A white fungus strain and its application

Abstract

The present invention belongs to the field of microbial technology, and specifically relates to a white fungus strain and its application. The white fungus strain is the white fungus JAUACW008 strain, which was deposited in the China Center for Type Culture Collection on April 19, 2024, with a deposit number of CCTCCM2024719. The white fungus JAUACW008 strain of the present invention can be used for the production of fungus products, large-scale cultivation in tropical areas where cassava and rubber are abundant, and fermentation applications of nutrients such as trehalose.

Description

Technical Field

[0001] The invention belongs to the technical field of microorganisms, and particularly relates to a white fungus strain and application thereof. Background Art

[0002] Auricularia is a major edible fungus species in my country, and generally refers to the black fungus, hairy fungus, wrinkled fungus, and brittle fungus that can be artificially cultivated. Hairy fungus is the second largest cultivated species of the genus Auricularia. White hairy fungus (Auricularia cornea) is a white mutant of hairy fungus under natural conditions and is rarely distributed in nature. In 2015, the team of Academician Li Yu of Jilin Agricultural University systematically bred a new variety of white hairy fungus, "Jade Fungus". The market demand for white hairy fungus has increased suddenly, and the output and planting area have also been greatly improved. Jade Fungus has become the main cultivated variety in the national white hairy fungus market. It is urgent to develop white hairy fungus germplasm resources and enrich white hairy fungus varieties.

[0003] The study found that the crude polysaccharide, essential amino acid and flavor amino acid content of white fungus are higher than those of white fungus. The existing white fungus varieties are single and the white fungus germplasm resources are few, which hinders the healthy development of the fungus industry. Therefore, it is urgent to develop new germplasm resources of white fungus. Summary of the invention

[0004] In order to solve the problem of the limited white Auricularia auricula germplasm resources in the prior art, the present invention develops a white Auricularia auricula strain with a morphology different from the existing main cultivated varieties, with obvious wrinkles on the ventral surface of the ear piece, enriching the white Auricularia auricula varieties. To achieve the above purpose, the present invention adopts the following technical scheme:

[0005] The present invention provides a white fungus strain, wherein the white fungus strain is white hairy fungus JAUACW008 Auricularia cornea JAUACW008, and the hairy fungus JAUACW008 strain was deposited in the China Center for Type Culture Collection on April 19, 2024, with a deposit number of CCTCCM2024719, and a deposit address: Wuhan, China.

[0006] The method for obtaining the above-mentioned Auricularia auricula JAUACW008 strain is as follows:

[0007] The present invention isolates and screens a new white fungus strain from white wild fungus fruiting bodies collected from discarded cassava straw in Baodao New Village, Danzhou City, Hainan Province in December 2018. After 6 generations of systematic breeding, a new strain JAUACW008 with stable and excellent traits such as early maturity, high yield, and ventral folds is screened. The new strain JAUACW008 is identified and named white fungus JAUACW008 strain, or white fungus strain JAUACW008. The total sugar content of the dry product of the white fungus JAUACW008 strain is 38-40 mg / g; the trehalose content is 54-58 mg / g.

[0008] The white Auricularia auriculariae JAUACW008 strain provided by the present invention has the characteristics of medium-early maturity, high yield and ventral wrinkles.

[0009] The present invention also provides a method for preparing a strain of the white fungus JAUACW008 strain, comprising the steps of inoculating the activated white fungus JAUACW008 strain into a PDA culture medium and culturing the strain in the dark at 24-26° C. for 7-10 days to obtain a mother strain of the white fungus JAUACW008 strain; and then expanding and culturing the mother strain of the white fungus JAUACW008 strain to obtain an original strain of the white fungus JAUACW008 strain, that is, a solid original strain of the white fungus JAUACW008 strain.

[0010] Among them, the conditions for expansion and cultivation are as follows: the mother strain of white Auricularia auricularia JAUACW008 is inoculated into the original strain culture medium and cultured in the dark at 24-26° C. for 30-40 days.

[0011] The original seed culture material is composed of the following materials by mass fraction: 78% of sawdust, 20% of wheat bran, 1% of lime, and 1% of gypsum; the water content of the original seed culture medium is 60%. Preferably, the original seed culture material is composed of the following materials by mass fraction: 78% of sawdust, 20% of wheat bran, 1% of lime, and 1% of gypsum.

[0012] The present invention also provides a cultivation method of the white Auricularia auricula JAUACW008 strain, comprising the following steps:

[0013] After the cultivation material is filled in the cultivation bag, it is sterilized and inoculated with the white fungus JAUACW008 strain, and cultured for 45 to 60 days at 24 to 26° C. and 40 to 55% air humidity. Under the above conditions, the mycelium can completely eat the material, fully decompose the matrix, and accumulate nutrients for growing fungus.

