A multi-color hymenophaga and a cultivation method thereof
By mixing and inoculating liquid spawn and optimizing culture conditions, the problems of complex operation and high cost in the cultivation of shimeji mushrooms have been solved, enabling the efficient production of multi-colored shimeji mushrooms, improving product quality and yield, and making it suitable for supermarkets, restaurants, and ready-to-cook meals.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- GUANGDONG XINGHE EDIBLE FUNGI TECH RES CO LTD
- Filing Date
- 2023-12-28
- Publication Date
- 2026-05-19
AI Technical Summary
In existing technologies, the cultivation methods for shiitake mushrooms have problems such as long spawn production time, complex operation, high cost, and difficulty in guaranteeing product quality and yield. In particular, when producing multi-colored shiitake mushrooms, it is difficult to achieve simultaneous fruiting of multiple colors in the same bottle.
By using liquid spawn, brown and white *Pleurotus eryngii* strains were mixed and inoculated in the same culture bottle in a specific ratio. By optimizing culture conditions and management methods, and avoiding the spraying of nutrient solution and blue light exposure, the growth of multicolored *Pleurotus eryngii* was achieved.
The process has been simplified, production costs have been reduced, product diversity and yield have been increased, the growth cycle has been shortened, and shiitake mushroom products with four colors have been produced, which are suitable for supermarkets, restaurants and ready-to-cook meals and other fields.
Smart Images
Figure CN117598155B_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the field of shiitake mushroom technology, specifically to a multi-colored shiitake mushroom and its cultivation method. Background Technology
[0002] In the current technology, due to the limitations of cultivation technology and other conditions between different varieties, it is impossible to grow different colored varieties in the same cultivation bottle during the cultivation of shiitake mushrooms. Therefore, the cultivation of multicolored shiitake mushrooms mostly relies on genetic modification. However, genetically modified foods are still controversial.
[0003] CN 108713446 A discloses a method for cultivating bicolor *Pleurotus eryngii*. This method involves adjusting the nutrient solution composition and culture conditions within a conventional cultivation method. The bicolor cultivation method can achieve simultaneous fruiting of white and brown *Pleurotus eryngii* after a cultivation cycle of 75-85 days and a fruiting period of 20-22 days. The method involves inoculating white and brown *Pleurotus eryngii* mother cultures into the same culture medium, taking half of each mother culture and inoculating half of each half into the same culture medium. However, it has the following problems:
[0004] 1. Using solid spawn, which is prepared separately and then inoculated together into a cultivation bottle, results in a long spawn preparation time, low inoculation efficiency, and inconvenience in operation. When half of each spawn is inoculated, most of the mushrooms will be Pleurotus ostreatus, with fewer Pleurotus eryngii.
[0005] Second: Nutrient solution needs to be sprayed during the cultivation period, increasing the cost of raw materials and labor management. Nutrient solution should be sprayed on the side of the brown shiitake mushroom every 2-3 days during the germination and mushroom cultivation periods.
[0006] Third: Irradiate the brown shiitake mushroom side with 200-300Lx blue light for 6-8 hours every day until harvest; the problem is that, with the same bottle opening diameter of 8-9 cm, it is difficult to avoid affecting the white mushroom side when spraying nutrient solution and irradiating blue light on the shiitake mushroom side, and it also increases the operation process and cost.
[0007] The above three problems are serious defects in the current technology, which directly affect the implementation of cultivation techniques and cannot guarantee the quality and yield of products, resulting in the fact that there are still no multicolored shiitake mushroom products on the market. Summary of the Invention
[0008] In view of this, the present invention provides a multicolored *Pleurotus eryngii* mushroom and its cultivation method. Two *Pleurotus eryngii* strains of different colors are inoculated into a cultivation bottle according to a specific ratio and inoculation method, producing *Pleurotus eryngii* fruiting bodies with multiple colors on the cap of the same mushroom.
