• Title, Summary, Keyword: Phytophthora infestans

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Selection of RAPD Markers for Phytophthora infestans and PCR Detection of Phytophthora infestans from Potatoes

  • Kim, Kyung-Su;Lee, Youn-Su
    • Journal of Microbiology
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    • v.39 no.2
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    • pp.126-132
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    • 2001
  • For rapid and secure differentiation of P. infestans from other Phytophthora species, two fragments obtained from randomly amplified polymorphic DNA (RAPD) profiles were selected as markers. Also, primers for in polymerase chain reaction (PCR) to detect P. infestans specifically were developed by analyzing the sequences of ITSII regions in rDNA of Phytophthora species. The primers, PISP-1 and ITS3 amplified a single. Fragment 450 bp of about in P. infestans, but not in other fungal or bacterial isolates. Annealing temperatures and template DNA quantities were varied for the optimization of PCR conditions. From the result of the PCR detection study, species-specific primers were selected under annealing temperatures ranging from 55$^{\circ}C$ to 61$^{\circ}C$, and template DNA levels ranging from 10 pg to 100 ng.

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Fungicide-Resistance and Mating Type of Phytophthora infestans Causing Potato Late Blight (감자역병균(Phytophthora infestans De Bary)의 약제저항성 및 교배형)

  • 이왕휴;소만서;최인영
    • Korean Journal Plant Pathology
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    • v.10 no.3
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    • pp.192-196
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    • 1994
  • Two hundred and fourty-seven isolates of Phytophthora infestans obtained from infested potato leaves from the spring of 1991 to the fall of 1993 from potato fields in various regions of Korea were tested for their fungicides resistances. A total of 20.9% isolates were not suppressed at 50 ppm of metalaxyl in 1991, but isolates from 1993 were suppressed at 50 ppm of metalaxyl. Ten resistant isolates and 10 susceptible isolates to metalaxyl were selected and tested against oxadixyl, fosetyl-Al, and phosphorous acid. Effectiveness of these chemicals were no better than that of metalaxyl. Dimethomorph suppressed all isolates at 1 $\mu\textrm{g}$/ml suggesting that it might be a potential chemical to control Phytophthora infestans. Mating types of all isolates from diseased leaves in 1993 turned out to be A2 type.

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Genetic DNA Marker for A2 mating type in Phytophthora infestans

  • Kim, Kwon-Jong;Lee, Youn-Su
    • Journal of Microbiology
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    • v.40 no.4
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    • pp.254-259
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    • 2002
  • The Phytophthora infestans requires two mating types for sexual reproduction. Amplified fragment length polymorphism (AFLP) was used to specifically detect different mating types of P. infestans. The AFLP primers E+AA (5'-GACTGCGTACCAATTCAA-3') and M+CAA (5'-GATGAGTCCTGAG-TAAC AA-3') detected a fragment that is specific in the A2 mating type of P. infestans. This fragment was cloned and sequenced. Based on the sequence data, PHYB-1 and PHYB-2 primer were designed to detect the A2 mating type of P. infestans. A single 347 bp segment was observed in the A2 mating type of P. infestans, but not in the A1 mating type of P. infestans or other Phytophthora spp. Identification of mating type was performed with phenotype (sexual reproduction) and genotype (CAPs marker) methods. Two factors, the annealing temperature and template DNA quantity, were investigated to determine the optimal conditions. Using mating type-specific primers, a unique band was obtained within annealing temperatures of 57$^{\circ}C$-62$^{\circ}C$ and DNA levels of 10pg-100 ng (data not shown).

Rapid and Accurate Species-Specific Detection of Phytophthora infestans Through Analysis of ITS Regions in Its rDNA

  • Kim, Kyoung-Su;Lee, Youn-Su
    • Journal of Microbiology and Biotechnology
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    • v.10 no.5
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    • pp.651-655
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    • 2000
  • Polymerase chain reaction (PCR) was used to specifically detect Phytophthora infestans by analyzing the sequences of the ribosomal internal transcribed spacer regions (ITS) in the rDNA of the Phytophthora species. Based on the sequence data, PISP-1 together with the ITS3 primer were used to detect p. infestans. A single ca. 450 bp segment was observed in P. infestans, but not in the other fungal or bacterial isolates. Two factors, the annealing temperature and template DNA quantity, were investigated to determine the optimal conditions. Using these species-specific primers, a unique band was obtained within annealing temperatures of $55^{\circ}C$-$61^{\circ}C$ and template DNA levels of 10 pg-100 ng.