[0014] Wherein, the cultivation material is composed of the following materials by mass fraction: 31.2-39.0% cassava straw, 46.8%-39.0 rubber sawdust, 20% wheat bran, 1% lime, 1% gypsum; the sum of the mass percentages of the above materials is 100%; the water content of the cultivation material is 55-58%. Preferably, the cultivation material is composed of the following materials by mass fraction: 31.2% cassava straw, 46.8% rubber sawdust, 20% wheat bran, 1% lime, 1% gypsum. According to the fact that the wild host of the white hairy fungus JAUACW008 strain is cassava straw, combined with the recycling of cassava straw resources and the demonstration and promotion of the JAUACW008 strain, cassava straw and rubber sawdust are preferentially selected as the main materials of the cultivation material. The yield of the white hairy fungus JAUACW008 strain cultivated with this formula is significantly higher than that of the traditional cultivation formula.

[0015] The size of the cultivation bag is 16.5cm*33~37cm. This specification of cultivation bag is suitable for short bag cultivation, which is convenient for promotion in the fungus production area.

[0016] The inoculation amount during inoculation is 10-15g sawdust stock / bag. According to the size of the inoculation hole and the inoculation amount, the inoculation amount is 10-15g sawdust stock / bag to ensure the inoculation quality, fast germination and save costs.

[0017] Punching: Use a puncher to punch holes evenly around the bag, with a length of 4-6mm, a depth of 4-5mm, and a spacing of 10-20mm. The single-piece rate of ear pieces obtained from cultivation is high, and the yield is high.

[0018] Hanging bag germination: After punching, wait for the mycelium to recover and then put it into the shed for germination. Hang 6 to 8 bags on each string of hanging wires (rope), with a spacing of 25 to 30 cm between each string, and 60 to 70 bags per square meter. The temperature is 16 to 22°C, the humidity is 65 to 85%, and the cultivation is carried out away from strong light for 5 to 7 days. Make the ear buds in the bag uniform and neat, and the ears will grow neatly at the end, which is more convenient for ear management.

[0019] It should be noted that hanging bags refers to hanging the bags with ropes or other connecting objects. The above-mentioned "hanging bags into the greenhouse" means transporting the bags from the mushroom room to the greenhouse and hanging them with ropes or other connecting objects.

[0020] "Primula" refers to the young stage of the fruiting body of Auricularia auricularia, where the hyphae are twisted to form a white colloid body.

[0021] Until the holes are completely covered with white hyphae or primordia appear.

[0022] When the primordium growth appears, the humidity is increased to 80-90%, the temperature is 16-22°C, and the ear pieces are differentiated. The ear pieces are managed by alternating dry and wet, and the space is sprayed with water 2-3 times a day, the humidity is 85-90%, the temperature is 16-24°C, and the scattered light is irradiated. After the ear pieces mature, the outer edge becomes thinner and tilts upward, and a small amount of spores are ejected, that is, the ear pieces are mature and can be harvested, and the white hairy fungus strain JAUACW008 is harvested. The obtained fruiting body ear pieces are thicker, the ear shape is good, and the yield is high.

[0023] The invention provides a white Auricularia auricula JAUACW008 strain. Meanwhile, the invention utilizes cassava straw and rubber sawdust as main culture materials to cultivate the strain, and screens out a culture (cultivation) material formula and a cultivation technology suitable for the growth of the strain.

[0024] The formula of 1 kg of the cultivation material is 0.312 kg of cassava straw, 0.468 kg of rubber sawdust, 0.2 kg of wheat bran, 0.01 kg of lime and 0.01 kg of gypsum; the water content of the cultivation material is 55%.

[0025] The key technology in the cultivation method is: the main materials of cassava straw and rubber sawdust are pre-wetted 10 hours in advance before making the mushroom bags, and wheat bran, lime and gypsum are added on the second day, and water is added and stirred evenly until the moisture content of the culture medium reaches 55%. The mixed culture medium is put into a cultivation bag to make a cultivation bag (mushroom bag), the mushroom bag adopts a 17×33 cm polyethylene plastic bag, is sterilized under high pressure for 2 hours, and is inoculated after the mushroom bag is cooled to room temperature; the mushroom bag is cultured at 25-28° C. in the early stage, 22-24° C. in the middle and late stages, and the mushroom bag is ripened for 10 days after it is full, and holes are punched for germination, the temperature is 15-20° C., the relative air humidity is 70-80%, and the primordium appears at the punching position for ear management, the greenhouse is 15-20° C., the relative air humidity is 85-90%, and the space is sprayed with water 2-3 times a day to maintain humidity and irradiate with scattered light; after the ear pieces are mature, the outer edges become thinner and tilt upwards, a small amount of spores are ejected, and the ear pieces are mature and can be harvested, that is, the white hairy wood ear JAUACW008 strain is harvested.