[0009] The technical solution of this invention is as follows:
[0010] A method for cultivating multicolored shiitake mushrooms includes the following steps:
[0011] S1. Brown and white *Pleurotus ostreatus* strains were inoculated into liquid culture medium and then fermented or shaken to prepare liquid spawn of white and brown *Pleurotus ostreatus*.
[0012] S2. Simultaneously inoculate brown and white liquid spawn of *Pleurotus eryngii* into the same liquid culture medium to prepare a mixed liquid spawn; in S2, the ratio of the amount of brown liquid spawn to white liquid spawn is 1:1 to 1:18; the amount of brown liquid spawn should not exceed the amount of white liquid spawn, otherwise almost all the fruiting bodies will be brown, and multicolored *Pleurotus eryngii* will not grow.
[0013] S3. Inoculate the mixed liquid spawn into the culture medium, and after a period of cultivation, carry out fruiting management to obtain the multicolored Pleurotus ostreatus fruiting bodies.
[0014] The white shiitake mushroom strain includes at least one of Xinghe 2, white shiitake mushroom WB-19, and white shiitake mushroom WB-86.
[0015] The brown shiitake mushroom strain includes at least one of shiitake mushroom XCXH-2 and shiitake mushroom strain WZ-29.
[0016] In one embodiment of the present invention, the white *Pleurotus eryngii* strain is Xinghe 2; the brown *Pleurotus eryngii* strain is *Pleurotus eryngii* XCXH-2.
[0017] Step S1 specifically involves inoculating the mother cultures of white and brown *Pleurotus ostreatus* strains into liquid culture medium, respectively, and incubating at 120-160 rpm. -1 The liquid spawn of white and brown *Pleurotus ostreatus* was obtained by shaking culture at 18-28℃ for 5-10 days.
[0018] The mother culture can be either a test-tube mother culture or a plate culture.
[0019] Step S2 specifically involves simultaneously inoculating the white and brown liquid spawn of *Pleurotus eryngii* from step S1 into the same liquid culture medium, and incubating at 120-160 rpm. -1 The mixed liquid bacterial culture was prepared by shaking culture at 18-28℃ for 5-10 days.
[0020] In steps S1 and S2, the liquid culture medium, by mass percentage, includes 0.3-0.5% sugar, 0.3-0.5% soybean meal, 0.05-0.1% potassium dihydrogen phosphate, 0.05-0.2% magnesium sulfate, and the remainder is water.
[0021] Step S3 specifically involves inoculating the mixed liquid spawn into the culture medium and culturing it at 24–26°C, relative humidity 65–75%, and CO2 concentration 3500 ppm for 75–90 days. After this, fruiting management is carried out to obtain the multicolored Pleurotus ostreatus fruiting bodies.
[0022] In step S3, the conditions for mushroom cultivation management are as follows:
[0023] For the first 1-5 days, it will be dark, with a temperature of 13℃~15℃, relative humidity of 85%~95%, and carbon dioxide concentration of 1000~2000ppm;
[0024] From day 6 to 22, maintain a temperature of 13℃ to 15℃ and a light intensity of 300 to 500 Lux. Turn on the shelf lights intermittently for 5 to 60 minutes each day and turn them off for 30 to 60 minutes. Harvest when the mushroom caps are nearly mature.
[0025] The culture medium for the cultivar comprises 36-40% cultivar substrate by weight percentage, with the remainder being water.
[0026] The culture medium formula for the cultivar includes, by weight percentage: 25-30% corn cob, 18-20% cottonseed hulls, 18-20% wheat bran, 15-17% beet pulp, 5-7% corn flour, 7-10% rice bran, 2-3% soybean meal, and 1-2% shell powder. The culture medium can also use commonly used cultivar formulas in existing technologies.
