Euphorbia mushroom strain, method for cultivating by utilizing flammulina velutipes mushroom dregs and application of euphorbia mushroom strain

Abstract

The invention discloses a yuan mushroom strain, a method for cultivating the yuan mushroom strain by utilizing flammulina velutipes mushroom dregs and application of the yuan mushroom strain, and belongs to the field of edible mushroom cultivation. The preservation number of the yuan mushroom strain is CGMCC (China General Microbiological Culture Collection Center) No.41571 The flammulina velutipes mushroom dregs are used as the cultivation material for cultivating the yuan mushrooms, and the use amount of the flammulina velutipes mushroom dregs and the conditions for cultivating the yuan mushrooms are optimized, so that resource utilization of wastes is realized; moreover, the growth vigor and the growth speed of yuan mushroom hyphae are also remarkably improved, the total biotransformation rate and the yield are improved, the growth cycle is shortened, and the quality of yuan mushrooms is improved. The method provides powerful guarantee for sustainable development of the yuan mushroom industry.

Description

Technical Field

[0001] The invention relates to the field of edible fungi cultivation, in particular to a enoki mushroom strain and a cultivation method and application thereof using Flammulina velutipes mushroom residue. Background Art

[0002] In my country, edible fungi are the fifth largest planting industry after grain, oil, fruit and vegetables. In recent years, with the rapid development of my country's edible fungi industry, especially the increasing factory production, the production scale of Flammulina velutipes has been expanding, and the amount of fungus residue has been increasing. It is reported that when 1 kg of edible fungi is produced, 5 kg of fungus residue waste will be generated. The treatment of fungus residue has increasingly become a bottleneck restricting the sustainable development of the industry. The conventional disposal method is to discard and pile it up, which will cause the following problems: ① The volume of fungus residue is large, occupying land resources and affecting the beauty of the environment; ② The fungus residue contains more pathogens and nematodes, etc. If it is directly piled up or used as fertilizer, it may cause the occurrence of diseases and insect pests; ③ Harmful gases such as methane and hydrogen sulfide produced during the decomposition of fungus residue will pollute the atmospheric environment; ④ During the stacking of fungus residue, water evaporation leads to the accumulation of soil salt, affecting soil quality.

[0003] However, mushroom residue is the waste produced after fermentation, fruiting and harvesting of edible fungus culture medium. It is not only rich in cellulose, hemicellulose and lignin, but also rich in a large amount of mycelium residual protein, fat, amino acids, minerals and fungal secondary metabolites, and is of great value for development and utilization.

[0004] The failure to use mushroom residues in a reasonable and efficient manner not only wastes a lot of resources, but also causes serious pollution to the ecological environment. In the current era of resource and energy shortages, timely and scientific disposal of mushroom residues to achieve an organic combination of economic, environmental and social benefits is of great practical significance.

[0005] In view of the above problems, the present invention proposes a method for cultivating enoki mushrooms using enoki mushroom residues. The method solves the problems existing in the traditional treatment process of enoki mushroom residues by optimizing the residue treatment process, and provides an efficient and environmentally friendly resource utilization method for the edible fungus industry. Summary of the invention

[0006] The purpose of the present invention is to provide a strain of enoki mushroom and a method and application of cultivating the strain using enoki mushroom residue, so as to solve the problems existing in the above-mentioned prior art. The present invention can not only effectively solve the problems existing in enoki mushroom residue by optimizing the enoki mushroom cultivation method, but also realize the recycling of resources, and can also improve the yield and quality of enoki mushrooms and shorten the growth cycle, etc., thereby providing a strong guarantee for the sustainable development of my country's edible fungus industry.

[0007] To achieve the above object, the present invention provides the following solutions:

[0008] The invention provides a Pleurotus ostreatus strain, and the preservation number of the Pleurotus ostreatus strain is CGMCC No.41571.