[0026] Compared with the prior art, the present invention has the following beneficial effects:

[0027] The present invention provides a white auricularia auricula strain and its application. The white auricularia auricula strain JAUACW008 provided by the present invention has the characteristics of medium-early maturity, high yield, and ventral wrinkles. At the same time, the present invention also develops a suitable cultivation material formula and cultivation method (technology) for the white auricularia auricula strain JAUACW008 produced by using cassava straw.

[0028] The white Auricularia auriculariae JAUACW008 strain provided by the invention has the potential to be developed into a new variety, enriches the varieties of white Auricularia auriculariae, and solves the problems of single production variety and few germplasm resources of white Auricularia auriculariae; the developed suitable cultivation material formula and cultivation technology can be quickly applied to the market and promoted on a large scale, thus saving raw material costs, recycling agricultural and forestry wastes, and achieving environmental protection and ecological protection. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 It is the antagonistic reaction diagram of the white hairy fungus JAUACW008 strain (test number AH2) and the control variety (test number AH0) in the present invention; wherein A is the front view of the antagonistic reaction mycelium; B is the back view of the antagonistic reaction mycelium.

[0030] Figure 2 It is a picture of the ear growth of the white hairy fungus JAUACW008 strain and the control variety in the present invention; wherein A is the ear growth of the control variety (experimental number AH0); and B is the ear growth of the white hairy fungus JAUACW008 strain (experimental number AH2).

[0031] Figure 3 This is the ITS phylogenetic tree of the Auricularia auriculariae JAUACW008 strain (experimental number AH2) in the present invention.

[0032] Figure 4 This is a photo of the wild fruiting body of the white Auricularia auricularia strain JAUACW008 in the present invention.

[0033] Figure 5 It is the mycelium of the white hairy fungus JAUACW008 strain in the present invention grown in the PDA culture medium. DETAILED DESCRIPTION

[0034] The present invention is described in detail below in conjunction with the accompanying drawings and specific examples, but should not be construed as limiting the present invention. Unless otherwise specified, the technical means used in the following examples are conventional means well known to those skilled in the art, and the materials, reagents, etc. used in the following examples, unless otherwise specified, can be obtained from commercial sources.

[0035] The culture medium used in the following examples is as follows:

[0036] PDA medium: 200 g potatoes, 20 g glucose, 10 g agar powder, and 1000 mL water.

[0037] Example 1

[0038] 1. Obtaining the strain of Auricularia auricula JAUACW008

[0039] 1. Strain acquisition:

[0040] White wild Auricularia auricularia fruiting bodies collected from discarded cassava straw in Baodao New Village, Danzhou City, Hainan Province in December 2018 ( Figure 4 ). Then the strains are obtained by tissue separation method. The specific method is as follows:

[0041] First, wipe the fruiting body of white wild Auricularia auricula with 75% alcohol cotton, dry the water, tear open the fruiting body of white wild Auricularia auricula to expose the sterile medulla inside, cut off a 0.3 cm tissue block, place it on PDA culture medium, place it in a 25°C incubator, and culture it in the dark. When the hyphae germinate, pick the 0.5 cm long mycelium at the tip of the hyphae, transfer it to a new PDA culture dish, and purify it to obtain a pure strain. The obtained strain is named white Auricularia auricula JAUACW008 strain, or white Auricularia auricula strain JAUACW008 ( Figure 5 ).

[0042] 2. Strain identification:

[0043] DNA was extracted from the mycelium of the white hairy fungus strain JAUACW008 using the CTAB method, and the ITS fragment was specifically amplified in the PCR reaction program using the universal primers ITS1 and ITS4 for fungi. The primer sequences are shown in Table 1, and the ITS-PCR reaction system is shown in Table 2.

[0044] Table 1 ITS-PCR primer sequences

[0045]

[0046] Table 2 ITS-PCR reaction system

[0047] Components Volume (μL) DNA template 1.0 2×TaqMasterMix 15.0 ITS1ForwardPrimer 1.0 ITS4ReversePrimer 1.0 Sterile double distilled water 12.0 Overall reaction system 30.0

[0048] The PCR reaction procedure is as follows:

[0049]

[0050] After ITS amplification, the PCR product was subjected to electrophoresis detection, and the agarose concentration during the detection was 1%. The PCR product was sequenced, and the sequencing work was completed by Shanghai Shenggong Biotechnology Co., Ltd., and then the ITS of the sample and the hairy fungus (Auricularia cornea) in GENBANK were compared by Blast. Identities = 525 / 528 (99%), Gaps = 1 / 528 (0%), and the analysis determined that the pure culture separated was hairy fungus mycelium, and it was named white hairy fungus JAUACW008 strain, or white hairy fungus strain JAUACW008.

