• 한국식물병리학회지
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• 제14권6호
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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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• 제30권4호
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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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• 제15권1호
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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.

• 아시안잔디학회지
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• 제25권2호
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• pp.171-176
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• 2011

동전마름병을 일으키는 Sclerotinia homoeocarpa는 한국에서 가장 일반적인 한지형 잔디 병원균이다. 경기도, 충청남도, 전라북도, 경상북도 등 다양한 지역에서 분리한 13개 균주로부터 유도된 nit 변이균주를 이용하여 의한 체세포화합성군(vegetative compatibility group, VCG) 평가가 이루어졌다. 돌연변이주는 질소동화작용에 근거하여 nit1, nit2, nit3, NitM 등 4가지 유형으로 분류되었다. 균주간에 돌연변이주 발생정도는 크게 차이가 있었으며 최대 15개의 변이주를 생산한 균주가 있는 반면에 전혀 형성하지 못한 균주도 존재하였다. 선택배지에서 4가지 돌연변이주 유형 중에 nit1과 NitM의 발생빈도(80%)가 높았으며, nit2와 nit3은 발생빈도가 매우 낮았다. 한 개의 큰 체세포화합성군 그룹과 4개의 작은 그룹이 18개 돌연변이주로부터 결정되었다. Nit 변이균주를 이용하여 의한 VCG 평가는 매우 효율적이었으며, 향후 한국의 Sclerotinia homoeocarpa균의 효율적 관리를 위한 생태학적 역학적인 면을 연구하기 위해서는 보다 많은 지역으로부터 분리된 균주의 평가가 필요하다.

• 한국식물병리학회지
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• 제12권4호
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• pp.395-401
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• 1996

국내의 고추, 사과, 포도, 스타티스(statice)에서 분리한 Glomerella cingulata 균주들의 병원성과 체세포화 합성군(vegetative compatibility group : VCG)과의 관계를 조사하였다. 사과, 포도, 스타티스에서 분리된 균주들은 전기주에 모드 병원성을 나타내었으나 고추에는 병원성이 없었다. 한편, 고추에서 분리된 균주들은 무상처접종시 병원성을 나타내지 못하였다. Nitrate nonutilizing(nit) mutant는 1.5%의 KClO\ulcorner을 첨가한 최소배지에서 선발하였다. nit mutnat들은 nit1, nit2, nit3, nitM의 4가지 표현형 중 하나에 속하였다. VCG는 표현형이 다른 nit mutan간의 대치배양을 통해 결정하였는데 사과, 포도, 스타티스에서 분리한 균주들간에는 같은 VCG를 나타낸 반면 고추에서 분리된 균주들은 다른 VCG로 나타났다. 따라서 G. cingulata 균주들의 체세포화합성은 병원성과 밀접하게 관련되어 있음을 알수 있었다. 또한 동일한 VCG에 속하며 기주가 다른 균주들의 대치배양시 heterokaryon을 형성했으나 자낭을 형성하지는 않았다.

• The Plant Pathology Journal
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• 제26권3호
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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.

• 한국식물병리학회지
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• 제14권2호
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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.

• 한국식물병리학회지
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• 제11권4호
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• pp.330-337
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• 1995

Forty-six isolates of Fusarium oxysporum collected from infected tomato plants and soils in greenhouses in Sedo, Chungnam and Angang, Kyeongbuk and 8 isolates of F. oxysporum f. sp. lycopersici from Japan and USA were used to determine vegetative compatibility groups (VCGs). Vegetative comaptibility was assessed on the basis of heterokaryon formation among nitrate nonutilizing mutants. All Korean isolates of F. oxysporum f. sp. lycopersici used in this study belonged to the same type of VCG (003) regardless of their geographic origin, cultivar and race, but they were incompatible with the foreign isolates of VCG 0030, 0031, 0032 and 0033. Based on the results, the Korean isolates of F. oxysporum f. sp. lycopersici were classified as a new VCG 003.

• The Plant Pathology Journal
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• 제22권3호
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• pp.222-229
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• 2006

Twenty-two isolates of Fusarium oxysporum f. sp. fragariae were obtained from diseased strawberry plants and their characteristics were investigated by vegetative compatibility group (VCG), random amplified polymorphic DNA (RAPD), and pathogenicity. Three major VCGs (A, B, and C) and one incompatible group were identified by nitrate reductase complementation test. The virulence pattern of the 22 isolates was studied in relation to four cultivars including Dochiodome, Red-pearl, Maehyang and Akihime. RAPD markers were used to determine genetic relationship, and created three major clusters among the 22 isolates of F. oxysporum f. sp. fragariae. Isolates belong to VCG-C were strongly pathogenic, and relatively high correlation was existed among VCG and RAPD, and virulence. In addition, VCG and RAPD pattern between pathogenic and non-pathogenic isolates were distinctly different.

• 한국식물병리학회지
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• 제12권1호
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• pp.33-40
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• 1996

우리나라 여러 딸기 재배지에서 전형적인 시들음 증상을 나타내는 이병식물에서 분리한 32개의 Fusarium oxysporum 균주들을 vegetative compatibility와 전기영동에 의한 동위효소상의 차이에 의해 분류하고 이들과 병원성과의 관계를 알아보았다. Nitrate nonutilizing(nit) mutant를 이용하여 vegetative compatibility group으로 분류해 본 결과 크게 A, B, C, D의 4개 그룹으로 나눌 수 있었으며 A그룹에는 15개 균주, B그룹에는 7개 균주, C와 D그룹에는 각각 2개 균주, 그리고 single VCG인 6개 균주가 존재하였다. 전기영동에 의한 esterase, catalase, acid phosphatase, leucin-aminopeptidase(LAP)의 동위효소상을 비교하여 본 결과 I, II, III, IV의 4개 그룹으로 나눌 수 있었으 I 그룹에는 18개 균주, II그룹에는 2개 균주, III그룹에는 6개 균주, IV그룹에는 6개 균주로 분류할 수 있었다. 또한, VCG의 A와 D그룹에 속하는 모든 균주들의 동위효소상의 I 그룹에 속하였으며 VCG B그룹의 7개 균주들 중에서 5개 균주가 동위효소상의 IV그룹, 7개 균주들 중에서 5개 균주가 동위효소상의 IV그룹, 나머지 2개 균주는 I 그룹과 III 그룹에 속하였으며 C그룹의 균주는 III과 IV 그룹에 속하였고, single VCGs들은 III 그룹에 4개 균주, II 그룹에 2개 균주가 속하므로 VCG와 동위효소상 간에는 밀접한 관계가 있음을 알 수 있었다. 4개 딸기 품종에 대해 병원성을 비교하여 본 결과 VCG A에서 선발된 2개 균주들은 보교조생에 대해 가장 병원성이 강하였으며 정보나 여홍에 대해서도 강한 병원성을 가지고 있었다. 반면에 B 그룹에서 선발된 2개 균주들은 4개 품종에 대해 병원성이 약하거나 거의 없었으며 C 그룹에서 선발된 1개 균주는 여홍에 대해 상대적으로 가장 강한 병원성을 가지고 있었다.