Morchella mother strain culture medium as well as preparation method and application thereof

Abstract

The invention relates to the technical field of edible fungus cultivation, and discloses a morchella mother strain culture medium and a preparation method and application thereof. According to the morchella mother strain culture medium provided by the invention, the content of corn flour and the content of monopotassium phosphate are remarkably increased, and experimental research shows that when the culture medium is used for culturing local morchella mother strains in Guizhou, the hypha growth speed can reach 9.54 mm / d at most and is remarkably higher than that of other reported morchella mother strain culture mediums; the highest dry weight of hyphae cultured for 12 days is 0.1113 g which is also remarkably higher than that of other culture media. That is, the research result shows that the culture medium for the morchella vulgaris mother strain provided by the invention can obviously improve the culture efficiency of the local morchella vulgaris mother strain in Guizhou.

Description

Technical Field

[0001] The invention relates to the technical field of edible fungus cultivation, and in particular to a Morchella mother culture medium and a preparation method and application thereof. Background Art

[0002] Morchella, also known as Morchella, belongs to the Ascomycetes, Peziaceae, Peziales, Morchellaceae, and Morchella genus. It is one of the precious and rare edible fungi in the world. Morchella is not only delicious, but also nutritious. It contains rich protein, amino acids, mineral elements, unsaturated fatty acids, and organic germanium, selenium and other elements necessary for the human body. It has extremely high medicinal and nutritional health value and is deeply loved by consumers. It is one of the four famous fungi in the world and has a very broad prospect for cultivation and promotion. At present, the cultivation and production areas of Morchella in China are mainly concentrated in Sichuan, Chongqing, Yunnan and other places, which are also the earliest areas of large-scale cultivation of Morchella in China. Other provinces and cities such as Guizhou, Hubei, Henan, Hebei, Shanxi, Gansu and other places also have a certain cultivation area.

[0004] Guizhou Province is rich in wild germplasm resources and is a region in China with relatively rich wild mushroom resources.

[0005] The development of the morel industry in Guizhou Province mainly faces the following problems: insufficient supply of high-quality strains and lack of industry standards. The morel strains in Guizhou Province are mainly introduced from scientific research institutes outside the province such as Sichuan, Yunnan, and Hubei. There are few mother strains of morels cultivated in the province, resulting in high costs for growers. When cultivating mother strains of morels suitable for the development of the local morel industry, the inventors found that the efficiency of cultivating Guizhou local morel mother strains using existing culture media is low, which is specifically manifested in slow mycelium growth and low mycelium dry weight. Summary of the invention

[0006] In view of this, the present invention provides a Morchella mother culture medium and a preparation method and application thereof.

[0007] In a first aspect, the present invention provides a Morchella mother culture medium, which comprises the following components in parts by weight:

[0008] 40-60 parts of potatoes, 6-9 parts of corn flour, 0.6-0.9 parts of potassium dihydrogen phosphate, 3-7 parts of glucose, 3.5-6.5 parts of agar powder, 0.1-0.4 parts of magnesium sulfate, and 200-350 parts of water.

[0009] In an optional embodiment, the Morchella mother culture medium comprises the following components in parts by weight:

[0010] 50 parts of potatoes, 7.5 parts of corn flour, 0.75 parts of potassium dihydrogen phosphate, 5 parts of glucose, 5 parts of agar powder, 0.25 parts of magnesium sulfate, and 250 parts of water.

[0011] In a second aspect, the present invention provides a method for preparing the above-mentioned Morchella mother culture medium, comprising the following steps:

[0012] (1) Place potatoes and corn flour in cold water, stir while cooking, and continue cooking for 15 to 20 minutes after boiling until the potatoes are crisp but not mushy, obtaining a solid-liquid mixture;

[0013] (2) taking the solid-liquid mixture and filtering it to obtain a filtrate;

[0014] (3) Take the filtrate, adjust the volume to the target volume, add potassium dihydrogen phosphate, glucose, agar powder and magnesium sulfate in target weight portions, stir and heat until completely dissolved.