[0051] The commercial properties of the above-mentioned white Auricularia auricularia strain JAUACW008 are: the fruiting body is flaky, white, wrinkled on the ventral side, and has cilia on the back. The fruiting body size is 5.43cm×4.46cm×0.086cm.

[0052] The white hairy Auricularia auricularia strain JAUACW008 obtained through the above isolation and identification was deposited in the China Center for Type Culture Collection on April 19, 2024, with the deposit number CCTCCM2024719.

[0053] 2. Research on the cultivation technology of white Auricularia auricula strain JAUACW008

[0054] In order to develop a suitable cultivation material formula and cultivation technology for the white Auricularia auricula strain JAUACW008, the present invention can be quickly applied to the market, save raw material costs, and improve the nutritional value of the white Auricularia auricula strain JAUACW008. The cultivation technology of the white Auricularia auricula strain JAUACW008 is studied, and a cultivation method of the white Auricularia auricula strain JAUACW008 is provided. The specific method is as follows:

[0055] 1. Preparation of mother stock:

[0056] The purified Auricularia auriculariae JAUACW008 strain was inoculated into PDA medium and cultured at 25° C. in the dark for 10 days to obtain the mother strain of Auricularia auriculariae JAUACW008 strain.

[0057] 2. Stock seed preparation:

[0058] The mother strain of Auricularia auriculariae JAUACW008 was propagated in a cultivation bag filled with original culture medium and cultured in a dark place at 25°C for 35 days. After the bag was full, the original strain of JAUACW008 was obtained and used for inoculation of the cultivation bag.

[0059] The formula composition of 1 kg of original seed culture material is 0.78 kg of sawdust, 0.20 kg of wheat bran, 0.01 kg of lime and 0.01 kg of gypsum; the water content of the original seed culture medium is 60%.

[0060] 3. Preparation of cultivars:

[0061] Put the prepared cultivation material into a cultivation bag with a size of 16.5cm*33cm, with a loading of 0.75kg per bag. After sealing, sterilize it at 121℃ for 2 hours, cool it to 25℃ and inoculate the cultivation bag. The inoculation amount is 15g / bag, seal the cultivation bag, and place it in the culture room at 25℃ for 50 days. After the mycelium of the cultivation bag fills the cultivation bag, carry out subsequent ear management. The inoculated cultivation bag is also called a fungus bag.

[0062] Among them, the formula composition of 1kg of cultivation culture medium is 0.312kg of cassava straw, 0.468kg of rubber sawdust, 0.2kg of wheat bran, 0.01kg of lime and 0.01kg of gypsum; the water content of the cultivation material is 55%.

[0063] 4. Punching and germination:

[0064] After the bags are full, continue to cultivate for 7 days to complete the mycelium after-ripening, and then punch holes for germination. The punching machine is used to punch holes in the bags. Each bag is punched with 200 holes, and the punching specifications are 5mm long and 5mm deep. After punching, wait for the mycelium to recover and then put the bags into the shed for hanging. Each string of hanging wires (ropes) hangs 6 bags, with a spacing of 30cm between each string, and 60 bags per square meter. The temperature is 22℃, the humidity is 65%, and it is cultured for 5 days away from strong light until the holes are completely covered with white mycelium or primordia appear. The temperature remains unchanged, the humidity is increased to 90%, and the scattered light intensity is maintained at 200Lux for more than 8 hours per day for germination, until the ear buds are neat and the ear management is carried out.

[0065] It should be noted that hanging bags refers to hanging the bags with ropes or other connecting objects. The above-mentioned "hanging bags into the greenhouse" means transporting the bags from the mushroom room to the greenhouse and hanging them with ropes or other connecting objects.

[0066] "Primula" refers to the young stage of the fruiting body of Auricularia auricularia, where the hyphae are twisted to form a white colloid body.

[0067] 5. Ear management:

[0068] When the primordium growth appears, the humidity in the greenhouse is raised to 85%, the temperature is 20℃, and the ear pieces are differentiated. The dry and wet alternating ear management is adopted: the space is sprayed with water 3 times a day, the humidity is 90%, the temperature is 22℃, and the scattered light is irradiated. When the ear pieces mature, the outer edge becomes thinner, tilts upward, and a small amount of spores are ejected, which means that the ear pieces are mature and can be harvested. The harvested mature ear pieces are the fruiting bodies of white hairy fungus JAUACW008.

[0069] Comparative Example 1

[0070] A cultivation method of white Auricularia auricula JAUACW008 strain comprises the following steps:

[0071] 1. Preparation of mother stock: same as in Example 1.

