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Characterization of Fusarium oxysporum f. sp. fragariae Based on Vegetative Compatibility Group, Random Amplified Polymorphic DNA and Pathogenicity

  • Nagarajan Gopal;Kang Sung-Woo;Nam Myeong-Hyeon;Song Jeong-Young;Yoo Sung-Joon;Kim Hong-Gi
    • The Plant Pathology Journal
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    • v.22 no.3
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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.

Vegetative Compatibility Grouping and Pathogenicity of Colletotrichum gloeosporioides Isolates from Different Host Plants

  • Ahn, Il-Pyung;Kim, Soonok;Im, Kyung-Hwan;Lee, Yong-Hwan
    • The Plant Pathology Journal
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    • v.19 no.6
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    • pp.269-273
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    • 2003
  • A total of 57 isolates of Colletotrichum gloeosporioides were recovered from diseased tissues of Hall's crab apple (Malus haliana), 3 cultivars of edible apple (M. pumila var. dulcissima), red pepper (Capsicum annum), and grapevine (Vitis vinifera) fruits. All isolates showed strong virulence on their own host plants. Isolates from edible apple exhibited high level of cultivar specificity in pathogenicity tests. Ten isolates from apple cultivar 'Fuji' were virulent on 'Jonathan' and 'Rall's Genet'. However, 12 isolates from 'Jonathan' and 'Rall's Genet' were not virulent on 'Fuji'. Among the 24 isolates from red pepper, only seven and two isolates were infective on edible apple and grapevine fruits, respectively. All six isolates from grapevine were only virulent on their own host. These isolates were grouped into five vegetative compatibility groups (VCGs), A, B, C, D, and E, by demonstrating heterokaryosis through complementation using nitrate-nonutilizing (nit) mutants. Among them, isolates belong to VCG-A and VCG-D accounted for 24 and 17 isolates; those in VCG-A exhibited wide host range involving Hall's crab apple, all three edible apple cultivars, and red pepper. On the other hand, isolates of VCG-D and VCG-E showed limited host range specific to red pepper and grapevine, respectively. Taken together, the data suggest that among C. gloeosporioides isolates, the concepts of pathotype and/or forma specialis may exist, and that three is a relationship between host specificity and VCG grouping among C. gloeosporioides isolates.

Vegetative Compatibility Groups and Pathogenicity Variation among Isolates of Fusarium oxysporum f.sp. melonis

  • Ahn, Il-Pyung;Lee, Yong-Hwan
    • 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.

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Differentiation of Fusarium oxysporum f. sp. fragariae Isolates by Random amplified Polymorphic DNA (RAPD) Analysis. (Random Amplified Polymorphic DNA(RAPD)를 이용한 딸기 시들음병균(Fusarium oxysporum f. sp. fragariae)의 분류)

  • 현재욱;박원목
    • Korean Journal Plant Pathology
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    • v.12 no.1
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    • pp.41-46
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    • 1996
  • 본 실험은 이병 딸기의 조직에서 분리 동정된 시들음병균(Fusarium oxysporum f. sp. fragariae) 균주들의 유?거 변이를 random amplified polymorphic DNA(RAPD) marker들을 이용하여 조사하였다. 총 24개의 딸기 시들음병 균주들의 DNA를 주형으로 하여 16개의 random 10-mer primer들을 사용하여 증폭시킨 결과 총 231개의 marker들을 이용하여 유전적 변이를 조사해 본 결과 크게 RAPD I과 RAPD II의 2개 그룹으로 나눌 수 있었다. RAPD I그룹에 속하는 균주는 VCG A에 속하는 Y1, K1, K2, K3, K4, N2, N3, N4-1, N6-1, N6-2, N8, N9, N10, M1-2-1 균주, VCG B에 속하는 M4-1 균주 그리고 VCG C에 속하는 N1, Y2 균주들이었고, RAPD II그룹에는 VCG B에 속하는 M1-1, M2-2-1, M2-4-2, M3-2, M3-3-2 균주와 VCG D에 속하는 N1 1 균주가 속하였다. 이들 2그룹 간에는 31%의 유사성이 있었다.

