Coralline tremella strain as well as culture method, identification primer and application thereof

Abstract

The invention provides a coralline tremella fuciformis strain as well as a culture method, an identification primer and application thereof. The coralline tremella fuciformis strain has the preservation number of CGMCC (China General Microbiological Culture Collection Center) NO.41327. A new coralline tremella fuciformis strain is separated and purified from tremella fuciformis sporocarp, the liquid fermentation polysaccharide content of the strain is more than 1.2 times of that of a conventional tremella fuciformis cultivation strain, and after domestication and cultivation, the sporocarp is coralline, small in ear base and good in ear piece elasticity, has a unique genetic background, and is suitable for industrial production. And a solid foundation is laid for breeding a new variety of tremella fuciformis with high quality, strong adaptability and obvious characters.

Description

Technical Field

[0001] The invention is used in the field of biotechnology and mainly relates to a coral-shaped Tremella fuciformis strain and a culture method thereof, an identification primer and an application thereof. Background Art

[0002] Tremella fuciformis Berk., also known as white fungus, belongs to the genus Tremella, class Trimellomycetes, order Tremella, order Tremella, phylum Basidiomycota, kingdom Fungi. Tremella is a traditional edible and medicinal fungus in my country. It is not only a well-known medicine in my country's medical treasure house, but also a tonic and banquet delicacy. It has developed into one of the most popular cultivated edible fungi varieties in my country.

[0003] The existing Tremella fuciformis cultivars are single, mainly the white Tremella fuciformis "Tr01" and the yellow Tremella fuciformis "Tr21" approved by Fujian Province in 2007. The above varieties have little difference in morphological characteristics during the production process, and the market segmentation is limited, resulting in a continuous decline in production efficiency. Breeding high-quality, highly adaptable, and distinctive varieties is of great significance to enriching the existing germplasm resources of Tremella fuciformis and promoting industrial development. Summary of the invention

[0004] In order to solve the above problems, the present invention discloses a coral-shaped Tremella fuciformis strain and a culture method, identification primers and uses thereof.

[0005] In order to achieve the above objectives, the present invention is implemented through the following technical solutions:

[0006] A coral-shaped tremella strain, the preservation number of the coral-shaped tremella strain is CGMCC NO.41327, and the strain is preserved in the China General Microbiological Culture Collection Center, the address of which is Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

[0007] A use of a coral-like Tremella strain, wherein the coral-like Tremella strain is as described in claim 1, and the coral-like Tremella strain is used for preparing Tremella polysaccharide.

[0008] Further improvement, the method for preparing Tremella polysaccharide is as follows:

[0009] Step 1, activating the Tremella binuclear yeast-like spores of the coral-like Tremella strain on a PDA solid culture medium, picking the activated strains with an inoculation loop, inoculating them into a PDA liquid culture medium, culturing them in a constant temperature shaker at 25° C. and 160 rpm, and transferring the seed liquid obtained after fermentation and cultivation for 7 days, transferring it to a PDA liquid fermentation medium at an inoculation amount of 1%, and culturing it in a shaker, and obtaining Tremella strain fermentation liquid on the 9th to 12th day of fermentation;

[0010] Step 2: Add three times the volume of 95% ethanol to the fermented broth of Tremella fuciformis, stir and shake thoroughly, and then place it in an environment of 4°C for alcohol precipitation for 1 day. After alcohol precipitation, centrifuge at 4000rpm for 15min, discard the supernatant, add ultrapure water and place it in an ultrasonic machine to fully dissolve the precipitate, ventilate and place it until the residual ethanol is completely volatilized, and obtain a Tremella fuciformis crude polysaccharide aqueous solution, place the Tremella fuciformis crude polysaccharide aqueous solution in a -35°C refrigerator for 24h to completely freeze, and then place it in a freeze dryer for freeze drying for 2 days to obtain Tremella fuciformis polysaccharide.

[0011] As a further improvement, the coral-like Tremella strain is used to produce binuclear yeast-like spores, mycelium or fruiting bodies.

[0012] A further improvement is that the coral-like Tremella strain is used as breeding material.