[0015] In an optional embodiment, in step (1), when placing potatoes and corn flour in cold water, the ratio of potatoes to water is: (40-60): (300-500).

[0016] In an optional embodiment, in step (2), the filtering refers to filtering using 1 to 4 layers of gauze.

[0017] In a third aspect, the present invention provides an application of the above-mentioned Morchella mother culture medium in the cultivation of Morchella mother culture.

[0018] In a fourth aspect, the present invention provides a method for cultivating a mother strain of Morchella edulis, comprising the following steps:

[0019] Take the above-mentioned Morchella mother culture medium, sterilize it, inoculate Morchella mycelium, and culture it in the dark under constant temperature conditions.

[0020] In an optional embodiment, the sterilization conditions include:

[0021] The temperature is 120-122°C, the pressure is 0.1-0.15MPa, and the time is 25-30min.

[0022] In an optional embodiment, the temperature of the constant temperature condition is 18-22°C.

[0023] In an optional embodiment, the Morchella is a Morchella native to Zunyi City, Guizhou Province, China.

[0024] The technical solution of the present invention has at least the following beneficial effects:

[0025] The Morchella mother seed culture medium provided by the present invention significantly increases the content of corn flour and potassium dihydrogen phosphate. Experimental studies have found that when the culture medium is used to culture the local Morchella mother seed in Guizhou, the mycelial growth rate can reach up to 9.54 mm / d, which is significantly higher than other reported Morchella mother seed culture media; the mycelial dry weight after 12 days of culture is up to 0.1113 g, which is also significantly higher than other culture media. That is, the research results show that the Morchella mother seed culture medium provided by the present invention can significantly improve the culture efficiency of the local Morchella mother seed in Zunyi City, Guizhou Province. DETAILED DESCRIPTION

[0026] The following examples are provided for a better understanding of the present invention, but are not intended to limit the best mode of implementation, nor to limit the content and protection scope of the present invention. Any product identical or similar to the present invention obtained by anyone under the inspiration of the present invention or by combining the features of the present invention with other prior arts shall fall within the protection scope of the present invention.

[0027] If no specific experimental steps or conditions are specified in the examples, the conventional experimental steps or conditions described in the literature in the field can be used. If no manufacturer is specified for the reagents or instruments used, they are all conventional reagent products that can be obtained commercially.

[0028] The present invention is further described in detail below in conjunction with specific embodiments. These embodiments should not be construed as limiting the scope of protection claimed by the present invention.

[0029] Example 1

[0030] The formula of the mother culture medium of Morchella in this embodiment is: 50g potato, 7.5g corn flour, 0.75g potassium dihydrogen phosphate, 5g glucose, 5g agar powder, 0.25g magnesium sulfate, and 250ml water.

[0031] Prepare the Morchella mother culture medium of this embodiment as follows:

[0032] (1) Weigh peeled and eyed potatoes and cut them into small pieces of the same size; then place the potatoes and corn flour in 400 mL of tap water, cook while stirring, and after boiling, turn to low heat and continue cooking for 15 to 20 minutes until the potatoes are crisp but not mushy, to obtain a solid-liquid mixture;

[0033] (2) taking the solid-liquid mixture obtained in step (1), filtering it through four layers of gauze to obtain a filtrate;

[0034] (3) Take the filtrate obtained in step (2), add water to make the volume to 250 mL, then add potassium dihydrogen phosphate, glucose, agar powder and magnesium sulfate, stir and heat until completely dissolved, to obtain the mother culture medium of Morchella of the present embodiment.

[0035] Example 2

[0036] The mother culture medium of Morchella was prepared according to the method of Example 1, except that the formula of the mother culture medium of Morchella in this embodiment was:

[0037] 40g potatoes, 9g corn flour, 0.6g potassium dihydrogen phosphate, 7g glucose, 3.5g agar powder, 0.4g magnesium sulfate, 200ml water.

