Red tricholoma matsutake culture medium using litchi branches as matrix and cultivation method thereof

Abstract

The invention discloses a tricholoma matsutake culture medium using litchi branches as a matrix and a cultivation method thereof. The tricholoma matsutake culture medium is prepared from the following components in percentage by weight: 30-50% of litchi sawdust, 30-50% of cottonseed hulls, 3-8% of bran, 3-8% of corncobs, 3-8% of rice husks, 1-5% of corn flour and 1-5% of lime. The method is characterized in that the litchi branches are added into the stock liquid culture medium for making the branch seeds for the first time, on one hand, the tricholoma matsutake strains can be trained to generate extracellular enzymes capable of utilizing litchi wood chips, macromolecules of the litchi branches can be rapidly decomposed, and on the other hand, nutrient elements contained in the litchi branches can promote growth of tricholoma matsutake hyphae more rapidly, and the growth rate of the tricholoma matsutake hyphae is increased. The branch seeds are large in inoculation area and high in hybrid resistance, the growth time of mycelium of the cultivation material can be shortened, and the fruiting rate is increased. The invention not only provides a solution for innocent treatment of the litchi branches, but also finds a new culture medium for red tricholoma matsutake culture.

Description

Technical Field

[0001] The present invention relates to the technical field of the cultivation of Stropharia rugosoannulata, and specifically, to a culture medium for Stropharia rugosoannulata using litchi branches as a substrate and a cultivation method thereof. Background Art

[0002] Stropharia rugosoannulata, scientifically known as Stropharia rugosoannulata Farlow ex Murrill, also known as Stropharia rugosoannulata, Stropharia rugosoannulata, and Stropharia vinaceoavellanea, is one of the top ten mushrooms in the international mushroom trading market and is also one of the mushroom species recommended by the Food and Agriculture Organization of the United Nations (FAO) for cultivation in developing countries. The mushroom color is wine red, the stalk is thick and the cap is plump. It has a fragrant taste, smooth and tender flesh and a crispy stalk, and has an excellent taste. Its fruit body is rich in high-quality mycelial protein, amino acids and various trace elements. It is a rare edible mushroom with extremely high nutritional value and has the reputation of "the meat among vegetables", and is deeply loved by consumers. It belongs to mesophilic bacteria and can be cultivated in most regions of China. In China, it is mainly cultivated with corn straw, wheat straw, rice straw, etc. Generally, broad-leaved tree sawdust is added to optimize the nutritional components of the cultivation material to extend the fruiting period. In recent years, with the rapid development of the Stropharia rugosoannulata industry in China, the required cultivation raw materials have increased accordingly. The rising price of traditional cultivation materials has restricted the development of this industry. In order to save cultivation costs and solve the problem of shortage of raw materials, exploring new cultivation substrates has gradually become a new trend.

[0003] Agricultural and forestry waste is an important biological resource. How to turn agricultural and forestry waste into "treasure", reduce environmental pollution and improve the ecological environment has become an important topic of concern. Litchi is the main subtropical fruit tree in China, mainly planted in regions such as Guangdong, Guangxi, and Hainan, and is an important cash crop in southern China. During the cultivation of litchi, it is necessary to prune the branches and leaves. Appropriate pruning can not only promote flower bud formation and flowering, but also improve the fruit quality. The pruning work of litchi trees is carried out in summer, autumn and winter, so a large amount of litchi branches are produced in the main litchi producing areas every year. The pruned litchi branches are resources that can be utilized, which contain a large amount of organic matter and nutrients. Yao Lixian et al. measured that the nitrogen content in the branches of the main litchi cultivars was 5.7 g / kg, the potassium content was 4.3 g / kg, the calcium content was 7.7 g / kg, and the phosphorus content was 0.7 g / kg. Therefore, using Stropharia rugosoannulata, which can decompose sawdust to provide nutrients for itself, to reasonably recycle a large amount of waste litchi branches can not only make full use of this forestry waste of litchi branches and reduce forestry non-point source pollution, but also realize the biological conversion and utilization of litchi waste.