[0009] The present invention also provides a method for cultivating enoki mushrooms, comprising the step of performing secondary cultivation on the enoki mushroom strain in a cultivation medium using enoki mushroom residue to replace part of sawdust, wherein the cultivation medium comprises the following raw materials in parts by weight: 35 parts of enoki mushroom residue, 45 parts of sawdust, 19 parts of bran and 1 part of lime.

[0010] Preferably, the cultivation method comprises the following steps:

[0011] Inoculating the Pleurotus eryngii strain onto a PDA slant for culture to obtain a mother strain;

[0012] Inoculate the mother strain into a liquid strain culture medium, shake and culture, and obtain a liquid strain;

[0013] The cultivation medium is bagged to form a fungus bag, and then the fungus bag is inoculated with the liquid fungus, and then transferred to a culture room for cultivation;

[0014] Fruiting and harvesting.

[0015] Preferably, after the PDA slant is inoculated with the PDA strain, it is cultured at 24° C. for 9 days. After the mycelium is fully grown, a test tube strain with dense, white, robust mycelium and no infection by any foreign bacteria is selected as the mother strain.

[0016] Preferably, the shaking culture conditions are: temperature of 24° C., rotation speed of 150 r / min, and constant temperature shaking culture for 7 days.

[0017] Preferably, each of the mushroom bags is filled with 1.0-1.5 kg of the cultivation medium.

[0018] Preferably, 20 bacteria bags are inoculated with every 400 mL of the liquid bacteria.

[0019] Preferably, the culture medium is cultured under the following conditions: 24-25°C on the 1st to 2nd day after inoculation; 20-22°C on the 3rd to 15th day after inoculation; 18-20°C on the 16th to 35th day after inoculation;

[0020] During the culture period, when the mycelium begins to germinate and grow, ventilation should be carried out once a day at noon for 1 hour each time; when the mycelium grows to more than 1 / 3 of the bag, ventilation should be carried out once a day in the morning, noon and evening for 1 hour each time.

[0021] Preferably, the Flammulina velutipes residue is processed as follows before use: pollution-free and mold-free Flammulina velutipes residue is selected, exposed to the sun, and then crushed into sawdust.

[0022] The present invention also provides the use of the mushroom strain or the cultivation method in any of the following:

[0023] (1) Application in improving the growth potential and growth rate of Pleurotus ostreatus mycelium;

[0024] (2) Application in increasing the yield of mushrooms;

[0025] (3) Application in shortening the cultivation cycle of mushrooms;

[0026] (4) Application in improving the bioconversion rate of mushrooms.

[0027] The present invention discloses the following technical effects:

[0028] (1) The present invention utilizes the residue of Flammulina velutipes as the cultivation material for cultivating enoki mushrooms, thereby realizing the resource utilization of waste. According to statistics, each ton of Flammulina velutipes residue can reduce carbon dioxide emissions by about 0.8 tons, while reducing land occupation and environmental pollution. The present invention also proves that when 35% of the residue of Flammulina velutipes is added to the cultivation material for enoki mushroom cultivation, the mycelium grows faster, which is 3 days earlier than the full bag days of sawdust cultivation, and the cultivation week can be shortened.

[0029] (2) Flammulina velutipes residue still contains certain nutrients and can serve as an excellent substrate for the growth of enoki mushrooms. Experiments have shown that when the amount of Flammulina velutipes residue added to the culture medium is 35%, the bioconversion rate of enoki mushrooms can reach more than 95.8%, and the fruiting body is the thickest, 0.85 mm thicker than the control group (CK), which is a more ideal mushroom shape.

