Astragalus sinicus plant and method for cultivating astragalus sinicus plant

Abstract

This invention belongs to the field of edible fungus cultivation technology. More specifically, it relates to a short-haired Auricularia auricula-judae strain, Gan Duanmaoer No. 1, and its propagation method. This invention provides a short-haired Auricularia auricula-judae strain (… Auricularia villosula This invention relates to a novel strain, "Gan Duanmaoer No. 1," and its efficient propagation technology system. This strain exhibits stable characteristics and strong fruiting ability, achieving systematic artificial cultivation of short-haired Auricularia auricula-judae for the first time. By optimizing the liquid culture medium, the mycelial ball density and biomass of this strain increased by 62.65% and 55.06%, respectively. Through log cultivation, Elaeocarpus decipiens, Pterocarya stenoptera, Ligustrum lucidum, and Sophora japonica were selected as the optimal substrates. Furthermore, a high-yield bag cultivation substrate was developed specifically for this strain, with a biological efficiency of 75%–83%, and a total yield of approximately 130g of fresh fruiting fruit per bag across three flushes. This invention overcomes the bottleneck of relying on wild harvesting for short-haired Auricularia auricula-judae, providing germplasm and technical support for industrialization and possessing significant application value.

Description

Technical Field

[0001] This invention belongs to the field of edible fungi cultivation technology. More specifically, it relates to a short-haired Auricularia auricula strain, Gan Duanmaoer No. 1, and its propagation method. Background Technology

[0002] Short-haired wood ear fungus ( Auricularia villosula It is mainly distributed in the tropical and subtropical regions of my country, often growing on the bark of living broad-leaved trees or fallen, decaying wood. It is a wood-decaying fungus with both edible and important medicinal value. Rich in polysaccharides, proteins, and various trace elements, it has physiological effects such as moisturizing the lungs, cleansing the intestines, and aiding weight loss, and has huge market potential. However, currently, the supply of short-haired wood ear fungus almost entirely relies on wild collection; there are no systematic reports on artificial cultivation, let alone industrialization. Its main bottlenecks are: the lack of stable, high-yielding strains with strong fruiting ability, and the lack of an efficient breeding and cultivation technology system.

[0003] Therefore, screening superior strains suitable for artificial cultivation from wild germplasm resources and establishing efficient and scalable breeding and cultivation techniques are of great ecological, economic and social significance for promoting the sustainable utilization of short-haired wood ear fungus resources, promoting the development of under-forest economy and upgrading the edible fungus industry. Summary of the Invention

[0004] The present invention aims to overcome the defects and shortcomings of existing technologies such as reliance on wild resources, lack of excellent production strains, low vitality of liquid spawn, and low efficiency of log and bag cultivation of Auricularia auricula-judae, and provides a strain of Auricularia auricula-judae, Gan Duanmaoer No. 1, and its breeding method.

[0005] The first objective of this invention is to provide a short-haired wood ear mushroom ( Auricularia villosula ) Gan Duanmao Ear No. 1 strain.

[0006] The second objective of this invention is to provide a substrate for bagging short-haired wood ear fungus.

[0007] The third objective of this invention is to provide a method for bag cultivation of short-haired wood ear fungus.

[0008] The fourth objective of this invention is to provide a method for preparing liquid cultures of the above-mentioned strains.

[0009] The fifth objective of this invention is to provide a method for cultivating the above-mentioned strains on logs.

[0010] The sixth objective of this invention is to provide the application of the above-mentioned bag substrate, the above-mentioned bag cultivation method, the above-mentioned method for preparing liquid spawn of the above-mentioned strain, and the above-mentioned method for log cultivation of the strain in the production of short-haired black fungus.

[0011] The above-mentioned objective of this invention is achieved through the following technical solution: This invention provides a short-haired wood ear fungus (Auricularia villosula The strain 1 of *Eurys nigra* was deposited at the China General Microbiological Culture Collection Center on June 11, 2024, with the accession number CGMCC No. 41333.

[0012] This invention provides fruiting bodies obtained by cultivating the above-mentioned short-haired Auricularia auricula-judae strain Gan Duanmaoer No. 1.

[0013] This invention provides mycelium and / or spores obtained from culturing the above-mentioned short-haired wood ear fungus strain Gan Duanmaoer No. 1.