[0004] The Russula lepida is a straw-decomposing mushroom. The cultivation raw materials mainly include rice straw, corn straw, wheat straw, soybean straw, water bamboo leaves and other crop straws, with bran, rice bran, etc. as auxiliary materials. However, there is no report on using litchi branches as cultivation materials. At the same time, when culturing the mother strain of Stropharia rugosoannulata, wheat grain medium or culture medium mainly composed of glucose, corn flour and soybean meal powder is generally used for cultivation. The obtained mother strain of Stropharia rugosoannulata has poor degradation ability for straw cellulose, resulting in a long straw degradation time or incomplete straw decomposition. Summary of the Invention

[0005] In view of the above problems, the present invention provides a Russula lepida culture medium using litchi branches as a substrate and its cultivation method. The cultivation method of the present invention can shorten the growth time of the mycelium of the cultivation material and improve the fruiting rate.

[0006] In order to achieve the above object, the present invention adopts the following technical solutions:

[0007] A Russula lepida culture medium using litchi branches as a substrate is made of the following components in weight percentage:

[0008] Litchi sawdust 30 - 50%, cottonseed hull 30 - 50%, bran 3 - 8%, corn cob 3 - 8%, rice husk 3 - 8%, corn flour 1 - 5%, lime 1 - 5%.

[0009] Preferably, it is made of the following components in weight percentage:

[0010] Litchi sawdust 40%, cottonseed hull 40%, bran 5%, corn cob 5%, rice husk 5%, corn flour 3%, lime 2%.

[0011] A cultivation method of Russula lepida includes the following steps:

[0012] (1) Preparation of litchi sawdust:

[0013] The trimmed litchi branches are dried in the sun and then broken into small sections 6 cm - 8 cm long, and further pulverized to obtain litchi sawdust with a particle size of 2 - 4 mm.

[0014] (2) Preparation of litchi branch seeds:

[0015] Take 2 - 3 mycelial blocks with a side length of about 5 mm from the activated mother strain plate of Russula lepida, inoculate them into the liquid original seed medium, and after the mycelium grows, put the sterilized litchi branches into the liquid original seed medium and let them stand for cultivation. Wait until the mycelium covers the litchi branches to obtain litchi branch seeds.

[0016] (3) Preparation of Russula lepida cultivation seeds:

[0017] According to weight percentage, mix 30 - 50% of litchi sawdust, 30 - 50% of cottonseed hulls, 3 - 8% of wheat bran, 3 - 8% of corncobs, 3 - 8% of rice husks, and 1 - 5% of corn flour evenly, then add 1 - 5% of lime, add water, stir for 20 - 30 min, control the water content to be 60% - 65%, it is appropriate that water droplets ooze out when the material is pinched tightly by hand, control the pH to be 7.0 - 7.5, then divide and pack into strain bags and collars. The strain bag specification is a polypropylene corner - folded bag of 18 cm × 36 cm × 0.005 cm, and the wet weight of each bag of material is 0.95 kg (equivalent dry weight is 0.35 kg), sterilize at 121 °C for 30 min, and reserve after naturally cooling to room temperature; inoculate the cultured litchi branch seeds into the cultivation material in the strain bag, and cultivate at a constant temperature of 21 °C until the mycelium fills the strain bag to make the red Tricholoma matsutake cultivation strain stick.

[0018] (4) Preparation and fermentation of the red Tricholoma matsutake cultivation material medium:

[0019] According to weight percentage, mix 30 - 50% of litchi sawdust, 30 - 50% of cottonseed hulls, 3 - 8% of wheat bran, 3 - 8% of corncobs, 3 - 8% of rice husks, and 1 - 5% of corn flour evenly to form a material pile, then add 1 - 5% of lime, add water, and stir evenly again, control the water content to be 60% - 65%, and then ferment naturally; make several ventilation holes with a pore diameter of 3 - 5 cm on the material pile of the red Tricholoma matsutake cultivation material medium; when the fermentation temperature of the material pile reaches 65 °C, maintain it for 24 hours, turn the pile once, add urea when turning the pile for the first time, adjust the water content to 65%, continue to ferment, turn the pile 3 - 8 times in total until the temperature of the material pile no longer rises; replenish water each time when turning the pile, so that when the fermentation ends, the water content of the cultivation material medium is 60%;

[0020] (5) Cultivation method:

[0021] Mix the fermented red Tricholoma matsutake cultivation material medium and litchi orchard soil in a volume ratio of 1:1 to obtain the covering soil material; take out the cultured red Tricholoma matsutake cultivation strain stick from the strain bag, bury the red Tricholoma matsutake cultivation strain stick with the covering soil material until the strain stick is invisible, control the thickness of the covering soil layer to be 3 - 5 cm, and then cover a layer of litchi fallen leaves on the surface for moisture preservation; keep the covering soil material moist, and spray - water in a mist form once every 3 days; when the temperature is relatively low, cover the covering soil layer with black non - woven fabric for heat preservation, moisture preservation, and shading; buds will appear 40 - 50 days after covering the soil.

