Fruiting body development and genetic analysis of somatic hybrids by protoplast fusion in edible fungi

식용버섯의 원형질체 융합체의 자실체 발생 및 유전분석

  • Yoo, Young Bok (Applied Microbiology Division, National Institute of Agricultural Science and Technology, RDA) ;
  • Kong, Won Sik (Applied Microbiology Division, National Institute of Agricultural Science and Technology, RDA) ;
  • Oh, Se Jong (Applied Microbiology Division, National Institute of Agricultural Science and Technology, RDA) ;
  • Jhune, Chang Sung (Applied Microbiology Division, National Institute of Agricultural Science and Technology, RDA) ;
  • Shin, Pyung Gyun (Applied Microbiology Division, National Institute of Agricultural Science and Technology, RDA) ;
  • Kim, Beom Gi (National Institute of Agricultural Biotechnology, RDA) ;
  • Kim, Gyu Hyun (Department of Biotechnology, Yonam College of Agriculture) ;
  • Park, Minsun (Department of Biochemistry and Molecular Biology, School of Medicine, Ajou University) ;
  • Min, Byung Re (Department of Biology, Sangmyung University)
  • 유영복 (농업과학기술원 응용미생물과) ;
  • 공원식 (농업과학기술원 응용미생물과) ;
  • 오세종 (농업과학기술원 응용미생물과) ;
  • 전창성 (농업과학기술원 응용미생물과) ;
  • 신평균 (농업과학기술원 응용미생물과) ;
  • 김범기 (농업생명공학연구원) ;
  • 김규현 (천안연암대학교 생물배양학과) ;
  • 박민선 (아주대학교 의과대학 생화학교실) ;
  • 민병례 (상명대학교 생물학과)
  • Published : 2004.09.30

Abstract

Somatic hybrids of inter-compatible and inter-incompatible strains were obtained by protoplast fusion. The fusion products between compatible strains, Pleurotus ostreatus and P. florida, formed heterokaryons, while fusants between incompatible strains such as P. cornucopiae + P. florida, P. ostreatus + Ganoderma applanatum, P. florida + Ganoderma lucidum, and P. ostreatus + Flammulina velutipes formed synkaryons that retained genes from both parents. The heterokaryons showed the same level of basidioma development. In contrast, the synkaryons showed unique characteristics including clamp connection formation at mitosis, either partner basidioma development, and abnormal segregation and recombination compared with inter-compatible strains. Synkaryons can be classified into homokaryoyic and heterokaryotic type. A comparison of somatic hybrids with compatible and incompatible strains was made using random amplified polymorphic DNA (RAPD) analysis. The heterokaryons between compatible species showed the same level of variability and contained both parental RAPD bands. In contrast, most of the synkaryons between incompatible species showed similarity to those of either parental bands and non-parental RAPD bands. Synkaryons can be classified into microgenome insertion type and macrogenome insertion type. A tetrapolar mating system was found among monospore isolates in somatic hybrids and wild type P. ostreatus. Homokaryons from each somatic hybrid combination were paired with tester homokaryons of the initial wild type of P. ostreatus. The changed mating types were identified in progenies. The pattern of mating type switching in somatic hybrids depends on compatibility of fusion partner. There are several factors related to the mechanism of clamp connection formation and fruiting body development of synkaryons. Of these,the major factor may be associated with self-fertility and mating type switching such as homokaryotic fruiting of wild type P. ostreatus. This review will discuss these aspects.

원형질체 융합에 의한 화합성 및 불화합성 종간 체세포 잡종을 얻었다. 화합성 종간인 Pleurotus ostreatus와 P. florida의 융합체는 이질핵체(heterokaryon)를 형성하였고, 불화합성 종간인 P. cornucopiae + P. florida, P. ostreatus + Ganoderma applanatum, P. florida + Ganoderma lucidum, 그리고 P. ostreatus + Flammulina velutipes는 합핵체(synkaryon)를 형성하였다. 이질이핵체는 동일한 양상의 자실체를 형성하는데 비해 합핵체는 유사분열상의 꺽쇠연결체 형성, 한쪽 친과 유사한 자실체 형성, 비정상적 유전형질 분리 및 유전자재조합 현상을 나타내었다. 화합성 및 불화합성 계통간 융합체의 RAPD 분석결과 화합성 종간 융합체는 동일한 DNA 패턴을 나타내었고, 불화합성 종간 융합체는 한쪽 친과 유사한 DNA 양상이면서 비양친 DNA 밴드도 형성하였다. 합핵체의 패턴은 microgenome insertion type과 macrogenome insertion type으로 구분되었다. 합핵체의 자실체 발생은 융합 모균주 양친의 자가임성에 의존하는데 이는 느타리의 동형핵체 자가임성과 유사한 양상이었고, 교배형 전환과 관련이 있는 것으로 사료된다. 여기서는 이러한 관점에서 논할 것이다.

Keywords