Cultivation and seedling raising method for seedling raising associated fungi of cremastra appendiculata

Abstract

The method comprises the following steps: S1, preparation of a mother strain culture medium: firstly, cleaning potatoes, peeling, cutting into blocks, adding water, boiling for 30 minutes, and filtering to remove the potatoes; adding agar powder, continuously decocting until boiling, then sequentially adding monopotassium phosphate, magnesium sulfate, peptone and VB1, uniformly stirring, decocting until boiling, turning off fire, cooling, adding glucose, supplementing water to 1L, sub-packaging, sterilizing at high temperature, taking out, and cooling to obtain a mother strain culture medium; and S2, obtaining the isolated species of the original strains of the sporotrichum fulvum: collecting the original strains of the sporotrichum fulvum, disinfecting, and placing on an inoculation workbench. According to the method, fungi capable of rapidly promoting germination of the cremastra appendiculata seeds form associated fungi suitable for germination of the cremastra appendiculata seeds after being cultured, cremastra appendiculata seedlings can be bred in a large scale in a short time through a specific seedling raising technology, the requirement of the market for cremastra appendiculata planting is met, and the market potential is huge.

Description

Technical Field

[0001] The invention relates to the technical field of mushroom cultivation, and in particular to a cultivation method of associated bacteria for cultivating seedlings of Sagittaria sagittifolia. Background Art

[0002] Mao Cigu (also known as Shan Cigu and Du Suanlan) is a precious Chinese medicinal material. Its wild species belong to the national second-level wild protected plants and are mainly distributed in the high-altitude mountainous areas of Yunnan, Guizhou and Sichuan. At present, the seedling cultivation of Mao Cigu is generally two methods: root bulb differentiation and direct seeding. However, the bulb differentiation time cycle is long and the quantity is limited, so it is difficult to obtain seedlings on a large scale. However, direct seeding using the companion bacteria method can breed Mao Cigu seedlings on a large scale. In general, if the companion bacteria seedling cultivation technology is not adopted, basically no seedlings will emerge or the emergence rate will be extremely low. At the same time, in the prior art, after cultivation, it is difficult to form a companion bacteria suitable for Mao Cigu seed germination, and then it is not easy to breed Mao Cigu seedlings on a large scale in a short period of time, so it is difficult to meet the market demand for Mao Cigu cultivation. Summary of the invention

[0003] The purpose of the present invention is to provide a cultivation and seedling cultivation method for companion bacteria of Trichosanthes sagittaria seedling cultivation, which can quickly promote the germination of Trichosanthes sagittaria seeds. After cultivation, companion bacteria suitable for the germination of Trichosanthes sagittaria seeds are formed. Through specific seedling cultivation technology, Trichosanthes sagittaria seedlings can be bred on a large scale in a short period of time, meeting the market demand for Trichosanthes sagittaria cultivation, and the market potential is huge.

[0004] In order to achieve the above object, the main technical solutions adopted by the present invention include:

[0005] A method for cultivating bacteria associated with the cultivation of Cnidium monnieri seedlings, comprising the following steps:

[0006] S1. Preparation of mother culture medium: first wash and peel potatoes, cut into pieces, add water and boil for 30 minutes, filter and remove potatoes; add agar powder, continue to boil until boiling, then add potassium dihydrogen phosphate, magnesium sulfate, peptone and VB1 in sequence, stir evenly, boil until boiling, turn off the heat and cool, add glucose, then add water to 1L and package, sterilize at high temperature, take out and cool to obtain mother culture medium;

[0007] S2. Acquisition of the original strain of yellow-haired radiata: Collect the original strain of yellow-haired radiata in nature, disinfect it, separate its cap and stipe on the inoculation workbench, transfer it to the test tube culture room for cultivation, and complete the original seed production after it grows full;

[0008] S3, the preparation method of the original seed culture medium and the seed culture medium for seedling cultivation is as follows: the sawdust, bran, sugar and gypsum are mixed evenly, water is added and stirred, after standing, high temperature and high pressure sterilization is taken out, and after cooling, the original seed culture medium and the seed culture medium for seedling cultivation are obtained;

[0009] S4. Preparation of mother strain: inoculate the separated isolate into the mother strain culture medium according to aseptic operation until the mycelium is fully grown to obtain the mother strain;

[0010] S5. Preparation of stock seed: inoculate the mother seed into the stock seed culture medium bag by aseptic operation until the mycelium is fully grown to obtain the stock seed;

[0011] S6. Preparation of cultivars: inoculate the original seeds into the cultivar culture medium bag according to aseptic operation until the mycelium is fully grown to obtain the cultivar of Sagittaria sagittifolia companion fungus.

