• Korean Journal Plant Pathology
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• v.14 no.6
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• pp.583-588
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• 1998

This study was conducted to investigate the potential of nitrate-nonutilizing mutants (nit mutants) in ecological studies of Fusarium disease of strawberry. Nit mutants of Fusarium oxysporum from strawberry were easily formed on chlorate-containing media. Nit mutants were assigned to three phenotypic classes, nit1, nit3, and NitM, on the basis of their growth on media containing one of the following five different nitrogen sources ; nitrate, nitrite, hypoxanthine, ammonium and uric acid. Frequency of nit mutation and proportion of three phenotypes of nit mutants depended on the isolate. Mutation rate was 45.6% and ranged from 15.0% to 95.0%. The frequency of nit1 mutants was higher than that of nit3 or NitM. The complementary reaction between nit1 and NitM was higher than that of other combination. There has been no complementary response observed between nit3 and nit3. The nit mutants showed similar growth pattern as the that of wild type isolate on potato sucrose agar and potato sucrose liquid media. Most of the mutants retained pathogenicity, and maintained their phenotypes even after two year preservation through subculture on slanted PSA at room temperature. Nit mutants were selctively isolated from infested soil and infected plants on the selective medium (MMCPA) containing potassium chlorate with their original phenotypes, while naturally occurring isolates of Fusarium oxysporum were not grow on the medium. On the contrary, nit mutants showed very slight growth on the medium (MMPA) containing nitrate as a sole nitrogen source, and therefore could be distinguished from wild type isolate.

• The Plant Pathology Journal
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• v.16 no.5
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• pp.247-251
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• 2000

Latent infections of healthy-appearing oaks of Hypoxylon atropunctatum complicates field studies by interfering with inoculation experiments to follow pathogenesis, fungal development and reproduction of this canker rot fungus. Mutants with unique and easily scorable phenotypes would be useful for inoculation studies. There is a broad range in the capacity of wild-type isolates to utilize nitrate as a sole nitrogen sources. Several types of nitrate-nonutilization mutants (nit1, Nit3, NitM) were selected from nitrate-utilizing wild-type isolates. Also, a few mutants of Hypoxylon atropunctatum were selected that could only grow poorly on basal medium supplemented with various nitrogen sources and even on yeast extract agar. These unknown mutants need to be characterized further. Nit mutants of Hypoxylon atropunctatum were readily selected, grew well and were recovered after inoculation into oak stems. These results suggest that nit mutants could be useful for inoculation studies in trees that contain latent infections.

• Asian Journal of Turfgrass Science
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• v.25 no.2
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• pp.171-176
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• 2011

Sclerotinia homoeocarpa, the causal agent of dollar spot, is one of the most common pathogens of cool season turfgrasses in South Korea. The vegetative compatibility group (VCG) assay was carried out using nitrate-nonutilizing (nit) mutants recovered from 13 South Korean isolates with various geographical origins. The mutants were divided into four phenotypic classes based on mutation loci associated with nitrogen assimilation: nit1, nit2, nit3, and NitM. The recovered number of nit mutants greatly varied among the isolates, ranging from 0 to 15 mutants. Of the mutants isolated, nit1 and nit2 mutants were most common (80%) while NitM and nit3 were relatively rare. One dominant and four minor VCGs were determined from 18 mutant isolates tested. To study population structures of Korean S. homoeocarpa isolates and increase our understanding of its ecological and epidemiological aspects for dollar spot management on turfgrass, more generated mutants should be tested with more diverse isolate collections.

• The Plant Pathology Journal
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• v.15 no.1
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• pp.53-56
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• 1999

Fifty-three isolates of Fusarium graminearum were obtained from corn and barley samples in several provinces of Korea. Gas chromatography-mass spectrometric analysis of trichothecenes produced by these isolates revealed that 37 and 16 isolates were nivalenol (NIV)- and deoxynivalenol (DON)-chemotypes, respectively. Two hundred and seventy-five nitrate-nonutilizing (nit) mutants were obtained from the isolates. Of these mutants, 187 were identified as nit1, nit3, or NitM, but 88 could not be identified as one of these classes. The highest frequency of nit mutant was nit1 (65%), followed by nit3 (20%) and NitM (15%). Higher frequency of NitM was observed in DON-chemotypes than in NIV-chemotypes. The mutants were used for vegetative compatibility group (VCG) analysis by examining heterokaryosis using complementary mutant pairs. No heterokaryon formation was observed among all 1,248 pairwise combinations, suggesting that all isolates tested belong to different VCGs. Higher frequency of self-incompatibility was observed in NIV-chemotypes than in DON-chemotypes. These results suggest that the like-lihood of asexual genetic recombination may be very low I F. graminearum under the field condition.