[0013] A method for culturing a coral-shaped Tremella fuciformis strain, wherein the coral-shaped Tremella fuciformis strain is as claimed in claim 1, and specifically comprises the following steps:

[0014] Step 1, preparing a mother culture medium: the formula of the mother culture medium is: 68 parts by weight of sawdust, 30 parts by weight of wheat bran, 1 part by weight of white sugar, and 1 part by weight of gypsum; sterilizing the prepared mother culture medium, inoculating the incense ash fungus in the middle of the plate, and inoculating the coral-like Tremella fuciformis strain 1 to 2 cm away from the incense ash fungus seed block. After 15 days of inoculation, it is observed that the hyphae cover the plate and form a "white hair ball" around the Tremella fuciformis inoculation block;

[0015] Step 2, preparing tremella cultivation material, the formula of tremella cultivation material is: 81 parts by weight of cottonseed hulls, 13 parts by weight of bran, 5 parts by weight of corn flour, and 1 part by weight of gypsum; after sterilizing the prepared tremella cultivation material, inoculating the "white hair ball" part into the center of a glass bottle, after growing for 10 days, the mycelium of the glass bottle covers the bottle mouth; after growing for 20 days, the mycelium of the glass bottle covers the bottle mouth, and an ear base begins to appear; after growing for 40 days, a tremella fruiting body is obtained.

[0016] The invention discloses an identification primer for a coral-shaped tremella strain, wherein the identification primers for the coral-shaped tremella strain are I1, I2, I4, I7, I13, I16 and I17; wherein the nucleotide sequence of I1 is: 5'-CTCCTCCTCCTCSC-3'; the nucleotide sequence of I2 is: 5'-BDBACAACAACAACAACA-3'; the nucleotide sequence of I4 is: 5CACCACCACCACSC-3'; the nucleotide sequence of I7 is: 5'-VDHTCGTCGTCGTCGTCG-3'; the nucleotide sequence of I13 is: 5'-DHBCGACGACGACGACGA-3'; the nucleotide sequence of I16 is: 5'-AGAGAGAGAGAGAGAGT-3'; the nucleotide sequence of I16 is: 5'-TCTCTCTCTCTCTCTCTGC-3';

[0017] Among them, B=G, C, T; D=A, G, T; H=A, C, T; V=A, G, C; S=G, C.

[0018] Compared with the prior art, the present invention has the following advantages:

[0019] The invention separates and purifies a new coral-shaped tremella strain from tremella fruiting bodies. The liquid fermentation polysaccharide content of the strain is more than 1.2 times that of a conventional tremella cultivation strain. After domestication and cultivation, the fruiting body is coral-shaped, the ear base is small, the ear pieces have good elasticity, and the strain has a unique genetic background, thus laying a solid foundation for breeding high-quality, highly adaptable, and obviously trait new tremella varieties. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 It is the coral-like Tremella Tr-1001 binuclear yeast-like spore;

[0021] Figure 2 It is the mycelium of coral-shaped Tremella fuciformis Tr-1001;

[0022] Figure 3 It is the UPGMA cluster diagram based on ISSR molecular markers;

[0023] Figure 4 is the glucose standard curve;

[0024] Figure 5 It is the coral-shaped Tremella Tr-1001 flat-plate mother strain;

[0025] Figure 6 It is the coral-shaped fruiting body of Tremella fuciformis Tr-1001.

[0026] Specific implementation method

[0027] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0028] It should be understood that the terms described in the present invention are only for describing a particular embodiment and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. The intermediate value in any stated value or stated range, and each smaller range between any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0029] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0030] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to those skilled in the art. The present invention description and examples are exemplary only.

[0031] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0032] Example 1 Isolation, purification and identification of strains

[0033] 1. Isolation and purification of strains

[0034] (1) Fruiting body collection: collected from dead wood in Gongzhuji Forest Park, Pubei County, Qinzhou City, Guangxi Zhuang Autonomous Region in June 2021. The strain was deposited in the General Microbiological Center of China Microbiological Culture Collection on June 3, 2024, with the deposit number CGMCC NO.41327.

[0035] (2) Tissue separation: Rinse the fruiting body three times with 75% alcohol, then rinse it three times with sterile water, and wipe dry. Take the base of the fruiting body and place it in a 30°C drying oven to dry for 10 days. Take the dried Tremella fuciformis base, place it in 75% alcohol for 1 minute for surface disinfection, then rinse it three times with sterile water, and use sterile filter paper to absorb the surface moisture. Use a sterile scalpel to cut the internal tissue of the Tremella fuciformis base, place it on a PDA culture dish and culture it at 25°C for 5 to 7 days to obtain Tremella fuciformis binucleate yeast-like spores ( Figure 1 ), streak purification three times with a sterile inoculating loop, and store the strain for future use.

