, based on their growth on minimal medium (Czapek solution agar) supplemented with NaNO2 or hypoxanthine. Complementary heterokaryons were observed in pairings between different phenotypes of nit mutants derived from compatible isolates, but not in self-fusions or pairings between incompatible isolates. The vigor of prototrophic growth varied with isolates and mutant phenotypes. Strong and continuous heterokaryons, as well as weak and spontaneous ones, formed depending on pairings of nit mutants. Stable heterokaryons between compatible isolates, but apoptotic reactions between incompatible isolates, were observed immediately after hyphal fusion under the epifluorescence microscope. The 21 isolates used in this study, which were previously assigned into 11 different vegetative compatibility groups (VCGs) based on the formation of a barrage zone at the contact site of paired isolates on complete medium (potato dextrose agar), were regrouped into five VCGs based on heterokaryon formation between nit mutants on minimal medium.


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