[0014] The present invention also provides a food or health product containing the above-mentioned short-haired wood ear fungus strain No. 1 and / or the above-mentioned fruiting bodies and / or the above-mentioned mycelium and / or spores.

[0015] This invention provides a substrate for bagging short-haired wood ear fungus, which, by dry weight percentage, contains the following components: 50-65% hardwood sawdust, 13-20% cottonseed hulls, 8-20% wheat bran, 0-20% corn cob, 0.5-1.5% quicklime, and 0.5-1.5% gypsum.

[0016] Specifically, the short-haired fungus is the aforementioned short-haired fungus ( Auricularia villosula ) Gan Duanmao Ear No. 1 strain.

[0017] Optionally, the above-mentioned bag material substrate, by dry weight percentage, contains the following components: 65% hardwood sawdust, 13% cottonseed hulls, 20% wheat bran, 1% quicklime, and 1% gypsum.

[0018] Optionally, the above-mentioned bag material substrate, by dry weight percentage, contains the following components: 50% hardwood sawdust, 20% cottonseed hulls, 8% wheat bran, 20% corn cobs, 1% quicklime, and 1% gypsum.

[0019] Preferably, the above-mentioned bag material substrate contains the following components by dry weight percentage: 20-40% maple wood chips or 40-50% Elaeocarpus decipiens wood chips, 0-30% miscellaneous wood chips, 13-20% cottonseed hulls, 8-20% wheat bran, 0-20% corn cob, 0.5-1.5% quicklime and 0.5-1.5% gypsum.

[0020] As an alternative implementation, the above-mentioned bag substrate contains the following components by dry weight percentage: 20-40% maple wood chips, 10-30% miscellaneous wood chips, 13-20% cottonseed hulls, 8-20% wheat bran, 0-20% corn cobs, 0.5-1.5% quicklime, and 0.5-1.5% gypsum.

[0021] As an alternative implementation, the above-mentioned bag substrate contains the following components by dry weight percentage: 20-40% maple wood chips, 10-30% miscellaneous wood chips, 20% cottonseed hulls, 8% wheat bran, 20% corn cobs, 1% quicklime, and 1% gypsum.

[0022] As an alternative implementation, the above-mentioned bag substrate contains the following components by dry weight percentage: 40-50% Elaeocarpus decipiens sawdust, 0-10% miscellaneous sawdust, 13-20% cottonseed hulls, 8-20% wheat bran, 0-20% corn cob, 0.5-1.5% quicklime, and 0.5-1.5% gypsum.

[0023] As an alternative implementation, the above-mentioned bag substrate contains the following components by dry weight percentage: 40-50% Elaeocarpus decipiens sawdust, 0-10% miscellaneous sawdust, 20% cottonseed hulls, 8% wheat bran, 20% corn cob, 1% quicklime, and 1% gypsum.

[0024] As an alternative implementation, the moisture content of the above-mentioned bag material substrate is 60-65%.

[0025] This invention provides a method for cultivating short-haired wood ear fungus using bag substrate, which is described above for the cultivation of short-haired wood ear fungus.

[0026] As an alternative implementation method, the bag cultivation method for short-haired wood ear mushrooms includes the following steps: S1. Pack the above-mentioned substrate into bags and sterilize the bags; S2. Inoculate 10-15 mL of the above-mentioned strain into the mushroom bag and incubate at 26-28℃. After the mycelium has fully grown into the mushroom bag, continue to ripen for 5-7 days. S3. Cut open the mushroom bag. The mushroom bags will produce fruiting bodies at 18~25℃ and 85~95% relative humidity. Then carry out harvesting and management.

[0027] Optionally, the bag is a polypropylene bag. The polypropylene bag has dimensions of 120mm × 240mm × 5mm.

[0028] Specifically, the sterilization process involves sterilizing at 121°C and 0.1 MPa for 3-4 hours.

[0029] Specifically, the incubation period at 26-28°C described in step S2 is 30-40 days.

[0030] Optionally, the culture bags in step S3 are cultured under diffused light conditions for more than 8 hours, with a light intensity of 300~500 Lux.

