Shiitake mushroom strain 'Shanghai Xiang F7' and intensive production method thereof

Abstract

The invention relates to a shiitake mushroom strain 'Shanghai Xiang F7' suitable for an intensive production mode and a cultivation method for factory-like stick manufacturing and facility fruiting of the shiitake mushroom strain 'Shanghai Xiang F7'. The 'Shanghai Xiang F7' variety has the characteristics of short mushroom age, easiness in fruiting, neat fruiting, obvious moisture, low sporocarp malformation rate, good commodity and the like, is more suitable for an intensive production mode of'factory-like stick preparation and facility-like fruiting ', can realize the industrial production advantages of high efficiency, standardization and high quality of factory-like stick preparation in the mode, and also has the advantages of low production cost and the like. The production mode has the characteristics of low cost, energy conservation, long fruiting period and high quality, is higher in risk resistance, higher in economic benefit and more suitable for shiitake mushroom production in China at the present stage, and easily embodies the effect of combining farmers with farmers.

Description

Technical Field

[0001] The present invention relates to the technical field of mushroom cultivation, and more particularly to a shiitake mushroom strain "Huxiang F7" and an intensive production method thereof. Background Art

[0002] Shiitake mushrooms are the most produced and consumed edible fungi in my country. China is a major producer of edible fungi. According to statistics from the China Edible Fungi Association, the total output of edible fungi nationwide in 2023 reached 43.3417 million tons (fresh products, the same below), a year-on-year increase of 2.64%. The total output value in 2022 reached 396.557 billion yuan, a year-on-year increase of 2.02%. The edible fungi industry has the "Five Non-Competition" characteristic: it does not compete with humans for grain, land, fertilizer, time, or resources. Shiitake mushrooms are considered my country's "national mushroom." With low investment and rapid returns, the industry has reached a scale of hundreds of billions of yuan and plays a significant role in my country. Traditional shiitake mushroom stick bag production primarily relies on small, family-run workshops. Preparation, stick making, sterilization, and inoculation are all performed manually by individual households. This is time-consuming, labor-intensive, inefficient, and subject to significant waste, making standardization and scalability difficult. The stick cultivation process primarily involves natural spawning, which is susceptible to extreme weather conditions like high temperatures, leading to stick burn, high contamination rates, and severe stick rot. With rising labor costs, an aging population of mushroom farmers, and increasing environmental protection requirements in my country, the traditional family-run workshop-based shiitake mushroom production model is struggling to adapt to the needs of the country's mushroom industry.

[0003] Therefore, this field urgently needs to develop shiitake mushroom cultivation methods and their supporting cultivation strains suitable for my country's current national conditions, so as to achieve high-efficiency, low-cost and standardized production needs and support the transformation and upgrading of my country's shiitake mushroom industry. Summary of the Invention

[0004] The purpose of the present invention is to provide a shiitake mushroom production method and its supporting cultivation strains suitable for the current national conditions in my country, so as to achieve high-efficiency, low-cost and standardized production requirements and support the transformation and upgrading of my country's shiitake mushroom industry.

[0005] In a first aspect of the present invention, a shiitake mushroom strain suitable for intensive production is provided. The strain is classified and named as shiitake mushroom "Huxiang F7" Lentinula edodes, and is preserved in Guangdong Provincial Microbiological Culture Collection Center with a preservation number of GDMCC No: 66428 and a preservation date of May 28, 2025.

[0006] In a specific embodiment, the bacterial colony biological characteristics of the shiitake mushroom strain are as follows: the mycelium colony is white in color, the mycelium grows rapidly, and generally can cover the plate in 7-8 days at 23°C; the mycelium is flocculent and relatively loose; the mycelium grows neatly with relatively rounded edges; the aerial mycelium is less, short, and relatively sparse.

[0007] In a specific embodiment, the mycelial microstructure of the shiitake mushroom strain is as follows: the mycelial diameter is between 3.4-4.5 μm, the thickness is uniform, the mycelial diameter is colorless and transparent, the mycelial diameter is tubular, the mycelial diameter is relatively small, the mycelial wall is smooth and slightly thick, and the mycelial wall has a distinct lock-like structure.

[0008] In a specific embodiment, the fruiting body characteristics of the shiitake mushroom strain are as follows: the fruiting body is solitary and round; the cap is brown, the cap diameter is 6.33±0.57 cm, the mushroom shape grade is medium to large, the average single mushroom weight is 26.8±2.72 g, and the top of the cap longitudinal section is convex; the scales on the surface of the cap are light brown and medium in size; the stipe is columnar, the stipe length is 5.22±0.47 cm, which is medium in length; the surface hairs are abundant and light brown; the gills are regularly arranged and flat, and the fruiting body deformity rate is low.

[0009] In a second aspect of the present invention, a molecular marker identification method for the Lentinus edodes strain according to the first aspect of the present invention is provided, comprising culturing and collecting mycelium, extracting genomic DNA, amplifying specific fragments by PCR, detecting by electrophoresis, and displaying specific bands;

[0010] The specific nucleotide sequences of the specific fragment PCR amplification primers Lefp_id001 to Lefp_id007 are:

[0011] The forward sequence of primer Lefp_id001 is shown in SEQ ID NO: 1: 5′-ACGGATGGAGAGACACAGGA-3′, and the reverse sequence is shown in SEQ ID NO: 2: 5′-TGCCCCACTTACTCTCAACC-3′;

[0012] The forward sequence of primer Lefp_id002 is shown in SEQ ID NO: 3: 5′-CCTTTCTCAAAAGCGGACTG-3′, and the reverse sequence is shown in SEQ ID NO: 4: 5′-GGGAGTGGGTTGTTTGGATA-3′;

[0013] The forward sequence of primer Lefp_id003 is shown in SEQ ID NO: 5: 5′-CGGATGTTATGCACTGGAAG-3′, and the reverse sequence is shown in SEQ ID NO: 6: 5′-CGTACGGTTGGACATTTGAA-3′;

[0014] The forward sequence of primer Lefp_id004 is shown in SEQ ID NO: 7: 5′-AGCCTTTCACAGTCAGCTCG-3′, and the reverse sequence is shown in SEQ ID NO: 8: 5′-GTCAACGGAGGGAAACAGAG-3′;

[0015] The forward sequence of primer Lefp_id005 is shown in SEQ ID NO: 9: 5′-GTAGCACTCCTCATACAACC-3′, and the reverse sequence is shown in SEQ ID NO: 10: 5′-ACCAAATGTCACAGCACAGG-3′;

[0016] The forward sequence of primer Lefp_id006 is shown in SEQ ID NO: 11: 5′-ACATTGGCGAAGGCTGTACG-3′, and the reverse sequence is shown in SEQ ID NO: 12: 5′-CCCTTTTGCCCTATAAGGCG-3′;

[0017] The forward sequence of primer Lefp_id007 is shown in SEQ ID NO: 13: 5'-AACAGTAACCTGTGCACTGC-3', and the reverse sequence is shown in SEQ ID NO: 14: 5'-CATGATCAGATCACACAGCG-3'.