• The Plant Pathology Journal
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• v.26 no.3
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• pp.216-222
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• 2010

Fusarium proliferatum is the causal agent of stalk and root rot disease of maize, foot rot disease of rice, basal and root rot disease of onion and knife cut disease of sugarcane in Iran. In recent years, incidence and severity of these diseases have been increased in Iran. Fifty seven F. proliferatum single-spore isolates collected from diseased maize, rice, onion and sugarcane plants at different areas were used to study genetic diversity by determination of vegetative compatibility groups (VCGs). Chlorate-resistant nitrate non-utilizing (nit) mutants were recovered from selected isolates of F. proliferatum and used in complementation tests. All isolates in which both nit1 and NitM (or nit3) mutants were recovered, demonstrated self-compatibility. Vegetative compatibility tests by pairing nit mutants identified 30 VCGs among 57 isolates. Twenty-three isolates belonged to singlemember VCGs and the remaining 34 isolates, belonged to other seven multimember VCGs. Segregation of F. proliferatum isolates obtained from various area and host plants into different VCGs in Iran is reported for the first time. In this study, none of isolates obtained from rice complemented with any other isolates from onion and sugarcane and, non complementation occurred between onion and sugarcane isolates. Also, only one complementation occurred between one isolate of maize and one isolate of sugarcane and rice. Thus, a correlation between VCGs grouping and host preferences was founded. It is concluded that natural populations of F. proliferatum in Iran are probably genetically divergent and include isolates representing a potential risk for disease development.

• The Plant Pathology Journal
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• v.27 no.2
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• pp.116-119
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• 2011

Pyrenophora graminea, the causal agent of leaf stripe disease, is an economically important pathogen of barley found worldwide. Forty-four isolates of diverse geographical origin within Syria were grouped into vegetative compatibility groups (VCGs) by demonstrating heterokaryosis by complementation tests using nitrate nonutilizing (nit) mutants. All isolates were grouped into three VCGs-1-A, 1-B and 1-C. No self-incompatibility was observed in any of the isolates tested. VCG 1-A was the most common group within growing regions in Syria and proved to be the most virulent of the VCGs identified. These data indicate that the level of virulence in P. graminea is related to VCG.

• The Plant Pathology Journal
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• v.30 no.4
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• pp.384-396
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• 2014

Vegetative compatibility groups (VCGs) are determined for many fungi to test for the ability of fungal isolates to undergo heterokaryon formation. In several fungal plant pathogens, isolates belonging to a VCG have been shown to share significantly higher genetic similarity than those of different VCGs. In this study we sought to examine the relationship between VCG and genetic similarity of an important cool season turfgrass pathogen, Sclerotinia homoeocarpa. Twenty-two S. homoeocarpa isolates from the Midwest and Eastern US, which were previously characterized in several studies, were all evaluated for VCG using an improved nit mutant assay. These isolates were also genotyped using 19 microsatellites developed from partial genome sequence of S. homoeocarpa. Additionally, partial sequences of mitochondrial genes cytochrome oxidase II and mitochondrial small subunit (mtSSU) rRNA, and the atp6-rns intergenic spacer, were generated for isolates from each nit mutant VCG to determine if mitochondrial haplotypes differed among VCGs. Of the 22 isolates screened, 15 were amenable to the nit mutant VCG assay and were grouped into six VCGs. The 19 microsatellites gave 57 alleles for this set. Unweighted pair group methods with arithmetic mean (UPGMA) tree of binary microsatellite data were used to produce a dendrogram of the isolate genotypes based on microsatellite alleles, which showed high genetic similarity of nit mutant VCGs. Analysis of molecular variance of microsatellite data demonstrates that the current nit mutant VCGs explain the microsatellite genotypic variation among isolates better than the previous nit mutant VCGs or the conventionally determined VCGs. Mitochondrial sequences were identical among all isolates, suggesting that this marker type may not be informative for US populations of S. homoeocarpa.