[0072] 2. Stock seed preparation: same as Example 1.

[0073] 3. Preparation of cultivars: The basic steps are the same as those in Example 1, except that the formula of the cultivar culture medium is different. The formula composition of 1 kg of cultivar culture medium in Example 1 is: 0.312 kg of cassava straw, 0.468 kg of rubber sawdust, 0.2 kg of wheat bran, 0.01 kg of lime, and 0.01 kg of gypsum; the water content of the culture medium is 55%. The formula composition of 1 kg of cultivar culture medium in this comparative example is: 0.78 kg of miscellaneous sawdust, 0.20 kg of wheat bran, 0.01 kg of lime, and 0.01 kg of gypsum; the water content of the original seed culture medium is 60%.

[0074] It should be noted that the cultivation medium is also called the cultivation medium.

[0075] 4. Punching and accelerating germination: same as in Example 1.

[0076] 5. Ear management: same as in Example 1.

[0077] In order to illustrate the effects of Example 1 and Comparative Example 1, we selected the fruiting bodies of white Auricularia auricula JAUACW008 harvested from Example 1 and Comparative Example 1, respectively, and detected the nutritional components therein using anthrone colorimetry and HPLC. The specific detection steps are as follows:

[0078] (1) Preparation of glucose standard curve: Take 6 20mL test tubes, number them, and prepare a series of standard glucose solutions of different concentrations according to the data in Table 1. Add 0.5mL of anthracene reagent (see Table 3) to each tube, then slowly add 5mL of concentrated H2SO4, shake, open the stopper of the test tube, boil it in a boiling water bath for 10 minutes, take it out and cool it to room temperature, perform colorimetry at a wavelength of 620nm, measure the optical density (OD) of each tube solution, and use the standard glucose content as the horizontal axis and the optical density value as the vertical axis to draw a standard curve.

[0079] Table 3 Standard glucose solution dosage table

[0080]

[0081]

[0082] (2) Determination of total sugar: Weigh 0.1 g of the powder of the fruiting body sample of white Auricularia auricula JAUACW008 harvested in Example 1 and Comparative Example 1, add about 3 mL of water, grind into a homogenate in a mortar, transfer to a conical flask, and rinse the mortar 3 times with about 12 mL of distilled water, and the eluate is also transferred to the conical flask. Then add 10 mL of 6 mol / L hydrochloric acid to the conical flask, stir evenly, and hydrolyze in a boiling water bath for half an hour. After cooling, neutralize with 10% NaOH solution to neutralize the pH. Then dilute to 100 mL with distilled water, filter, take 10 mL of the filtrate, dilute to 100 mL with distilled water, and dilute to 1000 times the total sugar hydrolyzate.

[0083] Among them, the measured absorbance value is calculated by the formula:

[0084] Sugar content of sample (g / 100g dry weight) = sugar content obtained from standard curve (μg) × dilution factor × 100 / sample weight (g) × 10 6。

[0085] The regression equation of the standard curve is y = 0.698x + 0.278 (R 2 =0.992), and the total sugar content of the fruiting body sample liquid of Auricularia auricula-judae JAUACW008 harvested in Example 1 and Comparative Example 1 was calculated by combining the standard curve and the above calculation formula.

[0086] The results obtained from the above calculations are shown in Table 4.

[0087] (1) Preparation of trehalose standard solution: Weigh 0.5 mg of trehalose-6-phosphate standard substance into a 10 mL volumetric flask, dissolve it in 80% by volume acetonitrile and make up to volume as a standard stock solution; dilute the standard stock solution in stages with 80% by volume acetonitrile to prepare standard solutions with trehalose-6-phosphate standard concentrations of 25 μg / mL, 12.5 μg / mL, 6.25 μg / mL, 3.1254 μ / mL, 1.5627 μg / mL, 0.78125 μg / mL, and 0.390625 μg / mL, respectively.

[0088] (2) Trehalose determination: Weigh 0.1 g of the powder of the fruiting body sample of white Auricularia auricula JAUACW008 harvested in Example 1 and Comparative Example 1 respectively into a 50 mL centrifuge tube, add 25 mL of deionized water, mix well, and heat and extract for 60 min in a constant temperature water bath at 80°C. After extraction, put it in a 4°C refrigerator, centrifuge it at 12000×g for 20 min after cooling, take the supernatant, filter it with a 0.22 μm microporous filter membrane, and use the filtrate for HPLC analysis. Set 3 replicates for each sample.

[0089] Among them, the chromatographic conditions of HPLC are: COSMOSIL Sugar-D chromatographic column (4.6mm×250mm, 5μm); column temperature 30°C; mobile phase is acetonitrile-water (70:30, V / V) solution, flow rate 1.0mL / min; injection volume 10μL; drift tube temperature 85°C; nebulization temperature 70°C; air flow rate 1.6L / min; analysis time 10min; using the external standard method, according to the formula: trehalose content (μg / g) = (trehalose concentration x sample volume) / sample mass, the trehalose content in the sample is calculated.

