Strain of hebeloma crustaceum, strain preparation and application

Abstract

A strain of Inocybe geophylla (HFM-NT1) Coprinellus radians HFM-NT1 is preserved in China Center for Type Culture Collection (CCTCC), the address is Room 211, Wuhan University, Wuchang District, Wuhan City, Hubei Province, China, the preservation number is CCTCC NO: M 2026193, and the preservation time is January 22, 2026. The Inocybe geophylla HFM-NT1 of the present application has a wide temperature adaptation range and can grow normally at 10-30 DEG C, has strong resistance to miscellaneous ability, and can cover Trichoderma in mycelium growth and significantly inhibit the growth of mold when co-cultured with Trichoderma. When HFM-NT1 is used as the symbiotic fungus of Xerocomus chrysenteron, the mycelium infection cycle is accurately matched with the whole development process of Xerocomus chrysenteron seeds in a mountainous low-temperature environment, efficiently promotes the germination of Xerocomus chrysenteron seeds in a low-temperature and open environment, and effectively shortens the germination cycle and seedling cycle of Xerocomus chrysenteron seeds.

Description

Technical Field

[0001] This invention relates to the field of microbial cultivation technology, specifically to a strain of Coprinus radiata, its preparation, and its application. Background Technology

[0002] Radiated Devil's Umbrella ( Coprinellus radians Coprinus comatus belongs to the kingdom Fungi, phylum Basidiomycota, class Agaricales, order Agaricales, family Coprinus truncataceae, genus Coprinus. It is a widely distributed saprophytic fungus, mainly found in decaying wood, fallen leaves, and soil rich in organic matter. It has a strong ability to degrade lignocellulose and is highly adaptable to its environment. As a fungus with both saprophytic and symbiotic characteristics, Coprinus comatus has been proven to have multiple applications.

[0003] In the field of orchid propagation, *Coprinus radiata* is an important symbiotic germination fungus for rare medicinal orchids such as *Cymbidium goeringii* and *Cremastra appendiculata*. Orchid seeds are tiny and lack endosperm reserves; under natural conditions, they must rely on symbiotic fungi to provide carbon, nitrogen, and mineral nutrients to break dormancy and complete protocorm development. Current research shows that *Coprinus radiata* isolated from the rhizosphere of wild *Cymbidium goeringii* can continuously supply nutrients for seed development by penetrating the seed coat, inducing embryonic cell swelling, and forming a mycelial mass symbiotic structure. Simultaneously, it secretes auxins and gibberellins to regulate protocorm differentiation. Under optimal conditions, the symbiotic germination rate of *Cymbidium goeringii* seeds can reach approximately 75%, making it one of the core functional strains for the artificial propagation of orchids. Currently, several *Coprinus radiata* strains have been publicly reported for use in the propagation of *Cremastra appendiculata* and *Cymbidium goeringii* seedlings, providing technical support for the artificial conservation of rare medicinal orchids.

[0004] Although *Coprinus radiata* has been researched and applied in this field, existing publicly available strains still have significant technical defects that severely limit their large-scale application, especially in open environments such as orchid seedling cultivation in simulated wild conditions and understory cultivation, where they struggle to maintain stable efficacy. First, *Coprinus radiata* exhibits weak antagonism against *Trichoderma*, making it difficult to resist *Trichoderma* contamination. Its inhibition rate is generally low at normal temperatures, and its antibacterial activity further diminishes at low temperatures, making it unable to maintain stable efficacy in *Trichoderma*-contaminated environments. Second, its mycelial growth rate is insufficient, resulting in weak niche competition. Existing conventional *Coprinus radiata* strains typically grow only 8-12 mm of mycelium per day at a suitable temperature of 25℃, lower than the 10-15 mm growth rate of *Trichoderma*. At low temperatures of 10-15℃, the mycelial growth rate further decreases, making it difficult for the mycelium to rapidly colonize the substrate and occupy niches. Especially in open-field seedling cultivation, spores of other fungi easily invade through the air and substrate. Target strains, due to their inferior growth, cannot form a dominant flora and are easily suppressed by other fungi. Therefore, they can only be used in strictly sterilized substrates and aseptic environments, significantly increasing the cost and operational barriers to large-scale seedling cultivation. Third, performance degradation is significant at low temperatures, making them unsuitable for applications in cool mountainous areas. In the main orchid-producing areas of southwestern mountains, represented by Guizhou, the altitude is mostly between 1000 and 1800 m, and the average temperature during the seedling stage is often between 10 and 20℃. Existing *Coprinus radiata* strains show a significant decrease in mycelial viability, infectivity, and antibacterial ability within this temperature range. They cannot efficiently promote seed germination and are unable to resist contamination by other fungi, resulting in low success rates and poor stability in understory, simulated wild seedling cultivation. This fails to meet the application requirements for orchid seedling propagation in open, low-temperature environments. Therefore, developing a strain of Coprinus radiata with rapid mycelial growth, strong antagonism against Trichoderma and other miscellaneous fungi, the ability to co-culture with Trichoderma and form a growth advantage, and the ability to stably promote germination at low temperatures is a technical problem that urgently needs to be solved in this field. Summary of the Invention