[0012] The above-mentioned method for cultivating the bacteria associated with the seedling cultivation of Trichosanthes kirilowii, wherein the mother culture medium formula in S1 is prepared using the following components in weight percentage: 180-220 g / L of peeled potatoes, 20 g / L of glucose, 1.5 / L of potassium dihydrogen phosphate, 0.5 g / L of magnesium sulfate, 1-1.5 g / L of peptone, 0.05-0.1 mg / L of VB1 and 20-22 g / L of agar, water, 1000 ml.

[0013] In the above-mentioned method for cultivating bacteria associated with the cultivation of C. sagittatum seedlings, the sterilization temperature in S1 is controlled at 110-120° C., and the sterilization time is controlled at 20-30 min.

[0014] In the above-mentioned method for cultivating the bacteria associated with the cultivation of Trichosanthes kirilowii seedlings, the original strain of Actinopsis pilosula in S2 selects the stipe as the object, cuts it into rice-sized particles with a sharp scalpel, and then quickly transfers it to the prepared culture medium with an inoculation needle, and the inoculation is completed after sealing.

[0015] The above-mentioned method for cultivating the bacteria associated with the cultivation of Trichosanthes kirilowii seedlings, wherein the formulas of the original seed culture medium and the seedling cultivation culture medium in S3 are prepared using the following components in weight percentage: 77% to 78% sawdust, 19% to 20% bran, 1% to 2% white sugar and 1% to 2% gypsum.

[0016] In the above-mentioned method for cultivating bacteria associated with the cultivation of C. sagittatum seedlings, the standing time in S3 is 40 to 60 minutes, the sterilization temperature is controlled at 110 to 130° C., and the sterilization time is controlled at 2 to 3 hours.

[0017] A method for growing seedlings of associated bacteria of Sagittaria sagittifolia seedlings, which is applied to the associated bacteria of Sagittaria sagittifolia seedlings grown by any of the above-mentioned methods for growing associated bacteria of Sagittaria sagittifolia seedlings, comprises the following steps:

[0018] P1. Selection of nursery plots: nursery plots are generally selected in loose and permeable land that is not prone to water accumulation, well-ventilated and shaded, and convenient for management and safety protection to reduce investment costs;

[0019] P2. Land preparation: plow the land, remove weeds and stones, and dig furrows and ridges to form seedbeds;

[0020] P3. Selection of seedling raising time: The best time for seedling raising is generally from January to April (slightly different according to the climate conditions in different regions); the temperature is 14-25℃, and the relative humidity is 65%-75%; the most ideal temperature is 16-25℃, and the humidity is best controlled at 70%;

[0021] P4. The sowing standard is: 1600 catties of associated bacteria and 800-1000 fruit clips per mu of the nursery bed. Raising seedlings under this standard is the best ratio for germination.

[0022] P5. Seedling bed management: Keep the seedbed moist before germination. When the weather is dry, pay attention to watering to keep moisture and cool down. The relative humidity of the air should reach 70% to 75%. Prevent bacteria infection caused by high temperature and high humidity;

[0023] P6. Management during the seedling period: 6-8 days after the seeds of the Cibotium fruit clips are planted, the mycelium penetrates the wood and contacts the fruit clip seeds to begin germination. After 40 days, rice-sized dragon eggs begin to appear. After 60 days, the dragon eggs enter a rapid growth stage, which consumes most of the nutrients of the wood under the seedbed. At this time, a large area of ​​mushrooms (white coprinus mushrooms and yellow-radiated hairy mushrooms) will grow in the seedbed at the same time as the dragon eggs consume nutrients. During this period, artificial nutrition needs to be supplied to the seedbed to ensure the normal growth of the dragon eggs. After 120 days, the seedbed begins to emerge, and then a large area of ​​seedlings will emerge one after another;

[0024] P7. Transplanting of seedlings: Select fertile, loose and breathable sandy soil, plant in boxes, dig furrows and make ridges. The ridge height is 20-25 cm, the distance between boxes is 30-40 cm, the distance between seedlings is 12-15 cm, and 20,000-40,000 plants are planted per mu. After transplanting the seedlings, wait for the seedlings to grow new roots, then apply well-rotted farmyard manure to promote the growth of the seedlings. It will reach commercial-grade sale standards in two years.