[0027] Compared with the prior art, the present invention has the following beneficial effects:
[0028] 1. Due to antagonistic interactions between different strains, this invention selects strains XCXH-2 and Xinghe No. 2 from different *Hylocereus undatus* strains through antagonistic experiments. Furthermore, it optimizes the inoculation method and ratio. Compared to existing technologies, the entire management process does not require nutrient solution spraying. Using the method of this invention, only routine management is needed; there is no need to spray nutrient solution or expose to blue light. This solves the problems of multiple, long, and complex seed production processes and inoculation steps when two colors of *Hylocereus undatus* are grown in one production bottle, while simultaneously improving yield and quality. It allows a single *Hylocereus undatus* mushroom to contain four colors, achieving a cultivation cycle of approximately 85 days, a growth cycle of approximately 21 days, and a bioconversion rate of approximately 110%.
[0029] 2. By adjusting the ratio of brown and white liquid spawn, different trial production needs can be met; the proportion of brown and white fruiting body caps can be adjusted and controlled when the fruiting bodies emerge. However, the ratio of brown to white liquid spawn should be controlled between 1:1 and 1:18. When the amount of white liquid spawn is less than that of brown liquid spawn, the proportion of dark brown and light brown will be 100%, and it will be almost impossible for white fruiting bodies to grow.
[0030] 3. By adopting the method of this invention, a product with four colors on a single shiitake mushroom can be produced. It can be used in the food industry such as supermarkets, catering, and pre-cooked meals. Compared with the traditional single-color shiitake mushroom, the colors are rich and diverse, which can further enhance the color diversity of dishes. It also provides the market with a brand-new multi-colored shiitake mushroom product, which will be more popular with consumers. Moreover, the preparation process is simple, the growth cycle is short, the production capacity is high, and the cost is low.
[0031] 4. The multi-colored shiitake mushroom product prepared by this invention has a production process that is basically the same as that of existing bottle-cultured industrialized shiitake mushrooms. No additional factory construction or workshop renovation is required for production. The process is simple and highly stable. Furthermore, liquid spawn can be used to replace traditional solid spawn in the spawn preparation process, which optimizes the spawn production process, reduces production costs, reduces labor in spawn production, and improves spawn quality and automated production capabilities. Attached Figure Description
[0032] Other features, objects, and advantages of the present invention will become more apparent from the following detailed description of non-limiting embodiments with reference to the accompanying drawings:
[0033] Figure 1 This is a graph showing the antagonistic experiment results of Example 1;
[0034] Figure 2 This describes the fruiting performance of the solid-seed hybrid inoculation method in Example 2;
[0035] Figure 3 This is a comparison image of the fruiting performance of experimental group 11 in Example 3 with that of the fruiting bodies of Pleurotus eryngii XCXH-2 and Xinghe 2 inoculated alone.
[0036] Figure 4 The fruiting performance of experimental groups 9, 10, 11, 12, and 13 in Example 3, and the comparison of fruiting bodies of *Pleurotus eryngii* XCXH-2 and *Xinghe 2* inoculated alone.
[0037] Figure 5 This describes the batch cultivation performance of liquid seed cross-grafting method B in Example 2;
[0038] Figure 6 This refers to the fruiting performance of experimental group 11 in Example 3 on day 20;
[0039] Figure 7 This refers to the fruiting performance of experimental group 13 in Example 3 on day 20;
[0040] Figure 8 This is a diagram of sub-entity 14 in test group 3 of Example 3. Detailed Implementation
[0041] The present invention will be described in detail below with reference to embodiments. These embodiments will help those skilled in the art to further understand the present invention, but do not limit the invention in any way. It should be noted that those skilled in the art can make several adjustments and improvements without departing from the concept of the present invention. These all fall within the protection scope of the present invention.
[0042] In the following embodiments:
[0043] Source of strain:
[0044] True Plum Mushroom XCXH-2 ( Hypsizygus marmoreus *Pleurotus eryngii* XCXH-2: Preservation number CCTCCM2020068, is a brown *Pleurotus eryngii*.
[0045] Hypsizygus marmoreus Galaxy No. 2: Collection number: GDMCC NO:61908, is a white true mushroom;
[0046] Pleurotus ostreatus strain WZ-29 ( Hypsizygus marmoreus WZ-29): Its preservation number is GDMCC NO: 60276, brown shiitake mushroom.
