Korean Journal of Breeding Science (한국육종학회지)

The Korean Society of Breeding Science (한국육종학회)
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Variation of Amylose Content Using dsRNAi Vector by Targeting 3'-UTR Region of GBSSI Gene in Rice

GBSSI 유전자 3'UTR 영역의 발현 억제 dsRNAi 벡터를 이용한 아밀로스함량 조절 벼 개발

  • 박향미 (농촌진흥청 국립식량과학원) ;
  • 최만수 (농촌진흥청 국립식량과학원) ;
  • 천아름 (농촌진흥청 국립식량과학원) ;
  • 이정희 (농촌진흥청 국립식량과학원 춘천출장소) ;
  • 김명기 (농촌진흥청 국립식량과학원) ;
  • 김연규 (농촌진흥청 국립식량과학원) ;
  • 신동범 (농촌진흥청 국립식량과학원) ;
  • 이장용 (농촌진흥청 국립농업과학원) ;
  • 김율호 (농촌진흥청 국립식량과학원)
  • Received : 2010.11.10
  • Published : 2010.12.31
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Abstract

The amylose content of starch is a major factor in the texture of cooked cereal grains. Therefore, down-regulation of amylose synthesis is one of the alternative method to improve eating quality of rice. We developed transgenic rice plants designed to suppress granule-bound starch synthase I(GBSSI) gene using RNA interference(RNAi) technology. Transgenic plants with RNAi vector containing the 3'-UTR region of GBSSI showed a lower amylose content in rice endosperm than that of wild-type. The range of amylose content was 5.9~9.0% in the transgenic plants, whereas that of wild-type was 17.7~18.0%. Transgenic rices showed the decrease of short chain and the increase of long chain by analyzing chain length distribution of amylopectin in the endosperm. In the SEM micrographs, we found that compound starch granules in whole grains of the wild-type rice were readily split during fracturing, while the starch granules in RNAi-transgenic lines showed small voluminous, non-angular rounded bodies.

본 연구에서는 RNAi 기작을 이용하여 식미에 중요한 영향을 미치는 아밀로스 함량을 다양화하기 위해 GBSSI 유전자의 3'-UTR 부위를 targeting하여 dsRNA를 생성시킬 수 있는 운반체를 제작하고, 벼에 형질전환 하였다. 작성된 형질전환체들을 대상으로 $I_2$-KI 용액 반응과 아밀로스 함량을 분석한 결과, $I_2$-KI 용액에 대한 반응은 waxy 타입으로 나타났으나 아밀로스 함량은 찰벼와 저아밀로스 벼 사이에 해당되는 범위를 보였다. 원품종과 형질전환체 간의 아밀로펙틴 사슬 분포의 차이를 비교한 결과, 단쇄에 해당되는 A1과 B1 사슬의 분포는 감소한 반면, 중쇄에 해당되는 B2와 장쇄인 B3 사슬의 분포는 다소 증가하였으며, B3 사슬의 분포비율은 사용된 품종에 따라 약간의 차이를 보였다. 배유 단면의 전자현미경적 구조를 비교한 결과, 원품종에 비해 형질전환체의 전분립 크기가 작아지고 쪼개짐의 형태가 완만한 굴곡을 보였다. 이러한 결과를 바탕으로, RNAi 기술을 이용하여 다양한 아밀로스 함량이 조절된 형질전환 벼를 개발하기 위해서는 targeting 부위를 결정하는 것이 하나의 중요한 전략이 될 수 있음을 확인하였다.

Keywords

Acknowledgement

Supported by : 국립식량과학원

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