[0025] In the above-mentioned method for raising seedlings of the companion bacteria of the cultivation of the seedlings of the Trichosanthes, the width of the seedbed in P2 is 90 to 100 centimeters, the height of the ridge is 20 to 25 centimeters, the seedbed is best when it is high in the middle and low on both sides (pig back shape), and the interval between the ridges is 30 to 40 centimeters.

[0026] In the above-mentioned method for raising seedlings of the companion bacteria of the seedling raising of the Sagittaria sagittifolia, the P6 needs to maintain sufficient moisture when the seedlings emerge, and the optimal humidity is 68-72%. The seedbed is supplemented with nutrients every 9-11 days to ensure that the seedlings are healthy and strong.

[0027] In the above-mentioned method for raising seedlings of Agaricus sagittifolia, the best time for transplanting the seedlings in P7 is from October to March of the following year.

[0028] Compared with the prior art, the advantages and positive effects of the present invention are:

[0029] 1. The present invention can quickly promote the germination of Trichosanthes seeds. After cultivation, the fungi can form companion bacteria suitable for the germination of Trichosanthes seeds. Through specific seedling raising technology, Trichosanthes seedlings can be bred on a large scale in a short period of time to meet the market demand for Trichosanthes cultivation.

[0030] 2. In the present invention, the original strain separation is inoculated into the mother culture medium according to aseptic operation until the mycelium is full, and the mother culture is obtained; the mother culture is inoculated into the original culture medium until the mycelium is full, and the original culture is obtained; the original culture is inoculated into the cultivation base bag of the cultivated culture until the mycelium is full, and the cultivated culture is obtained, which is the companion bacteria of the hairy mushroom seedling cultivation. The companion bacteria and the hairy mushroom fruit clip are directly planted on the seedling bed according to the seedling cultivation technology, and the seedling bed is carefully managed. After the processes of mycelium growth, growing dragon eggs, germination, and seedling management, it takes about 6 months to complete the artificial cultivation of hairy mushroom seedlings. This method can artificially cultivate yellow (white) hairy actinia as a companion bacteria for the cultivation of Chinese medicinal material hairy mushroom seedlings, which greatly improves the efficiency of hairy mushroom seedling cultivation and opens up a new path for large-scale cultivation and breeding of hairy mushroom seedlings. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] The drawings described herein are used to provide a further understanding of the present application and constitute a part of the present application. The illustrative embodiments of the present application and their descriptions are used to explain the present application and do not constitute an improper limitation on the present application. In the drawings:

[0032] Figure 1 The present invention is a flow chart of the cultivation of Actinocampus erythrocephala in the method for cultivating the associated bacteria of Sagittaria sagittifolia seedlings;

[0033] Figure 2 The present invention is a flow chart of the cultivation of Actinobacillus eryngii in the cultivation method of the associated bacteria for cultivating the seedlings of Sagittaria sagittifolia;

[0034] Figure 3 The present invention is a flow chart of the method for cultivating the seedlings of the companion bacteria of the Cnidium monnieri. DETAILED DESCRIPTION

[0035] The following will describe the implementation methods of the present application in detail with the help of accompanying drawings and examples, so that the implementation process of how the present application uses technical means to solve technical problems and achieve technical effects can be fully understood and implemented accordingly.