[0047] White Pleurotus eryngii WB-19: The strain was obtained by isolating fruiting body tissue from "Yulongdong" seafood mushrooms purchased from the market.
[0048] White Pleurotus eryngii WB-86: The strain was obtained by isolating fruiting body tissue from “Shennong Baixue” seafood mushroom purchased from the market.
[0049] Among them, the tissue isolation method is referenced in "Key Points of Edible Fungus Tissue Isolation Method for Seed Production. Lü Zhimin and Zhong Xiuying, Journal of Xichang University (Natural Science Edition), 2001, No. 4".
[0050] Preparation Example 1
[0051] 1. Preparation of PDA culture medium and petri dishes
[0052] 1.1 PDA culture medium
[0053] Formula: Use 200g potato, 20g glucose, and 20g agar per 1L of culture medium.
[0054] Autoclave at 121°C for 20 minutes. After sterilization, pour the sterilized plate into a clean bench to prepare a plate culture medium. Alternatively, while still hot, dispense the contents into test tubes and sterilize. Fill the test tubes with about one-fifth to one-quarter of their height. Seal the test tubes with cotton plugs and autoclave at 121°C for 20 minutes. After sterilization, place the plate on a slant to prepare a slant culture medium for later use.
[0055] 1.2 Preparation of Petri Dishes
[0056] Test tube cultures of strains *Pleurotus eryngii* XCXH-2, Xinghe 2, WZ-29, WB-19, and WB-86 were inoculated into the plate culture medium prepared in step 2.1.1 and cultured at 22-25℃ in the dark until the mycelium completely covered the plates, thus obtaining plate cultures of strains *Pleurotus eryngii* XCXH-2, Xinghe 2, WZ-29, WB-19, and WB-86 for later use.
[0057] 2. Liquid culture medium
[0058] Formula: By weight percentage, it includes 0.3% white sugar, 0.3% soybean meal powder, 0.05% potassium dihydrogen phosphate, 0.05% magnesium sulfate, and the balance is water.
[0059] Preparation method: Prepare liquid culture medium according to the ratio, dispense into Erlenmeyer flasks, and autoclave at 121℃ and 101KPa for 30 minutes for later use; 300 ml of the liquid culture medium is prepared into 500 ml Erlenmeyer flasks (preparing liquid culture medium No. 1); 700 ml of the liquid culture medium is prepared into 1000 ml Erlenmeyer flasks (preparing liquid culture medium No. 2).
[0060] 3. Original culture medium
[0061] Original culture medium: by weight percentage, it consists of 36% original culture medium and the remainder is water.
[0062] The original culture medium consists of, by weight percentage, 65% sawdust, 20% rice bran, and 15% wheat bran.
[0063] Preparation method of original culture medium: Prepare the original culture medium according to the ratio and put it into the original culture bottle. Make a hole in the middle to the bottom of the bottle (hole diameter 18mm). Sterilize at 121℃ and 0.13MPa for 120 minutes. The original culture bottle is 850cc and the amount of culture medium used in each bottle is 500g.
[0064] 4. Culture medium for cultivation seeds
[0065] The culture medium for the cultivar consists of 36% cultivar substrate by weight, with the remainder being water.
[0066] Cultivation substrate formula: by weight percentage, it includes 30% corn cob, 20% cottonseed hull, 18% wheat bran, 15% beet pulp, 5% corn flour, 7% rice bran, 3% soybean meal, and 2% shell powder.
[0067] Preparation method of cultivation medium: Prepare the cultivation medium according to the ratio and put it into the cultivation bottle (make a hole in the middle to the bottom of the bottle with a diameter of 18mm). Sterilize at 121℃ and 0.13MPa for 120 minutes. The cultivation bottle size is 850cc and the amount of culture medium used in each bottle is 580g.
[0068] Example 1: Strain Screening
[0069] 1. Plate Antagonism Test
[0070] The test method was based on NY / T 1845 Differential Identification of Edible Fungi Strains and Antagonistic Reactions.