[0031] As an alternative implementation, the harvesting management in step S3 is as follows: harvest when the ear flaps have extended to 3-5cm, stop spraying water after harvesting and allow the fungus to grow for 3-5 days, and then proceed to the next flush of ear flap management.

[0032] This invention provides a method for preparing liquid bacterial cultures of the above-mentioned strains, wherein the strains are inoculated into a liquid culture medium and cultured to obtain liquid bacterial cultures; the liquid culture medium comprises the following components: 150-250g potato extract, 18-22g glucose, 8-12g tryptone, 1.2-1.8g potassium dihydrogen phosphate, 0.8-1.5g magnesium chloride, and 1000mL water.

[0033] Specifically, the method for preparing the potato extract is as follows: take 150-250g of potatoes, add 800mL of water and boil for 30 minutes, cool and then filter the extract with 6 layers of medical gauze.

[0034] As an alternative implementation, the liquid culture medium is prepared as follows: 200g potato extract is mixed with 20g glucose, 10g tryptone, 1.5g potassium dihydrogen phosphate and 1.0g magnesium chloride, and the volume is adjusted to 1000mL with distilled water, and the pH is adjusted to 7.5~8.5.

[0035] As an alternative implementation, in the above-mentioned liquid bacterial strain preparation method, the culture is carried out at 26~30℃ and 180~220 r / min for 9~13 days.

[0036] This invention provides a method for cultivating the above-mentioned strain on logs, wherein the strain is cultivated on logs, and the variety of the logs is *Elaeocarpus decipiens* (Elaeocarpus spp.). Elaeocarpus glabripetalus ), maple ( Pterocarya stenoptera ), Privet ( Ligustrum lucidum ), Chinese scholar tree ( Styphnolobium japonicum Any one of them.

[0037] As an alternative implementation method, the above-mentioned log cultivation method includes the following steps: (1) Select deadwood from Elaeocarpus decipiens, Pterocarya stenoptera, Ligustrum lucidum and Sophora japonica to make logs, and then punch holes and sterilize the logs; (2) Inoculate the liquid strain of the above strain into the logs treated in step (1) and incubate at 26~28℃; (3) After the mycelium has fully grown on the wood segment, the wood segment is cultured for ear formation at 20~25℃ and 80~95% relative humidity.

[0038] As an alternative implementation, the drilling process in step (1) is as follows: the log is drilled with holes spaced 4-6 cm apart and the row spacing is 4-6 cm apart.

[0039] As an alternative implementation scheme, in step (1), after the log is perforated, it is soaked in 1% quicklime water for 2-3 days.

[0040] As an alternative implementation, the sterilization process in step (1) is as follows: after the logs are put into bags, they are sterilized at 121°C and 0.1 MPa for 3-4 hours.

[0041] As an alternative implementation scheme, the specific method of inoculation in step (2) is as follows: the liquid strain of the above strain is inoculated into the holes on the log and sealed with sterilized corn cob.

[0042] Specifically, in step (2), the culture is carried out in the dark, and the culture period is 30 to 40 days.

[0043] Specifically, in step (3), the light conditions for cultivation are that the daily diffused light and dark time each account for 50%, and the light intensity is 300~500 lux.

[0044] Specifically, in step (3), the ventilation conditions for cultivation are: ventilating 3 to 5 times a day, for 20 to 0 minutes each time.

[0045] The application of the above-mentioned bag substrate, the above-mentioned bag cultivation method, the above-mentioned liquid spawn preparation method, or the above-mentioned log cultivation method in the production of short-haired black fungus should also be within the scope of protection of this invention.

[0046] The present invention has the following beneficial effects: This invention provides a short-haired wood ear fungus with excellent agronomic traits ( Auricularia villosula The new strain “Gan Duanmaoer No. 1” and its supporting high-efficiency breeding method.

[0047] This invention marks the first systematic artificial propagation of short-haired Auricularia auricula-judae. Addressing the industry bottleneck of long-term reliance on wild collection and the lack of stable, high-yielding strains, this invention successfully isolated, purified, and preserved the stable and high-yielding strain "Gan Short-haired Auricularia auricula-judae No. 1" (CGMCC No. 41333), laying a germplasm foundation for the artificial cultivation and industrialization of short-haired Auricularia auricula-judae.