[0030] (3) The present invention takes only 47-50 days from inoculation to the end of the first wave of mushroom harvesting, and has a short production cycle, high biological conversion rate, high degree of automation, low labor cost, good production efficiency, and easy cultivation and management. The present invention provides a new method for cultivating shiitake mushrooms with high efficiency and high yield. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0032] Figure 1 The culture characteristics of the Pleurotus eryngii strain on the plate; the left and right pictures are the observation results of the front and back plates respectively;

[0033] Figure 2The results of electrophoresis detection; the left figure is an enlarged view of 100-3000bp Ladder-K, and the first and second lanes of the right figure are 100-3000bp Ladder-K and amplified samples, respectively;

[0034] Figure 3 It is the result of liquid bacterial culture bottle culture;

[0035] Figure 4 This is a picture of Flammulina velutipes residue;

[0036] Figure 5 This is a picture of the mushroom bag for the ingredients of Flammulina velutipes;

[0037] Figure 6 This is a picture of the fruiting body of the mushroom bag;

[0038] Figure 7 This is a picture of the fruiting body of wild Agaricus sphaerocephala. DETAILED DESCRIPTION

[0039] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0040] It should be understood that the terms described in the present invention are only for describing a particular embodiment and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. The intermediate value in any stated value or stated range, and each smaller range between any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0041] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0042] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to those skilled in the art. The present invention description and examples are exemplary only.

[0043] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0044] Example 1

[0045] 1. Source of strains

[0046] The mushroom strain of the present invention is isolated from wild mushroom fruiting bodies (such as Figure 7 As shown in the figure, the strains were obtained by tissue separation, and PDA medium was selected as the culture medium. Specifically, it includes strain isolation, morphological feature identification and molecular identification.

[0047] 1.1 Isolation of bacterial strains

[0048] ①Prepare PDA culture medium with the following formula: 200g potato, 20g glucose, 20g agar, 1000mL water. Wash and peel the potato, weigh 200g, slice or dice (1cm 3 ) into a pot, add 1000mL of water into the pot, cook while stirring, cook until it breaks when pressed with chopsticks or a glass rod, filter with 4 to 8 layers of gauze, make the filtrate volume up to 1000mL, put it back into the pot, add agar, stir continuously until it is completely dissolved, add glucose, stir while adding, until it is completely dissolved, and the culture medium is ready.

[0049] ② Bottling: Pour the cooked PDA culture medium into a conical flask and seal it with a sealing film (cotton plugs can also be used); at the same time, wrap the number of culture dishes to be used with newspapers.

[0050] ③Sterilization: Place the sealed Erlenmeyer flask and wrapped culture dish in an autoclave, adjust to 121℃, and autoclave for 30 minutes.

[0051] ④ Ultraviolet sterilization: After the sterilization is completed and the pressure of the sterilizer drops to zero, partially open the lid of the sterilizer, dry the cotton plugs and newspapers, leave the flask in the sterilizer, put the culture dish, scalpel, needle-nosed tweezers, and alcohol lamp on the clean bench, and sterilize with ultraviolet light for 30 minutes. After the sterilization is completed, turn off the ultraviolet lamp, turn on the fan for 5 minutes, take the culture medium in the flask to the clean bench, disinfect the surface of the flask with alcohol, and prepare to pour the plate.

[0052] ⑤ Pour the plate: In the clean bench, first wipe your hands with 75% alcohol, then turn on the alcohol lamp, remove the sealing film of the triangular flask near the flame, place the mouth of the triangular flask near the flame, open the sterilized plate, and pour the culture medium into the plate (the holding plate method and the stacking plate method can be used), pour about 10mL on each plate (be careful not to pour the culture medium on the wall of the plate to avoid contamination. The amount of culture medium is determined according to the function and culture time).

[0053] ⑥Put the poured plate on the clean bench for later use (usually use it as soon as possible to avoid contamination). Turn off the alcohol lamp and clean the clean bench. After solidification, take out several culture media sealed with sealing film and culture them at 37℃ to observe whether they are completely sterilized.

[0054] ⑦ Tissue separation: Place the bacterial tissue to be separated on a clean bench, wipe hands and mushrooms with 75% alcohol to disinfect the surface, and light an alcohol lamp. Cut or tear the mushrooms from the base of the tissue (if cutting with a knife, the blade should be burned and sterilized first), use a sterilized blade to cut rice-sized mushroom flesh from the cross section, pick it up with pointed tweezers, quickly transfer it to the prepared flat culture medium, and place it in the center (note that the blade and tweezers should be cooled in the air after burning to prevent the mycelium from being scalded to death), mark it, and apply sealing film.