[0005] The purpose of this invention is to provide a strain of *Coprinus radiata*, which has a fast mycelial growth rate, strong resistance to contamination, and can be co-cultured with *Trichoderma*. The mycelial growth covers *Trichoderma*, making it less susceptible to contamination by other fungi when planting seedlings in a simulated wild environment under forest cover.

[0006] Another objective of this invention is to provide a method for preparing a tertiary strain of *Coprinus radiata*. The prepared strain has a short cultivation cycle and low energy consumption.

[0007] The present invention also provides specific applications of the above-mentioned strains.

[0008] The objective of this invention is achieved through the following technical solution: A strain of *Coprinus radiata*, characterized in that: the strain is classified as *Coprinus radiata* (…). Coprinellus radians)HFM-NT1 is deposited at the China Center for Type Culture Collection (CCTCC), Room 211, Wuhan University, Wuchang District, Wuhan City, Hubei Province, China, with accession number CCTCC NO: M 2026193, and deposited on January 22, 2026.

[0009] The HFM-NT1 strain of *Coprinus radiata* has a wide temperature adaptability range, growing normally from 10-30℃, and is not easily limited by geographical location or environmental conditions, making it suitable for a wide range of applications.

[0010] The mycelium grows vigorously, with dense and robust hyphae that gradually turn from grayish-white to yellow, radiating outwards in a radial pattern, consistent with the typical hyphal morphology of Coprinus radiata.

[0011] The HFM-NT1 variety of *Coprinus radiata* exhibits good resistance to contamination. When co-cultured with *Trichoderma*, the mycelium can grow normally, and after 5 days of culture, the mycelium will cover *Trichoderma*.

[0012] The above-mentioned HFM-NT1 of *Coprinus radiata* promotes the growth of *Sagittaria sagittifolia* (… Cremastra appendiculata Application in seed germination of Rhododendron simsii.

[0013] Furthermore, the aforementioned *Coprinus radiata* HFM-NT1 strain controls *Trichoderma* species in orchid seedling cultivation. Trichoderma Application of miscellaneous bacteria (spp.).

[0014] Furthermore, the Trichoderma contaminants include *Trichoderma viride* (…). Trichoderma viride Trichoderma harzianum ( Trichoderma harzianum At least one of the following: The control refers to the strain forming a growth advantage in the co-culture system by inhibiting the growth of Trichoderma mycelium and spore germination, thereby avoiding contamination of the seedling substrate by Trichoderma.

[0015] A method for preparing a tertiary strain of *Coprinus radiata*, characterized by comprising the following steps: (1) Transfer the HFM-NT1 strain of Coprinus radiata to PDA medium and incubate in a constant temperature incubator at 25℃ in the dark for 4-5 days until the mycelium fully grows on the medium as the mother culture of HFM-NT1 of Coprinus radiata; (2) The mother culture plate of the radiata coprinus 'HFM-NT1' from step (1) was inoculated into the liquid culture medium in a shake flask and cultured in a full-temperature shaking incubator at 25℃ and 200 r / min for 3 days to obtain the shake flask culture. (3) Add the same liquid culture medium as in step (2) to the fermenter. After sterilization, use the flame ring protection method to inoculate the HFM-NT1 shake flask strain of *Coprinus radiata* into the fermenter at an inoculation rate of 0.12%-0.15%. Incubate at 25°C for 3 days to obtain the liquid strain for production.

[0016] Furthermore, the PDA culture medium consists of 20% potato, 2% glucose, 2% agar, and 1 L of water.

[0017] Furthermore, the liquid culture medium is prepared by adding 2% glucose, 0.3% peptone, 0.3% yeast extract, 0.1% potassium dihydrogen phosphate, 0.1% magnesium sulfate, 1 L of water, and 1-2 drops of defoamer.