[0048] This invention significantly improves mycelial ball density (by 62.65%), biomass (by 55.06%), and optimizes mycelial ball morphology (by more uniform and denser diameter) by optimizing the liquid culture medium formula, providing a highly viable and highly consistent liquid primary culture for subsequent inoculation of logs or bags.

[0049] In addition, the present invention systematically screened deadwood from 100 tree species and identified *Elaeocarpus decipiens* (…). Elaeocarpus glabripetalus ), maple ( Pterocarya stenoptera ), Privet (Ligustrum lucidum ) and Chinese scholar tree ( Styphnolobium japonicum It is the most suitable high-quality substrate for the cultivation of short-haired wood ear mushrooms on logs, with a yield of 35-45 pieces per log segment.

[0050] In bag cultivation, this invention innovatively replaces conventional miscellaneous wood chips with sawdust from high-quality logs (Elaeagnus decipiens, Pterocarya stenoptera, and Sophora japonica), and develops a high-yield cultivation formula and supporting process for the "Gan Duanmao Ear No. 1" strain. The biological efficiency (bioconversion rate) of this optimized formula is 75%~83%, and the fresh ear yield reaches 130g / bag (total of 3 flushes), which is significantly better than the conventional formula, and has the advantages of both high yield and efficient resource utilization.

[0051] In summary, this invention not only provides a new high-yield strain of short-haired Auricularia auricula-judae, but also constructs a complete and efficient breeding technology system covering liquid spawn preparation, log cultivation, and bag cultivation. It has significant economic and social value for promoting the transformation of short-haired Auricularia auricula-judae from wild collection to large-scale artificial cultivation, promoting the development of under-forest economy, and upgrading the specialty edible fungi industry. Attached Figure Description

[0052] Figure 1 The mycelial growth of *Gan Duanmao Er* No. 1 after tissue isolation and purification.

[0053] Figure 2 The effect of liquid culture prepared for the liquid fermentation of Gan Duanmaoer No. 1.

[0054] Figure 3 This is a diagram showing the fruiting effect of *Earring rotundifolia* No. 1 section wood cultivation.

[0055] Figure 4 This is a picture showing the ear emergence effect of the Gan Duanmaoer No. 1 variety cultivated in bag substrate.

[0056] Figure 5 The fruiting effect of adding Gan Duanmao Er No. 1 to a mixture of maple / Elaeocarpus decipiens sawdust and miscellaneous sawdust. Detailed Implementation

[0057] The present invention will be further described below with reference to the accompanying drawings and specific embodiments, but the embodiments do not limit the present invention in any way. Unless otherwise specified, the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in this technical field.

[0058] Unless otherwise specified, all reagents and materials used in the following examples are commercially available.

[0059] The liquid culture medium in Example 1 contains the following components: 18-22g glucose, 8-12g tryptone, 1.2-1.8g potassium dihydrogen phosphate, 0.8-1.5g magnesium chloride, 150-250g potato juice, 1000mL water, and pH 7.5-8.5. The sterilization conditions for the liquid culture medium are 121℃ and 0.1MPa for 30min.

[0060] PDA solid culture medium: 200g potato juice, 20g glucose, 20g agar, and water to a final volume of 1000mL.

[0061] The methods for determining mycelial ball density, mycelial biomass, and mycelial ball diameter are referenced in "Wang Yu, Huang Liang, Peng Mingmeng, et al. Optimization of liquid fermentation culture conditions for white enoki mushroom [J]. Northern Horticulture, 2018, (20): 148-153."

[0062] Example 1: Isolation and Identification of the Short-haired Auricularia auricula strain Gan Duanmaoer No. 1 The strain was collected on March 25, 2023, from the bark of a healthy *Elaeocarpus decipiens* tree in the Arboretum of Jiangxi Agricultural University. Auricularia villosula The fruiting bodies, after tissue isolation, were inoculated onto PDA solid medium and cultured at a constant temperature of 28°C in the dark. By repeatedly picking single hyphal tips for transfer, a pure culture was finally obtained, which is the mother culture for production.