[0036] At the same time, the ash fungus (Annulohypoxylon stygium) corresponding to the Tremella fuciformis strain was isolated. After the surface of the ear wood was disinfected with 75% disinfectant alcohol, the surface part was scraped off with a sterile scalpel, and a rice-sized wood block of the black part below the surface was placed on a PDA culture medium dish for cultivation at 25°C for 3 to 5 days to obtain the ash fungus. The purification was repeated 3 times, and the strain was preserved for later use.

[0037] 2. Inducing Tremella mycelium

[0038] Preparation of the extract of the ash fungus: accurately weigh 200 g of the sawdust culture medium containing the ash fungus (waste Tremella fuciformis sticks), add 1000 ml of water, boil for 20 minutes, filter through 8 layers of non-woven fabric, and add ddH2O to make up to 1000 mL.

[0039] Tremella mycelium germination promoting medium (1L): 10g maltose, 10g sucrose, 2g peptone, 1.5g dipotassium hydrogen phosphate, 1g ammonium sulfate, 0.5g manganese sulfate, 200ml of ash mushroom extract, 20g agar powder, dilute to 1000ml with ddH2O, natural pH, and autoclave at 121℃ for 25min.

[0040] The purified Tremella binuclear yeast-like spores were inoculated into Tremella mycelium germination promotion medium and cultured at 24°C. Tremella mycelium germination could be observed after 10 days, and Tremella mycelium with a diameter of about 3 cm could be obtained after 30 days. Figure 2 ). Take the tip of Tremella fuciformis hyphae, repeat purification 3 times on Tremella fuciformis hyphae germination promotion medium, and preserve the strain for later use.

[0041] 3. Strain identification

[0042] The genomic DNA of Tremella fuciformis binuclear yeast-like spores, Tremella fuciformis mycelium, and Rhizopus fuciformis mycelium were extracted using the Kangwei Century new plant genomic DNA extraction kit. The ITS gene of the strain was amplified by PCR and sent to Shanghai Sangon Biotechnology Co., Ltd. for sequencing analysis. The primers used to amplify the ITS gene are as follows:

[0043] The ITS sequence primers are: ITS4 (5'-TCCTCCGCTTATTGATATGC-3'), ITS5 (5'-GGAAGTAAAAGTCGTAACAAGG-3').

[0044] Amplification reaction system: PCR StarMix 12.5μL, ITS4 / ITS5 1μL each, DNA template 1μL, ddH2O supplemented to 25μL.

[0045] The reaction program was: 95°C for 4 min; 95°C for 30 s, 55°C for 50 s, 72°C for 1 min, 35 cycles; 72°C for 10 min.

[0046] By using the BLAST function of NCBI, sequence alignment was performed in the GeneBank database and it was found that the Tremella fuciformis binuclear yeast-like spores and Tremella fuciformis mycelium obtained by the present invention had the highest homology with Tremella fuciformis, which was 98.94%. Thus, the strain was determined to be Tremella fuciformis Berk. and named Tr-1001. The mycelium of Annulohypoxylon stygium obtained by the present invention had the highest homology with Annulohypoxylon stygium, which was 99.88%. Thus, the strain was determined to be Annulohypoxylon stygium and named An-1001.

[0047] Example 2 Genetic diversity analysis of coral-shaped Tremella fuciformis strain Tr-1001

[0048] Strain collection: 15 cultivated and wild Tremella germplasm resources were collected for testing in Fujian, Guangxi, Henan and Anhui. Tremella strains A, B, C, E, G, H, I, and J are factory-cultivated strains from Guangxi, Tremella strains C and F are factory-cultivated strains from Fujian, K is an agriculturally cultivated strain from Henan, L is a wild strain from Guangxi, M is a wild strain from Anhui, and N and O are wild strains from Fujian. See Table 1 for details.

[0049] Table 1 Information of tested strains

[0050]

[0051]

[0052] Extraction of mycelium genomic DNA: Select about 20 mg of mature, clean, and uncontaminated mycelium for the extraction of Tremella fuciformis genomic DNA. Use a sterilized bamboo stick to scrape off the surface layer and place it in a 1.5 ml centrifuge tube. Number it for later use. Put it in liquid nitrogen and take it out for rapid grinding. Extract mycelium genomic DNA according to the instructions of the new plant genomic DNA extraction kit, with 3 replicates for each strain. . Use 1.2% agarose gel electrophoresis to detect DNA purity and concentration, and store it in a -20℃ refrigerator.