• The Plant Pathology Journal
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• v.16 no.4
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• pp.227-230
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• 2000

A total of 90 isolates of Fusarium oxysporum f.sp. melonis, the causal agent of oriental melon (Cucumis melo var. makuwa) wilt, was isolated from symptomatic tissues of oriental melon from 4 provinces in Korea. These isolates were grouped into vegetative compatibility groups (VCGs) by demonstrating heterokaryosis through complementation using nitrate nonutilizing (nit) mutants. No self-incompatibility was observed in any of isolates. All isolates were grouped into 3 VCGs ; A, B, and C. iSOLATES BELONGING TO VCG A and VCG B accounted for 87% and 91% of the fungal population collected in 1991 and 1993, respectively. As the increment of cultivation period in the same field, the proportion of isolates belonging to VCG B increased whereas that of isolates belonging to VCG A decreased. Mean virulence of a total population increased as the increment of cultivation period in the same field. Isolates belonging to VCG B showed the highest increment of virulence. These data suggest that replanting of a host plant in the same field may cause increase of virulence in the pathogens. Furthermore, virulence of F. oxysporum f.sp. melonis isolates is related to the VCGs.

• Korean Journal Plant Pathology
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• v.14 no.2
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• pp.145-149
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• 1998

Four nonpathogenic isolates of Fusarium oxysporum isolated from spinach showed suppressive effect on the occurrence of the Fusarium wilt of spinach caused by F. oxysporum f. sp. sprinaciae, among which NF01 controlled the disease most effectively. And NF01 was not pathogenic to tomato, cucumber, radish and spinach. This isolate was further tested for the biological control of the disease. The isolate was not inhibitory to the growth of the pathogen on potato sucrose agar medium, however the Fusarium wilt disease occurred less by drenching spore suspension of the nonpathogenic isolate. The control effect of the isolate was higher at lower inoculum level of the pathogen than at the higher inoculum level, and in the pretreatments than the simultaneous treatment of the isolate with the pathogen inoculation. The nit mutants of the isolate were easily formed on chlorate containing media, and was reisolated selectively as nit mutant from infected soil and plants. The reisolation rate of the isolate as opposed to pathogen was high at preinoculated soil and plants relative to the simultaneous inoculation of the isolate with the pathogen.

• The Plant Pathology Journal
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• v.17 no.5
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• pp.281-289
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• 2001

We analyzed 88 strains of Gibberella fujikuroi (Analmorph: Fusarium section Liseola) from maize in Korea for mating population, mating type, fumonisin production vegetative compatibility, and random amplified polymorphic DNA (RAPD) patterns. We found 50 strains that were MATA-2, 22 that were MATA-1, 1 that was MATD-1, and 15 that were not reproducibly fertile with any of the mating type testers. Of the 50 MATA-2, 15 were female fertile, while 10 of the 22 MATA-1 strains were female fertile. A total of 1,138 nitrate non-utilizing (nit) mutants were recovered from a total of 88 strains. These strains were grouped into 39 vegetative compatability groups (VCGs) by demonstrating heterokaryosis between nit mutants. A single maize ear could be infected by more than one VCG of F. moniliforme. RAPD analysis measured genetic diversity among 63 strains of F. moniliforme. Several VCGs were distinguished by RAPD fingerprinting patterns. Most strains produced significant levels of fumonisins. However, 6 MATA-2 strains from a single VCG produced higher levels of fumonisin $\textrm{B}_3$ than that of fumonisin $\textrm{B}_1$ or $\textrm{B}_2$. From these data, we concluded that most Korean strains of F. moniliforme associated with maize belonged to mating population A and produced significant levels of fumonisins. Futhermore, RAPD analysis could differentiate strains associated with different VCGs.