[0018] In a specific embodiment, the primer Lefp_id001 produces a 203 bp band, the primer Lefp_id002 produces a 236 bp band, the primer Lefp_id003 produces a 214 bp band, the primer Lefp_id004 produces a 270 bp band, the primer Lefp_id005 produces two bands of 346 bp and 378 bp, the primer Lefp_id006 produces two bands of 262 bp and 277 bp, and the primer Lefp_id007 produces a 284 bp band. The simultaneous detection of the seven primers obtains DNA marker bands of corresponding sizes, which are the markers of the shiitake mushroom "Huxiang F7".

[0019] In a specific embodiment, the PCR amplification reaction system includes: 10 μL of 2×Es Premix TaqTM, 1 μL each of forward primer and reverse primer (10 μmol / L), 2 μL of template DNA (20-30 ng / μL), and ddH2O added to 20 μL.

[0020] In a specific embodiment, the PCR amplification reaction procedure is: 94°C for 5 min; 94°C for 30 s, 55-60°C (preferably 58°C or 60°C) for 45 s, 72°C for 30 s, 35 cycles; 72°C for 7 min; and storage at 4°C.

[0021] In a third aspect of the present invention, there is provided an intensive production method of the shiitake mushroom strain of the first aspect of the present invention, comprising the following steps: preparing raw materials, preparing mushroom sticks and high-pressure sterilization, mechanical inoculation, factory-scale cultivation, and facility-based mushroom fruiting management;

[0022] The preparation of raw materials includes preparing bacterial strains and cultivation substrates;

[0023] The preparation of bacterial strains includes preparing solid bacterial strains of "Huxiang F7";

[0024] The formula of the cultivation matrix comprises:

[0025] (a) Stock formulation: PDA medium containing 200 g peeled potatoes, 20 g glucose, 18-20 g agar, and 1000 mL water;

[0026] (b) a cultivar formulation comprising the following ingredients in a mass-to-volume ratio: sawdust 78%, wheat bran 20%, sucrose 1%, gypsum 1%, a moisture content of 60%-65%, and a pH of 5.5-6.5;

[0027] (c) A mushroom log production formula comprising the following ingredients in a mass-to-volume ratio:

[0028] (c1) oak sawdust 75-85wt% (preferably 78wt%), bran 15-25wt% (preferably 20wt%), gypsum 1-3wt% (preferably 2wt%), moisture content 50%-55%, pH 6-7 before sterilization;

[0029] (c2) oak sawdust 40-50wt% (preferably 48wt%), fruit sawdust 20-40wt% (preferably 30wt%), bran 15-25wt% (preferably 20wt%), gypsum 1-3wt% (preferably 2wt%), water content 53%-58%, pH 6-7 before sterilization.

[0030] In a specific embodiment, the oak sawdust or fruit sawdust is crushed into a particle size of 0.5 cm-1.0 cm and is composted and fermented for more than 10 days.

[0031] In a specific embodiment, the cultivation matrix further comprises rice bran, corn flour, lime and calcium carbonate as auxiliary materials, and the moisture content of the auxiliary materials does not exceed 14%.

[0032] In a specific embodiment, the preparation of bacterial sticks and sterilization comprises the following steps:

[0033] (a) using an automatic bagging machine to fill the cultivation substrate into bags, and mechanically tying the bags to obtain material sticks;

[0034] (b) stacking the material rods into a sterilization container and performing high-pressure sterilization;

[0035] The high-pressure sterilization temperature is 115-120° C., and the sterilization time is 300-1000 min.

[0036] In a specific embodiment, the bagging requirements include: based on a 17 cm modified polyethylene bag, the weight of the cultivation substrate filled in is 2790-2810 g, so that the material sticks are compacted.

[0037] In a specific embodiment, after the mechanical tying, 1-2 exhaust holes are pierced near the inoculation hole, and a breathable tape is affixed to prepare a material stick.

[0038] In a specific embodiment, the material rods are stacked on a sterilization rack, with the upper and lower layers of material rods placed in a herringbone shape. The exhaust holes of the bottom layer of material rods need to be tilted 45° downward, and the exhaust holes of the upper layer of material rods face directly downward. The exhaust holes are all facing the gap to avoid being blocked.

[0039] In a specific embodiment, the packed material sticks should be sterilized in time, and the time from adding water and stirring to the completion of bagging and starting sterilization should be controlled within 4 hours.

[0040] In a specific embodiment, the mechanical inoculation comprises the following steps:

[0041] (1) The high-pressure sterilized rods are transferred to a 10,000-level cleanroom for the first cooling and the second cooling; wherein,

[0042] The first cooling is carried out by introducing fresh air with high efficiency filtration to cool the rod to 40-55°C;

[0043] The second cooling is carried out by using a refrigeration fan to quickly cool the material bar to below 26°C;

[0044] (2) Transfer the cooled rods to the inoculation room and use an inoculation machine for inoculation;

[0045] The inoculation step comprises: using a fully automatic shiitake mushroom solid inoculator to inoculate the solid spawn onto a material stick to obtain a mushroom stick; inoculating 3-4 holes with the solid spawn, with an inoculation depth of 3.5 cm-5 cm and an inoculation weight of 15 g-20 g per hole.

[0046] In a specific embodiment, the factory culture comprises the following steps:

[0047] (1) Early culture stage of bacterial sticks: Within 13-15 days after inoculation, the bacterial sticks are stacked on the culture rack and placed in a 10,000-class purified pre-culture room. The temperature of the pre-culture room is controlled at 20°C-21°C and the humidity is 70%-80%. Culture is carried out in the dark. Mycelium germination is promoted in a clean environment and the inoculation port is sealed.

[0048] (2) Mid-stage of bacterial stick cultivation: After 13-15 days of bacterial stick cultivation, when the diameter of the inoculated hole is 6cm-8cm, the bacterial sticks are transferred to the shelves for "tic-tac-toe" stacking or grid racks for cultivation, with a density of 130-150 sticks / m 2 The diameter of the fungus ring is 10cm-15cm. When mycelium can be seen on the back of the inoculation hole, the first puncture is performed manually or mechanically. 10-15 holes are punctured at each inoculation hole, with a hole depth of 2.0cm-2.5cm.

[0049] About 5 days after the mushroom sticks are full of bags, use a puncture machine to perform a second puncture, with about 150 holes per stick, the hole depth is 5.0 cm-7.0 cm, the puncture holes should be arranged neatly and the spacing should be even; in the middle stage of the mushroom stick culture, the mushroom sticks should be cultured in the dark because the heat generated by the mushroom sticks is high, especially 1-2 days after puncture, the stick temperature is 2℃-4℃ higher than the room temperature, so the culture room temperature needs to be strictly controlled, and the room temperature should be controlled at 21℃-22℃;

[0050] (3) Late culture stage of bacterial sticks: After the second puncture for oxygenation, light is provided to promote the color change of bacterial sticks;

[0051] The intensity of the illumination is 50 lux to 200 lux, the light quality of the light is selected from white light (RBG) or blue light (B), and the illumination time is no less than 12 hours per day.

[0052] In a specific embodiment, during the color change culture of the bacterial sticks, the temperature of the bacterial sticks is 2°C-3°C higher than the room temperature, and the room temperature is controlled at 21°C-22°C;

[0053] After the color change culture of the bacterial sticks is completed, the respiration of the bacterial sticks is weakened and the heat generation is reduced. The stick temperature of the bacterial sticks is 0.5°C-1°C higher than the room temperature, and the room temperature is controlled at 23-24°C.

