Peroxiredoxin 유전자 발현 산화스트레스 내성 형질전환 고구마의 선발
Selection of transgenic sweetpotato plants expressing 2-Cys peroxiredoxin with enhanced tolerance to oxidative stress
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산화스트레스에 내성을 지닌 형질전환 고구마 식물체를 개발하 기 위해서 산화스트레스에 의해 발현이 강하게 유도되는 SWPA2 프로모터 또는 CaMV 35S 프로모터에 2-Cys peroxiredoxin (Prx) 유 전자가 발현되도록 연결한 형질전환 벡터 (pSP-K, pEP-K)를 제작 한 후, 각각 Agrobacterium 매개로 형질전환 하였다. 카나마이신 저 항성 배발생 캘러스로부터 체세포배발생 과정을 거쳐 100 mg/L kanamycin이 포함된 MS 배지에서 소식물체로 발달하였다. Southern 분석으로 외래 유전자가 안정적으로 고구마 게놈 내로 삽입되었음 을 확인하였다. 형질전환 고구마 잎 조직을 대상으로 20 μM methyl viologen에 대한 내성 검정을 조사하여 형질전환 고구마 식 물체가 비형질전환 식물체 또는 벡터 대조구 식물체 보다 40% 정 도 높은 산화스트레스에 대한 내성을 보여주었다. 선발된 형질전 환 식물체는 저온, 건조 등의 여러 가지 환경스트레스 내성검정에 이용될 것이며 향후 복합재해 내성 고구마 계통육성에 이용될 수 있을 것으로 기대된다
Abstract
In order to develop transgenic sweetpotato plants [Ipomoea batatas (L.) Lam. cv. Yulmi] with enhanced tolerance to oxidative stress, we constructed transformation vectors expressing 2-Cys peroxiredoxin (Prx) gene under the control of the stress-inducible SWPA2 or enhanced 35S promoter (named as SP or EP). Transgenic sweetpotato plants were attempted to generate from embryogenic calli using an Agrobacterium-mediated transformation system. Embryogenic calli gave rise to somatic embryos and then converted into plantlets on MS medium containing 100 mg/L kanamycin. Transgenic plants were regenerated in the same medium. Southern blot analysis confirmed that the Prx gene was inserted into the genome of the plants. To further study we selected the transgenic plant lines with enhanced tolerance against methyl viologen (MV). When sweetpotato leaf discs were subjected to methyl MV at 20 μM, transgenic plants showed about 40% higher tolerance than non-transgenic or empty vector-transformed plants.
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