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Really cornucopiae, also known as white jade, sea fresh mushroom, is a nutritious edible Chief medicinal rare edible fungus, is also popular in Japan in recent years, favored by consumers of edible fungi treasures. Morphological appearance of the mushrooms, crisp texture, taste delicious, and has now become one of the important varieties of the national mushroom factory production. True cornucopiae cultivation cycle is long (120-150d), cultivated under harsh conditions, poor resistance, uneven product quality problems, to some extent limit the development of large-scale factory production. Laccase degradation of lignin, such as the role of oxidative degradation of phenolic compounds, inhibit bacteria, improve fruiting quality. Higher laccase activity the versicolor and no paint the activity Hypsizigus mushroom as experimental material, enzymology and protoplast fusion technology combining versicolor protoplasts by high temperature inactivation and true cornucopiae protoplasm fusion, resulting in high-paint activity, cornucopiae fine varieties. Mushroom species from nine different varieties Hypsizigus mycelial growth rate, mycelium dense, growing, fast growth, mycelium dense, growing strong screened white jade plates as the original strain; from 9 different species of Ganoderma species by comparing the strains in the RB bright blue-PDA discoloration circles and discoloration speed, filtering out the the discolored ring Japanese discoloration speed strong versicolor as the original strain. And then the white jade screened and versicolor as a starting strain on them protoplast preparation, regeneration experiments, the following conclusions: of white jade protoplast preparation and regeneration of the best conditions: 2% wall enzyme solution for 3h, RM2 for regeneration culture group; versicolor protoplast formation and regeneration of the best conditions: 3% of the dissolved wall enzymatic hydrolysis 3h, RM2 for regeneration medium. Subsequently, the prepared versicolor protoplasts with 55. C water bath for 45min inactivated protoplasts with white jade 1:1 mixture at 32 ℃ PEG (4000) as the fusion agent deal with 25min. Fusion strains grown on regeneration medium using the RB-PDA flat panel color technology to screen out strong the discolored ring Japanese discoloration speed for strain RH01-RH04. Then compare them with the original strain of white jade and versicolor laccase activity differences, rescreening high laccase fusion strains RH01. Aggregate strain RH01-RH04 the antagonistic experiments, the fusion strains RH01-RH04 incompatibility that exists between them with the original strain between white and versicolor has obvious antagonistic phenomenon, preliminary identification belonging to the integration of the new strain . RAPD technique and ISSR the technology further analysis, based on the amplification of DNA profiles, suggesting that the fusion strains RH01 ~ RH04 kinship nearly white, the same versicolor genetic differences. Stability after the integration of the strain RH01 ~ RH04 continuous adapter 5 generations RH01 best stability. The experiments show that the fusion of the growth characteristics of the strain RH01: RH01 mycelial morphology between starting observation the 16x40 times the ordinary microscope, the hyphae are also differences between the strains of white jade and versicolor;; fungal resistance to Comparison, RH0l the antagonistic circle, indicating it's on fungal resistance to the strongest. Biological characteristics of the test results: fusion strains RH01 medium optimization results showed vigorous growth in the PDA carrot medium and strong vitality, dense, white mycelium; the fusion strains RH01 optimal growth temperature of 26. C; optimum pH value of 7.0; RH01 seed bottle mycelial growth rate of 0.31cm / d; fruiting experiments show that the production cycle is shorter than white jade, yield an average of 2.62g / bottle.
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