[0018] The present invention has the following technical effects: The *Coprinus radiata* HFM-NT1 fungus of this invention has a wide temperature adaptability range, growing normally from 10 to 30°C. It exhibits strong resistance to contamination and a significant advantage in inhibiting mold growth. When co-cultured with *Trichoderma*, the mycelial growth completely covers the *Trichoderma*, making it less susceptible to contamination by other fungi when planted under forest cover. Using HFM-NT1 as a symbiotic fungus for *Coprinus radiata* precisely matches the entire mycelial infection cycle with the seed development process of *Coprinus radiata* in the low-temperature environment of mountainous areas. It efficiently promotes the germination of *Coprinus radiata* seeds in low-temperature, open environments and effectively shortens the germination and seedling stages. Attached Figure Description

[0019] Figure 1 These are images of the fruiting bodies of the HFM-NT1 *Ulva radiata* strain of the present invention, collected in the field.

[0020] Figure 2 The images show the morphological characteristics of the *Coprinus radiata* HFM-NT1 strain of the present invention. Image a shows the *Coprinus radiata* HFM-NT1 strain of the present invention growing on a PDA plate; image b shows the hyphae of the *Coprinus radiata* HFM-NT1 strain of the present invention under a 40x microscope.

[0021] Figure 3 Images of the HFM-NT1 *Coprinus radiata* strain cultured at different temperatures according to the present invention.

[0022] Figure 4 This image shows the co-culture of HFM-NT1 *Coprinus radiata* and *Trichoderma* on a PDA plate.

[0023] Figure 5 This is a picture of the HFM-NT1 *Coprinus comatus* fungus bag of the present invention fully colonized.

[0024] Figure 6 The images show the effects of the HFM-NT1 radiated capillaris var. radiata on the growth of *Sagittaria sagittifolia* at different stages; a) is an image showing how the HFM-NT1 radiated capillaris var. radiata promotes the formation of protocorms from *Sagittaria sagittifolia* seeds; b) is a photograph of *Sagittaria sagittifolia* seed germination; c) is a photograph of commercial *Sagittaria sagittifolia* seedlings. Detailed Implementation

[0025] The present invention will be specifically described below through embodiments. It should be noted that the following embodiments are only used to further illustrate the present invention and should not be construed as limiting the scope of protection of the present invention. Those skilled in the art can make some non-essential improvements and adjustments to the present invention based on the above description.

[0026] Example 1 Isolation and identification of strains 1. Isolation of strains Fruiting bodies of wild fungi collected in the forests of Baiyan Group, Chuanyan Village, Yangchang Town, Dafang County, Bijie City, Guizhou Province (such as...) Figure 1 The mycelium was obtained through isolation and purification, and the specific method is as follows: After rinsing the fruiting bodies of wild strains with sterile water to remove surface impurities, they were then disinfected with 75% alcohol. The fruiting bodies were cut open along the middle on a sterile ultra-clean workbench, and the internal mycelial blocks were inoculated into PDA medium and cultured in a constant temperature incubator at 25°C. After the mycelia germinated, the mycelia were promptly transferred to a new PDA medium for purification and culture. The mycelia were purified twice to obtain pure mycelia.

[0027] 2. Identification of strains After morphological identification, the results are as follows: Figure 2 As shown ( Figure 2 a represents the hyphal morphology of *Coprinus radiata* on a PDA plate. Figure 2 b shows the hyphal morphology of *Coprinus radiata* under a 40x microscope. The obtained hyphae grow vigorously on the plate, with dense and robust hyphae. The hyphae gradually change from grayish-white to yellow and radiate outwards, showing a radial texture, which is consistent with the hyphal characteristics of *Coprinus radiata*.

[0028] Molecular identification: This invention involves extracting genomic DNA from the *Coprinus radiata* strain and performing LSU-PCR amplification. The universal primer sequences are: 5'-ACCCGCTGAACTTAAGC-3', 5'-TCCTGAGGGAAACTTCG-3'. The nucleotide sequence of the obtained DNA fragment is shown in SEQ ID NO.1. BLAST homology comparison of this sequence in the NCBI database showed that the strain sequence has 100% similarity to *Coprinus radiata*, thus identifying it as *Coprinus radiata*. Coprinellus radians ).

[0029] The strain was designated HFM-NT1 and then deposited at the China Center for Type Culture Collection (CCTCC), located at Room 211, Wuhan University, Wuchang District, Wuhan City, Hubei Province, China. The accession number is CCTCC NO: M2026193, and the deposit date is January 22, 2026.