[0038] Example 3

[0039] The mother culture medium of Morchella was prepared according to the method of Example 1, except that the formula of the mother culture medium of Morchella in this embodiment was:

[0040] 60g potatoes, 6g corn flour, 0.9g potassium dihydrogen phosphate, 3g glucose, 6.5g agar powder, 0.1g magnesium sulfate, 350ml water.

[0041] Comparative Example 1

[0042] The Morchella mother culture medium was prepared according to the method of Example 1, except that the formula of the Morchella mother culture medium of this comparative example was:

[0043] 50g potatoes, 1.56g corn flour, 0.16g potassium dihydrogen phosphate, 5g glucose, 4.38g agar, 0.13g magnesium sulfate, 250ml water.

[0044] Comparative Example 2

[0045] The Morchella mother culture medium was prepared according to the method of Example 1, except that the formula of the Morchella mother culture medium of this comparative example was:

[0046] 50g potatoes, 0.75g corn flour, 0.1g potassium dihydrogen phosphate, 5g glucose, 5g agar, 0.25g magnesium sulfate, 250ml water.

[0047] Comparative Example 3

[0048] The Morchella mother culture medium was prepared according to the method of Example 1, except that the formula of the Morchella mother culture medium of this comparative example was:

[0049] 50g potatoes, 1.75g ​​corn flour, 0.2g potassium dihydrogen phosphate, 5g glucose, 5g agar, 0.25g magnesium sulfate, 250ml water.

[0050] Comparative Example 4

[0051] The formula of the Morchella mother culture medium in this comparative example is: 50g potato, 5g glucose, 5g agar, 0.5g potassium dihydrogen phosphate, 0.125g magnesium sulfate, 0.25g peptone, and 250ml water.

[0052] The mother culture medium of Morchella in this comparative example was prepared as follows:

[0053] Weigh the peeled and eyed potatoes, add 400 mL of sterile water and cook for 25 min, filter with four layers of gauze, take the filtrate and add sterile water to make up to 250 mL, then add the remaining components, stir and heat until dissolved.

[0054] Comparative Example 5

[0055] The formula of the Morchella mother culture medium in this comparative example is: 50g potato, 5g glucose, 12.5g sawdust, 5g agar, and 250ml water.

[0056] The mother culture medium of Morchella in this comparative example was prepared as follows:

[0057] Weigh the peeled and eyed potatoes, add 400 mL of sterile water together with sawdust and cook for 25 min. Filter with four layers of gauze, take the filtrate and add sterile water to make up to 250 mL, then add the remaining components, stir and heat until dissolved.

[0058] Comparative Example 6

[0059] The formula of the Morchella mother culture medium in this comparative example is: 50g potato, 5g glucose, 12.5g bran, 5g agar, and 250ml water.

[0060] The mother culture medium of Morchella in this comparative example was prepared as follows:

[0061] Weigh the peeled and eyed potatoes, add 400 mL of sterile water together with the bran and cook for 25 min. Filter with four layers of gauze, take the filtrate and add sterile water to make up to 250 mL, then add the remaining components, stir and heat until dissolved.

[0062] Comparative Example 7

[0063] The formula of the Morchella mother culture medium in this comparative example is: 25g potato, 5g glucose, 25g humus, 5g agar, and 250ml water.

[0064] The mother culture medium of Morchella in this comparative example was prepared as follows:

[0065] Weigh the peeled and eyed potatoes, add 400 mL of sterile water and humus soil and cook for 25 minutes, filter with four layers of gauze, take the filtrate and add sterile water to make up to 250 mL, then add the remaining components, stir and heat until dissolved.

[0066] Comparative Example 8

[0067] The formula of the Morchella mother culture medium in this comparative example is: 60g fresh pine needles, 5g glucose, 5g agar, 0.25g peptone, 0.125g yeast powder, 5g agar, and 250ml water.

