Establishment of Transgenic Nicotiana benthamiana by using S-Adenosylmethionine Synthetase (SAMS) Gene Isolated from Miscanthus sinensis
Background : Temperature is major factor for growth plant. Recently, because of global warming, abnormal temperature included drought, deluge, sudden temperature change and heavy snow damaged crops in the world. In Korea, crops have been sensitive to low temperature on early growth stage, e.g. fruit tree and ginseng, were damaged owing to sudden heavy snow and cold on Spring. Therefore, recently interest in cold resistance crops were increased in demand rapidly. This study was performed to establish transgenic Nicotiana benthamiana by transforming cold resistant gene related to cold tolerance S-adenosylmethionine synthetase (SAMS) isolated from Miscanthus sinensis. Methods and Results : Total RNA was extracted from leaves of M. sinensis using Trizol assay and isolated MsSAMS. Isolated MsSAMS was insert into SacⅠ- XbaⅠ sites of pMBP1 vector. The vector was transformed to Agrobacterium tumefaciens strain LBA4404 by DH5α. A. tumefaciens with binary plasmid were selected at YEP medium supplemented with kanamycin. Cut leaves of tobacco were co-cultured with selected A. tumefaciens. Co-cultured leaves was grown on regeneration medium for a month at dark condition, and transferred to at light condition. Regeneration shoot from callus were excised and transferred to root-induction medium. Approximately, 58% of leaves explant produced callus. Nearly, 30% of callus had shoot and approximately, 94% of shoots were rooted in root-induction medium. Conclusion : We established an efficient transformation system of N. benthamiana transformed by using MsSAMS gene related to cold tolerance isolated from M. sinensis. We may use the produced transgenic plants to prevent damages carried by cold.