[0022] The technical principle of the present invention: Red Tricholoma matsutake is an aerobic fungus. The present invention uses liquid culture to prepare branch seeds. Adding litchi branches to the culture medium can domesticate and stimulate the red Tricholoma matsutake fungus to produce extracellular enzymes that can utilize litchi branches. The produced enzymes can make the rigid structure of the branches become loose, so that the produced cellulase - like enzymes can quickly decompose macromolecules in the litchi branches into small - molecule substances that can be absorbed and utilized by the red Tricholoma matsutake mycelium, promoting the growth of the mycelium.

[0023] The cultivation medium of the present invention uses litchi sawdust, which contains a large amount of inorganic salt components and can be used as the source of inorganic salts and nitrogen nutrition for Rhizopogon rubescens. Moreover, the inoculation area of the branch species is large, the germination points are numerous, and the anti-contamination ability is strong, which can shorten the growth time of the mycelium of Rhizopogon rubescens in the cultivation medium.

[0024] When cultivating with soil covering in the present invention, litchi orchard soil is added. Its texture is soft, it has a high water holding rate, and its pH is 5.5 - 6.5, which can promote the formation of fruiting bodies.

[0025] Compared with the prior art, the present invention has the following beneficial effects:

[0026] The innovation of the present invention lies in that for the first time, litchi branches are added to the original seed liquid medium to make branch species. First, it can domesticate the Rhizopogon rubescens strain to produce extracellular enzymes that can utilize litchi sawdust, which can rapidly decompose the macromolecules of litchi branches. Second, the nutrient elements contained in litchi branches can more quickly promote the growth of Rhizopogon rubescens mycelium. Moreover, the inoculation area of the branch species is large and the anti-contamination ability is strong, which can shorten the growth time of the mycelium in the cultivation medium and improve the fruiting rate. The present invention not only provides a solution for the harmless treatment of litchi branches, but also finds a new cultivation substrate for the cultivation of Rhizopogon rubescens. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 For the well-cultivated Rhizopogon rubescens litchi branch species in Example 1;

[0028] Figure 2 For the mycelium diagram of the miscellaneous sawdust fungus stick in Comparative Example 1;

[0029] Figure 3 For the mycelium diagram of the litchi sawdust fungus stick in Example 1;

[0030] Figure 4 For the mycelium growth picture of the soil-covered cultivation for 15 days in Example 1;

[0031] Figure 5 For the growth situation of the Rhizopogon rubescens fruiting body after 45 days of soil-covered cultivation in Example 1. DETAILED DESCRIPTION OF THE INVENTION

[0032] Example 1:

[0033] A cultivation method of Rhizopogon rubescens, comprising the following steps:

[0034] (1) Preparation of litchi sawdust:

[0035] The trimmed litchi branches are dried in the sun and then broken into small sections 6 cm - 8 cm long, and further crushed to obtain litchi sawdust of 2 - 4 mm;

[0036] (2) Preparation of litchi branch species:

[0037] Take 2 - 3 agar blocks with a side length of about 5 mm from the activated mother culture plate of Russula vinosa, inoculate them into the liquid original seed medium. After the mycelium grows, put the sterilized litchi branches into the liquid original seed medium and let it stand for cultivation. Wait until the mycelium covers the litchi branches to obtain the litchi branch seeds, as Figure 1 shown.

[0038] The formula of the original seed medium is: 200 g of potato (peeled) (boil in water and take the filtrate to make up the volume to 1000 mL), 20 g of glucose, 2.0 g of yeast extract, 1.0 g of fish peptone, 1.0 g of potassium dihydrogen phosphate, 0.5 g of calcium chloride, 0.5 g of potassium sulfate, 0.5 g of magnesium sulfate, 2 mg of 2,4 - dichlorophenoxyacetic acid, pH 6.5 - 7.0, sterilize at 121 °C for 30 min.