[0071] The specific procedures for the antagonistic experiment are as follows: *Pleurotus eryngii* strains XCXH-2, Xinghe 2, WZ-29, WB-19, and WB-86 were inoculated and cultured on PDA plates according to Table 1, with inoculation sites 3 cm apart. The plates were incubated at 23℃ for 18 days. The results of the antagonistic experiment are shown in Table 1 and... Figure 1 As shown.
[0072] Table 1
[0073]
[0074] 2. Comparative experiment on fruiting rate
[0075] Fruiting tests were conducted on the following mushroom varieties (XCXH-2, Xinghe 2, WZ-29, WB-19, and WB-86) according to the experimental groups in Table 2.
[0076] Specifically, the steps include the following:
[0077] Step 1: Prepare liquid inoculum for each strain separately. Specifically, take three 6mm × 6mm inoculation blocks from each of the following strains (XCXH-2, Xinghe 2, WZ-29, WB-19, and WB-86) and inoculate them into the No. 1 liquid culture medium prepared in Preparation Example 1. Incubate on a shaker at 22-23℃ (140 rpm). -1 After 7 days of cultivation, liquid cultures of strains *Pleurotus ostreatus* XCXH-2, Xinghe 2, WZ-29, WB-19, and WB-86 were obtained.
[0078] Step 2: Following the combination method in Table 2, take 15 ml of liquid culture of strain A and 15 ml of liquid culture of strain B respectively, and inoculate them into the same bottle of liquid culture medium No. 2 prepared in step 2 of Preparation Example 1. Incubate in a shaker at 22-23℃ (140 rpm). -1 After 7 days of cultivation, a mixed liquid seed was obtained.
[0079] Step 3: Inoculate the mixed liquid inoculum (20ml) into the culture medium (Step 1.3 in Preparation Example 1), with 32 bottles inoculated for each experimental group, and test according to the fruiting results.
[0080] The fruiting test process conditions are as follows:
[0081] 1) Cultivation management in culture bottles: After inoculation, culture at 24-26℃, relative humidity 65-75%, and CO2 concentration controlled below 3500ppm. After the mycelium has fully grown, continue mycelial maturation culture for a total of 80-85 days before scratching.
[0082] Scratching refers to removing the overgrown and old mycelia from the surface of the substrate at the bottle opening after the mycelia have fully matured.
[0083] 2) Management of mushroom production in cultivation bottles
[0084] After scratching, during the first 1-5 days of growth, maintain darkness, a temperature of 13-15℃, humidity of 85%-95%, and a carbon dioxide concentration of 1500-2500ppm. From the 6th to the 22nd day, control the light exposure intermittently, turning it on for 5-60 minutes and off for 60 minutes each day, with a light intensity of 300-500Lx, a temperature of 13-15℃, a carbon dioxide concentration of 1500-2500ppm, and an indoor humidity of 90-98%. Harvest when the mushroom cap diameter is 1.2-1.8cm and the mushrooms are nearly mature.
[0085] The results are shown in Table 2.
[0086] Table 2
[0087]
[0088] Example 2: Cultivation Method Optimization
[0089] Test strains: The *Pleurotus ostreatus* XCXH-2 plate strain prepared in step 2 of Preparation Example 1 and the *Xinghe 2* plate strain.
[0090] 1. Solid-seed cross-grafting method
[0091] 1.1 Preparation of original seeds
[0092] The *Pleurotus eryngii* strain XCXH-2 and strain Xinghe No. 2, prepared in step 3 of Preparation Example 1, were aseptically inoculated onto the original culture medium prepared in step 3 of Preparation Example 1. Six 8mm × 8mm inoculation blocks were inoculated onto each bottle, covering the entire surface of the medium. After inoculation, the medium was incubated at 21–23℃ and 65–75% relative humidity. After the mycelium had fully grown, mycelial maturation culture was continued for a total of 40–45 days to obtain solid original cultures of *Pleurotus eryngii* strain XCXH-2 and strain Xinghe No. 2, respectively.
