논문 상세보기
pp.534-539
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 b
‘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+/Ca2+ 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.