[0063] The mother culture block was inoculated into sterile liquid culture medium and cultured in shake flasks at 26–30℃ and 180–220 rpm for 9–13 days to obtain the liquid spawn (original culture). The resulting liquid spawn had a mycelial ball density of 13,000–14,000 cells / L, a mycelial ball diameter of 2.4–2.6 mm, and a mycelial biomass of 15.0–18.0 g / L. This fermentation broth is the liquid spawn for *Auricularia auricula-judae*.

[0064] Based on morphological observation and ITS sequence analysis, the isolated strain was confirmed to be *Auricularia auricula-judae* (short-haired wood ear fungus). Auricularia villosula This strain was named "Gan Duanmaoer No. 1". The mycelial growth of the strain after tissue isolation and purification is as follows: Figure 1 As shown. This strain was deposited at the China General Microbiological Culture Collection Center (CGMCC) on June 11, 2024, with accession number CGMCC No. 41333.

[0065] Example 2: Preparation method of Gan Duanmaoer No. 1 liquid bacterial strain I. Preparation of Culture Medium 1. Experimental group culture medium Weigh 200g of peeled potatoes, add water and boil to extract the juice. After cooling, collect the juice. Separately add 20g of glucose, 10g of tryptone, 1.5g of potassium dihydrogen phosphate and 1.0g of magnesium chloride. Make up to 1000mL with distilled water, adjust the pH to 8.0, stir thoroughly to dissolve, and dispense into 250mL Erlenmeyer flasks, with each flask containing 100mL of liquid.

[0066] 2. Control group culture medium (basal PDB medium, potato glucose broth) Boil 200g of peeled potatoes to extract the juice, add 20g of glucose, and bring the volume to 1000mL. Set the pH to natural and dispense the juice in the same way as the experimental group.

[0067] II. Experimental Methods The "Gan Duanmaoer No. 1" mycelium blocks (approximately 5mm) that had been pre-cultured on PDA plates were placed on the plates. 2 The cultures were inoculated into Erlenmeyer flasks of the experimental and control groups, and cultured at 28°C and 200 r / min for 9 days to obtain liquid cultures.

[0068] Method for determining mycelial ball density: After shaking the liquid bacterial solution well, extract 1 mL of the solution and count the number of samples. Calculate the corresponding magnification after amplification to obtain the mycelial ball density. Repeat three times and take the average value.

[0069] Method for determining mycelial biomass: Mycelial biomass is measured by mycelial dry weight. Liquid inoculum is filtered through an 80-mesh sieve, washed three times with distilled water, and dried in a 60℃ oven to constant weight before being measured.

[0070] Mycelial ball diameter determination: Take 1 mL of liquid bacterial solution, randomly select 10 mycelial balls that are closely connected and arranged in a straight line, measure their total length, calculate the average diameter of the mycelial balls, and take the average value of 3 measurements.

[0071] III. Experimental Results The effect image of the liquid strain prepared by liquid fermentation of Gan Duanmaoer No. 1 is shown in the figure. Figure 2 As shown.

[0072] In the liquid strains obtained in the experimental group, the mycelial ball density was 1350±114.64 particles / 100mL, the mycelial ball diameter was 2.54±0.14mm, and the dry mycelial biomass was 1.56±0.04g / 100mL.

[0073] Compared with the control group of basic PDB medium, the optimized medium in this example increased mycelial ball density by 62.65%, decreased mycelial ball diameter by 34.70%, and increased dry mycelial biomass by 55.06%.

[0074] The results showed that the optimized formula significantly improved the morphological characteristics and biomass accumulation efficiency of mycelial balls during the liquid fermentation of Auricularia auricula-judae.

[0075] Example 3: Cultivation of Gan Duanmao Ear No. 1 logs I. Log Cultivation Process The short-haired wood ear liquid spawn prepared in Example 1 (using the experimental group culture medium) was inoculated onto the treated logs, followed by mycelial culture, fruiting management, and harvesting.

[0076] II. Log Cultivation Methods 1. Preparation of logs Deadwood from 100 tree species was collected from the arboretum of Jiangxi Agricultural University.

[0077] The deadwood was treated as follows: Using an electric saw, the deadwood was cut into uniform lengths. Holes were drilled into the surface of each section using a 1.5 cm diameter drill bit, with a hole spacing of 5 cm and a row spacing of 6 cm. The drilled sections were then completely immersed in a 1% (w / v) lime solution for 2 days, with the lime solution changed every 12 hours. After immersion, the sections were placed in high-temperature resistant polypropylene bags and sterilized by autoclaving at 121°C and 0.1 MPa for 4 hours.