[0054] In a specific embodiment, the facility-based mushroom production management includes the following steps:

[0055] (1) Constructing a facility-based mushroom shed: The facility-based mushroom shed is 2-3 m high (preferably 2.4 m), with galvanized pipes forming the shed frame, and plastic film and sunshade nets covering the shed frame. The shed is narrow from north to south and long from east to west to facilitate air circulation;

[0056] The facility-based mushroom shed has built-in multi-layer cultivation racks, which are used to place the mushroom sticks. The cultivation racks have the following structure: 5-6 layers, a layer spacing of 0.3-0.4 m, a bottom layer height of 0.15-0.2 m, a rack width of 0.40-0.45 m, two rows of cultivation racks in the middle, a row of cultivation racks on each side, and a width of the operating aisles on the left and right sides of 0.6-0.7 m.

[0057] In a specific embodiment, several dry and wet thermometers are hung at different locations of the facility-based mushroom shed to monitor and control the temperature and humidity of the mushroom shed at any time. The temperature of the mushroom shed is preferably controlled at 8-25°C, and the humidity is preferably controlled at 70%-90%. The mushroom shed is surrounded by an open area of ​​2 m to facilitate ventilation.

[0058] (2) Remove the mushroom sticks from the bags after cultivation. Keep the temperature in the fruiting shed at 8-20℃ and the humidity at 80%-90%. Spray water to moisten the mushroom sticks. After 5-6 days, buds will appear. Thin out the buds and leave 12-15 buds. Harvest after 4-6 days.

[0059] After harvesting, the temperature is controlled at 20-25°C, the humidity is 70-80%, and the CO2 concentration is 500-1500ppm. The mushrooms are allowed to rest for 15-20 days. Then, the temperature is controlled at 12-20°C, the humidity is 80-90%, and the CO2 concentration is 500-1500ppm to stimulate bud formation. Water is sprayed to humidify the mushroom sticks. After bud formation and mushrooms are produced, the second wave of harvesting is carried out. Repeat the above operation after harvesting for 3-5 waves of harvesting.

[0060] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features described in detail below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be listed here one by one.

[0061] Compared with the prior art, the present invention has the following technical effects:

[0062] The variety "Huxiang F7" has the characteristics of short mushroom age, uniform fruiting, obvious tide times, low fruiting body deformity rate and good commercial quality. It is more suitable for the intensive production model of "factory-based stick making and facility-based mushroom production". Under this model, it can not only achieve the industrial production advantages of high efficiency, standardization and high quality of factory-based mushroom stick preparation, but also achieve the characteristics of low cost, energy saving, long fruiting cycle and high quality of facility-based mushroom production. The production has stronger risk resistance, higher economic benefits, and is easier to reflect the effect of linking farmers and leading farmers, which is more suitable for my country's current shiitake mushroom production. BRIEF DESCRIPTION OF THE DRAWINGS

[0063] Figure 1What is shown is the InDel marker fingerprint of the strain "Huxiang F7" of the present invention, wherein M represents marker, numbers 1-7 represent Lefp_id001~Lefp_id007 respectively, and the bands indicated by the arrows are the positions of the bands corresponding to different primers.

[0064] Figure 2 Shown is a flow chart for the breeding of the shiitake mushroom "Huxiang F7" strain of the present invention.

[0065] Figure 3 Shown are the results of the relevant characteristics of the shiitake mushroom "Huxiang F7" strain of the present invention, wherein A is a colony biological characteristic diagram of the shiitake mushroom "Huxiang F7", B is a mycelium microstructure diagram of the shiitake mushroom "Huxiang F7", with a scale of 50um, and C is a fruiting body characteristic diagram of the shiitake mushroom "Huxiang F7".

[0066] Figure 4 Shown is the main production flow chart of the intensive production of shiitake mushroom "Huxiang F7" of the present invention.

[0067] Figure 5 Shown is a diagram of the stacking of pretreated sawdust in an embodiment of the present invention.

[0068] Figure 6 Shown are pictures of the mushroom sticks after mid-term culture puncture in an embodiment of the present invention, wherein A is the result of the first puncture, and B is the result of the second puncture.

[0069] Figure 7 Shown is a diagram of the three-dimensional mushroom production effect of a greenhouse facility in an embodiment of the present invention.

[0070] Figure 8 Shown is a diagram of the mushroom production effect of a simple greenhouse in an embodiment of the present invention. DETAILED DESCRIPTION

[0071] To make the purpose, technical solutions, beneficial effects, and significant improvements of the embodiments of the present invention more clear, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, all the embodiments described are only some embodiments of the present invention, not all embodiments; based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0072] the term

[0073] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0074] As used herein, the term “including” or “comprising” encompasses “comprising,” “consisting mainly of,” “consisting essentially of,” and “consisting of;” “consisting mainly of,” “consisting essentially of,” and “consisting of” are subordinate concepts of “containing,” “having,” or “including.”

[0075] Intensive production

[0076] The intensive production of shiitake mushroom sticks can centrally purchase large-scale machinery and equipment, and provide supporting environment-controlled spawning conditions, which greatly improves the production efficiency of mushroom sticks, standardizes the production standards of mushroom sticks, and is conducive to the promotion of improved varieties and methods.

[0077] Intensive shiitake mushroom production is a modern agricultural production model that achieves efficient, high-yield, high-quality, and sustainable shiitake mushroom production by centrally allocating land, capital, facilities, equipment, varieties, and technologies. It employs a "factory-based stick production and facility-based fruiting" model. Under this model, shiitake mushroom factory-based stick production companies can centrally purchase large-scale facilities and equipment, build standardized production plants, and provide environmentally controlled spawning conditions. This significantly improves the efficiency and standardization of spawning stick production. Furthermore, facility-based fruiting can extend the fruiting period suitable for shiitake mushrooms based on local climatic conditions through environmental control in greenhouses, achieving low-cost, high-quality shiitake mushroom production. This model also facilitates the promotion of improved varieties and methods. The core of intensive production lies in leveraging scale, standardization, mechanization, and scientific management methods to minimize investment while improving resource utilization and production efficiency, reducing production costs and enhancing industry competitiveness.

[0078] Shiitake mushroom strain "Huxiang F7"

[0079] The present invention relates to an InDel marker fingerprint of a shiitake mushroom strain "Huxiang F7" and an intensive production method. The fingerprint consists of 7 pairs of InDel markers developed based on whole-genome sequencing of shiitake mushrooms.

[0080] The intensive production method includes two main links: "factory stick making" and "facility-based mushroom fruiting". The strain and production method can be used to realize factory stick making in the production and cultivation stages of shiitake mushroom sticks. In the later fruiting stage, low-cost facility-based mushroom fruiting can be carried out according to local conditions based on the ecological environment and facility and equipment conditions of each place. This production method can realize a modern shiitake mushroom production technology system with unified procurement of raw materials, automated production of mushroom sticks, environmentally controlled fermentation, and facility-based mushroom fruiting as the core for large-scale shiitake mushroom production. It can not only realize the industrial production advantages of high efficiency, standardization, and high quality in factory-based mushroom stick preparation, but also realize the characteristics of low cost, energy saving, long fruiting cycle, and high quality in facility-based mushroom fruiting. It can effectively reduce production costs, improve the quality of mushroom stick production, make production more resistant to risks, have higher economic benefits, and more easily reflect the effect of linking farmers and leading farmers, and is more suitable for my country's current shiitake mushroom production.