[0036] Example 1

[0037] Cultivation of Actinocampus erythrorhizon:

[0038] Please refer to Figure 1 to Figure 2 As shown, an embodiment of the present invention provides a method for cultivating bacteria associated with the cultivation of Cnidium monnieri seedlings, comprising the following steps:

[0039] S1. Preparation of mother culture medium: The mother culture medium formula is prepared by using the following components in weight percentage: 180-220 g / L of peeled potatoes, 20 g / L of glucose, 1.5 / L of potassium dihydrogen phosphate, 0.5 g / L of magnesium sulfate, 1-1.5 g / L of peptone, 0.05-0.1 mg / L of VB1, 20-22 g / L of agar, and 1000 ml of water. The preparation method of the mother culture medium is as follows: first, wash and peel the potatoes, cut them into pieces, add water and boil for 30 minutes, filter and remove the potatoes; add agar powder, continue to boil until boiling, then add potassium dihydrogen phosphate, magnesium sulfate, peptone and VB1 in sequence, stir evenly, boil until boiling, turn off the heat and cool, add glucose, then add water to 1 L and then package, sterilize at 115°C for 20-30 minutes, take out and cool to obtain the mother culture medium;

[0040] S2. Obtaining the original strain of yellow-haired radiata: Collect the original strain of yellow-haired radiata in nature, disinfect it, separate its cap and stipe on the inoculation workbench, select the stipe as the object, cut it into rice-sized particles with a sharp scalpel, and then quickly transfer it to the prepared test tube culture medium with an inoculation needle, seal it and complete the inoculation. Transfer it to the test tube culture room for cultivation, and complete the original seed production after it grows full;

[0041] S3, the formula of the original seed culture medium and the seed culture medium for seed cultivation are prepared by using the following components in weight percentage: 77% to 78% of sawdust, 19% to 20% of bran, 1% to 2% of sugar and 1% to 2% of gypsum. The preparation method is: mix the sawdust, bran, sugar and gypsum evenly, add water and stir, let stand for 40 to 60 minutes, sterilize at 120℃ for 2 to 3 hours, take out, and cool to obtain the original seed culture medium and the seed culture medium for seed cultivation;

[0042] S4. Preparation of mother strain: inoculate the separated isolate into the mother strain culture medium according to aseptic operation until the mycelium is fully grown to obtain the mother strain;

[0043] S5. Preparation of stock seed: inoculate the mother seed into the stock seed culture medium bag by aseptic operation until the mycelium is fully grown to obtain the stock seed;

[0044] S6. Preparation of cultivars: inoculate the original seeds into the cultivar culture medium bag according to aseptic operation until the mycelium is fully grown to obtain the cultivar of Sagittaria sagittifolia companion fungus.

[0045] Example 2

[0046] Cultivation of Actinobacillus leucophylla:

[0047] Please refer to Figure 1 to Figure 2As shown, an embodiment of the present invention provides a method for cultivating bacteria associated with the cultivation of Cnidium monnieri seedlings, comprising the following steps:

[0048] S1. Obtaining the original strain isolated species of white-haired Actinobacteria: Collect white-haired Actinobacteria (white little devil agaric) in nature for spore separation to obtain the original strain isolated species. The specific operation method is: after the white little devil agaric is disinfected, spore separation is performed in the inoculation box, and the strain is suspended above the culture medium that has been cultured in advance, 3 to 5 cm away from the surface of the culture medium. After the strain matures, the spores will be scattered on the surface of the culture medium. After twelve hours, the spores begin to germinate new mycelium. After seven days, the mycelium will cover the entire surface of the culture medium. At this time, the strong part of the mycelium is transferred to the inoculation bottle or test tube culture medium on the sterile workbench. After it is full, the mother strain is completed;

[0049] S2, the formula of the original seed culture medium and the seed culture medium for seed cultivation are prepared by using the following components in weight percentage: 77% to 78% of sawdust, 19% to 20% of bran, 1% to 2% of sugar and 1% to 2% of gypsum. The preparation method is: mix the sawdust, bran, sugar and gypsum evenly, add water and stir, let stand for 40 to 60 minutes, sterilize at 120℃ for 2 to 3 hours, take out, and cool to obtain the original seed culture medium and the seed culture medium for seed cultivation;

[0050] S3. Preparation of mother strain: inoculate the separated isolate into the mother strain culture medium according to aseptic operation until the mycelium is fully grown to obtain the mother strain;

[0051] S4. Preparation of stock: inoculate the mother stock into the stock culture medium bag according to aseptic operation until the mycelium is fully grown to obtain the stock;

[0052] S5. Preparation of cultivars: inoculate the original seeds into the cultivar culture medium bag according to aseptic operation until the mycelium is fully grown to obtain the cultivar of Sagittaria sagittifolia companion fungus.

