055 Transformation of Chinese Cabbage Glutathione Reductase (GR) Gene into Lettuce (Lactuca sativa L.) with Microparticle Bombardment

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  • Depts. of 1Horticulture2 Animal Science and Biotechnology, Kyungpook National Univ., Taegu 702-701, Korea; 3Dept. of Horticulture, Sangju National Univ., Sangju 742-711, Korea

Lettuce (Lactuca sativa L.) were transformed using microparticle bombardment with two different genes, alpha-glucuronidase (GUS) gene and Chinese cabbage Glutathione Reductase (GR) gene. The adventitious shoots of cotyledonary explant from 4-day-old seedlings were formed (46.7%) in MS basal media supplemented with 5.0 μm IAA and 1.0 μm 2ip. When 1100 psi helium pressure, 9 target distance, and coating with tungsten 10 microparticles were used and explants were treated with osmoticum-conditioning medium (0.6M sorbitol/mannitol), 4 h prior to and 16 h after bombardment, it was identified by GUS assay that these conditions were the most efficient for transformation of foreign genes into cotyledon tissue of lettuce with particle bombardment. PCR confirmed that the band observed in the transgenic plants were originated from T-DNA tranfer with strong hybridization. The genomic Southern analysis showed that the 1.5-kbp fragment was hybridized with radiolabeled 1.5-kbp GR probe. To know whether the expression of the GR gene can be stably maintained in the next generation, when T2 selfing seeds that were obtained from the transformed mother plants were sowed on MS medium supplemented with 200 μm kanamycin, 70% of seedlings were revealed resistance to kanamycin.

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