[0081] The present invention provides an InDel marker fingerprint of the "Huxiang F7" strain suitable for intensive production of Lentinus edodes. The fingerprint consists of seven pairs of InDel markers developed based on InDel marker primers for insertion / deletion fragments in the Lentinus edodes genome. The amplification band pattern is good and highly reproducible. Detailed information on the markers is shown in Table 1.

[0082] Table 1: InDel marker primer information list

[0083]

[0084] PCR amplification reaction system: 2×Es Premix TaqTM 10 μL, forward primer, reverse primer (10 μmol / L) 1 μL each, template DNA (20-30 ng / μL) 2 μL, supplemented with ddH2O to 20 μL.

[0085] PCR amplification reaction program: 94°C for 5 min; 94°C for 30 s, 55 / 58 / 60°C for 45 s, 72°C for 30 s, 35 cycles; 72°C for 7 min; and storage at 4°C.

[0086] Electrophoresis detection process of PCR products: 7 μL of the product obtained by the above PCR amplification was spotted on an agarose gel with nucleic acid dye added for electrophoresis. The volume percentage concentration of the agarose gel was 3%, the electrophoresis buffer was 1xTAE, the voltage was 90 V, the current was 300 mA, the power was 100 W, the electrophoresis was performed for 5 h, and the gel imaging system was used to take pictures and analyze the results.

[0087] The present invention also provides an application of the InDel marker fingerprint map of Lentinus edodes F7 strain, which is to use 7 pairs of InDel marker primers developed by using the insertion / deletion fragments of the Lentinus edodes genome to perform InDel marker amplification on the Lentinus edodes strain, and the corresponding size bands are obtained, which are Lentinus edodes F7 strains; Figure 1 As shown, M represents Marker, 1-7 represent primers Lefp_id001~007, among which,

[0088] The primer Lefp_id001 produced a 203 bp band,

[0089] The primer Lefp_id002 produced a 236 bp band,

[0090] The primer Lefp_id003 produced a 214 bp band,

[0091] The primer Lefp_id004 produced a 270 bp band,

[0092] The primer Lefp_id005 produced two bands of 346 bp and 378 bp,

[0093] The primer Lefp_id006 produced two bands of 262 bp and 277 bp,

[0094] The primer Lefp_id007 produced a 284 bp band,

[0095] The DNA marker bands of corresponding sizes were obtained by simultaneous detection of 7 primers, which are the markers of the shiitake mushroom "Huxiang F7".

[0096] Production facilities and equipment

[0097] The site selection for industrial mushroom stick production generally requires flat terrain, convenient irrigation and drainage, and easy transportation. The mushroom stick production plant must have a wood chip yard, raw material warehouse, mixing area, bagging area, sterilization area, cooling area, inoculation area, culture area, cold storage, boiler room, and air conditioning room. These areas are typically semi-enclosed.

[0098] Factory-made mushroom sticks

[0099] Factory-based shiitake mushroom stick production utilizes modern shiitake stick production equipment and technology to produce shiitake mushroom sticks (culture medium) on a large scale and in a standardized manner. Compared to traditional manual stick production, this method offers advantages such as high efficiency, consistent quality, and low contamination, making it suitable for large-scale shiitake mushroom stick production.

[0100] Raw material preparation and production formula

[0101] The cultivation medium is the basis of shiitake mushroom mycelium, providing a suitable growth environment and carbon source, nitrogen source and mineral elements for the growth of shiitake mushroom mycelium. The physical and chemical properties of the medium, such as particle size, air permeability, pH value and water content, are also important factors affecting the growth of shiitake mushrooms. Factory-based stick making can realize the unified procurement and processing of raw and auxiliary materials under mechanized and large-scale production. Combined with the characteristics of production seeds, environmental control of spawning and facility-based mushroom production requirements, new raw materials suitable for this model are developed and utilized, which facilitates the standardized preparation of raw materials.

[0102] Large-scale factory-made mushroom sticks generally require that the sawdust be uniformly crushed into a particle size of 0.5 cm-1.0 cm and then piled and fermented for more than 10 days to soften the sawdust, increase its water holding capacity, and reduce the probability of micropore formation when bagging using a bagging machine, thereby improving the yield of the sticks.

[0103] The main auxiliary materials include wheat bran, rice bran, corn flour, gypsum, lime, and calcium carbonate. These must be fresh, clean, dry, free of insects, lumps, mildew, odor, and impurities, with a moisture content not exceeding 14%. Recommended recipes: Recipe 1: 78% oak sawdust, 20% wheat bran, 2% gypsum, 50%-55% moisture content, pH 6-7 before sterilization; Recipe 2: 48% oak sawdust, 30% fruit sawdust, 20% wheat bran, 2% gypsum, 53%-58% moisture content, pH 6-7 before sterilization.

[0104] When selecting a cultivation formula, factors such as raw material source, fruiting pattern, and market demand should be considered. Domestic fruiting patterns typically produce 5-6 waves of mushrooms. Formula 1 produces mushroom sticks with strong fruiting potential and high yields, making it suitable for fruiting in self-built production bases and supplying mushroom sticks to meet domestic market demand. Formula 2 produces mushroom sticks with a yield concentrated in the first 2-3 waves, making it suitable for factory-based and year-round fruiting, primarily targeting the needs of the international market, and generally producing 2-3 waves of mushrooms.

[0105] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are intended to illustrate the present invention only and are not intended to limit the scope of the present invention. The experimental methods in the following examples, for which specific conditions are not specified, are generally performed under conventional conditions such as those described in Sambrook et al., Molecular Cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or according to the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are calculated by weight. The experimental materials and reagents involved in the present invention can be obtained from commercial sources unless otherwise specified.

[0106] Example 1: Breeding of Lentinus edodes "Huxiang F7" strain

[0107] "Huxiang F7" is a hybrid bred variety. Its parents are "Huxiang F3", a mushroom factory cultivation variety bred by Shanghai Academy of Agricultural Sciences, and "0912", a main cultivated variety in northern my country (see the breeding process for the details). Figure 2 ).

[0108] "Huxiang F3" is a special variety selected and bred by the Shanghai Academy of Agricultural Sciences for the "briquetting" model of factory-based shiitake mushroom production. It has now become one of the main cultivated varieties of factory-based shiitake mushrooms in China and is widely used in production areas such as Henan, Shandong, Hubei, and Hebei. This variety has the advantages of uniform fruiting, obvious tides, and high bioconversion rate of two-tide mushrooms, but it has defects such as a slightly long fungus age and occasional deformities. The energy consumption is relatively high under factory-based production conditions, especially in the summer production process in the southern region; the "0912" strain is mainly used in Hebei, Liaoning, Inner Mongolia and other places. This variety has the advantages of a short fungus age, a relatively round mushroom shape, and a low deformity rate, but it has the disadvantages of a low fruiting temperature and a large ventilation requirement. The energy consumption is relatively high for factory-based production in summer. To further develop varieties suitable for intensive production with shorter spawn ages, higher temperatures during the fruiting period, uniform fruiting, and a low deformity rate, thereby reducing energy consumption in both spawn production and fruiting facilities and increasing the proportion of high-quality mushrooms, the team used single-spore hybridization to improve the "Huxiang F3" variety by introducing the "0912" variety. The resulting hybrid progeny resulted in the new "Huxiang F7" variety. Compared to its two parents, "Huxiang F7" has a spawn age of 85-90 days, over five days shorter than "Huxiang F3," and a temperature profile over 3°C higher than "0912." It can consistently fruit at temperatures between 18-22°C, with distinct tides, uniform fruiting, and a fruiting body deformity rate below 1%.