[0068] The mother culture medium of Morchella in this comparative example was prepared as follows:

[0069] Weigh fresh pine needles, add 400 mL of sterile water and boil for 25 min, filter with four layers of gauze, take the filtrate and add sterile water to make up to 250 mL, then add the remaining components, stir and heat until dissolved.

[0070] Comparative Example 9

[0071] The formula of the Morchella mother culture medium in this comparative example is: 50g raw pear, 5g glucose, 5g agar, 0.25g potassium dihydrogen phosphate, 0.25g peptone, and 250ml water.

[0072] The mother culture medium of Morchella in this comparative example was prepared as follows:

[0073] Weigh peeled and cored raw pears, cut into pieces, add 400 mL of sterile water and cook for 25 min, filter with four layers of gauze, take the filtrate and add sterile water to make up to 250 mL, then add the remaining components, stir and heat until dissolved.

[0074] Comparative Example 10

[0075] The formula of the Morchella mother culture medium in this comparative example is: 50g apple, 5g glucose, 5g agar, 0.25g potassium dihydrogen phosphate, 0.25g peptone, and 250ml water.

[0076] The mother culture medium of Morchella in this comparative example was prepared as follows:

[0077] Weigh peeled and cored apples, cut into pieces, add 400 mL of sterile water and cook for 25 min, filter with four layers of gauze, take the filtrate and add sterile water to make up to 250 mL, then add the remaining components, stir and heat until dissolved.

[0078] Experimental Example 1

[0079] The Morchella mother culture medium prepared in Examples 1-3 is numbered 1-3, and the Morchella mother culture medium prepared in Comparative Examples 1-10 is numbered D1-D10.

[0080] Preparation of strains: Fruiting bodies with typical characteristics of Morchella edulis that are healthy, growing well, 70% to 80% mature, and identified as Morchella edulis are collected in the wild in Loushanguan, Zunyi City. After separating them, a 3mm*3mm tissue block at the junction of the cap and the stipe is inoculated into the mother culture medium of Implementation 1, and placed in an incubator at 20°C to grow into a mother strain. The purified test tube slant strains (source: strain 1 (Morchella edulis collected above), strain 2 (Morchella edulis from Hubei)) are inoculated onto a 90mm PDA plate, cultured at 20°C in the dark, and set aside after the hyphae are fully grown.

[0081] Each of the above culture media was placed in a conical flask, sterilized in an autoclave at 121°C and 0.1MPa for 30 minutes, and after sterilization, the culture media were poured into a flat plate (90 cm) in a clean bench (5 replicates for each strain for each formula), and after cooling, the pre-cultured Morchella mycelium blocks without foreign bacteria and with good growth were inoculated on the above 13 mother culture medium plates with a sterilized 6mm puncher, and placed upside down in a 20°C incubator for dark culture. The mycelium growth state, growth, colony color, and density of each strain were observed every 24 hours, and the diameter of the colony was measured and recorded by the cross method. The results are shown in Tables 1 and 2, wherein Table 1 shows the mycelium growth status of strain 1 in different culture media, and Table 2 shows the mycelium growth status of strain 2 in different culture media.

[0082] Table 1 Mycelial growth of strain 1 in different culture media

[0083]

[0084] Note: +++ indicates dense mycelium growth, ++ indicates relatively dense mycelium growth, + indicates sparse mycelium growth; letters represent significant differences, the same letters indicate no significant differences, and the earlier the letters are, the more significant the differences are.

[0085] Table 2 Mycelial growth of strain 2 in different culture media

[0086]

[0087]

[0088] Note: +++ indicates dense mycelium growth, ++ indicates relatively dense mycelium growth, + indicates sparse mycelium growth; letters represent significant differences, the same letters indicate no significant differences, and the earlier the letters are, the more significant the differences are.

