| Armillaria spp., belongs to Basidiomycotina, Agaricales, Tricholomataceae, Armillaria. It is distributed all over the continents and mainly in HiLongjiang, JiLin, LiaoNing, NeiMenggu in China. Armillaria spp. is a kind of famous medicinal fungus symbiotic with Chinese herb "Gastrodia elata Blume"(Tianma). Many experiments showed that the mycelia and fermented broth of Armillaria spp. had the function of cerebrum protection and nerve regulation, such as hypnogenesis, anti-convulsion and anti-anoxia. It also has bioactivities such as increasing the blood flow rate in cerebral and cardiovascular vessel and strengthening immune regulation. So great attentions were paid to study bioactivity and chemical components,develope series of preparation of Armillaria spp. and study the submerge fermentation technology since 1970s. And lots of achievements were obtained.There are abundant chemical components in Armillaria spp. Through the systemic study on the mycelia and fermented broth of Armillaria spp., many chemical compositions have been found, such as polysaccharide, sesquiterpenoids, purine and so on. The physical-chemical properties and bioactivities of those compounds were also studied. As wild TianMa resource is limited in our country. Although it had been cultivated successfully, it still needs Armillaria spp. as germination and nutrition donor. Tianma is hard to satisfy the need of healthcare and clinic use because of its selective techniques, high producing cost and price. It is an effective new way to use Armillaria spp. and its fermented products to replace "TianMa" in order to solve lack-supply and protect wild "TianMa" resource. With years of relentless efforts of scientists and medical enterprises, Armillaria spp. preparations as "NaoXinshu" Oral Liquid, Armillaria spp. pill and Armillaria spp. syrup work a comparatively good cure. Only "NaoXinshu" Oral Liquid one part was earned more than 600millions RMB annually in our country. The Armillaria spp. preparations, taking NaoXinshu Oral Liquid for example, just belongs to the Drug preparationâ…¢, because they used Armillaria spp. mycelia and fermented broth as main materials, which did not have clear effective part and active components. What's more, there are still several adverse factors: no unified strains of Armillaria spp. no regulated fermentation technology and control, low effective yield, no clear monitoring aims for products and preparation quality. Not only do all factors above put great impact on the producing, developing and preparation of Armillaria spp., but also restrain products advance.This paper was the most significant part of "Study on Armillaria spp. submerges fermentation technology and its functional food" which was a science and technology developing plan project of JiLin Province in 2001 conducted by the author. Its chief target was to screen strains with favorable growing characters and soporific function from wild Armillaria spp. in ChangBai Mountain through fermentation and animal trials. The target of this thesis is to clear effective pot with hypnogenesis function, and yield rate of it as chief index, in order to optimize the submerged fermentation technology to realized maximization of the yield rate. The research results as follows:1. 16 Armillaria spp. strains were separated from "TianMa" and the mycelia of Armillaria spp. collected from different areas in ChangBai Mountain, which were named Ar-02, Ar-03, Ar-04, Ar-07, Ar-08, Ar-09, Ar-11, Ar-13, Ar-16, Ar-17, Ar-20, Ar-21, Ar-25, Ar-26, Ar-29, Ar-31 after evaluation respectively. Taking the Armillaria spp. strain which was used to produce "NaoXinshu" Oral Liquid (At-00) as a control, many researches has been done to got information about sprouting time of mycelia and rhizomorph, morphological characters, the mycelia shape at liquid culture, the color and luster of the fermented broth, the fluorescence intensity, biomass of the products and so on. These ascertained the growing characters of the strains. And then 8 strains with favorable growing characters (Ar-02, Ar-03, Ar-07, Ar-08, Ar-09, Ar-11, Ar-13, Ar-31) were screened by the indexes, the strains' growing character and biomass.2. The optimal culture conditions were screened respectively for the 8 Armillaria spp. strains through liquid submerged fermentation study. On the base, the 8 Armillaria spp. strains were cultured enlarging, by using "NaoXinshu" Oral Liquid as a positive control and animal trail for soporific function. Favorable strain Ar-07 with definite soporific function and highest biomass was finally screened. Molecular biological identification was also done, and strain Ar-07 Was confirmed to be Armillaria ostoyae Romagn at last. The optimum culture conditions for Ar-07 was screened and determined through single factor experiment and orthogonal experiment, for example, optimal initial pH value, volume of culture, inoculating method, inoculating volume, cultural temperature, and time.3. After layers of separating and extracting, the extracting part with definite soporific function in Ar-07 fermented broth was ascertained through animal trials.4. Armillaria spp. liquid submerged fermentation technology was optimized through orthogonal experiment and kinetics study. The mycelia yield obtained by the optimized technique parameters was higher 18.30ï¼…than that of normal fermentation; the yield of soporific function extracting part was higher 21.64ï¼…. The optimum technology confirmed by this research was proved reasonable and reliable through analysis test and animal trial.Since collecting Armillaria spp. samples, separating the strains, screening favorable strains, ascertaining the extracting part and optimizing fermentation technology, the whole course was first systemically studied in this paper. Innovations as follows:1. Favourable strain Armillaria spp. Ar-07 was finally screened through lots of experiments for its better growing characters, high biomass, definite potency and steady activation. And this strain was confirmed to be Armillmraia ostoyae Romagn by molecular biological identification method. This provided steady and reliable strain for Armillaria spp. preparation production and exploration.2. The ascertain of Ar-07 extracting part by submerge fermentation and animal trials, provided reference to optimize fermentation technology and monitor the quality for fermented products, and laid foundation for the secondary exploration of Armillaria spp. preparations.3. The culture conditions and control parameter were optimized and expanding trials were done. The yield rate of mycelia raised 18.3ï¼…and the valid yield rate of the extracting part with soporific function in broth increased 21.64ï¼…. So reliable technology was prepared for the development of Armillaria spp. liquid submerged fermentation.4. The smashing and homogenizing inoculation method confirmed. It has proved that could raise the biomass and save the inoculation amount (save 40ï¼…). Obviously, this method is innovative. It can be practiced in the fungus liquid submerged fermentation after further perfecting.The innovative achievement obtained in this paper prepared the ground for the further study in Armillaria spp. fermented characters, chemical components analysis of fermented products, separating and extracting the effective part and active monomer and Armillaria spp. products applying. The research also provided scientific and proper technology and favorable Armillaria spp. strain for developing series of products. There was certain reference value to Armillaria spp. standard fermentation producing and technique monitoring. The results had great applying value and immediate significance in increasing the quality and stability of Armillaria spp. preparation. It could enhance the second development of"NaoXinshu" Oral Liquid and development of the Third Generation Functional Food, and improve the industrial secondary exploration of edible and pharmaceutical mushroom.
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