• The Plant Pathology Journal
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• 제15권5호
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• pp.287-294
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• 1999

A technique based on the polymerase chain reaction (PCR) for the specific detection of genus Phytophthora and Phytophthora cryptogea-P. drechsleri complex group was developed using nucleotide sequence information of ribosomal DNA (rDNA) regions. The internal transcribed spacers (ITS) including 5.8S were sequenced for P. cryptogea-P. drechsleri complex group and its related species. Two pairs of oligonucleotide primers were designed. Primer pair ITS1/Phy amplified ca. 240 bp fragment in 12 out of 13 specie of Phytophthora, but not in Pythium spp., Fusarium spp.and Rhizoctonia solani. Primer pair rPhy/Pcd amplified 549 bp fragment only in P. cryptogea-P. drechsleri complex group, but not in other Phytophthora spp.and other genera. Specific PCR amplification using the primers was successful in detecting Phytophthora and P. cryptogea-P. drechsleri complex group in diseased plants.

• 한국균학회지
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• 제38권1호
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• pp.40-47
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• 2010

Phytophthora 속의 핵(ypt 유전자, rDNA-IGS region) 및 미토콘드리아(Cox 유전자, $\beta$-tubline 유전자, EF1A 유전자) 내에 존재하는 5가지 유전자 영역을 이용하여 국내 Phytophthora 속 14종의 유전적 다양성을 분석하였다. 국내 Phytophthora 속은 외국의 Phytophthora 속과 동일한 clade를 형성하였으나, 외국의 Phytophthora 속과 마찬가지로 본 연구에서도 분자생물학적 분류와 형태학적 분류와는 연관성을 찾기 어려웠다. 기존에 보고된 국내 P. palmivora KACC 40167 균주의 그룹이 국내에서 보고된 분류체계와 일치하지 않아 추후 재검토가 필요하였다. P. cryptogea-P. drechsleri complex group 내 국내 P. cryptogea KACC 40161 균주와 P. drechsleri KACC 40195 균주는 서로 94% 이상의 유사도를 보여 재동정이 필요하였으며, P. parasitica와 P. nicotianae간의 유사도가 99% 이상으로 나타나 이 두 종간에 통일된 종명이 요구된다. 또한 현재 분자계통학상 5그룹으로 구분된 국내 Phytophthora 속을 외국균주들과 비교하여 10개의 그룹으로 새롭게 재분류하였다. 이러한 결과들은 국내 Phytophthora 속의 유전적 다양성 연구를 위해 유용한 자료가 될 것이다.

• The Plant Pathology Journal
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• 제15권4호
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• pp.228-235
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• 1999

Genetic diversity of ninety-five Korean isolates of Phytophthora was investigated on the basis of PCR-RFLP of ribosomal DNA. The isolates were previously identified as following fifteen species by mycological and cultural characteristics; P. boehmeriae, P. cactorum, P. cambivora, P. capsici, P. cinnamoni, P. citricola, P. citrophthora, P. cryptogea, P. drechsleri, P. erythroseptica, P. infestans, P. megasperma, P. nicotianae, P. palmivora and P. sojae. The regions of small subunit (SSU) and internal transcribed spacer (ITS) of rDNA were amplified with primer pair, NS1 and ITS4, by polymerase chain reaction (PCR) and digested with nine restriction enzymes. P. boehmeriae, P. cactorum, P. cambivora, P. capsici, P. cinnamomi, P. citricola, P. citrphthora, P. infestans, P. nicotianae and P. palmivora showed specific band patterns for each species. However, P. sojae and P. erythroseptica presented identical band patterns and P. cryptogea, P. drechsleri and P. megasperma were divided into six groups, which were not compatible with delineation of the species. A group originated from cucurbits showed distinct band patterns from other groups, but the other five groups were closely related within 96.0% similarity, forming one complex group. Consequently, Korean isolates of Phytophthora were divided into thirteen genetic groups and each group was readily differentiated by comparing digestion patterns of AvaII, HaeIII, MboI, HhaI and MspI. Therefore, PCR-RFLP of rDNA using the five enzymes can be used to differentiate or identify the Phytophthora species reported in Korea so far.

• The Plant Pathology Journal
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• 제16권1호
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• pp.29-35
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• 2000

The internal transcribed spacer regions (ITS I, 5.8S and ITS II) of the ribosomal DNAs were amplified from Korean isolates of Phytophthora spp. and sequenced to characterize them. Sequences from 33 isolates previously identified as P. boehmeriae, P. cactprum, P. cambivora, P. capsici, P. cinnamomi, P. erythroseptica, P. infestans, P. megasperma, P. melonis, P. nicotianae, P. palmivora and P. sojae were compared with published sequences, and a phylogenetic tree was produced. All isolates belonging to 10 species, P. cactorum, P. cambivora, P. capsici, P. cinnamomi P. citricola, P. infestans, P. nicotianae, P. palmivora and P. sojae were clearly clustered into published isolates of each species above 97% bootstrap value. Cucurbits isolates of Phytophthora previously identified as either P. melonis or P. drechsleri showed distinct evolutionary lineages from the P. megasperma was closely related to isolates of P. cryptogea-P. drechsleri showed distinct evolutionary lineages from the P. cryptogea-P. drechsleri complex group, indicating that P. melonis is a valid species. A Korean isolate of P. megasperma was closely related to isolates of P. erythroseptica showed distant genetic relationship with published isolates of P. erythroseptica (CBS 956.87). It is probable that the two Korean isolates could be genetically different from foreign isolates or misidentified. A grouping of species according to ITS sequence divergence matched, to some degree, the broad classification based on type of papilla. However, a separation of semi-papillate species and papillate species was not wvident in this study.