[0039] (3) Preparation of Russula vinosa cultivated seeds:

[0040] According to the weight percentage, mix 40% of litchi sawdust, 40% of cottonseed hulls, 5% of bran, 5% of corncobs, 5% of rice husks and 3% of corn flour evenly, then add 2% of lime, add water, and stir for 20 - 30 min. Control the water content to be 60% - 65%, with water droplets oozing out when squeezing the material by hand being appropriate. Control the pH to be 7.0 - 7.5. Then pack it into the strain bags and put on collars. The strain bag specification is a polypropylene folded - corner bag of 18 cm × 36 cm × 0.005 cm. The wet weight of each bag of material is 0.95 kg (equivalent to a dry weight of 0.35 kg), sterilize at 121 °C for 30 min, and reserve it after naturally cooling to room temperature; inoculate the cultivated litchi branch seeds into the cultivation material in the strain bag, and cultivate at a constant temperature of 21 °C until the mycelium fills the strain bag to make the Russula vinosa cultivated seed stick.

[0041] (4) Preparation and fermentation of Russula vinosa cultivation material medium:

[0042] According to the weight percentage, mix 40% of litchi sawdust, 40% of cottonseed hulls, 5% of bran, 5% of corncobs, 5% of rice husks and 3% of corn flour evenly to form a material pile, then add 2% of lime, add water, and stir evenly again. Control the water content to be 60% - 65%, and then ferment naturally; make several ventilation holes with a pore diameter of 3 - 5 cm on the material pile of the Russula vinosa cultivation material medium; when the fermentation temperature of the material pile reaches 65 °C, keep it for 24 hours and turn the pile once. Add urea when turning the pile for the first time, adjust the water content to 65%, and continue fermentation. Totally turn the pile 3 - 8 times until the temperature of the material pile no longer rises; add water during each turning of the pile so that the water content of the cultivation material medium at the end of fermentation is 60%;

[0043] (5) Cultivation method:

[0044] Mix the fermented Russula brevipes cultivation medium with litchi orchard soil in a volume ratio of 1:1 to obtain the covering soil material; take out the cultivated Russula brevipes cultivation spawn bags from the spawn bags, and bury the Russula brevipes cultivation spawn bags with the covering soil material until the bags are no longer visible. The thickness of the covering soil layer is controlled at 3 - 5 cm, and then cover a layer of litchi leaves on the surface for moisturizing; keep the covering soil material moist, and conduct fog-like watering once every about 3 days. When the temperature is low, cover the covering soil layer with black non-woven fabric for heat preservation, moisture retention and shading. Observe the growth of mycelium after 15 days of soil covering cultivation, starting from Figure 4 It can be seen that the mycelium is thick, with strong growth potential and fast spreading speed. Observe the growth of Russula brevipes fruiting bodies after 45 days of soil covering cultivation, starting from Figure 5 It can be seen that the caps of the Russula brevipes fruiting bodies are plump, with uniform color, the stipes are straight, and the overall growth is good.

[0045] Control Example 1:

[0046] Referring to Example 1, replace the litchi sawdust with ordinary miscellaneous sawdust, and replace the Russula brevipes cultivation medium with a control cultivation medium, and the others are the same as in Example 1. The formula of the control cultivation medium: cottonseed hull 30%, miscellaneous sawdust 20%, wheat bran 18%, corn cob 30%, lime 2%.

[0047] The results are shown in Table 1 and Figures 2-3 as follows. Starting from Figure 2 It can be seen that the mycelium of the miscellaneous sawdust spawn bags in Control Example 1 is sparse and grayish-white in color; while in Example 1 as Figure 3 shown, the mycelium of the litchi sawdust spawn bags is dense and white in color. The growth rate of the mycelium of the litchi sawdust spawn bags is significantly faster than that of the miscellaneous sawdust spawn bags, and the anti-contamination ability of the mycelium of the litchi sawdust spawn bags is also stronger than that of the miscellaneous sawdust spawn bags. It can be seen that the cultivation method of the present invention can shorten the growth time of the mycelium of the cultivation species.