[0053] By adopting the technical solutions of Example 1 and Example 2, companion fungus cultivars cultivated by yellow-haired Actinomycetes and white-haired Actinomycetes are obtained. The two are different in the original strain isolation method. The original strain isolation species of yellow-haired Actinomycetes are obtained by tissue separation method, while the original strain isolation species of white-haired Actinomycetes are obtained by spore separation method. The production of mother species, original species and cultivated species of the two is basically the same. Both companion fungus cultivars have very good seedling cultivation effects on Trichosanthes kirilowii.

[0054] Example 3

[0055] Please refer to Figure 1 to Figure 2 As shown, a method for growing seedlings of a mushroom seedling-associated bacteria provided by an embodiment of the present invention comprises the following steps:

[0056] P1. Selection of nursery plots: nursery plots are generally chosen to be plots with loose soil, good water permeability, and not prone to water accumulation. Ventilated and shady locations are preferred, and places that are convenient for management and safety protection can reduce investment costs.

[0057] P2. Land preparation: After plowing the land, remove weeds and stones in the field, and dig furrows and ridges to form seedbeds. The width of the seedbed is 90-100 cm, and the height of the ridge is 20-25 cm. The best seedbed is high in the middle and low on both sides (pig back shape). The interval between ridges is 30-40 cm, which is convenient for daily management.

[0058] P3. Selection of seedling raising time: The best time for seedling raising is generally from January to April (slightly different according to the climate conditions in different regions). The temperature is 14-25℃, and the relative humidity is 65%-75%. The most ideal temperature is 16-25℃, and the humidity is best controlled at 70%.

[0059] P4. The sowing standard is: 1,600 kilograms of associated bacteria and 800-1,000 fruit clips per acre of the seedling bed. Raising seedlings under this standard is the best ratio for germination.

[0060] P5. Seedling bed management: Keep the seedbed moist before germination. When the weather is dry, pay attention to watering to keep moisture and cool. The relative humidity of the air should reach 70% to 75%. Prevent bacteria infection caused by high temperature and high humidity.

[0061] P6. Management during the seedling period: 6 to 8 days after the seeds of the Cibotium fruit clips are planted, the mycelium penetrates the wood and contacts the fruit clip seeds to begin to germinate. After 40 days, rice-sized dragon eggs begin to appear. After 60 days, the dragon eggs enter a rapid growth stage, which consumes most of the nutrients of the wood under the seedbed. At this time, a large area of ​​mushrooms (white small coprinus mushrooms and yellow radiata mushrooms) will grow in the seedbed at the same time as the dragon eggs consume nutrients. During this period, artificial nutrition needs to be supplied to the seedbed to ensure the normal growth of the dragon eggs. After 120 days, the seedbed begins to emerge, and then a large area of ​​seedlings will emerge one after another. At this time, it is necessary to maintain sufficient water. The optimal humidity is 68-72%. Supplementing nutrients to the seedbed once every 10 days can make the seedlings healthy and strong.

[0062] P7. Transplanting seedlings: The best time to transplant seedlings is from October to March of the following year. Choose fertile, loose and breathable sandy soil, plant in boxes, dig furrows and ridges, the ridge height is 20-25 cm, the distance between boxes is 30-40 cm, the distance between seedlings is 12-15 cm, and 20,000-40,000 plants are planted per mu. After the seedlings are transplanted, wait for the seedlings to grow new roots, and then apply decomposed farmyard manure to promote the growth of the seedlings. Generally, it takes two years to reach the commercial grade sale standard;

[0063] By adopting the above technical scheme, artificially cultivated seedlings can be successfully obtained, and the sub-plants have thick texture, developed root system, commercial traits reaching high-quality seedling level, and planting survival rate is high; at the same time, for the artificial cultivation of hairy arrowhead seedlings, not only can the market demand for seedlings be greatly met, but also people's excavation of wild hairy arrowheads can be greatly reduced, protecting wild resources from excessive excavation and damage by people, and can also give full play to its economic value, so as to achieve the purpose of helping farmers increase their income.