[0030] Example 2 Culture and anti-contamination verification of HFM-NT1 of *Coprinus radiata* at different temperatures (1) Culture at different temperatures: The HFM-NT1 strain of *Coprinus radiata* was cut into 5 mm pieces using a scalpel on a sterile operating table and inoculated into PDA medium. After inoculation, the culture was placed in a constant temperature incubator with temperature gradients of 10℃, 15℃, 20℃, 25℃, and 30℃ for 96 h. The diameter of hyphal growth was recorded using the streak method. The results of the temperature gradient culture are as follows: Figure 3 ( Figure 3 (The mycelial state after 4 days of culture) is shown in Table 1.

[0031] Table 1:

[0032] from Figure 3 As shown in Table 1, *Coprinus radiata* HFM-NT1 can grow at temperatures ranging from 10 to 30°C, demonstrating strong adaptability to environmental temperatures. At 10°C, germination is slow, taking 10 days to reach usability standards. The fastest mycelial growth occurs at 30°C, but the mycelium is relatively sparse during this temperature range.

[0033] The commonly used reference strains of *Coprinus radiata*, CICC 41797 and BNCC 341407, both from domestic scientific research and production, were selected as control strains. Under the same conditions as the strain HFM-NT1 of this invention, they were inoculated into PDA plates (90 mm plates, 20 mL culture medium) and cultured in the dark at constant temperature at 10℃ (simulating the seedling temperature in mountainous areas of Guizhou) and 25℃ (suitable temperature), respectively. Colony diameter was measured daily, and the average daily mycelial growth was calculated. Each group had three replicates. The results are shown in Table 2.

[0034] Table 2:

[0035] Note: The mycelial growth grade is as follows: "++++" indicates that the mycelium is dense, thick and uniform; "+++" indicates that the mycelium is relatively dense and grows evenly; "++" indicates that the mycelium is thin and sparse with few aerial mycelia; and "+" indicates that the mycelium is extremely thin and weak and can only be vaguely seen.

[0036] (2) Anti-contamination test: The plate confrontation method was used, with *Trichoderma viride* (…) Trichoderma virideTrichoderma (hereinafter collectively referred to as *Trichoderma*) was used as the test contaminant. *Coprinus radiata* HFM-NT1 and *Trichoderma* were inoculated separately on the same PDA plate and cultured at 15℃ and 25℃ respectively as experimental groups. *Trichoderma* inoculated alone on PDA plates served as the control group. On day 5, the colony diameter of *Trichoderma* was measured, and the inhibition rate of the target strain on *Trichoderma* mycelial growth was calculated. The mycelial growth status of the target strain and the degree of inhibition of *Trichoderma* in the co-culture system were observed. Simultaneously, CICC 41797 and BNCC 341407 were tested under the same conditions, with three replicates per group. The inhibition rate of each strain on *Trichoderma* was calculated using the following formula:

[0037] The results are shown in Table 3.

[0038] Table 3:

[0039] Note: The mycelial growth grade is as follows: "++++" indicates that the mycelium is dense, thick and uniform; "+++" indicates that the mycelium is relatively dense and grows evenly; "++" indicates that the mycelium is thin and sparse with few aerial mycelia; "+" indicates that the mycelium is extremely thin and weak and can only be vaguely seen; "-" indicates that the colony is completely covered by Trichoderma and no mycelia are visible.

[0040] The results showed that *Coprinus radiata* HFM-NT1 exhibited good contamination resistance; its mycelia grew normally when co-cultured with *Trichoderma*, and the mycelia covered *Trichoderma* after 5 days of culture. Figure 4 As shown in the photo (photograph taken at 15℃), the mycelium of HFM-NT1 maintained good growth even at the low temperature of 15℃. Due to its good antibacterial ability, HFM-NT1 combined with its competitive growth advantage at low temperatures achieved a dual effect of inhibiting the growth of Trichoderma at low temperatures, further increasing the inhibition rate compared to room temperature. In contrast, the growth of the control group was significantly inhibited by temperature at low temperatures, resulting in a further decrease in its inhibition rate against mold compared to room temperature.