[0055] ⑧Cultivation: Place the transferred culture dish in a constant temperature incubator at 25℃.

[0056] ⑨ Strain purification: When the mycelium germinates and grows to 1.5-2cm, prepare for strain purification. In the clean bench, use the sterilized inoculation spatula to pick up the mycelium tip culture medium from the flat culture medium, and quickly transfer it into the prepared test tube (the inoculation spatula should be cooled and operated under an alcohol lamp), and mark it.

[0057] ⑩ Cultivation: Place the transferred test tube in a 25℃ constant temperature incubator for cultivation. When the mycelium is fully grown, a pure strain of bacteria will be obtained.

[0058] 1.2 Morphological characteristics identification

[0059] The fruiting body is large and fleshy. It is white at the beginning and turns to light yellow or light brown after maturity. The fruiting body is irregularly spherical or elliptical, with a diameter of up to 10-20cm, and has dense thorn-like protrusions on the surface arranged in a radial pattern. The cap is hemispherical at the beginning and flat at the end. There are fine scales on the surface, with thin and wavy edges. The stipe is short and thick, about 1-2cm in diameter, connected to the cap, and sometimes not obvious. The gills are white, dense, unequal in length, and connected to the stipe.

[0060] Basidiospores are colorless, smooth, oval or nearly spherical, and are about (5-7) μm × (4-5) μm in size. Mycelium is white, fluffy, and about (2-4) μm in diameter. Mycelium grows faster on the culture medium. Mycelium is slender, and the colony is loose, spider-web-like or cotton-like. After 8 days, the diameter is 7 cm. The front side is dull, and the back side is light orange-yellow in pigmentation, as shown below. Figure 1 shown.

[0061] 1.3 Molecular identification

[0062] (1) Primer sequence

[0063] ITS1 (SEQ ID NO.2): 5'-TCCGTAGGTGAACCTGCGG-3';

[0064] ITS4-R (SEQ ID NO. 3): 5'-TCCTCCGCTTATTGATATGC-3'.

[0065] (2) DNA extraction

[0066] The DNA from the strain was extracted using the Ezup column fungal genomic DNA extraction kit (B518255).

[0067] (3) PCR amplification

[0068] The reaction system and reaction conditions are shown in Table 1 and Table 2, respectively.

[0069] Table 1 PCR reaction system

[0070]

[0071]

[0072] Table 2 PCR reaction conditions

[0073]

[0074] (4) PCR electrophoresis

[0075] The samples were electrophoresed using 1.5% agarose gel and 1×TAE electrophoresis buffer at 150 V and 100 mA for 20 min and then observed.

[0076] The electrophoresis results are as follows Figure 2 As shown, a single bright band appeared, which was consistent with the expected 623 bp (sequence shown in SEQ ID NO.1).

[0077] (5) Sequencing

[0078] The sequencing results were compared with the genes on NCBI, and the comparison results are shown in Table 3.

[0079] Table 3 Gene comparison results

[0080]

[0081]

[0082]

[0083] According to the above comparison results and combined with the morphological characteristics analysis, the strain isolated and screened by the present invention was identified as the genus Sarcomyxa, and was determined to be the delicious sticky mushroom, namely, Yuan Mushroom (Sarcomyxa edulis), and was named Yuan Mushroom 2016100112.