[0090] The results obtained from the above calculations are shown in Table 4.

[0091] The test results of the nutritional components of the fruiting bodies of white Auricularia auricularia JAUACW008 harvested in Example 1 and Comparative Example 1 are shown in Table 4.

[0092] Table 4 Nutritional components of fruiting bodies of Auricularia auricula JAUACW008 grown in different cultivation media

[0093]

[0094] As can be seen from Table 4, the total sugar and trehalose contents in the fruiting bodies of white Auricularia auricula JAUACW008 harvested by the cultivation method of Example 1 are significantly higher than the total sugar and trehalose contents in the fruiting bodies of white Auricularia auricula JAUACW008 harvested by the cultivation method of Comparative Example 1, indicating that the cultivation medium in Example 1 can significantly improve the quality of the fruiting bodies of white Auricularia auricula JAUACW008 when used to cultivate the white Auricularia auricula JAUACW008 strain; the cultivation medium in Example 1 has the effect of improving the quality of the fruiting bodies.

[0095] The above results show that the cultivation method of the white hairy Auricularia auricula strain JAUACW008 provided by the present invention has the effect of improving the nutritional quality of the JAUACW008 strain, such as the total sugar and trehalose content.

[0096] 2. Comparative study on the characteristics of white Auricularia auricularia strain JAUACW008 and cultivated Auricularia auricularia auricularia strain

[0097] In order to illustrate the performance of the white Auricularia auricula strain JAUACW008 provided by the present invention, the present invention selected the white Auricularia auricula strain JAUACW008 obtained by the cultivation method of the above-mentioned white Auricularia auricula strain JAUACW008 and the cultivated strain Auricularia auricula for comparative study of ear-producing characteristics.

[0098] Among them, the white hairy Auricularia auricularia strain JAUACW008 and the cultivated strain Auricularia auricularia were used as test strains, and the cultivated strain Auricularia auricularia was used as a control strain, which was derived from the Auricularia auricularia fruiting bodies purchased from the market, and pure strains were obtained through tissue separation.

[0099] The white Auricularia auricularia strain JAUACW008 was numbered AH2, and the cultivated Auricularia auricularia auricularia strain was numbered AH0. Antagonism test, ear-out test, and ITS sequence comparison were carried out respectively. The specific research results are as follows:

[0100] 1. Comparison of physiological characteristics between Auricularia auriculariae strain JAUACW008 and control strains

[0101] The most direct method to preliminarily identify different strains is to use physiological characteristics. The specific method is: take the white hairy fungus strain JAUACW008 and the control strain and inoculate them into the same PDA culture dish, with a distance of 2 cm between the blocks, and place them in a 25℃ incubator in the dark. When the hyphae of the two blocks extend to the junction, observe whether the antagonistic line appears. The results are shown in Figure 1 .

[0102] Depend on Figure 1 It can be seen that there is an obvious antagonistic line between the white hairy Auricularia auricularia strain JAUACW008 and the cultivated strain Auricularia auricularia, and it is preliminarily determined that these two strains have different physiological characteristics.

[0103] 2. Comparison of agronomic traits between the white hairy fungus strain JAUACW008 and the control strain

[0104] Through the ear test, the growth period, commercial characteristics, yield and other agronomic characteristics of the two strains were compared. The specific method is as follows:

[0105] (1) Production of ear fungus bags:

[0106] The formula composition of 1kg of cultivation culture medium is 0.78kg of miscellaneous sawdust, 0.20kg of wheat bran, 0.01kg of lime and 0.01kg of gypsum; the water content of the original seed culture medium is 60%.

[0107] The above cultivation medium is placed in a polypropylene cultivation bag of 17×33 cm in size, and each cultivation bag is filled with 300 g of the above cultivation medium. When filling, the wood chips are pressed tightly by hand, and no gaps are left at the bottom of the bag. The bag is tied with a rope and placed in an autoclave for sterilization at a temperature of 121°C, 0.12 kPa, and sterilization for 120 minutes.

[0108] (2) Inoculation: After the cultivation bag has cooled to room temperature, place it in a clean bench and sterilize it for 30 minutes before inoculating the white hairy Auricularia auricularia strain JAUACW008 and the cultivated Auricularia auricularia strain JAUACW008 into the bag. The inoculation process must be performed in accordance with aseptic operation.

[0109] Germ management:

[0110] I Reproductive period

[0111] We recorded the time of germination, primordium occurrence, and fruiting body maturity of the two strains.

