Establish Efficient Regeneration System Of Tissue Culture Of Alfalfa NFF2

Alfalfa is the world’s oldest cultivated and the most widely distributedperennial legume, which has played an important role in livestock production for along time. With the severely degraded grassland vegetation and pasture damage theecological environment, we can produce a large number of new varieties of alfalfafor livestock forage production to meet increasing needs. To solve this problem,there are fundamentally essential means of biotechnology-assisted breeding. Modernbiotechnology is one of the important ways to improve the alfalfa forage productionand the quality of genetic optimization, and the highly efficient and stableregeneration system is the precondition of using alfalfa biotechnology, alfalfaregeneration is highly affected by the genotype and environmental conditions, thealfalfa unstable high frequency regeneration is always the bottleneck of alfalfagenetic engineering development. In this article, currently the biotechnologyexperiment problems of alfalfa problems and future prospects are discussed, as wellas research alfalfa NFF2, in order to provide a reference for the study of geneticallyengineered of alfalfa regeneration mechanism and system aspects. Alfalfa NFF2iswidely grown and applications in Canada, but has not been established regenerationsystem for the effective organization, resulting in biotechnology researchapplications in alfalfa NFF2is restricted.Based on this, the test choose easily transfected Agrobacterium alfalfa NFF2asthe research object, using the three-part callus induction which derived from theNFF2sterile seedling cotyledons and hypocotyls with7days growth. The the leafand petiole30-50d after training, as well as part of the experimental fields takenfrom Dragon Lake experimental field, without pests leaves and petioles, theinfluence factors of Agrobacterium-mediated transformation system were studied theeffect of different mediums, hormones in different concentrations on callusinduction and the effect of different mediums on callus differentiation. Lay thefoundation for further alfalfa transgenic system building.Use the aseptic petioles with30-50days and petiole, which is the best callus induction explants. When the sterilization of explants in experimental fields with0.1%HgCl2with8min, which is the best way to experiment and leaf petioles fielddisinfection; explants were inoculated in the culture medium was first performed7ddark, you can add value and contribute to the differentiation of cells in the organwhen callus induction use improved SH2K additional1mg/L2,4-D and0.2mg/L KT,callus induction rate can reach91%, and shorten the cycle. Tests to select the bestmedium for callus subculture is BOi2Y; MSO for optimal medium callus;1/2MS forseedling medium.