Establishment And Optimization Of Genetic Transformation System On LycB Transgenic Potato

Potato(Solanum tuberosum.L)is one of the most important crops in the world.The quality improvement of potato get extensive attention all over the world.With the development of molecular biology,genetic engineering has become an important method to improve the quality of potato.Microtuber induction system was established for three kinds of potato with different maturity and induction rates reached to 100%.The stem regeneration rates were different from three cultivated varieties.The callus induction rates of Atlantic was up to 91.2%,Zaodabai and Dongnong 303 induction rates were 84.3%,71.4%.The microtuber regeneration rates of Atlantic and Zaodabai were 100%,Dongnong303 regeneration rate was 92.3%.Meanwhile,The optimized genetic transformation methods for three kinds of potato were establised.Lycopene ?-cyclase(LycB)catalyze the formation of two ?-rings.The lycopene?-cyclase gene from Lycium chinense was transformed to patato in this research.The target gene was transducted into explants via Agrobacterium-mediated transformation method.The stem conversion rates of Atlantic,Zaodabai and Dongnong 303 were28.1%,35.6%,36.2% separately.The microtuber conversion rate were 17.4%,45.6%,43.1%.Therefore,the microtubers are suitable explants compared with stems in Zaodabai and Dongnong 303,but the situation is opposite in Atlantic.HPLC was used to detect total carotenoids and ?-carotene in transgenic potato leaves of Zaodabai.The content of total carotenoid and ?-carotene was higher than the control.Total carotenoids content and ?-carotene of the LycB-L1 increased 13.4% and18.7% separately.Moreover,other ?-branch carotenoids increased in the transgenic lines,including zeaxanthin,antheraxanthin and violaxanthin.Vitrification phenomenon leads to physiological disorder of potato seedlings,meanwhile this phenomenon impact the field planting seriously.The concentration of plant growth regulators,ions and agar are important elements on the generation and reversion of vitrification of potato.The result demonstrated that hormones play an important role in the occurrence and recovery of vitrified process.The cause was explored through the physiological and biochemical experiments of the normal seedlings,the vitrified seedlings and the restored seedlings,The results indicated thatthe vitrification phenomenon is correlate with oxidative stress.