[0109] Lentinus edodes "Huxiang F7" is preserved in Guangdong Provincial Microbiological Culture Collection Center with the preservation number GDMCC No: 66428 and the preservation date of May 28, 2025.

[0110] The biological characteristics of the "Huxiang F7" strain are as follows: the mycelial colony is white in color, the mycelial growth rate is relatively fast, and it can generally cover the plate in 7-8 days at 23°C. The mycelial colony is flocculent and relatively loose, the mycelial growth is neat, the edges are relatively round, and the aerial mycelial colony is relatively few, short, and sparse. Figure 3 A).

[0111] The mycelial microstructure of the "Huxiang F7" strain is as follows: the mycelial diameter is between 3.4-4.5 μm, the thickness is uniform, the mycelial is colorless and transparent, the mycelial is tubular, with few branches, the mycelial wall is smooth, the wall is slightly thick, and the lock-like structure is obvious ( Figure 3 B).

[0112] The fruiting body characteristics of the strain "Huxiang F7" are as follows: the fruiting body is solitary and round; the cap is brown, with a cap diameter of 6.33±0.57 cm, the mushroom shape is medium to large, the average single mushroom weight is 26.8±2.72 g, and the top of the cap is convex in the longitudinal section; the scales on the cap surface are light brown and medium in size; the stipe is columnar, with a stipe length of 5.22±0.47 cm, which is medium in length; the surface has abundant light brown cilia; the gills are regularly arranged and flat, and the fruiting body deformity rate is low ( Figure 3C).

[0113] Example 2: Intensive production method of shiitake mushroom "Huxiang F7"

[0114] The main production process of the intensive production method of the present invention, which is applicable to the "factory-based stick making and facility-based mushroom fruiting" of shiitake mushroom "Huxiang F7", is as follows: Figure 4 As shown, it includes:

[0115] Raw material preparation, mechanized mixing, automatic stick making (factory stick making), high-pressure sterilization, forced cooling, mechanical inoculation, environmentally controlled culture (environmentally controlled fungus cultivation), mechanical puncture, color change culture, bagging and shelving, mushroom management and timely harvesting.

[0116] The intensive production method of shiitake mushroom "Huxiang F7" comprises the following steps:

[0117] (1) Production of Shiitake Mushroom "Huxiang F7"

[0118] 1) Preparation process of shiitake mushroom mother culture:

[0119] Mother seed formula (PDA medium): 200 g peeled potatoes, 20 g glucose, 19 g agar, and 1000 mL water.

[0120] Preparation: Peel the potatoes and boil them for 20 minutes. Filter the juice, dissolve it in glucose and agar, and aliquot into test tubes (15 x 150 mm), approximately 7 mL per tube. Seal the tubes with cotton plugs and sterilize them by autoclaving at 121°C for 30 minutes. While still hot, shape the tubes into a slant (approximately half the length of the test tube). When the PDA culture medium temperature drops below 20°C, use a flame-sterilized inoculation needle to remove a 5 mm x 5 mm inoculum block and quickly insert it into the center of the PDA slant. Place the inoculated tubes in a 23°C incubator in the dark for 8 days. Once mycelium has grown all over the slant, with neat edges and no contamination from other bacteria, they can be used for cultivation and inoculation.

[0121] 2) Cultivar preparation process:

[0122] Formula: 78% sawdust, 20% wheat bran, 1% sucrose, 1% gypsum, 63% water content, pH 5.7

[0123] Preparation: Mix the main ingredients (such as sawdust) and auxiliary materials in appropriate proportions. Add water and stir thoroughly. Test the moisture content (use your hands to squeeze the culture medium until water droplets appear between your fingers, but no water drips). The pH is ideal. Use 17cm × 33cm × 0.05cm denatured polyethylene plastic bags (Yingkou Zhengda Industrial Co., Ltd.) and pack approximately 500g of dry material (1.2kg wet weight) into each bag. Ensure the bag is tightly packed (flexible to the touch) to prevent damage to the bag walls. Seal the bag with a ring. Place the filled bag in an autoclave at 121°C for 2.5 hours. After sterilization, remove the bag while it's still hot and transfer it to a sterile room or inoculation box, where it will be cooled to below 25°C for inoculation. Open the bag near an alcohol lamp and use tweezers or dig out a jujube-sized mycelium block (with a small amount of culture medium) from the mother or original seed. Inoculate the bag with the bacteria and culture it in a culture room at 22-24°C and 60%-70% humidity. Check for contamination 10 days after inoculation. If the bag is contaminated, immediately remove it and remove it from the culture room. Cultivate the bag for 30 days until it is full and ready for use.

[0124] (2) Raw material preparation and production formula

[0125] Sawdust pretreatment process:

[0126] Crush the sawdust into 0.5 cm-1.0 cm particle size, spread the sawdust to a thickness of 30 cm-40 cm, water it until water flows out from the bottom, and then build a pile for fermentation for more than 10 days ( Figure 5 ), which is conducive to softening the sawdust, increasing the water holding capacity of the sawdust, and reducing the probability of micropores forming during bagging by the bagging machine, thereby improving the yield of rod making.

[0127] The main auxiliary materials for shiitake mushroom production are wheat bran, rice bran, corn flour, gypsum, lime and calcium carbonate. Wheat bran, rice bran and corn flour must be fresh, clean, dry, free of insects, lumps, mildew, odor and impurities, and the moisture content must not exceed 14%.

[0128] Production formula: oak sawdust 48%, apple sawdust 30%, bran 20%, gypsum 2%, moisture content 55%, pH 6.

[0129] The effects of sawdust pretreatment on the factory-scale production of shiitake mushrooms under the intensive production model of shiitake mushrooms are shown in Table 2.

[0130] Table 2: Effects of sawdust pretreatment on shiitake mushroom log production

[0131]

[0132] The results in Table 2 show that when the sawdust was pretreated, the microporosity was below 1% (0.8%), the contamination rate was below 3‰ (2.3‰), and the spawn consistency was high. When the sawdust was not pretreated, the microporosity was above 5% (5.8%), the contamination rate was above 5% (6.3%), and the spawn consistency was poor. Therefore, the sawdust was pretreated during the production process of the mushroom sticks of the present invention.

[0133] (3) Factory rod making and high-pressure sterilization

[0134] The mushrooms were bagged using a fully automatic double-barrel mushroom bagging machine manufactured by Zibo Longtai Machinery Technology Co., Ltd. The sticks were tightly packed, using 17 cm × 60 cm bags weighing 2800 ± 100 g of wet material. After filling, the bags were mechanically sealed. A vent hole was punctured near the inoculation hole and covered with breathable tape. When the sticks were stacked on the sterilization cart, the upper and lower layers of the sticks were arranged in a herringbone formation. The vent holes on the bottom layer of sticks were tilted downward at a 45° angle, while the vent holes on the upper layers faced downward, with both holes facing open to prevent blockage. Sterilization of the packed sticks was carried out promptly, starting from the beginning of bagging until the end, and ideally within 4 hours. Sterilization was performed using high-pressure steam at 0.11-0.14 MPa, at a temperature of approximately 118°C, for at least 330 minutes.