[0078] 2. Inoculation and mycelial culture Liquid inoculum (prepared in the same way as in Example 1, using the experimental group culture medium) was inoculated into the holes (about 70-80% full) on the sterilized logs, and sealed with sterilized corn cobs. The inoculated logs were then placed in a 26°C, light-proof incubation room for mycelial growth, with a cultivation period of 35-50 days, until the mycelium had completely covered the logs.

[0079] 3. Ear management and harvesting Once the mycelium has fully colonized the log and short-haired Auricularia auricula primordia are observed on the log, the bag is opened and the logs are transferred into the auriculation chamber for further cultivation. The auriculation chamber maintains approximately 50% light and 50% darkness daily (using diffused light), a temperature of 23°C, and a relative humidity of 80-95%. Ventilation is provided 3-5 times daily for 30 minutes each time to ensure sufficient oxygen supply and promote primordia differentiation. Harvesting can begin when the auricularia auricula have extended to 3-5 cm and their edges are slightly curled.

[0080] III. Ear-like effects of logs from different tree species The fruiting effect of *Earring rotundifolia* No. 1 log cultivation is shown in the following image. Figure 3 As shown.

[0081] After screening 100 tree species, the fruiting rate of some of the better-performing species is shown in Table 1. Among them, Elaeocarpus decipiens and Pterocarya stenoptera showed the best performance, with an average of 42.36±2.23 fruiting segments and 39.84±1.95 fruiting segments per log, respectively.

[0082] Table 1. Number of ear-like appendages produced by short-haired ear-like growth inoculated on logs of different tree species.

[0083] Note: Different lowercase letters represent the significance of differences between different treatments in the same column. p <0.05), the same applies below.

[0084] Example 4: Bag cultivation of Gan Duanmao Er No. 1 I. Bag Cultivation Process The short-haired black fungus liquid spawn prepared in Example 1 (using the experimental group culture medium) was inoculated into a spawn bag containing cultivation material, and mycelial culture, spawn bag opening, fruiting management and harvesting were carried out in sequence.

[0085] II. Bag Cultivation Method 1. Cultivation substrate formulation and preparation Eighteen different bag cultivation formulas were developed, with specific compositions shown in Table 2. The moisture content of all cultivation media was controlled between 60% and 65%. The prepared culture media were filled into polypropylene cultivation bags measuring 120mm × 240mm × 5mm, ensuring even compaction to avoid gaps or excessive density. After filling, the bags were placed in an autoclave and sterilized at 121℃ and 0.1MPa for 3–4 hours.

[0086] 2. Inoculation and Culture of Mushroom Bags Each bag was inoculated with 10-15 mL of liquid inoculum (prepared using the same method as in Example 1, but using the optimized culture medium from the experimental group). The inoculated bags were then transferred to a mycelium incubation room and cultured at 26°C in the dark for 30-40 days. After the mycelium had completely colonized the bags, a further ripening culture was carried out for 5-7 days to promote nutrient accumulation and enhance fruiting potential.

[0087] 3. Management of mushroom bag opening and fruiting stage After the mycelium matures, make "V"-shaped openings (4-6 openings per bag) on ​​the surface of the bag with a sterile blade. Then, place the bags upright on the shelf in the fruiting shed. The ambient temperature for fruiting should be controlled at 25℃, the relative humidity at 85-95%, and at least 8 hours of diffused light (300-500 Lux) per day. At the same time, strengthen ventilation to maintain fresh air and promote primordia differentiation.

[0088] 4. Harvesting Management Harvesting can begin when the ear lobes have stretched to 3-5 cm and the edges are slightly curled. Each bag can yield 2-3 flushes of ear lobes. After each flush, promptly remove any remaining ear base from the substrate surface, stop watering, and allow the mycelium to grow for 3-5 days before proceeding to the next flush management.

[0089] III. Biological Efficiency Analysis Biological efficiency calculation method: Fresh weight of ear pieces / Dry weight of substrate * 100% The fruiting effect of Gan Duanmao Er No. 1 bag cultivation is shown in the following figure. Figure 4 As shown.