[0093] 1.2. Cultivation
[0094] 20g of each of the solid primary cultures of *Pleurotus eryngii* strain XCXH-2 and strain Xinghe No. 2 were inoculated together in the same culture bottle for fruiting test, with 32 bottles cultivated each time.
[0095] The fruiting test process conditions are as described in Example 1.
[0096] Test results:
[0097] Mushroom bud differentiation stage: Brown and white mushroom buds are distributed at their respective inoculation sites, with a relatively clear color distinction;
[0098] Fruiting body growth period: The brown *Pleurotus eryngii* grows too quickly, covering most of the white *Pleurotus eryngii*, resulting in a low proportion of white products. For example... Figure 2 As shown.
[0099] 2. Liquid type mixing method A
[0100] Six 8mm × 8mm mycelial blocks were taken from each of the *Pleurotus eryngii* XCXH-2 and *Xinghe 2* culture plates and inoculated into the same bottle of liquid culture medium No. 1 (prepared in step 2 of Preparation Example 1). The culture was then placed in a shaker at 23°C and incubated at 140 rpm. -1 After 7 days of cultivation, the culture medium was inoculated into the culture medium (prepared in step 3 of preparation example 1) at an inoculation volume of 20 ml per bottle under aseptic conditions for fruiting test. 64 bottles were tested.
[0101] The fruiting test process conditions are as described in Example 1.
[0102] Test results:
[0103] The fruiting body cap has four colors: dark brown, light brown, milky white, and white. White and dark brown account for about 20%, while light brown and milky white account for about 60% and 20%, respectively.
[0104] 3. Liquid-type mixing method B
[0105] 3.1 Preparation of Pleurotus eryngii XCXH-2 liquid spawn: Take three 6mm × 6mm inoculation blocks from the Pleurotus eryngii XCXH-2 plate spawn (preparation method is described in step 1.2 of Preparation Example 1), inoculate them onto liquid culture medium No. 1 (prepared in step 2 of Preparation Example 1), and incubate them on a shaker at 22℃ (140 r·min). -1 5 days.
[0106] 3.2 Preparation of Xinghe 2 liquid inoculum: Three 6mm × 6mm inoculum blocks of Xinghe 2 (preparation method is described in step 1.2 of Preparation Example 1) were taken and placed in liquid culture medium No. 1 (prepared in step 2 of Preparation Example 1) at 22℃ on a shaker (140 r·min). -1 5 days.
[0107] 3.3 Under aseptic conditions, take 2 ml of *Pleurotus ostreatus* XCXH-2 liquid inoculum and 4 ml of *Xinghe No. 2* liquid inoculum, and inoculate them into the same No. 2 liquid culture medium (prepared in step 2 of Preparation Example 1). Incubate at 22℃ and 140 rpm. -1The culture was carried out on a shaker for 7 days to obtain a mixed liquid spawn. The mixed liquid spawn was then inoculated onto the culture medium for fruiting tests.
[0108] The fruiting test conditions and procedures are as described in Example 1.
[0109] Test results:
[0110] The fruiting body cap has four colors: dark brown, light brown, milky white, and white. White and dark brown account for approximately 30% of the total, while light brown and milky white account for approximately 50% and 20%, respectively. Figure 5 ).
[0111] 4. Solid-liquid bonding and mixing
[0112] 4.1. Under aseptic conditions, the *Pleurotus eryngii* strain XCXH-2 and strain Xinghe No. 2 prepared in step 3 of Preparation Example 1 were inoculated onto the original culture medium prepared in step 3 of Preparation Example 1. Six 8mm × 8mm inoculation blocks were inoculated into each bottle, covering the surface of the medium. After inoculation, the medium was incubated at 23℃ and 75% relative humidity. After the mycelium had fully grown, mycelial maturation culture was continued for a total of 40-45 days (depending on the degree of mycelial growth). Solid original cultures of *Pleurotus eryngii* strain XCXH-2 and strain Xinghe No. 2 were obtained.