[0053] ISSR polymorphism detection: 20 ISSR primers (Table 2) were synthesized by Shenggong Bioengineering (Shanghai) Service Co., Ltd. PCR reaction system (25uL): ExTaq Mix (2×) 12.5uL, primer 1uL, 50ng DNA template, and filled with ddH2O. PCR amplification program: 94℃ pre-denaturation 4min (94℃ denaturation 1min, 36-64℃ renaturation 1min, 72℃ extension 3min) × 35, 72℃ fill 7min. Finally, store at 4℃. Take 5μL of the amplified product, perform 1% agarose gel electrophoresis, 50V constant voltage for 1.5h, observe and photograph by UV gel imaging system. The obtained DNA bands were counted, and the amplified bands of each sample were recorded according to whether there were any or not. All strains without bands were recorded as 0, and clear bands were recorded as 1. A "0-1" matrix table was constructed and entered into Excel software. The Jaccard's genetic similarity coefficients among the Tremella fuciformis strains were calculated using NTSYS-pc2.02 software, and the cluster diagram of the 15 Tremella fuciformis strains was constructed using the unweighted pair group method arithmetic averages (UPGMA) method. Figure 3 ).

[0054] Table 2 ISSR primers and sequences

[0055]

[0056]

[0057] Note: B=G, C, T; D=A, G, T; H=A, C, T; V=A, G, C; S=G, C; Y=C, T.

[0058] Seven primers with rich polymorphism, clear bands and good repeatability were screened out from 20 ISSR primers, namely I1, I2, I4, I7, I13, I16 and I17. ISSR results showed that the genetic similarity coefficient of 15 Tremella strains was between 0.45 and 0.94, and when the similarity coefficient was 0.63, the 15 tested strains could be clustered into 3 categories. Among them, the three wild strains TR-22501, TR-22601 from Fujian and TR-22901 from Anhui were clustered into Class I; the factory-based substitute cultivation strains TR-2307B, TR-LZ, TR-2302, TR-2307S, TR-2202, TR-BY, TR-XY, TR-DY from Guangxi, the factory-based substitute cultivation strains TR-FJ, TR-HY from Fujian and the agricultural cultivation strain TR-HN from Henan were clustered into Class II; the wild strain TR-1001 from Guangxi was clustered into Class III alone. It can be seen that the genetic differences between different Tremella strains are large, the genetic background is rich, and the Tremella strain TR-1001 has a large genetic difference from all the collected strains and has a unique genetic background.

[0059] Example 3 Analysis of polysaccharide content in liquid fermentation of Tremella fuciformis strain Tr-1001

[0060] The polysaccharide content of liquid fermentation of coral-shaped Tremella fuciformis strain Tr-1001 was determined and compared with that of liquid fermentation of conventional Tremella fuciformis cultivation strain (TR-21) under the same conditions.

[0061] (1) Preparation of Tremella liquid fermentation polysaccharide samples: Activate Tremella binuclear yeast-like spores on PDA solid culture medium, pick the activated strains with an inoculation loop, inoculate them into PDA liquid culture medium, and culture them in a constant temperature shaker at 25°C and 160rpm. After 7 days of fermentation, the seed liquid can be transferred. Use a pipette to accurately measure the seed liquid according to the inoculation amount of 1% and transfer it to PDA liquid fermentation medium, culture it in a shaker, and take samples on the 9th and 12th days of fermentation to determine the polysaccharide content.

[0062] (2) Extraction of crude polysaccharides from Tremella liquid fermentation: Take 40 mL of Tremella fermentation liquid, add three times the volume of 95% ethanol, shake and stir thoroughly to allow the ethanol and fermentation liquid to fully contact, and place in an environment of 4°C for alcohol precipitation for 1 day. After alcohol precipitation, centrifuge at 4000rpm for 15min, discard the supernatant, add an appropriate amount of ultrapure water and place in an ultrasonic machine to fully dissolve the precipitate, place in a fume hood until the residual ethanol is completely evaporated, and obtain a crude Tremella polysaccharide aqueous solution. Place in a -35°C refrigerator to freeze completely for 24h, then place in a freeze dryer for lyophilization for 2 days to obtain a crude Tremella polysaccharide freeze-dried powder.

[0063] (3) Detection of crude polysaccharide content in Tremella fuciformis liquid fermentation: first take an appropriate amount of glucose standard, set the oven temperature to 60°C for drying, accurately weigh 10 mg of glucose dried to constant weight, add a small amount of ultrapure water to dissolve in a small beaker, and then pour into a 100-mL volumetric flask. Rinse the beaker with a small amount of ultrapure water and pour into the volumetric flask, repeat two to three times, and finally add ultrapure water to make up to 100 mL to obtain a 100 μg / mL glucose standard solution. Mark the solution and set aside.