[0112] Among them, the spawning time refers to the time between the second day after inoculating the fungus bag and punching the fungus bag.

[0113] The primordium occurrence time refers to the time from the second day after the inoculation of the fungus bag to the time when the primordium is produced.

[0114] The fruiting body maturity time refers to the time from the second day after the inoculation of the bag to the first harvest of the bag.

[0115] The results are shown in Table 5.

[0116] Table 5 Growth period of white Auricularia auricularia strains

[0117] strain Full bag period / d Primordial period / d Harvest period / d AH0 25 34 45 AH2 27 43 53

[0118] As shown in Table 5, the bag-filling period, primordium period and harvesting period of the white hairy Auricularia strain JAUACW008 are longer than those of the control strain.

[0119] II Output

[0120] a. Expansion degree of ear slices: Use a 0.01g electronic balance to weigh the dry ear slices (dried fruiting bodies) of the white hairy fungus strain JAUACW008 and the cultivated strain Auricularia auricula strain, respectively, and then soak the dry ear slices weighed in RO water for about 5 hours, take out the expanded ear slices and put them on filter paper to absorb excess water and weigh them again. This weighing is the wet weight of the ear slices, and repeat 5 times. Finally, the ratio between the wet weight and dry weight of the ear slices is calculated as the expansion degree.

[0121] b Yield: the mass (kg) of dry fungus produced per 100kg dry culture medium.

[0122] The results are shown in Table 6.

[0123] Table 6 Comparison of the degree of foaming and biological efficiency of white Auricularia auricularia strains

[0124] strain Foaming degree Output(kg) AH0 7.9 7.03 AH2 8.6 6.82

[0125] As shown in Table 6, the yield of the white Auricularia auricularia strain JAUACW008 is close to that of the control strain, but its foaming degree is higher than that of the control variety.

[0126] III Product Characteristics

[0127] Wrinkles on the abdomen and back of the ear: The fruiting bodies of the test strains were picked after they were mature, and then they were observed and recorded by visual inspection; The length of the ear: The fruiting bodies of the test strains were picked after they were mature, and then the length was measured and recorded with a vernier caliper.

[0128] Ear width: Pick the fruiting bodies of the test strain after they mature, and then measure and record them with a vernier caliper.

[0129] Ear thickness: After the fruiting bodies of the tested strains matured, they were picked and measured and recorded using a digital thickness gauge. All properties were measured 30 times. The results are shown in Table 7.

[0130] Table 7 Comparison of fruiting body size and wrinkles of white Auricularia auricularia strains

[0131] strain Ear length (mm) Ear width (mm) Ear thickness (mm) Ventral folds AH0 34.51 55.51 1.27 none AH2 44.14 53.94 0.86 have

[0132] It can be seen from Table 7 that the commercial properties of the white Auricularia auricularia strain JAUACW008 are quite different from those of the control strain, especially the ventral surface has more wrinkles, while the control strain has no wrinkles on the ventral surface.

[0133] The color of the fungus slices was quantitatively characterized using a 3nh spectrophotometer. The colors of the fresh and dry ears, back and ventral surfaces were measured, and repeated three times to obtain L* (brightness axis, 0 is black, 100 is white), a* (red-green axis, positive values ​​are red, negative values ​​are green, and 0 is neutral), and b* (yellow-blue axis, positive values ​​are yellow, negative values ​​are blue, and 0 is neutral). The chromaticity value of the fungus slices was calculated. The larger the chromaticity value, the brighter the color. Formula: Chromaticity value = (a*2+b*2)1 / 2. The results are shown in Table 8 and Figure 2 .

[0134] Table 8 Comparison of fruiting body colors of white Auricularia auricularia strains

[0135]

[0136] According to the colorimeter test of fresh ear color in Table 8, the whiteness of the fresh ear color and dry ear color of the control strain is low; the whiteness of the fresh ear color of the white hairy Auricularia auricula strain JAUACW008 is higher, which is different from the control strain. Figure 2 It can be seen that the white Auricularia auricularia strain JAUACW008 is white with a slightly yellowish color and has many wrinkles on the ventral surface.

[0137] IV ITS sequence alignment and cluster analysis

[0138] ITS sequences were used to construct a phylogenetic tree of Auricularia species, including two test strains and four Auricularia strains cited from the NCBI website, namely Auricularia cornea isolate B02, Auriculariacornea cultivar AB111, Auricularia cornea RD 010593, and Auricularia corneacultivarMY01. The alignment results are shown in Table 9.

[0139] Table 9 Genetic distance analysis table

[0140]

[0141] The ITS sequence of the white Auricularia auricularia strain JAUACW008 was compared with the ITS sequence of the reported white variant of Auricularia auricularia, and the phylogenetic tree was constructed using the software MEGA5. Figure 3 shown.