[0113] 4.2 Under aseptic conditions, 10g each of *Pleurotus eryngii* XCXH-2 and *Xinghe 2* solid spawn were inoculated into the same No. 2 liquid culture medium (prepared in step 2 of Preparation Example 1) and incubated at 22℃ and 140 rpm. -1 After 7 days of shaking culture, the culture medium was inoculated into the culture medium (prepared in step 3 of preparation example 1) for fruiting test, with a sample size of 32 bottles.
[0114] The fruiting test conditions and procedures are as described in Example 1.
[0115] Test results:
[0116] The fruiting body cap has four colors: dark brown, light brown, milky white, and white. White and milky white account for a small proportion, about 15%, while light brown and dark brown account for about 70% and 15%, respectively.
[0117] 5. Inoculate at the connection line of the petri dish.
[0118] The specific steps are as follows:
[0119] The *Pleurotus eryngii* XCXH-2 culture prepared in step 2 of Preparation Example 1 and the *Xinghe 2* culture were simultaneously inoculated on a plate culture medium 3 cm apart, with inoculation blocks measuring 6 mm × 6 mm. The inoculations were incubated at 23°C for 16 days. Mycelia from the fusion point (6 blocks measuring 8 mm × 8 mm) were then inoculated onto liquid medium No. 1 (prepared in step 2 of Preparation Example 1) and cultured on a shaker at 22°C (140 rpm). -1 After 7 days of cultivation, the culture medium (prepared in step 3 of preparation example 1) was used for fruiting test, with a sample size of 32 bottles.
[0120] The fruiting test conditions and procedures are as described in Example 1.
[0121] Test results: The fruiting performance was unstable. In the same bottle, some fruits were completely white, some were completely brown, and some fruiting bodies had caps with four colors: dark brown, light brown, milky white, and white.
[0122] summary:
[0123] This invention compares and studies several different inoculation methods and finds that liquid inoculation method B produces the best fruiting results. The test results show that the fruiting body caps have four colors: dark brown, light brown, milky white, and white. White and dark brown account for a small proportion, about 30%, while light brown and milky white account for about 50% and 20%, respectively.
[0124] Example 3: Experiment on the Dosage of Pleurotus eryngii XCXH-2 and Xinghe No. 2 Liquid Spawn and Fruiting Performance of Pleurotus eryngii Multicolor
[0125] This embodiment uses the liquid seed mixing method B of embodiment 2, and the fruiting test conditions and steps are the same as those in embodiment 1.
[0126] The specific steps are as follows:
[0127] 1. Preparation of liquid spawn of *Pleurotus eryngii* XCXH-2: Take three 6mm × 6mm inoculation blocks from the *Pleurotus eryngii* XCXH-2 plate spawn (preparation method is described in step 1.2 of Preparation Example 1) and inoculate them into liquid culture medium No. 1 (prepared in step 2 of Preparation Example 1). Incubate on a shaker at 22-23℃ (140 r·min). -1 ).
[0128] 2. Preparation of Xinghe 2 liquid inoculum: Take three 6mm×6mm inoculum blocks of Xinghe 2 (preparation method is described in step 1.2 of Preparation Example 1), place them in liquid culture medium No. 1 (prepared in step 2 of Preparation Example 1), and incubate them on a shaker at 22-23℃ (140 r·min). -1 ).
[0129] 3. After culturing on a shaker for 5 days, under aseptic conditions, take the inoculation amounts of *Pleurotus eryngii* XCXH-2 liquid spawn and *Xinghe No. 2* liquid spawn according to Table 3, and inoculate them into the same No. 2 liquid culture medium (prepared in step 2 of Preparation Example 1). Incubate at 22℃ and 140 rpm. -1 After 7 days of shaking culture, the culture medium was inoculated into the culture medium (prepared in step 3 of preparation example 1) for fruiting test, with 32 bottles as the sample size for each experimental group.
[0130] The fruiting test conditions and procedures are as described in Example 1.
