CuZnSOD와 APX를 엽록체에 발현시킨 산화스트레스 내성 형질전환 감자의 선발
Selection of Transgenic Potato Plants Expressing Both CuZnSOD and APX in Chloroplasts with Enhanced Tolerance to Oxidative Stress
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초록
산화스트레스에 내성을 지닌 형질전환 감자 식물체를 개발하기 위하여 산화스트레스에 의해 발현이 강하게 유도되는 SWPA2 프로모터에 CuZnSOD와 APX 유전자가 엽록체에서 동시에 발현되도록 연결한 형질전환 벡터 (pSSA-K)를 제작한 후 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 조직에 10 μM methyl viologen을 처리하여 산화스트레스 내성 검정을 조사한 결과 형질전환체는 MV에 대해 강한 내성을 지님을 확인하였다. 내성을 보인 개체 중에서 환경스트레스에 대한 내성 조사를 위하여 품종별로 2 개체씩 선발하였다. 선발된 식물체는 건조, 고온 등의 여러 가지 환경스트레스 내성검정에 이용될 것이며 향후 복합재해 내성 감자 품종이 개발될 수 있을 것으로 기대한다.
Abstract
In order to develop transgenic potato plants with enhanced tolerance to multiple stress, we constructed the transformation vector expressing both superoxide dismutase and ascorbate peroxidase genes in chloroplasts under the control of a stress-inducible SWPA2 promoter. Transgenic potato plants (cv. Superior and Atlantic) were generated using an Agrobacterium-mediated transformation system. Transgenic potato plants were regenerated on MS medium containing 100 mg/L kanamycin. Genomic Southern blot analysis confirmed the incorporation of foreign genes into the potato genome. When potato leaf discs were subjected to methyl viologen (MV) at 10 M, transgenic plants showed higher tolerance than non-transgenic or vector-transformed plants. To further study we selected the transgenic plant lines with enhanced tolerance against MV. These plants will be used for further analysis of stress-tolerance to multiple environmental stresses.
참고문헌 (21)
- Allen RD, "Dissection of oxidative stress tolerance using transgenic plants", 107, pp.1049~1054 (1995)
- Allen RD, "Use of transgenic plants to study antixodants defenses", 23, pp.473~479 (1997)
- Asada K, "The water-water cycle in chloroplasts: Scavenging of active oxygen an dissipation of excess photons", 50, pp.601~639 (1999)
- Asemota HN, "A fast, simple, and efficient miniscale method for the preparation of DNA from tissues of yam (Dioscorea spp.)", 1, pp.19~21
- Foyer CH, "Protection against oxygen radicals an important defense mechanism studied in transgenic plants", pp.507~523 (1994)
- Inze D, "Oxidative stress in plants", 6, pp.166~172 (1995)
- Kim KY,, "A novel oxidative stress-inducible peroxidase promoter from sweet potato: molecular cloning and characterization in transgenic tobacco plants and cultured cells", 51, pp.831~838 (2003)
- Kwon SY,, "Enhanced tolerances of transgenic tobacco plants expressing both superoxide dismutase and ascorbate peroxidase in chloroplasts against methyl viologen-mediated oxidative stress", 25, pp.873~882 (2002)
- Kwon SY,, "Enhanced stress-tolerance of transgenic tobacco plants expressing a human dehydroascorbate reductase gene", 160, pp.347~353 (2003)
- Lee HS,, "Molecular characterization and expression of a cDNA encoding copper", zinc superoxide dismutase from cultured cells of cassava (Manihot esculenta Crantz, 1999, pp.262, 807~814
- McKersie BD, "Water-deficit tolerance and field performance of transgenic alfalfa overexpression superoxide dismutase", 111, pp.1177~1181 (1996)
- Moon HJ,, "NDP kinase 2 interacts with two oxidative stress-activated MAPKs to regulate cellular redox state and enhances multiple stress tolerance in transgenic plants", 100, pp.358~363 (2003)
- Murashiga T, "A revised medium for rapid growth and bioassays with tobacco tissue cultures", 15, pp.473~497 (1962)
- Oberschall A,, "A novel aldose", aldehyde reductase protects transgenic plants against lipid peroxidation under chemical and drought stresses, 2000, pp.24, 437~446
- Roxas VP, "Overexpression of glutathione-S-transferase", glutathione peroxidase enhances the growth of transgenic tobacco seedlings during stress, 1997, pp.15, 988~991
- Slooten L,, "Factors affecting the enhancement of oxidative stress tolerance in transgenic tobacco overexpressing manganese superoxide dismutase in the chloroplasts", 107, pp.737~750 (1995)
- Tang Li, "Susceptibility of two cultivars to various environmental stresses", 30, pp.405~410 (2003)
- Tepperman JM, "Transformed plants with elevated levels of chloroplastic SOD are not more resistant to superoxide toxicity", 14, pp.501~511 (1990)
- Van Camp W,, "Enhancement of oxidative stress tolerance in transgenic tobacco plants overproducing Fe-superoxide dismutase in chloroplasts", 112, pp.1703~1714 (1996)
- Youm JW, "Production of VP6 gene into potato plants by Agrobacterium-mediated transformation and analysis of VP6 expression in transgenic potatoes", 29, pp.93~98 (2002)
- Yun BW, "Differential resistance to methyl viologen in transgenic tobacco plants that express sweetpotato peroxidases", 156, pp.504~509 (2000)
