The scientific and technological field of the project: agricultural bioengineering/traditional Chinese medicine resources. Medicinal fungi (mushrooms) are an important part of traditional Chinese medicine in the motherland. Modern biotechnology, biochemistry and pharmacological technologies have gradually revealed the main active ingredients in traditional medicinal fungi for preventing and treating diseases; among these active substances, there are many studies on polysaccharides, terpenoids, sterols, etc., and almost blank protein components. This project responds to the major national strategic needs in the modernization of traditional Chinese medicine and the field of national biotechnology. Taking Ganoderma lucidum as research objects, it uses modern biotechnology to explore its functional genes, and conducts systematic analysis, evaluation and basic research on industrial application of functional genes, and has achieved the following achievements: 1. Propose new ideas to use biotechnology methods to study the interrelationship between genes, proteins and pharmacological functions in Chinese herbal medicines; A technical platform for cloning, expression and functional verification of fungal immune regulatory proteins (FIPs) genes has been established. It was the first time that China Ganoderma lucidum (G. The fungal immunoregulatory protein genes FIP-gsi and FIP-gat in sinensis and G.atrum reveal the relationship between the structure and function of FIP-gsi and FIP-gat genes; Prokaryote expression technology was used to produce FIPs proteins, and a method for protein induction expression and purification was established. Through mouse spleen cell cytokine induction expression experiments, the immunoregulatory function of the protein was proved and its functional mechanism was analyzed. 2. Successfully used DNA shuffling technology to reorganize FIPs genes, laying the foundation for their in vitro production and synthesis and improvement of the biological activity of proteins. FIPs genes were divided into intra-generic groups and inter-generic groups based on their origin, and the two groups of genes were recombined at the molecular level using DNA shuffling technology to build a shuffled library. The shuffled library was screened using dot hybridization technology and specific antibodies. A total of 426 intra-generic recombinant strains and 412 inter-generic strains were screened. Mutants that met the expected reorganization were screened and verified, the protein was expressed and purified, and relevant biological activity experimental analysis and verification were carried out. It was found that the reorganization mutant could be expressed in bacteria and had good immunoregulatory activity. 3. Taking the recombinant rFIP-SN15 and Ganoderma lucidum rFIP-gat in the genus Ganoderma as examples, the anti-tumor effect and mechanism of action of this family of proteins were initially revealed. The cytotoxicity of rFIPs on different tumor cells was screened by MTT assay, and the direct inhibitory effect of rFIPs on glioma cells U-251 and breast cancer cells MDA-MB-231 was found. In further experiments on the inhibition of rFIPs on breast cancer cells MDA-MB-231, flow cytometry, gene chip technology, transcriptome sequencing technology, etc. were used to discover the effect of rFIPs on the proliferation and apoptosis of MDA-MB-231 cells, and initially revealed the possible way for rFIPs to induce apoptosis of MDA-MB-231 cells. 4. Regarding Cordyceps sinensis as ldquo; a unified microecosystem rdquo;, Illumina MiSeq high-throughput sequencing technology was used for the first time at home and abroad to analyze the dynamic changes of multi-bacterial coexistence and symbiosis of Cordyceps sinensis in different geographical locations during the growth and maturity process; The diversity of microorganisms in soil, Cordyceps sinensis, and bacterial membranes enclosing insect bodies was analyzed, revealing the composition and structure of the microbial population ldquo; Microecosystem rdquo;;; A new idea that the process of Cordyceps sinensis infecting insects is ldquo; multi-bacterial synergy rdquo; was proposed. The above-mentioned research published 46 SCI papers, which were cited by him 624 times in SCIE. He cited 259 times in the 8 representative works submitted by him in SCIE, including 1 highly cited paper on ESI; Trends Biotechnol, the top international biotechnology journal, has repeatedly cited the author's results. Published 1 national planning textbook "Introduction to Biotechnology", participated in the compilation of the international monograph "Edible and Medicinal Mushrooms: Technology and Application", and wrote the Cultivation of Ganoderma lucium; and participated in the compilation of 1 volume "Chinese Herbal Medicine Biotechnology". This research laid the foundation for the in vitro synthesis of protein active ingredients of Chinese herbal medicines, set a model for the application of biotechnology to study Chinese herbal medicines, and promoted the pace of biotechnology application in the modernization of traditional Chinese medicine in my country.