NDP Kinase 2 유전자를 도입한 산화스트레스 내성 형질전환 감자의 선발
Selection of Transgenic Potato Plants Expressing NDP Kinase 2 Gene with Enhanced Tolerance to Oxidative Stress
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복합스트레스 내성 유전자 NDP kinase 2 유전자를 도입시킨 형질전환 감자를 개발하기 위하여 이 유전자를 산화스트레스에 의해 발현이 강하게 유도되는 SWPA2 프로모터 또는 enhanced CaMV 35S 프로모터에 연결한 벡터를 제작한 후 각각 Agrobacterium 매개로 형질전환 하였다. 기관발생 경로에 의해 kanamycin 저항성 식물체를 재분화 시킨 후 Southern 분석으로 외래 유전자가 안정적으로 감자 게놈내로 삽입되었음을 확인하였다. 형질전환 감자 식물체의 잎 조직을 대상으로 10 μM methyl viologen에 대한 내성 검정을 조사하여 산화스트레스 내성 형질전환 감자 식물체를 2 개체씩 선발하였다. 선발된 식물체는 건조, 고온 등의 여러 가지 환경스트레스 내성 분석을 실시할 예정이며 이로부터 복합재해에 내성을 지닌 감자 품종을 개발할 수 있을 것으로 기대한다.
Abstract
Arabidopsis NDPK2 (AtNDPK2) is a key singaling component that regulate cellular redox state and known to enhance multiple stress tolerance when over-expressed in Arabidopsis plant (Moon et al. 2003). In order to develop transgenic potato plants with enhanced tolerance to multiple stresses, we placed an AtNDPK2 cDNA under the control of a stress-inducible SWPA2 promoter or enhanced CaMV 35S promoter. Transgenic potato plants (cv. Superior and Atlantic) were generated using an Agrobacterium-mediated transformation system and selected on MS medium containing 100 mg/L kanamycin. Genomic Southern blot analysis confirmed the incorporation of AtNDPK2 cDNA into the potato genome. When potato leaf discs were treated with methyl viologen (MV) at 10 M, transgenic plants showed higher tolerance to MV than non-transgenic or vector-transformed plants. The NDPK2 transgenic potato plants will be further used for analysis of stress-tolerance to multiple environmental stresses.
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