[0084] The above-mentioned Sarcomyxa edulis 2016100112 was deposited in the General Microbiology Center of China Culture Collection Administration on October 9, 2024, with the deposit number CGMCC No. 41571, and the deposit address is the Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

[0085] Embodiment 2 The cultivation method of mushroom

[0086] This embodiment is to carry out secondary cultivation of the enoki mushroom strain isolated in the above embodiment 1 to solve the problems currently faced by the treatment of Flammulina velutipes residue, realize the recycling of resources, effectively reduce production costs, and promote the sustainable development of agriculture. The present invention provides a method for cultivating enoki mushrooms with Flammulina velutipes residue, which has high economic benefits and is easy to cultivate and manage, and can alleviate the pollution to the environment caused by the accumulation of residue.

[0087] (1) Preparation and preservation of mother stock

[0088] Mother culture medium formula: Potato dextrose agar medium (PDA): 200 g potato, 20 g glucose, 20 g agar and 1 L water, natural pH.

[0089] Preparation and preservation: Wash and peel the potatoes, cut them into small pieces, boil them for 20 minutes (the glass rod will break when touched lightly), filter them, add 20g agar to the filtrate, stir continuously until the agar is completely dissolved, add glucose and stir evenly, then add water to make the volume 1L, divide them into test tubes, seal them and sterilize them in an autoclave at 121℃ for 30 minutes, then take out the test tubes and place them on an inclined surface, inoculate them in a sterile environment after cooling, and culture them at 24℃ after inoculation. After 9 days of full growth, select the test tube strains with dense, white, and strong mycelium growth and no infection by any other bacteria and store them in a 4℃ refrigerator.

[0090] (2) Liquid strain preparation

[0091] The formula of liquid culture medium is: 200g potato (peeled), 20g glucose, 1g potassium dihydrogen phosphate, 0.5g magnesium sulfate, 10g peptone, 5g yeast extract powder, 10mg vitamin B1 and 1L water, pH natural.

[0092] Preparation method: Boil the potatoes for 20 minutes and filter out the filtrate. Add other ingredients and mix evenly (without filtering), then dispense into Erlenmeyer flasks. The filling amount is 2 / 5 of the volume (the bottle volume is 1L). After sealing, sterilize in an autoclave at 121°C for 30 minutes. After cooling, complete the inoculation in an ultra-clean workbench, and then place the Erlenmeyer flask in a shaker. Set the shaker temperature to 24°C and the speed to 150r / min, and culture at a constant temperature for 7 days. The state of the bacterial balls in the bottle should be white, dense in number, with a large number of spherical mycelium suspended and evenly distributed, without floating, sinking, or stratification. Figure 3 shown.

[0093] (3) Collection and treatment of Flammulina velutipes residue

[0094] Select pollution-free and mold-free Flammulina velutipes residue, expose it to the sun to kill the remaining bacteria, and then grind it into sawdust (such as Figure 4 Place in a dry and cool place for later use.

[0095] The experimental formula was set up by replacing part of the sawdust with mushroom residue, and pure sawdust culture was used as the control. Four formulas were set up for each type of mushroom residue, as shown in Table 4.

[0096] Table 4 Test formula

[0097] Formula (%, mass percent) CK A B C D Mushroom residue 0 15 25 35 45 Wood chips 80 65 55 45 35 Bran 19 19 19 19 19 lime 1 1 1 1 1

[0098] (4) Production of mushroom bags

[0099] According to the predetermined cultivation material formula, use polypropylene plastic film bags to mix, mix and bag (such as Figure 5 As shown). Add water to these materials one day in advance and stir them evenly. The water content should meet the following conditions: If the compost is held tightly in your hand, it will have moisture but will not drip. If water drops, it means that the compost is too wet. You should open the pile to let the moisture evaporate, or add some dry compost. If the material is not lumpy and there is no water mark on the palm, it means that it is relatively dry. You should add water and then stir it to a moderate temperature. When stirring, mix thoroughly. The bag should be tight at the top and loose at the bottom. The surface should be smooth without wrinkles and the material surface should be flat. Each bag should be about 1.2kg. The material should be loaded and sterilized on the same day. The bags should not be left together overnight, otherwise they will become rancid.

