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각종 작물로부터 분리한 Rhizoctonia solani 균주의 RFLP 및 PCR-RFLP를 이용한 분자계통한 특성 구명
최혜선,신환성,김희종,김경수,우수진 한국미생물학회 1999 미생물학회지 Vol.35 No.3
Rhizoctonia solani 분류를 위해 균사의 anastomosis group과 grouprks의 유전적 차이의 분석을 위해 rDNA 의 PCR-RFLP 와 RFLP를 실시하였다. rDNA 의 PCR-RFLP 결과 R. solani 는 크게 5 group 으로 나뉘어졌다. 균주 번호 1번과 3번은 AG-5 그룹과 0.976의 높은 상동성을 보였고, 12번, 13번은 AG-2-2와 0.976의 상동성을 보였다. 균주번호 7,8,11,13,15 는 AG-1에 속하였다. 제한효소 HaeIII로 절단하였을 때 균주번호 4,5,7,8은 700 bp에서 하나의 밴드가 나타나 AG-1에 속하였고, 균주번호 9는 AG-2-1과 517 bp에서 하나의 밴드가 나타났다. 도한 제한효소 Msp I을 사용하여 southern blotting을 한 결과 더 다양한 절단양상을 보이며 AG 간의 차이가 나타났다. AG-2-1과 균주번호 9는 200 bp에서 하나의 밴드가 나타났다. 균주 3,4,5,10,11,13은 AG-1과 1kb에서 하나의 밴드로 나타났다. As a result of PCR-RFLP, the isolates used in this study were classified into five groups. Isolates 1 and 3 were included in AG-5 with 97% genetic similarity. Isolates 12 and 13 were included in AG-1 wilh 100% genetic similarily. Isolates 10 and AG-2-2 showed 97% similarity Isolates 7, 8, 11. 13, and 15 were included in AG-1. When isolates of 4, 5, 7 and 8 were restricted with Hae I. there was a single 700 bp fragment matched with AG-1. A 517 bp restriction fragment of isolate 9 was matched with AG-2-1. Based on the result of southem hybridization of genomic DNAs, all isolates restricted with Msp I showed more variable restriction differences than those restricted with Hae Ill. Isolates AG-2-1 and 9 showed 200 bp restriction fragment, and isolates 3 and AG-1 showed 1 kb restriction fragments.
Phytophthora species 의 분자유전학적 분류 및 RAPD fingerprinting 을 이용한 P . infestans - specific 분자마커의 선발
이민웅,김경수,신환성,김희종,우수진,함영일,신관용,이정운,김병섭,심재욱,이윤수 한국균학회 1999 韓國菌學會誌 Vol.27 No.6
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∼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∼0.99, and P. infestans isolate 3 from potato showed similarities of 0.93∼0.95 compared with other P. infestans. Among isolates of P. capsici, similarities of 0.77∼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 far 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.