[0142] Depend on Figure 3 It can be seen that in the constructed phylogenetic tree, when the genetic distance is 0.0019, it can be divided into two categories: AH2 is one category, and AH0 and other cited Auricularia auricularia strains are clustered into one category. It can be seen that there are certain genetic differences between the white Auricularia auricularia strain JAUACW008 and the existing Auricularia auricularia resources.

[0143] The above results indicate that the ear-producing characteristics and fruiting body agronomic traits of the white Auricularia auricula strain JAUACW008 provided by the present invention are significantly different from those of existing cultivated strains, and the strain is a new white Auricularia auricula strain.

[0144] The white Auricularia auriculariae JAUACW008 strain provided by the invention has the potential to be developed into a new variety, enriches the varieties of white Auricularia auriculariae, and solves the problems of single production variety and few germplasm resources of white Auricularia auriculariae; the developed suitable cultivation material formula and cultivation technology can be quickly applied to the market and promoted on a large scale, thus saving raw material costs, recycling agricultural and forestry wastes, and achieving environmental protection and ecological protection.

[0145] It should be noted that when the claims of the present invention involve numerical ranges, it should be understood that the two endpoints of each numerical range and any numerical value between the two endpoints can be selected. In order to avoid redundancy, the present invention describes a preferred embodiment.

[0146] Although the preferred embodiments of the present invention have been described, those skilled in the art may make other changes and modifications to these embodiments once they have learned the basic creative concept. Therefore, the appended claims are intended to be interpreted as including the preferred embodiments and all changes and modifications that fall within the scope of the present invention.

[0147] Obviously, those skilled in the art can make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if these modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include these modifications and variations.

Claims

1. A white fungus strain, characterized in that The white fungus strain is white fungus ( Auricularia cornea ) JAUACW008 strain, the white Auricularia auricula JAUACW008 strain was deposited in the China Center for Type Culture Collection on April 19, 2024, with the preservation number CCTCCM2024719.

2. A method for preparing the strain of Auricularia auricula-judae JAUACW008 according to claim 1, characterized in that: The activated Auricularia auriculariae JAUACW008 strain is cultured to obtain the mother strain of Auricularia auriculariae JAUACW008 strain; the mother strain of Auricularia auriculariae JAUACW008 strain is then expanded and cultured to obtain the original strain of Auricularia auriculariae JAUACW008 strain, that is, the solid original strain of Auricularia auriculariae JAUACW008 strain.

3. The preparation method according to claim 2, characterized in that: The activated Auricularia auriculariae JAUACW008 strain is inoculated into a PDA culture medium and cultured in the dark at 24-26° C. for 7-10 days to obtain the mother strain of Auricularia auriculariae JAUACW008 strain.

4. The preparation method according to claim 2, characterized in that: The conditions for the propagation culture are as follows: the mother strain of Auricularia auriculariae JAUACW008 is inoculated into the stock culture medium and cultured at 24-26°C in the dark for 30-40 days; The original seed culture medium is composed of the following materials in mass fractions: 78% of sawdust, 20% of wheat bran, 1% of lime and 1% of gypsum; the water content of the original seed culture medium is 60%.

5. A method for cultivating the white fungus JAUACW008 strain as claimed in claim 1, characterized in that: The following steps are involved: After the cultivation material is filled in the cultivation bag, it is sterilized, and the white fungus JAUACW008 strain is inoculated, and the culture is carried out for 45 to 60 days at 24 to 26° C. and 40 to 55% air humidity; Punching: Punch holes evenly around the cultivation bag; Hanging bag germination: After drilling, wait for the mycelium to recover and then put it into the shed for germination in hanging bags until the holes are completely covered with white mycelium or primordia appear; After the primordium is formed, the ear removal management is carried out; the humidity is increased to 80-90%, the temperature is 16-24° C., the culture is continued, and the ear pieces are harvested when they are mature, and the white hairy fungus JAUACW008 strain is harvested; The cultivation material is composed of the following materials in mass fractions: 31.2-39.0% cassava straw, 46.8-39.0% rubber sawdust, 20% wheat bran, 1% lime, and 1% gypsum; the sum of the mass percentages of the above materials is 100%; and the water content of the cultivation material is 55-58%.

6. The cultivation method according to claim 5, characterized in that: The method for continuing cultivation is: wait for the ear pieces to differentiate, spray water 2 to 3 times a day, humidity 85 to 90%, temperature 16 to 24°C, and irradiate with scattered light.

7. The cultivation method according to claim 5, characterized in that: The size of the cultivation bag is 16.5cm*33-37cm.

8. The cultivation method according to claim 5, characterized in that: The inoculation amount during inoculation is 10-15g sawdust stock / bag.

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