Agrobacterium Tumefaciens-mediated Genetic Transformation Of Narcissus Tazetta L. Var. Chinensis Using Hpt As Select Marker And With LYCB Gene

Chinese narcissus (Narcissus tazetta var. Chinensis) belongs to Amaryllidaceae,which is a variety of polyanthus. Chinese narcissus, as a traditional flower in SpringFestival, is popular for its freshness, elegance and attractive appearance. Nevertheless,there are some problems in Chinese narcissus production, such as limited species andflower colours, degeneration of the breed and long breeding cycle. At the same time,Chinese narcissus is triploid, it is difficult to breed by the hybridization. Geneticengineering will be a new effective way for improvement of Chinese narcissus.In this study, Zhangzhou narcissus 'Jinzhanyintai' were used as the material tooptimize the genetic transformation system based on the hpt gene selection system.Expression vector of LYCB was constructed and transformed into Chinese narcissus.The main results were as follows:1. Establishment of genetic transformation system of Zhangzhou narcissus usinghpt gene as the selectable marker.Experiment results showed that the condition of pre-cultured for6d, infected inAgrobacterium suspension (OD600=0.5) for10min, co-cultured in medium addedwith100μm·L^-1AS for3d, delayed selection for5d and selection medium addedwith5mg·L^-1BA,0.3mg·L^-1NAA and10mg·L^-1Hyg was benefit to enhancing thetransformation efficiency and the positive rate was up to11.11%.2. Plant expression vector construction and transformation of LYCB gene.GUS gene of pCAMBIA1301was replaced by LYCB gene by Double Digests. Onthe basis of genetic transformation system established, LYCB gene was transformdinto Chinese narcissus and resistant plantlets were obtained.