[0064] Add glucose solution to the test tubes as shown in Table 3. Use the phenol-sulfuric acid method to add 1 mL of 5% phenol solution to 6 test tubes, shake well while adding, then add 5 mL of concentrated sulfuric acid against the wall of the test tube and shake well. Try to add concentrated sulfuric acid within 1 minute after adding phenol to reduce reaction error. Place in a 40°C water bath for 30 minutes, take out and cool to room temperature, and measure the absorbance at a wavelength of 490 nm using an ultraviolet spectrophotometer. Draw a glucose standard curve with glucose concentration as the horizontal axis and absorbance as the vertical axis ( Figure 4 ).

[0065] Table 3 Glucose standard solution

[0066]

[0067] Weigh about 0.020g of Tremella fuciformis crude polysaccharide freeze-dried powder, add a small amount of ultrapure water, ultrasonically treat at 35°C for 20min to completely dissolve the crude polysaccharide, then dilute to 25mL and shake well, take 1mL and dilute 10 times, use the phenol-sulfuric acid method, take 1mL of polysaccharide dilution and add it to a test tube, add 1mL of 5% phenol and shake well, then add 5mL of concentrated sulfuric acid and shake evenly, after placing in a 40°C water bath for 30min, take out and let cool to room temperature, and measure the absorbance at 490nm with a UV spectrophotometer.

[0068] Calculation of polysaccharide content:

[0069]

[0070] M=W%×m1×1000

[0071] In the formula

[0072] W%: polysaccharide content;

[0073] m1: total weight of polysaccharide freeze-dried sample g;

[0074] m2: weight of freeze-dried Tremella fuciformis polysaccharide sample weighed during the determination;

[0075] m3: The weight of polysaccharide in μg obtained by checking the glucose standard curve in the Tremella fuciformis crude polysaccharide sample weighed during the determination;

[0076] M: total content of Tremella fuciformis crude polysaccharide in the sample, mg;

[0077] v1: volume of Tremella fuciformis crude polysaccharide solution mL;

[0078] v2: volume of sample solution during measurement, mL;

[0079] 10: Dilution factor.

[0080] As can be seen from Table 4, the polysaccharide content of the liquid fermentation of Tr-1001 is significantly higher than that of the liquid fermentation of Tr 21. The polysaccharide content of Tr-1001 is 1.29 times that of Tr 21 at 9 days of fermentation, and the polysaccharide content of Tr-1001 is 1.26 times that of Tr 21 at 12 days of fermentation.

[0081] Table 4 Tremella liquid fermentation polysaccharide content

[0082]

[0083] Example 4 Cultivation of coral-shaped Tremella fuciformis strain Tr-1001:

[0084] The mother culture medium of coral-shaped Tremella fuciformis strain Tr-1001 was prepared. The formula of the mother culture medium was: 68 parts of sawdust, 30 parts of wheat bran, 1 part of white sugar, and 1 part of gypsum. The prepared mother culture medium was placed in a 9 cm glass plate, sterilized at 121°C for 1 hour, and inoculated with incense ash fungus in the middle of the plate after cooling. Tremella fuciformis was inoculated 1 to 2 cm away from the incense ash fungus seed block. Three pieces of Tremella fuciformis were inoculated on each Tremella plate. After 15 days of inoculation, the mycelium was observed to cover the plate, forming a "white hair ball" ( Figure 5 ).

[0085] The formula of Tremella cultivation material is: 81 parts of cottonseed hulls, 13 parts of bran, 5 parts of corn flour, and 1 part of gypsum. Put the prepared Tremella cultivation material into a glass bottle, sterilize it at 121℃ for 2 hours, and inoculate the Tremella flat-plate mother culture "white hair ball" part into the center of the glass bottle after cooling. After 10 days of growth, the mycelium of the glass bottle will fill the bottle mouth; after 20 days of growth, the mycelium of the glass bottle will fill the bottle mouth and ear bases will begin to appear; after 40 days of growth, the Tremella fruiting body will grow ( Figure 6 ), the fruiting body is coral-shaped, the ear base is small, and the ear pieces are elastic.

[0086] The above is only a specific guiding implementation mode of the present invention, but the design concept of the present invention is not limited to this. Any non-substantial changes to the present invention using this concept shall be deemed as an infringement of the protection scope of the present invention.