[0090] The results of the biological efficiency analysis are shown in Table 2. The results show that the biological efficiency (i.e., biological conversion rate, defined as the percentage of fresh ear yield to dry material weight) of the 18 bag material formulations ranges from 45.85% to 65.76%. Among them, the formulations with the best biological efficiency are the formulations of group 7 and group 18, which can reach more than 65%.

[0091] Correlation analysis of the components and biological efficiency in various bag feed formulations led to the following conclusions: hardwood sawdust is a key component for ensuring high biological efficiency, with an appropriate addition ratio of 50%–65%; cottonseed hulls are suitable at a ratio of 13%–20%. Table 2. Composition of 218 bag cultivation formulations and their biological efficiency (%)

[0092] Note: Different lowercase letters represent significant differences (p<0.05) between different treatments in the same column.

[0093] Example 5: Further optimization of bag material composition Based on the research results of Example 4, the bag material formulation was further optimized. High-quality log sawdust was used to partially replace conventional mixed wood sawdust. The moisture content of all formulations was uniformly adjusted to 60%.

[0094] Formula 1 (Optimized Maple Leaf Formula, based on dry weight): 1. Maple wood chips 10%, miscellaneous wood chips 40%, cottonseed hulls 20%, wheat bran 8%, corn cobs 20%, quicklime 1%, and gypsum 1%.

[0095] 2. 20% maple wood chips, 30% miscellaneous wood chips, 20% cottonseed hulls, 8% wheat bran, 20% corn cobs, 1% quicklime, and 1% gypsum.

[0096] 3. Maple wood chips 30%, miscellaneous wood chips 20%, cottonseed hulls 20%, wheat bran 8%, corn cobs 20%, quicklime 1%, and gypsum 1%.

[0097] 4. Maple wood chips 40%, miscellaneous wood chips 10%, cottonseed hulls 20%, wheat bran 8%, corn cobs 20%, quicklime 1%, and gypsum 1%.

[0098] 5. Maple wood chips 50%, miscellaneous wood chips 0%, cottonseed hulls 20%, wheat bran 8%, corn cobs 20%, quicklime 1%, and gypsum 1%.

[0099] Formula 2 (Optimized version of Elaeocarpus decipiens, based on dry weight): 1. 10% Elaeocarpus decipiens sawdust, 40% miscellaneous sawdust, 20% cottonseed hulls, 8% wheat bran, 20% corn cob, 1% quicklime, and 1% gypsum.

[0100] 2. 20% Elaeocarpus decipiens sawdust, 30% miscellaneous sawdust, 20% cottonseed hulls, 8% wheat bran, 20% corn cob, 1% quicklime, and 1% gypsum.

[0101] 3. 30% Elaeocarpus decipiens sawdust, 20% miscellaneous sawdust, 20% cottonseed hulls, 8% wheat bran, 20% corn cob, 1% quicklime, and 1% gypsum.

[0102] 4. 40% Elaeocarpus decipiens sawdust, 10% miscellaneous sawdust, 20% cottonseed hulls, 8% wheat bran, 20% corn cob, 1% quicklime, and 1% gypsum.

[0103] 5. 50% Elaeocarpus decipiens sawdust, 0% miscellaneous sawdust, 20% cottonseed hulls, 8% wheat bran, 20% corn cob, 1% quicklime, and 1% gypsum.

[0104] A cultivation substrate formula consisting of 50% hardwood sawdust, 20% cottonseed hulls, 8% wheat bran, 20% corn cobs, 1% quicklime, and 1% gypsum was used as a control.

[0105] I. Bag Cultivation Method 1. Cultivation substrate formulation and preparation Pack the prepared culture medium into polypropylene cultivation bags measuring 120mm×240mm×5mm, 400g per bag. Compact the bags evenly during packing to avoid gaps or over-tightening. After packing, place the bags in an autoclave and sterilize at 121℃ and 0.1MPa for 3 hours.

[0106] 2. Inoculation and Culture of Mushroom Bags Inoculate each bag with 10-15 mL of liquid inoculum (preparation method as in Example 1, using the optimized culture medium from the experimental group). After inoculation, transfer the bags to a mycelium incubation room and culture at 26°C in the dark for 30-40 days. Once the mycelium has completely colonized the bags, continue the after-ripening culture for 5-7 days to promote sufficient nutrient accumulation in the mycelium.

