Korean Journal of Plant Tissue Culture (식물조직배양학회지)

The Korean Society of Plant Biotechnology (한국식물생명공학회)
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The Role of S RNase Associated with Gametophytic Self-Incompatibility in Tomato (Lycopersicon peruvianum)

토마토 자가불화합성에 관여하는 S RNase 유전자의 기능

  • 강나영 (대구효성카톨릭대학교 생명자원학부) ;
  • 김명희 (대구효성카톨릭대학교 생명자원학부) ;
  • 조규형 (동경대학교 생물공학과) ;
  • 신동일 (대구효성카톨릭대학교 생명자원학부) ;
  • 김달웅 (경북대학교 농학과) ;
  • 박희성 (대구효성카톨릭대학교 생명자원학부) ;
  • 정일경 (대구효성카톨릭대학교 생명자원학부)
  • Published : 2000.05.01
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Abstract

Lycopersicon peruvianum has a gametophytic self-incompatibility (GSI) mechanism controlled by a single genetic locus (S locus) with multiple alleles. S RNases, an allelic series of abundant stylar proteins, are products of the S locus in L. peruvianum and other Solanaceous plants. The $S_{11}$ RNase gene from L. peruvianum was introduced into a self-compatible (SC) species (Lycopersicon esculentum) to examine whether the expression pattern in the heterologous host mimics that in L. peruvianum. The resultant transgenic L. esculentum plants expressed the introduced gene highly in their styles, which is similar manner to the expresion in L. peruvianum. The $S_{11}$ RNase gene was expressed in the syle at a similar stage of flower development in both transgenic plants of L. esculentum and L. peruvianum without any morphological changes.

야생종 토마토의 자가불화합성에 관여하는 S RNase 유전자를 자가화합성인 재배종 토마토에 도입하여 형질전환 식물체를 만들었다. 재조합 Ti-plasmid pBI-S$_{11}$을 제작하여 Agrobacteria를 이용한 leaf disc transformation방법으로 재배종 토마토에 형질전환시켰으며, Southern분석을 통해 형질전환 식물체의 염색체 내에 자가불화합성 유전자가 안정된 구조로 도입되었는지 확인하였다. 형질전환 식물체의 화주 단백질을 이용하여 RNase활성 염색을 하여 화주 내 5단백질의 활성을 확인하였다. 또한 형질전환 식물체의 화주 단백질을 이용하여 western분석을 수행한 결과, 야생종 토마토 유래의 자가불화합성 유전자에 의해 5단백질을 정확하게 발현하는 식물체를 확인, 선발하였다. 선발된 형질전환 식물체의 화분, 화주, 화변, 잎, 줄기 등 5개의 조직을 western분석하여 5 RNase 유전자가 야생종 토마토에서와 유사한 형태로 화주조직에서만 높게 발현되며 형질전환된 재배종 토마토에서도 조직 특이적으로 발현한다는 것을 확인하였다. 위의 결과로 야생종 토마토에서 자가불화합성에 결정적인 영향을 미치는 S RNase를 재배종 토마토에 발현시켰을 때 자가불화합성을 나타내는 재배종 토마토의 표현형은 변화가 없다는 것과 어떠한 형태적인 변화도 없다는 사실을 확인하였다. 그리고 배우자체 자가불화합성 기작에는 화주쪽의 S RNase 이외에 화분쪽의 또 다른 기구가 있다는 것을 추측할 수 있었다.

Keywords

References

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