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A Genetic Marker Associated with the A1 Mating Type Locus in Phytophthora infestans

  • KIM KWON-JONG;EOM SEUNG-HEE;LEE SANG-PYO;JUNG HEE-SUN;KAMOUN SOPHIEN;LEE YOUN SU
    • Journal of Microbiology and Biotechnology
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    • v.15 no.3
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    • pp.502-509
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    • 2005
  • Sexual reproduction plays an important role in the biology and epidemiology of oomycete plant pathogens such as the heterothallic species Phytophthora infestans. Recent worldwide dispersal of A2 mating type strains of P. infestans resulted in increased virulence, gene transfer, and genetic variation, creating new challenges for disease management. To develop a genetic assay for mating type identification in P. infestans, we used the Amplified Fragment Length Polymorphism (AFLP) technique. The primer combination E+AT/M+CTA detected a fragment specific to A1 mating type (Mat-A1) of P. infestans. This fragment was cloned and sequenced, and a pair of primers (INF-1, INF-2) were designed and used to differentiate P. infestans Mat-A1 from Mat-A2 strains. The Mat A1-specific fragment was detected using Southern blot analysis of PCR products amplified with primers INF-1 and INF-2 from genomic DNA of 14 P. infestans Mat-A1 strains, but not 13 P. infestans Mat-A2 strains or 8 other isolates representing several Phytophthora spp. Southern blot analysis of genomic DNAs of P. infestans isolates revealed a 1.6 kb restriction enzyme (EcoRI, BamHI, AvaI)-fragment only in Mat-A1 strains. The A1 mating type-specific primers amplified a unique band under stringent annealing temperatures of $63^{\circ}C-64^{\circ}C$, suggesting that this PCR assay could be developed into a useful method for mating type determination of P. infestans in field material.

Anti-oomycete Activity of Furanocoumarins from Seeds of Psoralea corylifolia against Phytophthora infestans

  • Shim, Sang-Hee;Kim, Jin-Cheol;Jang, Kyoung-Soo;Choi, Gyung-Ja
    • The Plant Pathology Journal
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    • v.25 no.1
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    • pp.103-107
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    • 2009
  • In the course of a searching natural antifungal compounds from plant seeds, we found that the methanol extract of Psoralea corylifolia seeds showed potent control efficacy against tomato late blight caused by Phytophthora infestans and wheat leaf rust Puccinia recondita. Under bioassay-guided purification, we isolated two furanocoumarins, psoralen and isopsoralen, with anti-oomycete activity against P. infestans. By 1-day protective application, both compounds strongly reduced the disease development of P. infestans on tomato seedlings, but hardly controlled development of leaf rust on wheat seedlings. This is the first report on the anti-oomycete activity of P. corylifolia as well as that of psoralen and isopsoralen.

Physiological Races of Phytophthora infestans in Korea

  • Zhang, Xuan-Zhe;Kim, Byung-Sup
    • The Plant Pathology Journal
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    • v.23 no.3
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    • pp.219-222
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    • 2007
  • A total of the 261 Phytophthora infestans isolates collected from 2003 to 2005 in Korea were investigated for their physiological race composition. Among the isolates, we detected 18 physiological races and the dominant races were R0.1.3.5.6.10.11 and R0.1.3.5.6.7.10.11 with frequencies of 18.4% and 11.4%, respectively. All of the P. infestans races carried multiple virulence genes and showed virulence to the potato resistance genes R1, R3, R5, R6, R7, R10 and R11, but not to R8 and R9. Therefore, it is likely that the physiological races of P. infestans were diverse in Korea.

