The Selection Of Acceptor Strain For Tremella Fuciformis Heredity Transformation And Research On Fermentation Condition

The basidiomycete Tremella fuciformis Berk, or'white jelly mushroom'favored by Chinese people as a gourmet and medicinal mushroom. For the special history of life, it not only can be planted by people, but also can be fermented culture. It is an ideal model carrier for transformation. Before using it as the model carrier and a planting transgenic plant, we must do some research on the selection of strains, planting of yeast-like conidia, fermentation technology and the genetic ground. So, we conducted this reach from three aspects. First: The selection of acceptor strain for Tremella fuciformis heredity transformation; Second, the research on fermentation condition for the heredity transformation acceptor; Third the preliminary research on mating type of tremella fuciformis acceptor strain. The detailed results as follows:1 The selection of acceptor strain for Tremella fuciformis heredity transformationAccroding to the methords of morphological verification, molecular level, sensitivity degree of hygromycin, as well as biomass , we selected a strain which is sensitive to hygromycin and has the fastest speed from 19 strains of Tremella fuciformis. Basing on T0040, we planted it and achieved 3 pairs of sporidiums which could mate with each other And it would be the reliable material for the research on the genetic background of acceptor for dual gene transformation. To solve the problem that used the transgenetic conidium for planting experiment. We conducted the research on nutrition transformation condition.. We achieved two culture mediums which could induce the yeast-like conidium to the mycelium. Formula 10% bran, 10% extract of xiang huijun and saw dust Formula two maltose 10.0 g, glucose 10.0 g, agar 30.0 g,; second power of rice 500.0g, bran 500.0 g, bone dust 10.0 g, ammonium sulfate 1.0 g, calcium phosphate 10.0 g.2 The research on fermentation condition for the heredity transformation acceptorOptimization of cultivation medium is an important link to microorganism culture. Using the single factor test ,we analysed the influence of carbon source, nitrogen source, mineral element, vitamin, auxin. The result showed that they were significant influence to the biomass of conidium. According the experiment results, we selected the carbon source glucose, nitrogen source peptone, mineral element bitter salt, vitamin VB1 as the factors in the experiment. Utilizing response surface method to seek optimization, through this method, we can achieve the optimum component of the medial at a less number of experiments, and reduce experimental costs. In this paper, we used this method to achieve the high-density fermentation medium of the model strain of T0040:Glucose 33.91 g / L, peptone 8.29 g / L, magnesium sulfate 2.81 g / L, VB1 14.43 mg / L KH2PO4 0.5 g / L.3 The preliminary research on mating type of tremella fuciformis acceptor strainThe result of mating experiments between basidiospores showed that T1 and T26, T1 and T19, T4 and T13 could pair with each other. We conducted a preliminary research on the mating type of tremella fuciformis, using the degenerate PCR method to successfully clone the mitochondrial intermediate peptidase (MIP) gene. At the same time we also tried using degenerate PCR to clone the tremella fuciformis T0040 STE3-like pheromone receptor fragment, and predicted the struct and function by bioinformatics methods.All of these studies would pay the way for the research on genetic background the mating type factors of the tremella fuciformis.