[0041] Comparison of the effects of Trichoderma contamination on the germination of Sagittaria trifolia seeds: A humus-sawdust seedling substrate (60% humus, 30% sawdust, and 10% vermiculite) was used. Trichoderma spore suspension was artificially inoculated to simulate a contamination scenario. CICC 41797 was selected as the control group. The experiment was divided into four groups: (1) CICC 41797 uncontaminated group: CICC 41797 was inoculated in a substrate uncontaminated with Trichoderma; (2) CICC 41797 contaminated group: CICC 41797 was inoculated in a Trichoderma contaminated substrate; (3) HFM-NT1 uncontaminated group: HFM-NT1 was inoculated in a substrate uncontaminated with Trichoderma; (4) HFM-NT1 contaminated group: HFM-NT1 was inoculated in a Trichoderma contaminated substrate. Foam boxes were used for seedling cultivation. Each group used a total of 30 kg of substrate, with six replicates. Each foam box contained 5 kg of substrate, and the inoculation amount for both CICC 41797 and HFM-NT1 was 0.7 kg / box. Simultaneously, mature and plump *Sagittaria sagittifolia* seeds were sown, and cultured under controlled temperatures of 20±2℃, relative humidity of 80-85%, and diffused light. After 90 days, the number of germinations and substrate contamination were recorded. The results are shown in Table 4. For each group, one 0.12-0.15 g *Sagittaria sagittifolia* fruit clip was used, mixed with humus and *Coprinus radiata* spawn, and then divided into 6 equal portions for use.

[0042] Table 4

[0043] The table shows that the average number of germinations in the CICC 41797 uncontaminated group was 23 seedlings / box, while the average number of germinations in the contaminated group was 8 seedlings / box. In this invention, the average number of germinations in the *Coprinus radiata* HFM-NT1 uncontaminated group was 45 seedlings / box, while the average number of germinations in the contaminated group was 31 seedlings / box. Both average germination counts were higher than those of the CICC 41797 groups, and the average number of germinations in the contaminated group was higher than that of the CICC 41797 uncontaminated group. These results indicate that the *Coprinus radiata* HFM-NT1 in this invention can better promote the germination of *Trichoderma* seeds, resulting in a higher number of germinations. It also possesses good antibacterial ability, and combined with growth competition, it significantly inhibits the growth of *Trichoderma*, achieving an integrated function of "symbiotic germination promotion + miscellaneous fungal control."

[0044] Example 3 A method for preparing a tertiary strain of *Coprinus radiata* includes the following steps: (1) Transfer the HFM-NT1 strain of Coprinus radiata to PDA medium and culture it in a constant temperature incubator at 25℃ in the dark for 4-5 days until the mycelium fully grows on the medium as the mother culture of HFM-NT1 of Coprinus radiata. The PDA medium is composed of 20% potato, 2% glucose, 2% agar and 1 L water.

[0045] (2) Inoculate the master plate culture of Hebeloma radians HFM-NT1 obtained in step (1) into a shake flask liquid medium, and culture in a full-temperature shaking incubator at 25°C and 200 r / min for 3 d to obtain shake flask strains. The liquid medium is formulated as follows: 2% glucose, 0.3% peptone, 0.3% yeast extract powder, 0.1% potassium dihydrogen phosphate, 0.1% magnesium sulfate, 1 L of water, and 1-2 drops of defoamer to prepare the liquid medium; (3) Add the same liquid medium as that in step (2) into the fermenter, and after sterilization, access the shake flask strains of Hebeloma radians HFM-NT1 into the fermenter at an inoculum size of 0.12%-0.15% using the flame loop protection method, and culture at 25°C for 3 d to obtain the liquid strain for production.

[0046] Carry out cultivation and culture of the above liquid strains: (1) Preparation of cultivation bags: A solid cultivation medium is prepared according to the formula of 55% poplar sawdust, 21% corncob, 9% wheat bran, 5% cottonseed hull, 8% corn flour, 2% calcium bicarbonate, with a water content of 63%-65%. After mixing all components uniformly in proportion, the bags are made by a bag making machine, with a single bag weight of 1.45-1.5 kg and a water content of 63-65%. After the bags are made, they are placed in a sterilizer for sterilization. Among them, the corncob and cottonseed hull need to be pre-wetted in advance to prevent the water content from being too low after sterilization.

[0047] (2) Inoculation and cultivation: After the cultivation bags are cooled, the liquid strains are inoculated into the cultivation medium at an inoculum size of 70-80 mL per bag, and then placed in an aseptic workshop for constant temperature culture at 25°C for 12-15 d to make the bags full. The cultivation time is short, which effectively reduces the energy consumption cost and improves the warehouse turnover rate. The Hebeloma radians strains prepared by the above method have fast mycelium growth rate and strong vitality during cultivation, and the results are shown in Figure 5 .

