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Tulip (Tulipa gesneriana L.), is one of the world's leading cut flowers, also widely used in flower beds, patio and indoor green, loved by the people of all countries. Not long, but the tulips flowering to extend the flowering period to extend the viewing period and adjust the circulation plays an important role. How to prolong the life of the flowers has become experts in recent years, the main topic of the research by cloning tulip ACC oxidase (ACO) gene fragment will build into a hairpin structure with intron RNAi plant expression vector, in order to cause tulip ACC oxidase gene silencing by expression of this regulation tulip ethylene to control the aging of flowers, thus prolonging the flowering tulips, and establish RNAi technology platform. Mainly for the following three aspects: 1, the ACO Gene Cloning and plant RNAi expression vector construction based on tulip ACO genetic sequence, to design a pair of primers, the tulip varieties 'Los Angeles' petals cDNA as a template, expanding tulip added ACO gene fragment, which is connected to the pUCm-T vector analysis shows the results of sequencing the the ACO gene fragment cloned successfully. At both ends the the ACO gene fragment cut successively by reverse and forward respectively connected to the intermediate vector pBSin intron, constitute a the ACO gene with intron the hairpin (IHP structures); The hairpin structure was connected to the binary carrier pCAMBIA35S-1301, detected by restriction enzyme digestion and sequencing analysis to successfully build a plant expression vector containing the ACO gene issuing structure. 2, tulips Experimental Study on tissue culture the Tulip scales, stems, and Tori, petals, anthers and pistil explants, tissue culture research found that, tulips can direct organogenesis and indirect organogenesis two way to the formation of regenerated plants. Through research, screening a suitable medium hormone combination, succeeded in obtaining a tulip Regenerated. Primary culture stage, tulip stems, receptacle and pistil the best explants choice contamination rate below 8.6%, callus induction rate as high as 72% -100%. Tulip stems, and the receptacle and pistil endures Callus induction and proliferation of cultured medium was MS 0.5 mg · L-1 2,4-D, suitable for the induction of adventitious buds of these explants and callus differentiation medium was MS 0.2 mg · L-12 ,4-D 6.0 mg · L-1ZT. Rooting stage, the optimum medium was 1/2 MS 1.0 mg · L-1 NAA 2.0 g · L-1A.C. ACO gene fragment containing RNAi expression vector genetic transformation using the freeze-thaw method RNAi expression vector into Agrobacterium strain GV3101, tulip tissue culture and callus preliminary ACO gene fragment containing RNAi expression vector genetic transformation.
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