[0100] (5) Vaccination

[0101] Inoculation refers to the process of transplanting the original seed into a sterilized culture bag under aseptic conditions. It is an important part of the cultivation of shiitake mushrooms, so it is necessary to maintain good inoculation equipment, a clean environment, and thorough disinfection and dust reduction. When the temperature of the mushroom bag drops to 25℃, start inoculation. The mycelium of shiitake mushrooms grows slowly, so the inoculation amount is slightly larger and the volume is equal. Each bottle of original seed can be inoculated with 20 bags. Before inoculation, be sure to wash your hands with 75% alcohol cotton balls, and move gently to reduce the flow of air. Before the inoculation is completed, all doors and windows should be closed to reduce air pollution. In particular, it should be noted that inoculation should be carried out above the flame of the alcohol lamp. The original seed bottle should always be aimed at the sterile area above the flame, and the bag mouth should also face the alcohol lamp. When pulling out the cotton plug, do not pull it up hard, but pull it out by rotating it to avoid the air pressure in the bag from dropping and letting the outside air suddenly rush in. If the cotton plug is wet, it should be replaced with a sterilized dry cotton plug. All inoculation equipment must be strictly sterilized. Opening the bag, inoculating, sealing, and placing the bags are all done in a continuous flow. When the inoculation equipment comes into contact with the bacteria, it should be burned and disinfected again with an alcohol lamp. From opening the bag to sealing the bag, the time should be shortened as much as possible to reduce the chance of contamination by bacteria. After the work is completed, the bag should be removed immediately and the room should be cleaned. If it needs to be used again, it needs to be thoroughly disinfected again.

[0102] (6) Cultivation

[0103] Before placing the bags in the culture room, the room must be strictly sterilized and fumigated with Bijie. After inoculation, the bags should be placed on the shelf immediately. Be careful when handling them and do not grab the bag mouth directly with your hands, otherwise it is likely to cause contamination. The bags should be placed on the shelf at a distance of 1 cm to facilitate ventilation and cooling. After the mycelium grows and feeds, check the survival status of the strain. If there is contamination by foreign bacteria, remove it immediately for treatment, and pay attention to strengthening ventilation.

[0104] Temperature is a key factor in the growth and development of shiitake mushrooms, affecting the growth rate and strength of mycelium. 22-25℃ is the most suitable growth temperature for shiitake mushroom mycelium, with 25℃ being the best. Within 3 days after inoculation, the indoor temperature should be higher, generally controlled at 24-25℃, so as to better promote the growth of mycelium. The mycelium sprouts faster, grows faster, and has a lower contamination rate of foreign bacteria. 3-15 days after inoculation, it is most suitable to maintain a culture room temperature of 20-22℃. Within 16-35 days, the temperature is most suitable at 18-20℃. At this time, the mycelium has already taken an advantage. Even if the temperature is lower, the mycelium's own metabolism will increase, and the mycelium will grow rapidly and healthily.

[0105] Asparagus is an aerobic fungus, good ventilation facilities should be provided during cultivation to maintain air circulation. In addition, attention should be paid to air convection to ensure that there is no odor in the greenhouse. In the first 5-7 days of cultivation, if the temperature is not too high, ventilation is not required. When the mycelium begins to germinate and grow, ventilation should be carried out immediately, once at noon every day, for 1 hour each time. When the mycelium grows to more than 1 / 3 of the bag surface, the frequency of ventilation should be increased, once in the morning, noon and evening every day, for 1 hour each time. If necessary, the doors and windows can be ventilated once.

[0106] The mycelium of the shiitake mushroom needs to be protected from light during its growth. The mycelium is prone to aging under light, resulting in the production of primordia and mushrooms growing in half a bag or around the bag, which has an adverse effect on the later yield. Therefore, the shiitake mushroom room should be shaded, that is, it should be carried out under dim light.