[0135] In addition, the present invention also compares the effects of using an automatic bagging machine and a semi-automatic bagging machine on the efficiency of stick making. The results are shown in Table 3:

[0136] Table 3: Comparison of rod making efficiency

[0137]

[0138] From the results in Table 3, it can be seen that the efficiency of the automatic bagging machine is 800-100 sticks / hour, each automatic bagging machine only requires one person to complete, and the standard of the produced mushroom sticks is high; while the efficiency of the semi-automatic bagging machine is 500-600 sticks / hour, each semi-automatic bagging machine requires five people to complete, which greatly increases the labor cost, and the standard of the produced mushroom sticks is low. Therefore, the present invention adopts an automatic bagging machine to make sticks.

[0139] In addition, the inventors also compared the effects of high-pressure sterilization and normal-pressure sterilization on the sterilization efficiency of the rods. The comparison results are shown in Table 4:

[0140] Table 4: Comparison of sterilization efficiency

[0141]

[0142] The results in Table 4 show that high-pressure sterilization of the rod material takes 7 hours, with no water pockets and a very low contamination rate. In contrast, normal-pressure sterilization requires 72 hours, with a water pocket rate of approximately 3%-5% and a contamination rate of approximately 5%-8%. Therefore, the present invention adopts high-pressure sterilization for rod sterilization.

[0143] (4) Forced cooling and mechanical inoculation

[0144] Open the door of the autoclave promptly after sterilization is complete and remove the rods from the autoclave when the temperature drops to around 80°C. After removal, transfer the rods to a Class 10,000 cleanroom for primary and secondary cooling.

[0145] The first cooling process uses fresh air with high efficiency filtration to cool the rods to about 50℃.

[0146] The secondary cooling uses a refrigeration fan to quickly cool down the material stick to below 26 ° C, and then it is transferred to the inoculation room and inoculated using the shiitake mushroom stick solid inoculator produced by Shandong Jinboyang Automation Equipment Co., Ltd.

[0147] Solid culture (the preparation process for the culture medium has been described above) is generally inoculated into 3-4 wells, at a depth of 3.5 cm-5 cm, with 15 g-20 g in each well. After inoculation, the culture medium is moved to the culture room for cultivation.

[0148] In addition, the present invention also compares the effects of inoculation by inoculation machine and manual inoculation on the inoculation efficiency, and the results are shown in Table 5:

[0149] Table 5: Comparison of inoculation efficiency

[0150]

[0151] As shown in Table 5, the inoculation efficiency of the inoculator is 1000-1500 bags / hour, which is not affected by weather and has a very low contamination rate. However, the inoculation efficiency of manual inoculation is only 300-400 bags / hour, which is greatly affected by weather and has a contamination rate of 5%-10%. Therefore, the present invention adopts the inoculator for inoculation.

[0152] Environmentally controlled bacterial culture and mechanical puncture

[0153] In order to improve the quality of mushroom stick culture and increase the utilization rate of the culture room, mushroom stick culture generally adopts a "three-stage" culture.

[0154] 1) Early stage of culture of bacterial sticks: Within 15 days after inoculation, the bacterial sticks are stacked on culture racks and placed in a pre-culture room (10,000-grade purification). The temperature of the pre-culture room is controlled at 20°C-21°C and the humidity is 70%-80%. Culture is carried out in the dark. Mycelium germination is promoted in a clean environment, and the inoculation port is sealed.

[0155] 2) Mid-term culture of bacterial sticks: After 15 days of culture, when the diameter of the inoculated hole is 6cm-8cm, the bacterial sticks are transferred to the grid rack for culture, with a density of about 140 sticks / m 2 ; The diameter of the fungus ring is 15 cm, and a puncture is performed (small puncture, see Figure 6 A) Each inoculation port is punctured 15 times with a depth of 2.5 cm. Punctures can be made manually or by machine.

[0156] 5 days after the mushroom sticks are filled with bags, perform the second mechanical puncture (big thorn, see Figure 6 B) Puncture 150 holes per stick, 6.0 cm deep. The holes should be neatly arranged and evenly spaced. During this stage, the sticks generate a lot of heat, especially one to two days after puncturing, when the stick temperature is 2°C to 4°C higher than the room temperature. Therefore, the incubation room temperature must be strictly controlled, generally at 21°C to 22°C.

[0157] 3) Late-Stage Culture, from the completion of puncture to the maturity stage: After the second mechanical puncture, light should be provided to promote color change. Light intensity should be 50-200 lux, with white light (RBG) or blue light (B) being the preferred light quality. Daily illumination should be at least 12 hours. During color change, the stick temperature is generally 2-3°C higher than the room temperature, which should ideally be maintained at 21-22°C. After color change, the stick respiration and heat generation decrease, and the stick temperature is generally 0.5-1°C higher than the room temperature, which should ideally be maintained at 23-24°C.

[0158] Natural cultivation of shiitake mushroom spawn involves placing the inoculated spawn in a "tic-tac-toe" or "triangle" pattern within a mushroom shed under natural conditions. Temperature, humidity, light, and carbon dioxide concentration are controlled through insulation, shading, and ventilation. The goal is to maintain a suitable cultivation environment within this range, encouraging the establishment, growth, and physiological maturity of the shiitake mycelium. Because the natural cultivation environment is difficult to precisely control and is significantly influenced by the natural environment, particular attention should be paid to temperature fluctuations, especially during high temperatures in summer, to prevent "burning" of the spawn (when temperatures exceed 30°C, the mycelium will die and rot). Furthermore, ventilation can easily lead to a decrease in humidity, so water spraying is necessary to prevent dehydration. In actual production, management measures should be flexibly adjusted based on local climate characteristics and the specific characteristics of the cultivar to reduce contamination risks and maximize the success rate of spawn cultivation.

[0159] In addition, the present invention also compared the effects of the above-mentioned environmental control culture and natural culture conditions on the culture efficiency of the mushroom sticks, and the results are shown in Table 6:

[0160] Table 6: Comparison of culture efficiency

[0161]

[0162] As can be seen from the results in Table 6, using the above-mentioned environmental control culture conditions, the culture density of the mushroom stick culture link can reach 200 sticks / square meter, and the parameters such as temperature, humidity, light, and carbon dioxide concentration in the mushroom stick culture link can be automatically controlled. The influence of weather is minimal, the consistency of the mushroom stick culture is high, and it is suitable for year-round and standardized production. However, using natural culture conditions, the culture density is about 120 sticks / square meter. The temperature, humidity, light, and carbon dioxide concentration are difficult to accurately control, which easily causes high temperature "stick rot". The consistency of the mushroom stick culture is poor and it is greatly affected by weather, making it suitable for seasonal production. Therefore, the present invention adopts the above-mentioned environmental control culture.