[0048] Example 4 Application of Hebeloma radians HFM-NT1 strain The open forest land in Bijie City, Guizhou Province is selected for planting Cremastra appendiculata. The altitude of the planting site is above 1300 m, and the planting is started from March (the planting temperature is 10-15°C). A layered structure planting mode is adopted, and the specific operations are as follows: Planting the long-acting fungal material at the bottom layer: Before sowing, first remove the weeds and stones in the open forest land, dig 1m-wide sloping cultivation ridges along the terrain, and dig drainage ditches between the ridges to avoid water accumulation in the rainy season; place 15-30 cm large wood blocks in the ridges, and spread a layer of leaves and humus soil on the wood blocks.

[0049] Intermediate symbiotic strains: Break the HFM-NT1 spawn of Coprinus radiata into small pieces and arrange them neatly in the ridge, with each piece spaced 4-6 cm apart. Then sprinkle 1-1.5 cm of sawdust on the pieces to promote rapid mycelial connection, thereby reducing the amount of spawn used.

[0050] Top layer sowing and covering: Crush the HFM-NT1 *Coprinus radiata* spawn and mix it with humus. Then add the *Sagittaria sagittifolia* seeds and mix thoroughly until the seeds are evenly distributed in the mixture of spawn bags and humus. Evenly scatter the mixed seeds in the prepared ridges, cover with a layer of leaves and 2-3 cm of humus, and finally cover with mulch to retain warmth and moisture. This completes the sowing process. *Sagittaria sagittifolia* seeds are extremely small; take care to avoid wind during mixing to prevent a large number of seeds from being blown away.

[0051] By cultivating *Hylocereus undatus* using the above method, precise and slow-release fertilization is ensured during the seed germination and bulb enlargement stages, resolving the issues of nutrient supply interruption during the early stages of vigorous metabolism, the later stages of reduced vitality, and the bulb enlargement stage. Figure 6 As shown, protocorms form about one month after sowing, commonly known as dragon eggs. Figure 6 a); Germination occurs 3-4 months after sowing ( Figure 6 b). Commercial seedlings of *Gnaphalium affine* can be obtained in 7-8 months, with a yield of 220,000-250,000 seedlings per mu (approximately 0.067 hectares). Figure 6 c). In existing technologies, the time for the emergence of dragon eggs is generally 40-60 days, but this time varies among different strains, and some even take longer. Previously, the time to obtain commercial seedlings of *Coprinus radiata* var. *radiata* assisted in cultivation was approximately 10-12 months, with a yield of about 150,000-200,000 seedlings per acre.

[0052] In summary, the planting method of this invention results in a shorter seedling cultivation cycle for *Gnaphalium affine* and a higher yield per acre, effectively improving the economic benefits for growers. It also solves the technical problems of severe seedling size differentiation and low seedling yield in direct seeding under forest cover. The entire technical system has a low operational threshold, can be replicated on a large scale, and has significant value for industrial application.

Claims

1. A strain of *Coprinus radiata*, characterized by: The strain is classified as *Coprinus radiata* (Radiata). Coprinellus radians) HFM-NT1 is deposited at the China Center for Type Culture Collection (CCTCC), Room 211, Wuhan University, Wuchang District, Wuhan City, Hubei Province, China, with accession number CCTCC NO: M 2026193, and deposited on January 22, 2026.

2. The application of the radiate-haired coprinus HFM-NT1 as described in claim 1 in promoting the germination of Rhododendron seeds.

3. The control of Trichoderma genus (as described in claim 1) by HFM-NT1 of *Coprinus radiata* in orchid seedling cultivation. Trichoderma Application of miscellaneous bacteria (spp.).

4. A method for preparing a tertiary strain of *Coprinus radiata*, characterized in that, Includes the following steps: (1) Transfer the HFM-NT1 strain of Coprinus radiata to PDA medium and incubate in a constant temperature incubator at 25℃ in the dark for 4-5 days until the mycelium fully grows on the medium as the mother culture of HFM-NT1 of Coprinus radiata; (2) The mother culture plate of the radiata coprinus 'HFM-NT1' from step (1) was inoculated into the liquid culture medium in a shake flask and cultured in a full-temperature shaking incubator at 25℃ and 200r / min for 3 days to obtain the shake flask culture. (3) Add the same liquid culture medium as in step (2) to the fermenter. After sterilization, use the flame ring protection method to inoculate the HFM-NT1 shake flask strain of *Coprinus radiata* into the fermenter at an inoculation rate of 0.12%-0.15%. Incubate at 25°C for 3 days to obtain the liquid strain for production.

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