[0064] The working principle of the present invention is:

[0065] Cultivation of the associated fungi of Actinia serrata: First, search for the original fungi of Actinia serrata in nature, separate the tissues in the inoculation studio, and obtain the original strain isolate; then inoculate the isolate in the mother culture medium according to aseptic operation until the mycelium is fully grown to obtain the mother culture; then inoculate the mother culture in the fungus bag of the original culture medium according to aseptic operation until the mycelium is fully grown to obtain the original culture; then inoculate the original culture in the fungus bag of the cultivation culture medium according to aseptic operation until the mycelium is fully grown to obtain the associated fungus cultivation; finally, when the fruit clips of Sagittaria sagittifolia are mature and the nursery bed is ready, large-scale seedling cultivation can be carried out;

[0066] Cultivation of companion fungi of white-haired Actinia: First, the original fungus of white-haired Actinia in nature is separated from the spores in an inoculation box to obtain the original strain isolate; then the isolate is inoculated into the mother culture medium according to sterile operation until the mycelium is fully grown to obtain the mother culture; then the mother culture is inoculated into the fungus bag of the original culture medium according to sterile operation until the mycelium is fully grown to obtain the original culture; then the original culture is inoculated into the fungus bag of the cultivation culture medium according to sterile operation until the mycelium is fully grown to obtain the companion fungus cultivation; finally, when the fruit clips of the hairy arrowhead are mature and the seedling bed is ready, large-scale seedling cultivation can be carried out.

[0067] The above description shows and describes several preferred embodiments of the present invention, but as mentioned above, it should be understood that the present invention is not limited to the form disclosed herein, and should not be regarded as excluding other embodiments, but can be used in various other combinations, modifications and environments, and can be modified within the scope of the invention concept described herein through the above teachings or the technology or knowledge of the relevant field. Changes and variations made by those skilled in the art do not depart from the spirit and scope of the present invention, and should be within the scope of protection of the claims attached to the present invention.

Claims

1. A method for cultivating bacteria associated with the cultivation of Cnidium monnieri seedlings, characterized in that: The steps include: S1. Preparation of mother culture medium: first wash and peel potatoes, cut into pieces, add water and boil for 30 minutes, filter and remove potatoes; add agar powder, continue to boil until boiling, then add potassium dihydrogen phosphate, magnesium sulfate, peptone and VB1 in sequence, stir evenly, boil until boiling, turn off the heat and cool, add glucose, then add water to 1L and package, sterilize at high temperature, take out and cool to obtain mother culture medium; S2. Acquisition of the original strain of yellow-haired radiata: Collect the original strain of yellow-haired radiata in nature, disinfect it, separate its cap and stipe on the inoculation workbench, transfer it to the test tube culture room for cultivation, and complete the original seed production after it grows full; S3, the preparation method of the original seed culture medium and the seed culture medium for seedling cultivation is as follows: the sawdust, bran, sugar and gypsum are mixed evenly, water is added and stirred, after standing, high temperature and high pressure sterilization is taken out, and after cooling, the original seed culture medium and the seed culture medium for seedling cultivation are obtained; S4. Preparation of mother strain: inoculate the separated isolate into the mother strain culture medium according to aseptic operation until the mycelium is fully grown to obtain the mother strain; S5. Preparation of stock: inoculate the mother stock into the stock culture medium bag according to aseptic operation until the mycelium is fully grown to obtain the stock; S6. Preparation of cultivars: inoculate the original seeds into the cultivar culture medium bag according to aseptic operation until the mycelium is fully grown to obtain the cultivar of Sagittaria sagittifolia companion fungus.

2. The method for cultivating the bacteria associated with the cultivation of Sagittaria sagittifolia seedlings according to claim 1, characterized in that: The mother culture medium formula in S1 is prepared using the following components in weight percentage: 180-220 g / L of peeled potatoes, 20 g / L of glucose, 1.5 / L of potassium dihydrogen phosphate, 0.5 g / L of magnesium sulfate, 1-1.5 g / L of peptone, 0.05-0.1 mg / L of VB1 and 20-22 g / L of agar, water, 1000 ml.

3. The method for cultivating the bacteria associated with the cultivation of the seedlings of Radix Sagittifolia according to claim 2, characterized in that: The sterilization temperature in S1 is controlled at 110-120° C., and the sterilization time is controlled at 20-30 minutes.