[0131] Test results:
[0132] Table 3
[0133]
[0134] in, Figure 3 This is a comparison image of the fruiting performance of experimental group 11 in Example 3 with that of the fruiting bodies of Pleurotus eryngii XCXH-2 and Xinghe 2 inoculated alone.
[0135] Figure 4 The fruiting performance of experimental groups 9, 10, 11, 12, and 13 in Example 3, and the comparison of fruiting bodies of *Pleurotus eryngii* XCXH-2 and *Xinghe 2* inoculated alone.
[0136] Figure 6 The fruiting performance of experimental group 11 on day 20;
[0137] Figure 7 The fruiting performance of experimental group 13 on day 20;
[0138] Figure 8 This is a diagram of the 14 sub-entities in the experimental group.
[0139] The specific embodiments of the present invention have been described above. It should be understood that the present invention is not limited to the specific embodiments described above, and those skilled in the art can make various modifications or variations within the scope of the claims, which do not affect the essence of the present invention.
Claims
1. A cultivation method for multicolored shiitake mushrooms, characterized in that, Includes the following steps: S1. Brown and white *Pleurotus ostreatus* strains were inoculated into liquid culture medium and cultured by shaking to prepare liquid spawn of white and brown *Pleurotus ostreatus*. S2. Brown and white liquid spawn of Pleurotus ostreatus are simultaneously inoculated into the same liquid culture medium to prepare a mixed liquid spawn. The ratio of brown shiitake mushroom liquid spawn to white shiitake mushroom liquid spawn is 1:1 to 1:
18. S3. Inoculate the mixed liquid spawn into the culture medium, and after a period of cultivation, perform fruiting management to obtain the multicolored shiitake mushroom fruiting bodies. The white shiitake mushroom strain is Hypsizygus marmoreus Galaxy 2; Collection number: GDMCC NO:61908; The brown shiitake mushroom strain is Hypsizygus marmoreus XCXH-2; accession number CCTCC M2020068; Step S1 specifically involves inoculating the mother cultures of white and brown *Pleurotus eryngii* strains into liquid culture medium, respectively, at 120-160 rpm. -1 The liquid spawn of white and brown *Pleurotus ostreatus* was obtained by shaking culture at 18-28℃ for 5-10 days.
2. The method for cultivating multicolored shiitake mushrooms according to claim 1, characterized in that, Step S2 specifically involves simultaneously inoculating the white and brown liquid spawn of *Pleurotus eryngii* from step S1 into the same liquid culture medium, and incubating at 120-160 rpm. -1 The mixed liquid bacterial culture was prepared by shaking culture at 18-28℃ for 5-10 days.
3. The method for cultivating multicolored shiitake mushrooms according to claim 1, characterized in that, In steps S1 and S2, the liquid culture medium, by mass percentage, includes 0.3-0.5% sugar, 0.3-0.5% soybean meal, 0.05-0.1% potassium dihydrogen phosphate, 0.05-0.2% magnesium sulfate, and the remainder is water.
4. The method for cultivating multicolored shiitake mushrooms according to claim 1, characterized in that, Step S3 specifically involves inoculating the mixed liquid spawn into the culture medium and culturing it at 24–26°C, relative humidity 65–75%, and CO2 concentration 3500 ppm for 75–90 days. After that, fruiting management is carried out to obtain the multicolored Pleurotus ostreatus fruiting bodies.
5. The method for cultivating multicolored shiitake mushrooms according to claim 1, characterized in that, In step S3, the conditions for mushroom cultivation management are as follows: For the first 1-5 days, it will be dark, with temperatures ranging from 13°C to 18°C, relative humidity from 85% to 95%, and carbon dioxide concentration from 1000 to 2000 ppm. From day 6 to 22, maintain a temperature of 13℃ to 18℃ and a light intensity of 300 to 500 Lux. Turn on the shelf lights intermittently for 5 to 60 minutes each day and turn them off for 30 to 60 minutes. Harvest when the mushroom caps are nearly mature.
6. A multicolored Pleurotus ostreatus obtained by the cultivation method according to any one of claims 1-5.