Claims

1. A coral-shaped Tremella fuciformis strain, characterized in that The preservation number of the coral-shaped Tremella fuciformis strain is CGMCCNO.41327.

2. A use of a coral-shaped Tremella fuciformis strain, characterized in that: The coral-like Tremella strain is as described in claim 1, and the coral-like Tremella strain is used for preparing Tremella polysaccharide.

3. The use of the coral-shaped Tremella fuciformis strain as claimed in claim 2, characterized in that, The method for preparing Tremella polysaccharide is as follows: Step 1, activating the Tremella binuclear yeast-like spores of the coral-like Tremella strain on a PDA solid culture medium, picking the activated strains with an inoculation loop, inoculating them into a PDA liquid culture medium, culturing them in a constant temperature shaker at 25° C. and 160 rpm, and transferring the seed liquid obtained after fermentation and cultivation for 7 days, transferring it to a PDA liquid fermentation medium at an inoculation amount of 1%, and culturing it in a shaker, and obtaining Tremella strain fermentation liquid on the 9th to 12th day of fermentation; Step 2: Add three times the volume of 95% ethanol to the fermented broth of Tremella fuciformis, stir and shake thoroughly, and then place it in an environment of 4°C for alcohol precipitation for 1 day. After alcohol precipitation, centrifuge at 4000rpm for 15min, discard the supernatant, add ultrapure water and place it in an ultrasonic machine to fully dissolve the precipitate, ventilate and place it until the residual ethanol is completely volatilized, and obtain a Tremella fuciformis crude polysaccharide aqueous solution, place the Tremella fuciformis crude polysaccharide aqueous solution in a -35°C refrigerator for 24h to completely freeze, and then place it in a freeze dryer for freeze drying for 2 days to obtain Tremella fuciformis polysaccharide.

4. The use of the coral-shaped Tremella fuciformis strain as claimed in claim 2, characterized in that The coral-like Tremella fuciformis strain is used for preparing binuclear yeast-like spores, mycelium or fruiting body.

5. The use of the coral-shaped Tremella fuciformis strain as claimed in claim 2, characterized in that, The coral-like Tremella strain is used as breeding material.

6. A method for culturing a coral-shaped Tremella fuciformis strain, characterized in that: The coral-like Tremella fuciformis strain as claimed in claim 1 specifically comprises the following steps: Step 1, preparing a mother culture medium: the formula of the mother culture medium is: 68 parts by weight of sawdust, 30 parts by weight of wheat bran, 1 part by weight of white sugar, and 1 part by weight of gypsum; sterilizing the prepared mother culture medium, inoculating the incense ash fungus in the middle of the plate, and inoculating the coral-like Tremella fuciformis strain 1 to 2 cm away from the incense ash fungus seed block. After 15 days of inoculation, the mycelium is observed to cover the plate and form a "white hair ball" around the Tremella fuciformis inoculation block; Step 2, preparing Tremella cultivation material, the formula of Tremella cultivation material is: 81 parts by weight of cottonseed hulls, 13 parts by weight of bran, 5 parts by weight of corn flour, and 1 part by weight of gypsum; after sterilizing the prepared Tremella cultivation material, inoculating the "white hair ball" part into the center of the glass bottle, after growing for 10 days, the mycelium of the glass bottle covers the bottle mouth; after growing for 20 days, the mycelium of the glass bottle covers the bottle mouth, and ear bases begin to appear; after growing for 40 days, Tremella fruiting bodies are obtained.

7. An identification primer for a coral-shaped Tremella fuciformis strain, characterized in that: The identification primers of the coral-shaped Tremella fuciformis strain are I1, I2, I4, I7, I13, I16 and I17; wherein the nucleotide sequence of I1 is: 5'-CTCCTCCTCCTCSC-3'; the nucleotide sequence of I2 is: 5'-BDBACAACAACAACAACA-3'; the nucleotide sequence of I4 is: 5CACCACCACCACSC-3'; the nucleotide sequence of I7 is: 5'-VDHTCGTCGTCGTCGTCG-3'; the nucleotide sequence of I13 is: 5'-DHBCGACGACGACGACGA-3'; the nucleotide sequence of I16 is: 5'-AGAGAGAGAGAGAGAGT-3'; the nucleotide sequence of I16 is: 5’-TCTCTCTCTCTCTCTGC-3’; wherein, B = G, C, T; D = A, G, T; H = A, C, T; V = A, G, C; S = G, C.

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