4. The method for cultivating the bacteria associated with the cultivation of Sagittaria sagittifolia seedlings according to claim 3, characterized in that: The original strain of Actinocystis spp. in S2 is selected from the stipe as the object, which is cut into rice-sized particles with a sharp scalpel, and then quickly transferred to the prepared culture medium with an inoculation needle, and the inoculation is completed after sealing.

5. The method for cultivating the bacteria associated with the cultivation of Sagittaria sagittifolia seedlings according to claim 4, characterized in that: The formulas of the original seed culture medium and the seedling cultivation culture medium in S3 are prepared by using the following components in percentage by weight: 77% to 78% of sawdust, 19% to 20% of bran, 1% to 2% of white sugar and 1% to 2% of gypsum.

6. The method for cultivating the bacteria associated with the cultivation of Sagittaria sagittifolia seedlings according to claim 5, characterized in that: The standing time in S3 is 40 to 60 minutes, the sterilization temperature is controlled at 110 to 130° C., and the sterilization time is controlled at 2 to 3 hours.

7. A method for growing seedlings of associated bacteria of Sagittaria sagittifolia, which is applied to the method for growing associated bacteria of Sagittaria sagittifolia seedlings as described in claims 1 to 6, characterized in that: The steps include: P1. Selection of nursery plots: nursery plots are generally selected in loose and permeable land that is not prone to water accumulation, well-ventilated and shaded, and convenient for management and safety protection to reduce investment costs; P2. Land preparation: plow the land, remove weeds and stones, and dig furrows and ridges to form seedbeds; P3. Selection of seedling raising time: The best time for seedling raising is generally from January to April (slightly different according to the climate conditions in different regions); the temperature is 14-25℃, and the relative humidity is 65%-75%; the most ideal temperature is 16-25℃, and the humidity is best controlled at 70%; P4. The sowing standard is: 1600 catties of associated bacteria and 800-1000 fruit clips per mu of the nursery bed. Raising seedlings under this standard is the best ratio for germination. P5. Seedling bed management: Keep the seedbed moist before germination. When the weather is dry, pay attention to watering to keep moisture and cool down. The relative humidity of the air should reach 70% to 75%. Prevent bacteria infection caused by high temperature and high humidity; P6. Management during the seedling period: 6-8 days after the seeds of the Cibotium fruit clips are planted, the mycelium penetrates the wood and contacts the fruit clip seeds to begin germination. After 40 days, rice-sized dragon eggs begin to appear. After 60 days, the dragon eggs enter a rapid growth stage, which consumes most of the nutrients of the wood under the seedbed. At this time, a large area of ​​mushrooms (white coprinus mushrooms and yellow-radiated hairy mushrooms) will grow in the seedbed at the same time as the dragon eggs consume nutrients. During this period, artificial nutrition needs to be supplied to the seedbed to ensure the normal growth of the dragon eggs. After 120 days, the seedbed begins to emerge, and then a large area of ​​seedlings will emerge one after another; P7. Transplanting of seedlings: Select fertile, loose and breathable sandy soil, plant in boxes, dig furrows and make ridges. The ridge height should be 20-25 cm. The distance between boxes should be 30-40 cm, and the distance between seedlings should be 12-15 cm. Plant 20,000-40,000 plants per mu. After transplanting the seedlings, wait for the seedlings to grow new roots, then apply well-rotted farmyard manure to promote the growth of the seedlings. The seeds will reach commercial-grade sale standards in two years.

8. The method for growing seedlings of a mushroom seedling-associated bacteria according to claim 7, characterized in that: The width of the seedbed in P2 is 90 to 100 cm, the height of the ridge is 20 to 25 cm, and the best seedbed is high in the middle and low on both sides (pig back shape), with a spacing of 30 to 40 cm between ridges.

9. The method for growing seedlings of a mushroom seedling-associated bacteria according to claim 8, characterized in that: When the P6 seedlings emerge, it is necessary to maintain sufficient moisture, the optimal humidity is 68-72%, and the seedbed is supplemented with nutrients every 9-11 days to ensure that the seedlings are healthy and strong.

10. The method for growing seedlings of a mushroom seedling-associated bacteria according to claim 9, characterized in that: The best time for transplanting the seedlings in P7 is from October to March of the following year.

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