The Korean Journal of Mycology (한국균학회지)

The Korean Society of Mycology (한국균학회)
권호 표지

Phylogenetic Relationships Among Pleurotus species Inferred from Sequence Data of PCR Amplified ITS II Region in Ribosomal DNA

rDNA의 ITS II 부위의 염기서열분석에 의한 느타리버섯 종간의 근연관계

  • Bae, Shin-Churl (Molecular Genetics Division, National Agricultural Science and Technology Institute) ;
  • Seong, Ki-Young (Molecular Genetics Division, National Agricultural Science and Technology Institute) ;
  • Lee, Shin-Woo (Molecular Genetics Division, National Agricultural Science and Technology Institute) ;
  • Go, Seung-Joo (Molecular Genetics Division, National Agricultural Science and Technology Institute) ;
  • Eun, Moo-Young (Molecular Genetics Division, National Agricultural Science and Technology Institute) ;
  • Rhee, In-Koo (Department of Agricultural Chemistry, Kyungpook National University)
  • 배신철 (농업과학기술원 분자유전과) ;
  • 성기영 (농업과학기술원 분자유전과) ;
  • 이신우 (농업과학기술원 분자유전과) ;
  • 고승주 (농업과학기술원 분자유전과) ;
  • 은무영 (농업과학기술원 분자유전과) ;
  • 이인구 (경북대학교 농과대학 농화학과)
  • Published : 1996.06.30
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Abstract

This study was carried out to identify the phylogenetic relationship among several isolates of Pleurotus species by comparing ITS II region of ribosomal DNA(rDNA) repeat unit. Two primers from ribosomal DNA sequences were chosen to amplify the specific internal transcribed spacer (ITS) II region of Pleurotus spp. The exact ITS II region with an unique band from six species of Pleurotus genus could be amplified using the two primers taken from at the 3'-end of 5.8S rDNA and 5'-end of 28S rDNA. Six representative species of the Pleurotus genus were easily characterized according to the length differences of ITS II region. Furthermore, within P. ostreatus species, different sizes of ITS II region could be observed in the isolates of ASI 2025 and ASI 2095 although they were classified as P. ostreatus by the conventional observation. The nucleotide sequence analyses of PCR-amplified ITS II region indicated that the isolates ASI 2025 and ASI 2095 were different from other Pleurotus spp. When the nucleotide sequences of six Pleurotus species were compared, three typical ITS II regions were highly variable especially at both ends of this region. The phylogenetic tree obtained by the Neighbor program of Felsenstein PHYLIP package with all the nucleotide sequence of Pleurotus spp. indicated that P. ostreatus, P. florida, P. sajor-caju and P. eryngii were closely related to one phylogenetic branch and P. cystidious was related to other branch with P. cornucopiae. The isolates ASI 2025 and 2038, however, were not closely related to any other Pleurotus spp. and formed their own individual branches.

느타리 버섯속 6종 23균주에 대한 rDNA의 ITS II 부위의 DNA 염기서 열을 비교 분석함으로서 종간 및 종내의 근연관계를 조사하였다. rDNA의 ITS II 부위를 증폭하고자 5.8S rDNA의 3'말단 부위와 285 rDNA의 5'말단 부위에 두개 프라이머를 이용하여 PCR증폭을 행하였다. 느타리버섯 6종의 게놈 DNA로 부터 ITS II 부위를 증폭한 결과 종에 따라 밴드 길이에 차이가 있었으며 ITS II 부위의 길이 차이로 느타리 버섯 6종을 구분할 수 있었다. 같은 종내 개체간 구분을 위하여 느타리 버섯 6종 23균주의 PCR 증폭 결과는 느타리버섯(P. ostreatus) ASI 2096, ASI 2025와 노랑 느타리 버섯(P. cornucopiae) ASI 2038 균주외에 동일 종내 균주의 ITS II 길이는 동일한 양상을 보였다. 느타리 버섯 6종 및 ASI 2095, ASI 2025 그리고 ASI 2038에 대한 ITS II 부위의 염기서열을 비교해 보면 염기서열의 변이가 ITS II 부위가 시작되는 부분과 말단 부분에서 주로 존재하였다. ASI 2095와 ASI 2025 균주들은 동일 종내 느타리 버섯(P. ostreatus, ASI 2001) 균주와 ASI 2038와 ITS II 부위의 DNA 염기서열에 차이를 보였다. 이들 염기서열을 기초로 하여 Neighbor program을 이용한 균주간 유연관계는 느타리 버섯(P. ostreatus), 사철 느타리 버섯(P. florida), 여름 느타리 버섯(P. cornucopiae) 그리고 맛 느타리 버섯(P. eryngii)들이, 노랑 느타리 버섯(P. cornucopiae)과 호고 느타리 버섯(P. cystidious)이 각각 서로 가까운 유연관계를 나타내었으나 ASI 2025와 ASI 2038 균주 들은 특이한 계통분지를 나타내었다.

Keywords

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