Korean Journal of Breeding Science (한국육종학회지)

The Korean Society of Breeding Science (한국육종학회)
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Introduction of CAX1 into 'Hongro' Apple via Agrobacterium tumefaciens

CAX1 유전자가 도입된 사과 '홍로' 형질전환체

  • Kim, Jeong-Hee (Fruit Research Division, National Institute of Horticultural & Herbal Science) ;
  • Shin, Il Sheob (Fruit Research Division, National Institute of Horticultural & Herbal Science) ;
  • Cho, Kang-Hee (Fruit Research Division, National Institute of Horticultural & Herbal Science) ;
  • Kim, Se Hee (Fruit Research Division, National Institute of Horticultural & Herbal Science) ;
  • Kim, Dae-Hyun (Fruit Research Division, National Institute of Horticultural & Herbal Science) ;
  • Hwang, Jeong Hwan (Fruit Research Division, National Institute of Horticultural & Herbal Science)
  • 김정희 (농촌진흥청 국립원예특작과학원 과수과) ;
  • 신일섭 (농촌진흥청 국립원예특작과학원 과수과) ;
  • 조강희 (농촌진흥청 국립원예특작과학원 과수과) ;
  • 김세희 (농촌진흥청 국립원예특작과학원 과수과) ;
  • 김대현 (농촌진흥청 국립원예특작과학원 과수과) ;
  • 황정환 (농촌진흥청 국립원예특작과학원 과수과)
  • Received : 2010.11.05
  • Published : 2010.12.31
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Abstract

'Hongro' is early-mid maturing cultivar with good quality like 'Tsugaru' and it has not preharvest drop. The CAX1 gene was introduced into Korean apple cultivar 'Hongro' by Agrobacterium tumefaciens LBA4404 harboring pBI121 to obtain transgenic apple with enhanced Ca level. The CAX1 gene playing the role of $H^+/Ca^{2+}$ transporter from Arabidopsis thaliana increases Ca concentration in several plants. Regenerated transgenic lines were confirmed by polymerase chain reaction (PCR) analysis and Southern blot analysis of genomic DNA for the existence of CAX1 gene. Southern blot analysis of 'Hongro' transformants showed that two putative transgenic lines were integrated with CAX1 gene in genomic DNA. The CAX1 comparative expression levels of two transgenic lines were higher than that of non-transformant evaluated by comparative quantification analysis using a real-time PCR. These two lines were multiplied in vitro, and micro-grafted on apple rootstocks 'M.9' in the isolated greenhouse. Since two years after micro-grafting, the fruits came into bearing. Compared to Ca level of the non-transgenic 'Hongro', that of the CAX1 transgenic 'Hongro' in the flesh and leaves was higher.

사과 국내 육성 품종인 '홍로'의 형질전환 체계를 확립하고 칼슘이 강화된 '홍로' 형질전환체를 육성하기 위하여 CAX1 유전자 전환을 실시하였다. 접종에 사용되는 절편체는 기내에서 증식 배양 중인 신초의 유엽을 사용하는 경우가 생육기 수체의 유엽을 사용하는 것보다 높은 재분화율을 보여 효과적이었다. 항생제가 첨가된 재분화 선발 배지에서 재분화된 신초들을 대상으로 PCR을 실시한 결과 5개체에서 유전자 도입을 확인할 수 있었고, 이들을 Southern blot 분석한 결과 2개체에서 안정적으로 유전자가 도입되었음을 확인할 수 있었다. 또한 유전자의 정량적 발현 분석을 위해 real-time PCR을 실시한 결과 대조구에 비해 형질전환체 모두에서 CAX1 유전자의 상대적 발현량이 높게 나타났다. 형질 전환체의 칼슘 분석 결과 기내 증식 잎과 결실된 과실의 과육과 잎에서 칼슘 함량이 대조구에 비해 높게 나타났다.

Keywords

References

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