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Vegetative Compatibility, Isozyme Polymorphisms and Pathogenicity of Isolates of Fusarium oxysporum f. sp. fragariae (딸기 시들음병균(Fusarium oxysporum f. sp. fragariae)의 균주간 Vegetative Compatibility, 동위효소 Polymorphism과 병원성)

  • 현재욱;김순옥;박원목
    • Korean Journal Plant Pathology
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    • v.12 no.1
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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.

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Distribution and Molecular Characteristics of Vibrio vulnificus Isolated from Seawater Along the Gadeok Island Coast (가덕도 연안 해수에서 Vibrio vulnificus의 분포 및 분리균주의 병원성 유전자 특성)

  • Oh, Hee-Kyung;Jeong, Hee-Jin;Kim, Young-Mog
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.53 no.5
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    • pp.688-693
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    • 2020
  • Vibrio vulnificus is a Gram-negative marine bacterium known to cause septicemia. This study was conducted to investigate the distribution of V. vulnificus along the coast of Gadeok Island in Korea and to determine the molecular characteristics of isolated strains sampled between March and November 2019 from seawater. The strains were mostly detected between July and September, when the average water temperature and average salinity were 22.2-26.2℃ and 14.2-29.9 psu, respectively. V. vulnificus was not detected in seawater below 15℃. In September, the highest population of V. vulnificus was observed at 2,100 MPN (most probable number)/100 mL, attributable to decreased salinity from heavy rains. In addition, the detection rate of V. vulnificus was higher at the sampling station near the Nakdong River. Virulence-related genes were also identified among the isolates, such as vvhA (97.1%), viuB (44.1%), and vcgC (57.4%). In particular, viuB and vcgC were only observed in V. vulnificus isolated from June to September, when the detection rate was high and water temperature was above 20℃, suggesting the role of seasonal characteristics.

Vegetative Compatibility Groups and Virulence Variation Among Isolates of Pyrenophora graminea

  • Arabi, Mohammad Imad Eddin;Jawhar, Mohammad
    • 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.

Detection Characteristics and Antimicrobial Resistance of Vibrio vulnificus Isolated from Sea Water Along the Gadeok Island Coast (가덕도 연안 해수에서 분리된 Vibrio vulnificus의 검출 특성 및 항생제 내성)

  • Oh, Hee-Kyung;Jeong, Hee-Jin;Jeong, Geum-Jae;Shin, Hye-Young;Shin, Joong Ho;Jung, Won-Kyo;Oh, Junghwan;Kim, Young-Mog
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.54 no.6
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    • pp.912-917
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    • 2021
  • We investigated the distribution, molecular characteristics, and antimicrobial resistance of Vibrio vulnificus isolated from seawater at Gadeok Island, the Republic of Korea between June to October. Interestingly no isolates were detected between December to February. The detection rate of V. vulnificus was high (80-100%) from July to September 2019 and from June to September 2020. This coincided with the relatively low salinity of the seawater, which ranged from 7.8-29.9 practical salinity units for that period. Additionally, V. vulnificus had a high detection rate at sampling stations near the Nakdong river. The detection rates of virulence genes, such as vvhA, viuB, and vcgC, among the isolates were 97.1%, 44.1%, and 57.4% in 2019 and 100%, 43.0%, and 50.0% in 2020, respectively. Notably, viuB and vcgC were detected in V. vulnificus isolated between June to October when water temperature was above 20℃. The antimicrobial susceptibility analysis of 80 isolates revealed that most of the strains were susceptible to most antimicrobial agents. However, some isolates showed intermediate resistance to cefepime (18.8%), cefoxitin (58.8%), and erythromycin (22.5%). Of note, 3.8% of the tested strains were resistant to cefoxitin. The minimum inhibitory concentration of highly cefoxitin-resistant strains was determined to be less than 32 ㎍/mL.