[0163] (6) Facility-based three-dimensional mushroom production

[0164] The "Huxiang F7" mushroom cultivation system is being grown in a multi-story facility, a three-dimensional production system. The shed is 2.4 meters high, constructed from galvanized pipes and covered with plastic film and shade netting. It is narrow from north to south and long from east to west, facilitating air circulation. It features six layers of multi-story cultivation racks for spawning mushroom sticks, each 0.4 meter apart. The bottom layer is 0.2 meters high and 0.45 meters wide, with two rows in the center and one row on each side. The left and right working paths are 0.7 meters wide. Thermometers are hung in various locations on the shed to monitor and control temperature and humidity. A 2-meter open area is maintained around the shed to facilitate ventilation.

[0165] After the mushroom sticks have been fermented, they are unbagged and placed on the shelves for fruiting. Newly bagged mushroom buds and young mushrooms less than 2-3 cm in diameter are still very delicate and require heat preservation and moisturizing during bud development. The temperature during bud development should be 8-20°C, with a humidity of approximately 80%-90%. The ideal humidity for the growth of "Huxiang F7" is 50%-68%, with an optimal range of 50%-55%. The temperature should range from 8-22°C, with an optimal range of 12-16°C. The maximum daily temperature in the greenhouse should be around 20°C, with a daytime and nighttime temperature difference of at least 10°C, for optimal growth, development, and cracking of "Huxiang F7."

[0166] "Huxiang F7" can be harvested when the fruiting bodies are 7-8 months old and the mushroom membrane has not yet cracked. Harvesting generally requires choosing suitable weather conditions. Before rainy weather arrives, when the relative humidity is above 90% for a long time, "Huxiang F7" should be picked appropriately in advance to avoid quality degradation due to high humidity and poor storage and transportation resistance. When picking, be careful not to leave small pieces of mushroom stems on the mushroom sticks to prevent the stems from rotting and causing mold contamination of the mushroom sticks.

[0167] In addition, the present invention also compares the above-mentioned facilities greenhouse three-dimensional mushroom production ( Figure 7 ) and simple greenhouses ( Figure 8 ) The effect of leaning against the mushrooms on the efficiency of mushroom production. The results are shown in Table 7:

[0168] Table 7: Comparison of fruiting efficiency

[0169]

[0170] As can be seen from the results in Table 7, the above-mentioned facility greenhouse three-dimensional fruiting method achieved a fruiting density of 200 sticks / square meter, which was rarely affected by weather, and a yield of 800-900 kg / stick. The facility greenhouse had good control over environmental factors such as temperature and humidity, and could be used for fruiting for 9-12 months each year. However, the simple greenhouse leaning against the fruiting method achieved a fruiting density of only 50 sticks / square meter, which was greatly affected by weather, and a yield of 600-700 kg / stick. Since the simple fruiting shed had poor control over environmental factors such as temperature and humidity, the time available for fruiting each year was generally short (3-6 months). Therefore, the present invention adopted the above-mentioned facility greenhouse three-dimensional fruiting method.

[0171] Comparative Example 1: Intensive production of Shiitake mushroom "L808"

[0172] The experimental procedures for Comparative Example 1 and Example 2 are essentially the same, except that the shiitake mushroom variety used in this comparative example is L808, a major domestically cultivated shiitake mushroom variety (a nationally approved variety, sourced from the National Edible Fungi Germplasm Resource Bank (Shanghai)). The comparative production results for the shiitake mushroom varieties "Huxiang F7" and "L808" of the present invention are shown in Table 8:

[0173] Table 8

[0174]

[0175] As can be seen from the results in Table 8, under the intensive production method of the present invention, compared with the "L808" strain, the shiitake mushroom variety "Huxiang F7" of the present invention has a shorter spawning time of only 85-90 days and fewer fruiting waves, only 2-3 times. It also has a higher yield per stick (650-800 kg / stick). The variety also has a shorter fruiting cycle (2-3 fruiting waves per batch of mushroom sticks generally take about 2 months), resulting in more batches available for fruiting each year and lower production costs.

[0176] Based on the above results, compared with the "L808" strain, the mushroom variety "Huxiang F7" of the present invention has a fungal age of 85-90 days and a fruiting temperature of about 20°C. The fruiting cycle of each batch of production is short. Combined with the environmental control of the facility-based mushroom greenhouse, it can achieve low-cost fruiting time and longer mushroom production in China. Therefore, more batches of production can be carried out each year, the production efficiency is higher, and it is more suitable for intensive production.

[0177] The above describes the preferred embodiments of the present invention. It should be understood that the present invention is not limited to the above-mentioned specific embodiments, and the devices and structures that are not described in detail should be understood to be implemented in a common manner in the art; any technician familiar with the art can use the above-mentioned disclosed methods and technical contents to make many possible changes and modifications to the technical solutions of the present invention without departing from the scope of the technical solutions of the present invention, or modify them into equivalent embodiments of equivalent changes, which does not affect the essential content of the present invention. Therefore, any simple modifications, equivalent changes and modifications made to the above embodiments based on the technical essence of the present invention that do not depart from the content of the technical solutions of the present invention are still within the scope of protection of the technical solutions of the present invention.

Claims

1. A shiitake mushroom strain suitable for intensive production, characterized in that: The strain is classified and named as Lentinus edodes "Huxiang F7", and is preserved in the Guangdong Provincial Microbiological Culture Collection Center with the preservation number GDMCC No: 66428 and the preservation date of May 28, 2025.

2. A molecular marker identification method for Lentinus edodes strains according to claim 1, characterized in that: It includes the cultivation and collection of mycelium, extraction of genomic DNA, PCR amplification of specific fragments, electrophoresis detection and display of specific bands; The specific nucleotide sequences of the specific fragment PCR amplification primers Lefp_id001 to Lefp_id007 are: The forward sequence of primer Lefp_id001 is shown in SEQ ID NO: 1: 5′-ACGGATGGAGAGACACAGGA-3′, and the reverse sequence is shown in SEQ ID NO: 2: 5′-TGCCCCACTTACTCTCAACC-3′; The forward sequence of primer Lefp_id002 is shown in SEQ ID NO: 3: 5′-CCTTTCTCAAAAGCGGACTG-3′, and the reverse sequence is shown in SEQ ID NO: 4: 5′-GGGAGTGGGTTGTTTGGATA-3′; The forward sequence of primer Lefp_id003 is shown in SEQ ID NO: 5: 5′-CGGATGTTATGCACTGGAAG-3′, and the reverse sequence is shown in SEQ ID NO: 6: 5′-CGTACGGTTGGACATTTGAA-3′; The forward sequence of primer Lefp_id004 is shown in SEQ ID NO: 7: 5′-AGCCTTTCACAGTCAGCTCG-3′, and the reverse sequence is shown in SEQ ID NO: 8: 5′-GTCAACGGAGGGAAACAGAG-3′; The forward sequence of primer Lefp_id005 is shown in SEQ ID NO: 9: 5′-GTAGCACTCCTCATACAACC-3′, and the reverse sequence is shown in SEQ ID NO: 10: 5′-ACCAAATGTCACAGCACAGG-3′; The forward sequence of primer Lefp_id006 is shown in SEQ ID NO: 11: 5′-ACATTGGCGAAGGCTGTACG-3′, and the reverse sequence is shown in SEQ ID NO: 12: 5′-CCCTTTTGCCCTATAAGGCG-3′; The forward sequence of primer Lefp_id007 is shown in SEQ ID NO: 13: 5'-AACAGTAACCTGTGCACTGC-3', and the reverse sequence is shown in SEQ ID NO: 14: 5'-CATGATCAGATCACACAGCG-3'.

