Molecular breeding of herbicide resistant transgenic plants with bromoxynil specific nitrilase gene

Bromoxynil 특이성 nitrilase 유전자를 이용한 제초제 저항성 형질 전환 식물의 분자육종

Bromoxynil is an antidicot herbicide widely used on cereal crops and has a short half life in the soil. A bxn gene, encoding a specific nitrilase that converts bromoxynil to its primary metabolite 3,5-dibromo-4-hydroxybenzoic acid, was inserted in plant binary vector pGA482, and then introduced into tobacco and lettuce plants via Agrobacterium mediated leaf-disc transformation method. Transgenic plants with the bxn gene were selected by kanamycin and regenerated to whole plants. The regenerated transgenic plants were determined level of expression of bxn gene by Northern blot analysis. Leaf-disc analysis and pot-assay confirmed that the transgenic tobacco and lettuce plants were resistant to high doses of bromoxynil.

Bromoxynil은 쌍떡잎 특이적 제초제로써 폭 넓게 이용되고 있으며 반감기가 매우 짧다. Bromoxynil을 3,5-dibromo-4-hydroxybenzoic acid로 분해하는 nitrilase를 암호화한 bxn 유전자를 식물 벡터인 pGA482에 도입하고 Agrobacterium과의 동시배양을 통해 담배와 상추에 형질전환하였다. Kanamycin을 이용해 형질전환 식물체를 선별하고 완전한 식물체로 분화시켰다. Northern hybridization을 통해 bxn 유전자의 발현정도를 검정하고 liaf-disc와 pot assay를 통해 형질전환 식물체가 고농도의 bromoxynil에 저항성을 보임을 확인하였다.