[0089] Experimental Example 2

[0090] Measurement of mycelium dry weight: First, dry the filter paper in an oven to constant weight, weigh the filter paper with a 1 / 10,000 electronic balance, then place the filter paper in a Büchner funnel of corresponding size, melt the culture medium into a culture solution using a microwave oven, pour all the culture solution from the conical flask into a Büchner funnel for filtration, and then filter and wash the filter paper twice with distilled water, put the filter paper and the mycelium attached to the filter paper into a constant temperature drying oven (50°C) and dry to constant weight, weigh the filter paper after drying with a 1 / 10,000 electronic balance, and subtract the filter paper mass before filtration from the filter paper mass after filtration to obtain the mycelium dry weight. The results are shown in Tables 3 and 4, where Table 3 is the mycelium dry weight after culturing in different culture media for strain 1, and Table 4 is the mycelium dry weight after culturing in different culture media for strain 2.

[0091] Table 3 Analysis of variance of mycelium dry weight of Morchella species 1 (mycelium grown for 12 days)

[0092]

[0093] Note: Letters represent significant differences. The same letters indicate that the difference is not obvious. The earlier the letter order, the more obvious the difference.

[0094] Table 4 Variance analysis of mycelium dry weight of Morchella species 2 (mycelium grown for 12 days)

[0095]

[0096]

[0097] Note: Letters represent significant differences. The same letters indicate that the difference is not obvious. The earlier the letter order, the more obvious the difference.

[0098] Obviously, the above embodiments are merely examples for the purpose of clear explanation, and are not intended to limit the implementation methods. For those skilled in the art, other different forms of changes or modifications can be made based on the above description. It is not necessary and impossible to list all the implementation methods here. The obvious changes or modifications derived therefrom are still within the scope of protection of the invention.

Claims

1. A Morchella mother culture medium, characterized in that: In parts by weight, it includes the following components: 40-60 parts of potatoes, 6-9 parts of corn flour, 0.6-0.9 parts of potassium dihydrogen phosphate, 3-7 parts of glucose, 3.5-6.5 parts of agar powder, 0.1-0.4 parts of magnesium sulfate, and 200-350 parts of water.

2. The Morchella mother culture medium according to claim 1, characterized in that In parts by weight, the Morchella mother culture medium comprises the following components: 50 parts of potatoes, 7.5 parts of corn flour, 0.75 parts of potassium dihydrogen phosphate, 5 parts of glucose, 5 parts of agar powder, 0.25 parts of magnesium sulfate, and 250 parts of water.

3. The method for preparing the Morchella mother culture medium according to claim 1 or 2, characterized in that: The steps include: (1) Place potatoes and corn flour in cold water, stir while cooking, and continue cooking for 15 to 20 minutes after boiling until the potatoes are crisp but not mushy, obtaining a solid-liquid mixture; (2) taking the solid-liquid mixture and filtering it to obtain a filtrate; (3) Take the filtrate, adjust the volume to the target volume, add potassium dihydrogen phosphate, glucose, agar powder and magnesium sulfate in target weight portions, stir and heat until completely dissolved.

4. The preparation method according to claim 3, characterized in that: In step (1), when placing potatoes and corn flour in cold water, the ratio of potatoes to water is: (40-60): (300-500).

5. The preparation method according to claim 3, characterized in that: In step (2), the filtering refers to filtering using 1 to 4 layers of gauze.

6. Application of the Morchella mother seed culture medium according to claim 1 or 2 in the cultivation of Morchella mother seed.

7. A method for cultivating a mother strain of Morchella, characterized in that: The steps include: The mother culture medium of Morchella described in claim 1 or 2 is sterilized, inoculated with Morchella mycelium, and cultured in the dark under constant temperature conditions.

8. The cultivation method according to claim 7, characterized in that: The sterilization conditions include: The temperature is 120-122°C, the pressure is 0.1-0.15MPa, and the time is 25-30min.

9. The cultivation method according to claim 7, characterized in that: The temperature of the constant temperature condition is 18-22°C.

10. The cultivation method according to any one of claims 7 to 9, characterized in that: The morels are native to Zunyi City, Guizhou Province, China.

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