3. The molecular marker identification method according to claim 2, wherein The primer Lefp_id001 produces a 203 bp band, the primer Lefp_id002 produces a 236 bp band, the primer Lefp_id003 produces a 214 bp band, the primer Lefp_id004 produces a 270 bp band, the primer Lefp_id005 produces two bands of 346 bp and 378 bp, the primer Lefp_id006 produces two bands of 262 bp and 277 bp, and the primer Lefp_id007 produces a 284 bp band. The seven primers are detected simultaneously to obtain DNA marker bands of corresponding sizes, which are the markers of the shiitake mushroom "Huxiang F7".

4. An intensive production method of the shiitake mushroom strain according to claim 1, characterized in that: The process includes the following steps: preparing raw materials, preparing mushroom sticks and high-pressure sterilization, mechanical inoculation, factory cultivation, and facility-based mushroom production management; Wherein, the preparation of raw materials includes preparing bacterial strains and cultivation substrates; The preparation of bacterial strains includes preparing solid bacterial strains of "Huxiang F7"; The formula of the cultivation matrix comprises: (a) Stock formulation: PDA medium containing 200 g peeled potatoes, 20 g glucose, 18-20 g agar, and 1000 mL water; (b) a cultivar formulation comprising the following ingredients in a mass-to-volume ratio: sawdust 78%, wheat bran 20%, sucrose 1%, gypsum 1%, a moisture content of 60%-65%, and a pH of 5.5-6.5; (c) A mushroom log production formula comprising the following ingredients in a mass-to-volume ratio: (c1) oak sawdust 75-85wt% (preferably 78wt%), bran 15-25wt% (preferably 20wt%), gypsum 1-3wt% (preferably 2wt%), moisture content 50%-55%, pH 6-7 before sterilization; (c2) oak sawdust 40-50wt% (preferably 48wt%), fruit sawdust 20-40wt% (preferably 30wt%), bran 15-25wt% (preferably 20wt%), gypsum 1-3wt% (preferably 2wt%), water content 53%-58%, pH 6-7 before sterilization.

5. The intensive production method according to claim 4, characterized in that: The preparation of the bacterial sticks and high-pressure sterilization comprise the following steps: (a) using an automatic bagging machine to fill the cultivation substrate into bags, and mechanically tying the bags to obtain material sticks; (b) stacking the material rods into a sterilization container and performing high-pressure sterilization; The high-pressure sterilization temperature is 115-120° C., and the sterilization time is 300-1000 min.

6. The intensive production method according to claim 4, characterized in that: The mechanical inoculation comprises the following steps: (1) The high-pressure sterilized rods are transferred to a 10,000-level cleanroom for the first cooling and the second cooling; wherein, The first cooling is carried out by introducing fresh air with high efficiency filtration to cool the rod to 40-55°C; The second cooling is carried out by using a refrigeration fan to quickly cool the material bar to below 26°C; (2) Transfer the cooled rods to the inoculation room and use an inoculation machine for inoculation; The inoculation step comprises: using a fully automatic shiitake mushroom solid inoculator to inoculate the solid spawn onto a material stick to obtain a mushroom stick; inoculating 3-4 holes with the solid spawn, with an inoculation depth of 3.5 cm-5 cm and an inoculation weight of 15 g-20 g per hole.

7. The intensive production method according to claim 4, characterized in that: The factory culture comprises the following steps: (1) Early culture stage of bacterial sticks: Within 13-15 days after inoculation, the bacterial sticks are stacked on the culture rack and placed in a 10,000-class purified pre-culture room. The temperature of the pre-culture room is controlled at 20°C-21°C and the humidity is 70%-80%. Culture is carried out in the dark. Mycelium germination is promoted in a clean environment and the inoculation port is sealed. (2) Mid-stage of bacterial stick cultivation: After 13-15 days of bacterial stick cultivation, when the diameter of the inoculated hole is 6cm-8cm, the bacterial sticks are transferred to the shelves for "tic-tac-toe" stacking or grid racks for cultivation, with a density of 130-150 sticks / m 2 The diameter of the fungus ring is 10 cm-15 cm. When mycelium can be seen on the back of the inoculation hole, the first puncture is performed manually or mechanically. 10-15 holes are punctured at each inoculation hole, with a hole depth of 2.0 cm-2.5 cm. About 5 days after the mushroom sticks are full of bags, use a puncture machine to perform a second puncture, puncture about 150 holes per stick, the hole depth is 5.0cm-7.0 cm, the puncture holes should be arranged neatly and the spacing should be even; in the middle stage of the mushroom stick culture, the mushroom sticks should be cultured in the dark because the heat generated by the mushroom sticks is high, especially 1-2 days after puncture, the stick temperature is 2℃-4℃ higher than the room temperature, so the culture room temperature needs to be strictly controlled, and the room temperature should be controlled at 21℃-22℃; (3) Late culture stage of bacterial sticks: After the second puncture for oxygenation, light is provided to promote the color change of bacterial sticks; The intensity of the light is 50 lux-200 lux, the light quality of the light is selected from white light (RBG) or blue light (B), and the illumination time is no less than 12 hours per day.

8. The intensive production method according to claim 7, characterized in that: During the color change culture of the bacterial sticks, the temperature of the bacterial sticks is 2°C-3°C higher than the room temperature, and the room temperature is controlled at 21°C-22°C; After the color change culture of the bacterial sticks is completed, the respiration of the bacterial sticks is weakened and the heat generation is reduced. The stick temperature of the bacterial sticks is 0.5°C-1°C higher than the room temperature, and the room temperature is controlled at 23-24°C.

9. The intensive production method according to claim 4, characterized in that: The facility-based mushroom production management includes the following steps: (1) Constructing a facility-based mushroom shed: The facility-based mushroom shed is 2-3 m high (preferably 2.4 m), with galvanized pipes forming the shed frame, and plastic film and sunshade nets covering the shed frame. The shed is narrow from north to south and long from east to west to facilitate air circulation; The facility-based mushroom shed has built-in multi-layer cultivation racks, which are used to place the mushroom sticks. The cultivation racks have the following structure: 5-6 layers, a layer spacing of 0.3-0.4 m, a bottom layer height of 0.15-0.2 m, a rack width of 0.40-0.45 m, two rows of cultivation racks in the middle, a row of cultivation racks on each side, and a width of the operating aisles on the left and right sides of 0.6-0.7 m.

10. The intensive production method according to claim 9, characterized in that: The facility-based mushroom production management further comprises the following steps: (2) Remove the mushroom sticks from the bags after cultivation. Keep the temperature in the fruiting shed at 8-20℃ and the humidity at 80%-90%. Spray water to moisten the mushroom sticks. After 5-6 days, buds will appear. Thin out the buds and leave 12-15 buds. Harvest after 4-6 days. After harvesting, the temperature is controlled at 20-25°C, the humidity is 70-80%, and the CO2 concentration is 500-1500ppm. The mushrooms are allowed to rest for 15-20 days. Then, the temperature is controlled at 12-20°C, the humidity is 80-90%, and the CO2 concentration is 500-1500ppm to stimulate bud formation. Water is sprayed to humidify the mushroom sticks. After bud formation and mushrooms are produced, the second wave of harvesting is carried out. Repeat the above operation after harvesting for 3-5 waves of harvesting.

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