[0107] As shown in Table 5, when the content of fungus residue exceeds 35%, the growth of mycelium changes from thick to thin, and the color of mycelium changes from white to dark white; in terms of density, the mycelium of groups CK and C is dense, group A is dense, and the rest are sparse; for the average growth rate of mycelium, the difference significance analysis shows that there is no significant difference between groups A and C and the control group CK, and the growth rate of group C is the fastest, which is 0.04 mm / d faster than the control group and 1.55 mm / d faster than the slowest group D. The number of days to fill the bag is significantly shortened, indicating that the formula of group C is the best formula.

[0108] Table 5 Growth potential and growth rate of Pleurotus eryngii mycelium in different formulas

[0109]

[0110] (7) Fruiting (such as Figure 6 shown) and harvested

[0111] The temperature is maintained between 10-25℃, preferably 15-18℃. During the mushroom bud formation period, the fruiting body primordium is formed at a low temperature of 10-15℃ and under the stimulation of light. Ventilation and cooling are required in the early stage to increase light and promote the formation of mushroom buds. The light should be sufficient and the air humidity should be maintained between 85%-95%. Do not spray directly to avoid causing the mushroom buds to shrink, rot or even die, which will also affect the development of the mushroom buds and the yield and quality of the shiitake mushrooms. When the color of the mushroom buds slowly changes from yellow to yellow-brown, a small amount of water mist can be sprayed. During this period, the cap gradually increases and the color becomes lighter. This is the period when the mushroom body grows most rapidly. Therefore, a large amount of water should be sprayed on it. At the same time, the oxygen demand of the fruiting body increases, and the daily ventilation frequency should be increased. After the shiitake mushroom fruiting body matures, its cap stretches, the edge changes from inward curling to backward curling, the cap turns yellow, and is covered with white spores. Harvest when the fruiting body is about to disperse the spores, and stop watering the day before picking.

[0112] As can be seen from Table 6, after incision, the time of primordium formation in group C was 1 day earlier than that in CK, and the time of mushroom fruiting was 4 days earlier than that in CK.

[0113] Table 6 The formation of primordium and mushroom in different formulas

[0114]

[0115] As can be seen from Table 7, when the amount of mushroom residue reached 45%, the edges of the fruiting bodies began to curl from smooth; the fruiting bodies of groups B and C were slightly larger than those of the control group, and group C was the thickest, and had a stronger fleshy texture, 0.85mm thicker than the control group (CK), which was a more ideal mushroom shape.

[0116] Table 7 Morphology of Pleurotus eryngii fruiting bodies under different cultivation formulas

[0117]

[0118]

[0119] The first wave of mushrooms is 15 days after mushroom induction. At this time, the cap is expanded to 70% mature, the edge is slightly rolled inward, the color is bright, and the texture is fresh. Picking late will cause the fruiting body to open and roll outward, and the color is dull. When picking, pick the mushroom feet clean to avoid contamination during the next wave of mushrooms. After harvesting, first place the mushroom bag to wait for the material surface to dry for 2 days, and then carry out the above steps of mushroom induction. After 6 days of mushroom induction, mushroom buds will emerge, and then the above steps can be carried out for the second wave of mushroom management. Usually, the total time from the first wave of mushroom picking to the second wave of mushroom picking is 18 days.

[0120] As can be seen from Table 8, the average yield of the first wave of the test formula C was the highest (261.4 g), and the total bioconversion rate of the two waves was 95.8%, which was slightly lower than that of the control group (CK). However, combined with the fact that the fruiting time was 4 days earlier than the control, overall, the performance of group C was outstanding.

[0121] Table 8 Statistics of harvesting of Yuanmo Mushroom and Liangchao Mushroom under different cultivation formulas

[0122]

[0123] Example 3

[0124] The cultivated mushrooms of Example 2 are subjected to any of the following processing:

[0125] (1) Preservation: After harvesting, fresh mushrooms are clustered, single-sliced, semicircular, fan-shaped, light yellow, and tender. If not stored properly, they will turn brown, soften, change color, become sticky, and even rot and deteriorate. In order to maintain their excellent condition, they are packaged in low-temperature fresh-keeping packaging (the packaging room temperature is 12°C) and stored at 4°C to 6°C for 25-30 days.

