Study on Genetic Transformation of LfMADS1 Gene into Lilium ** formosanum' Raizen No.1'

Abstract

Abstract: In order to create pollen-free double flowers, the in vitro bulblet scales of Lilium × formosanum ‘Raizen No. 1’were taken as explants, the effects of Kan and Carb on regeneration from bulblet scales were studied at first. Then a orthogonal test were conducted to screening such factors as pre-culture time, bacterial concentration, infection time and co-culture time, and in this way the transformation system was optimized. Through this system, LfMADS1 gene was transferred into bulblet scales mediated by Agrobacterium, and the transgenic plants were molecularly identified. The results indicated that Kan 100 mg/L was optimal to screen adventitious buds of Kan-resistant；Carb 500 mg/L was able to control growth of Agrobacterium. There are some key points include pre-culture for 1d, immersion of explants into bacterial suspension(OD 600 0.8)for 10 min, and co-cultivate for 5 d in the optimized transformation system. Four transgenic lines were obtained through this system identified by PCR, in which the exogenous gene had successfully integrated into genome of one line confirmed by PCR-Southern. These transformed lilies can be a new germplasm for the breeding of pollen-free or double flower lilies.

Key words: Bulblet scales, Agrobacterium-mediated transformation, Kanamycin, Carbenicillin, Orthogonal test

Li Yunhua, Liu Qing, Liu Qinglin. Study on Genetic Transformation of LfMADS1 Gene into Lilium ** formosanum' Raizen No.1'[J]. Biotechnology Bulletin, 2014, 0(3): 86-93.

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