[0107] 3. Management of mushroom bag opening and fruiting stage After the mycelium matures, make "V"-shaped openings (4-6 openings per bag) on ​​the surface of the bag with a sterile blade. Then, place the bags upright on the shelf in the fruiting shed. The ambient temperature for fruiting should be controlled at 25℃, the relative humidity at 90% or higher, and at least 10 hours of diffused light (300-500 Lux) per day should be provided. At the same time, ventilation should be strengthened to maintain fresh air and promote primordia differentiation.

[0108] 4. Harvesting Management Harvest when the ear-like structures are fully expanded and reach a diameter of approximately 5 cm. After each harvest, promptly remove any remaining ear-like structures from the substrate surface, stop watering, and allow the mycelium to mature for 3 days before proceeding with the management of the next harvest.

[0109] II. Experimental Results The fruiting results of adding Gan Duanmao Er No. 1 fruiting agent to a mixture of maple / Elaeocarpus decipiens sawdust and miscellaneous sawdust are as follows: Figure 5 As shown.

[0110] The optimized formula was used in a bag cultivation experiment, and the biological efficiency was analyzed according to the method in Example 4. The results are shown in Tables 3 and 4. The results showed that the average yield of fresh ear fungus per bag reached 130g, and the biological efficiency (fresh ear fungus weight / dry material weight × 100%) reached 75%~83%, which was significantly better than the traditional hardwood sawdust control group (67.36%, p<0.05). This indicates that high-quality log sawdust (especially Elaeocarpus decipiens and Pterocarya stenoptera) can effectively improve the bioconversion efficiency of the substrate and has significant application value.

[0111] Table 3. Effects of different proportions of maple wood chips replacing miscellaneous wood chips on biological efficiency (%)

[0112] Note: *, ** and *** indicate significant correlation with the control at the 0.05, 0.01 and 0.001 levels, respectively, and the same applies below.

[0113] Table 4. Effects of different proportions of *Elaeocarpus decipiens* wood chips replacing hardwood chips on biological efficiency (%)

[0114] The above embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above embodiments. Any changes, modifications, substitutions, combinations, or simplifications made without departing from the spirit and principle of the present invention shall be considered equivalent substitutions and shall be included within the protection scope of the present invention.

Claims

1. A method for bag cultivation of short-haired wood ear fungus, characterized in that, A method for cultivating short-haired wood ear mushrooms using a bag substrate; the bag substrate, by dry weight percentage, contains the following components: 20-40% maple sawdust or 40-50% Elaeocarpus decipiens sawdust, 0-30% miscellaneous wood sawdust, 13-20% cottonseed hulls, 8-20% wheat bran, 0-20% corn cob, 0.5-1.5% quicklime, and 0.5-1.5% gypsum; The short-haired wood ear fungus is short-haired wood ear fungus ( Auricularia villosula The strain 1 of *Eurys nigra* was deposited at the China General Microbiological Culture Collection Center on June 11, 2024, with the accession number CGMCC No. 41333.

2. A method for cultivating short-haired wood ear fungus on logs, characterized in that, Short-haired wood ear fungus is cultivated using logs, the variety of which is *Elaeocarpus decipiens* (Elaeocarpus spp.). Elaeocarpus glabripetalus ), maple ( Pterocarya stenoptera ), Privet ( Ligustrum lucidum ), Chinese scholar tree ( Styphnolobium japonicum Any one of the following; The short-haired wood ear fungus is short-haired wood ear fungus ( Auricularia villosula The strain 1 of *Eurys nigra* was deposited at the China General Microbiological Culture Collection Center on June 11, 2024, with the accession number CGMCC No. 41333.

3. The application of the bag cultivation method of claim 1 or the log cultivation method of claim 2 in the production of short-haired black fungus; The short-haired wood ear fungus is short-haired wood ear fungus ( Auricularia villosula The strain 1 of *Eurys nigra* was deposited at the China General Microbiological Culture Collection Center on June 11, 2024, with the accession number CGMCC No. 41333.

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