Diversity of VC and incidence of hypovirulence-associated ds-RNAs in the chestnut blight fungus Cryphonectria parasitica in Korea

  • Byeongjin Cha;Jinyoung Lim;Ju, Young-Jik;Kim, Dae-Hyuk
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.23-23
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    • 2003
  • Chestnut blight, caused by Cryphonectria parasitica, is the most destructive disease of American and European chestnut trees. A total of 672 C prasitica was isolated from blight lesion on chestnut twigs, which were collected from major chestnut plantations all over Korea in 1999. Isolation rates were over 30% in Kyunggj-, Kyongnam-, and Chonnam-do. The highest isolation rate was 37.4% and recorded in Kyongnam-do. On the other hand, Chonbuk-do had the lowest isolation rate as 13.5%. In grouping of C parasitica by colony shape and color, yellow colony with irregular margin were the most dominant colony type with a frequency of 65.2%. When the 672 isolates were inoculated on the chestnut twigs, 380 isolates (56.5%) caused lesions larger than the standard virulent isolate EP155-2, while 158 isolates (23.4%) caused smaller lesions than the standard hypovirulent isolate UEP-1. In Bavendamm test that determines phenol oxidase activity, 97.1% of all the isolates resulted the same or darker discoloration than EP155-2, and only 12.2% resulted the same or lighter discoloration than UEP-1. In the vegetative compatibility (VC) tests, total 670 isolates were divided into 121 VC groups (VCGs). Kyongnam-, Chonnam-, and Chungnam-do, the three principal chestnut plantation area, had 49, 33, and 27 VCGs, respectively. Among the VCGs, the biggest VCG, KR-VC104, was composed of 164 isolates and the second biggest VCG had 62 isolates. But, 64 of 121 VCGs consisted of sole member. More than 65.8% of KR-VC104, was isolated from the three provinces, Kyongnam-, Kangwon-, and Chungbuk-do. In KR-VC104, 62.8%, 59.1%, and 85.9% of the isolates looked like virulent in colony type, pathogenicity test, and Bavendamm test. In ds-RNA detection tests using cellulose chromatography, 77 of total 650 isolates were ds-RNA positive and detected ds-RNA segments were approximately 12kb, 3kb, 2.7kb, 2kb, and 1.8kb in size. Among the 77 isolates, 46 isolates had 12kb and 25 isolates had 12kb and 2.7kb. Other 6 Isolates had small ds-RNA segments. Kyongnam-, Chonnam-, and Chungnam-do had 43, 16, and 5 ds-RNA positive isolates, respectively. Among the 121 VCGs, only 29 VCGs had ds-RNA positive isolates.(중략)

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Profiles of Virulence Genes and Antimicrobial Resistance of Vibrio vulnificus Strains Isolated from Seawater and Fisheries (해수 및 수산물에서 분리한 Vibrio vulnificus의 병원성 유전자 및 항균제 내성)

  • Park, Kwon-Sam;Cho, Eui-Dong;Kim, Hee-Dai
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.54 no.6
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    • pp.918-926
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    • 2021
  • We isolated 28 Vibrio vulnificus strains from seawater and fisheries and investigated the positive rate of eight virulence genes. Additionally, we evaluated the susceptibility of these strains to 25 antimicrobials. The positive rates of fur, vvhA, tcp, rtxA, vcgC, viuB, vvp, and acfA were 100, 92.9, 92.9, 67.9, 64.3, 25.0, 14.3, and 7.1%, respectively. A disk diffusion susceptibility test revealed that, all the investigated strains had the highest resistance to amoxicillin and oxacillin, followed by that to streptomycin (96.4%), cefoxitin (92.9%), clindamycin (82.1%), amikacin (67.9%), vancomycin (46.4%), nalidixic acid (7.1%), penicillin G (7.1%), and ampicillin (3.6%). Moreover, they were susceptible to 10 other antimicrobials, including cefotaxime, chloramphenicol, erythromycin, gentamicin, and rifampicin. Notably, amoxicillin, oxacillin, and streptomycin had average minimum inhibitory concentrations of 132.6, 603.4, and 23.1 ㎍/mL against V. vulnificus, respectively. These observations provide new insights regarding the necessity for sanitation of commercial fisheries and can potentially, help reduce the risk posed by fisheries contaminated with bacteria resistant to antimicrobials.