SCAR Marker Linked with A1 Mating Type Locus in Phytophthora infestans

  • Zhang Xuan-Zhe;Seo Hyo-Won;Ahn Won-Gyeong;Kim Byung-Sup
    • Journal of Microbiology and Biotechnology
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    • v.16 no.5
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    • pp.724-730
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    • 2006
  • A sequence characterized amplified region (SCAR) marker, which was tightly linked with the A1 mating type locus in Phytophthora infestans, was developed. During the random amplified polymorphic DNA-based phylogenic studies of 33 isolates of P infestans collected from year 2002 to 2004, we found an A1 mating type-specific DNA fragment. This 573-bp DNA fragment was generated only in the genomic DNA of the A1 mating types, when OPC-5 primer was used. Based on the specific DNA sequence, we designed the primer sets for generating the A1 mating type-specific 569-bp DNA fragment. When 33 genomic DNAs of P. infestans were subjected to PCR amplification using different primer combinations, the A1 mating type-specific DNA was amplified, when LB-1F and LB-2R primers were used. The specific 569-bp DNA fragment was generated only from all 18 A1 strains, but not from 15 A2 mating type strains. These results corresponded to the mating type discriminating bioassay of 33 isolates of P. infestans. Therefore, the primer combination of LB-1F/LB2R was chosen as a SCAR marker. Overall, this study indicates that the SCAR marker could be developed into a useful tool for mating type determination of P. infestans.

Molecular Genetic Classification of Phytophthora Species and P. infestans-specific Marker Selection by RAPD Fingerprinting (Phytophthora species의 분자유전학적 분류 및 RAPD fingerprinting을 이용한 P. infestans-specific 분자마커의 선발)

  • Kim, Kyoung-Su;Shin, Whan-Sung;Kim, Hee-Jong;Woo, Su-Jin;Ham, Young-Il;Shin, Kwan-Yong;Lee, Jeong-Oon;Kim, Byung-Sup;Shim, Jae-Ouk;Lee, Min-Woong;Lee, Youn-Su
    • The Korean Journal of Mycology
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    • v.27 no.6
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    • pp.394-398
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    • 1999
  • Taxonomic and genetic analysis of Phytophthora species belonging to six different morphological groups (GI, GII, GIII, GIV, GV, GVI) was conducted using RAPD method. Amplified fragments ranged $0.3{\sim}3.2$ kb in their molecular weights. Among total of 145 bands, there were 109 polymorphic bands. Seven isolates of P. infestans showed high similarities of $0.92{\sim}0.99$, and P. infestans isolate 3 from potato showed similarities of $0.93{\sim}0.95$ compared with other P. infestans. Among isolates of P. capsici, similarities of $0.77{\sim}0.86$ were observed and they were grouped in 80% level. P. cinnamomi and P. cryptogea isolates which belonging to group GVI showed very similar RAPD fingerprinting pattern. Primers OPA-04, OPA-17, OPA-18, OPA-19, and OPB-12 showed high level of differences among the tested isolates in major bands and molecular weights. The similarity between the isolates was 0.67. P. megasperma and P. sojae in group GV showed similarity of 0.65. These two isolates showed big differences in single major band in reactions with primers OPA-08, OPA-17, and OPA-19. Phytophthora-specific and P. infestans-specific molecular markers were also selected with one of the random primers tested. In reaction with primer OPA-20, all the genus Phytophthora showed common band at 600 bp, and all the P. infestans isolates showed specific band at 680 bp. These markers can be useful for identification of Phytophthora speices or P. infestans. As a result, P. infestans isolated from tomato and/or potato can easily be differentiated from other Phytophthora species with this primer.

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Genetic variation of Phytophthora infestans by RAPD analysis

  • Lee, Yun-Soo;Jeong young Song;Kim, Nam-Kyu;Nam Moon;Park, Hye-Jin;Kim, Hong-Gi
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • pp.116.2-117
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    • 2003
  • Late blight, caused by Phytophthora infestans, is one of the most destructive disease on potato and tomato cultivation. To analysis genetic diversity P. infeatans isolates were collected from potato and tomato fields in Korea. These pathogens contained both Al and A2 mating type with metalaxyl-resistant and sensitive isolates. Polymorphisms showed base on RAPD (Random Amplified Polymorphic DNA) in both potato and tomato isolates of P. infestans. Cluster analysis showed high level genetic variation in potato isolates of P. infestans than tomato isolates. P. infestans isolates were observed genetic diversity among them but not grouped among isolates related mating type and metalaxyl response. These results exhibited that P. infestans isolates showing genetic difference among them were distributed in Korea.

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