[0048] Table 1 Effects of cultivation media made of different sawdusts on mycelium growth

[0049]

[0050] Table 2 Table of agronomic traits of fruiting bodies

[0051]

[0052]

[0053] * Biological conversion rate = fresh weight of fruiting body (g) / dry weight of cultivation material (g) × 100%.

Claims

1. A Russula lepida culture medium using litchi branches as a substrate, characterized in that It is made of the following components by weight percentage: Litchi wood chips 30 - 50%, cottonseed hulls 30 - 50%, wheat bran 3 - 8%, corncobs 3 - 8%, rice husks 3 - 8%, corn flour 1 - 5%, lime 1 - 5%.

2. The Pleurotus rufescens culture medium using litchi branches as a substrate according to claim 1, wherein It is made of the following components by weight percentage: Litchi wood chips 40%, cottonseed hulls 40%, wheat bran 5%, corncobs 5%, rice husks 5%, corn flour 3%, lime 2%.

3. A cultivation method of Russula lepida, characterized in that It includes the following steps: (1) Preparation of litchi wood chips: The trimmed litchi branches are dried in the sun and then broken into small sections 6 cm - 8 cm long, and further crushed to obtain litchi wood chips of 2 - 4 mm. (2) Preparation of litchi branch seeds: Take 2 - 3 pieces of mycelial blocks about 5 mm on the side from the activated mother culture plate of Rhizopogon rubescens, inoculate them into the liquid original seed medium, and after the mycelium grows, put the sterilized litchi branches into the liquid original seed medium, and let it stand for cultivation. Wait until the mycelium covers the litchi branches to obtain litchi branch seeds. (3) Preparation of Rhizopogon rubescens cultivated seeds: According to the weight percentage, mix 30 - 50% of litchi wood chips, 30 - 50% of cottonseed hulls, 3 - 8% of wheat bran, 3 - 8% of corncobs, 3 - 8% of rice husks and 1 - 5% of corn flour evenly, then add 1 - 5% of lime, add water, and stir for 20 - 30 min. Control the water content to be 60% - 65%, and it is appropriate that water droplets ooze out when the material is squeezed tightly by hand. Control the pH to be 7.0 - 7.5, and then sub-pack into the strain bags and collars. The strain bag specification is a polypropylene folded corner bag of 18 cm × 36 cm × 0.005 cm, and the wet weight of each bag of material is 0.95 kg, which is equivalent to a dry weight of 0.35 kg. Sterilize at 121 °C for 30 min, and reserve it after naturally cooling to room temperature; inoculate the cultivated litchi branch seeds into the cultivation material in the strain bag, and cultivate at a constant temperature of 21 °C until the mycelium fills the bag to make the Rhizopogon rubescens cultivated seed stick. (4) Preparation and fermentation of the Rhizopogon rubescens cultivation material medium: According to the weight percentage, mix 30 - 50% of litchi wood chips, 30 - 50% of cottonseed hulls, 3 - 8% of wheat bran, 3 - 8% of corncobs, 3 - 8% of rice husks and 1 - 5% of corn flour evenly to form a material pile, then add 1 - 5% of lime, add water, and stir evenly again. Control the water content to be 60% - 65%, and then ferment naturally; make several ventilation holes with a pore diameter of 3 - 5 cm on the material pile of the Rhizopogon rubescens cultivation material medium; when the fermentation temperature of the material pile reaches 65 °C, keep it for 24 hours and turn the pile once. Add urea when turning the pile for the first time, adjust the water content to 65%, and continue to ferment. Turn the pile 3 - 8 times in total until the temperature of the material pile no longer rises; replenish water every time when turning the pile, so that when the fermentation ends, the water content of the cultivation material medium is 60%. (5) Cultivation method: Mix the fermented Russula brevipes cultivation medium and litchi orchard soil in a volume ratio of 1:1 to obtain the covering material; take out the cultivated Russula brevipes cultivation spawn bags from the spawn bags, and bury the Russula brevipes cultivation spawn bags with the covering material until the spawn bags are no longer visible. The thickness of the covering layer is controlled at 3-5 cm, and then cover a layer of litchi leaves on the surface for moisture retention; keep the covering material moist and water in a mist every 3 days; when the temperature is low, cover the covering layer with black non-woven fabric for heat preservation, moisture retention and shading; primordia will appear 40-50 days after covering the soil.

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