[0126] (2) Drying: Under the premise of ensuring product quality, natural or artificial conditions are used to promote the evaporation of water in fresh mushroom bodies to reduce the water content to below 13%. Drying can be selected at a temperature of 50°C. Too high a drying temperature will lead to nutrient loss and the fruiting body will turn brown and look unsightly.

[0127] In summary, the strain of the Agaricus velutipes screened and isolated by the present invention can be used as a cultivation medium in which part of the sawdust is replaced by the Flammulina velutipes residue, which can significantly improve the growth potential and growth rate of Agaricus velutipes mycelium, shorten the cultivation period of Agaricus velutipes, increase the yield of Agaricus velutipes, and improve the quality of Agaricus velutipes.

[0128] SEQ ID NO.1:

[0129] .

[0130] The embodiments described above are only descriptions of the preferred modes of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all fall within the protection scope determined by the claims of the present invention.

Claims

1. A strain of Trichosanthes viridis, It is characterized in that The deposit number of the Pleurotus eryngii strain is CGMCC No.41571.

2. A method for cultivating shiitake mushrooms, It is characterized in that The method comprises the step of performing secondary cultivation on the enoki mushroom strain of claim 1 using a cultivation medium in which the enoki mushroom residue replaces part of the sawdust, wherein the cultivation medium comprises the following raw materials in parts by weight: 35 parts of enoki mushroom residue, 45 parts of sawdust, 19 parts of bran and 1 part of lime.

3. The cultivation method according to claim 2, It is characterized in that The cultivation method comprises the following steps: Inoculating the Pleurotus eryngii strain onto a PDA slant for culture to obtain a mother strain; Inoculate the mother strain into a liquid strain culture medium, shake and culture, and obtain a liquid strain; The cultivation medium is bagged to form a fungus bag, and then the fungus bag is inoculated with the liquid fungus, and then transferred to a culture room for cultivation; Fruiting and harvesting.

4. The cultivation method according to claim 3, It is characterized in that The PDA slant was inoculated with the described Pleurotus eryngii strain and cultured at 24° C. for 9 days. After the mycelium was fully grown, the test tube strain with dense, white, robust mycelium and no infection by any foreign bacteria was selected as the mother strain.

5. The cultivation method according to claim 3, It is characterized in that The shaking culture conditions are: temperature of 24° C., rotation speed of 150 r / min, and constant temperature shaking culture for 7 days.

6. The cultivation method according to claim 3, It is characterized in that Each of the mushroom bags is filled with 1.0-1.5 kg of the cultivation medium.

7. The cultivation method according to claim 3, It is characterized in that Every 400 mL of the liquid culture was inoculated into 20 bacteria bags.

8. The cultivation method according to claim 3, It is characterized in that The culture medium culture conditions are: 1-2 days after inoculation, the temperature is 24-25°C; 3-15 days after inoculation, the temperature is 20-22°C; 16-35 days after inoculation, the temperature is 18-20°C; During the culture period, when the mycelium begins to germinate and grow, ventilation should be carried out once a day at noon for 1 hour each time; when the mycelium grows to more than 1 / 3 of the bag, ventilation should be carried out once a day in the morning, noon and evening for 1 hour each time.

9. The cultivation method according to claim 2, It is characterized in that The Flammulina velutipes residue is processed as follows before use: pollution-free and mold-free Flammulina velutipes residue is selected and exposed to the sun, and then crushed into sawdust.

10. Use of the mushroom strain according to claim 1 or the cultivation method according to claims 2-9 in any of the following: (1) Application in improving the growth potential and growth rate of Pleurotus ostreatus mycelium; (2) Application in increasing the yield of mushrooms; (3) Application in shortening the cultivation cycle of mushrooms; (4) Application in improving the bioconversion rate of mushrooms.

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