A morchella preservation medium, a preparation method thereof and application thereof

Abstract

The present application relates to a kind of morchella preservation medium, by 5-7 parts of soil, 1-2 parts of sawdust, 1 part of wheat, 1-2 parts of chaff and 6 parts-6.5 parts of water are made of.The present application scientifically selects the composition and proportion of soil, sawdust, chaff, wheat, morchella preservation medium is prepared, for morchella preservation, without sealing, can be preserved for more than 7 years, improve enzyme activity in morchella culture, for field experiment can improve the yield of morchella, mushroom density and fruit body maturation period, significantly better than traditional preservation method, and simple operability is strong, low energy consumption, can be used for industrial production.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of microorganisms, in particular to a morchella conica preservation medium, a preparation method thereof and an application thereof. BACKGROUND

[0002] Morchella is a kind of edible fungus belonging to Ascomycota, Pezizomycetes, Pezizales, Morchellaceae and Morchella, and is known as one of the four wild famous fungi, together with truffle, pine mushroom and chicken oil fungus. Morchella is rich in crude protein, crude fat, carbohydrates, various amino acids and other nutrients, and has the functions of protecting liver, reducing blood lipids, enhancing immunity, resisting tumors, resisting oxidation, digesting and stomach, resisting fatigue and resisting bacteria, and can be used as raw materials to develop food flavoring ingredients, nutritional supplements and nutritional health products.

[0003] The cultivation of Morchella is a high-risk industry. The degradation of Morchella strain is one of the main factors leading to a significant decrease in fruiting body yield, so it is desirable to preserve high-quality strains for a long time. Existing preservation methods include the following: (1) repeated subculturing of multiple batches of strains, with the longest strain life being about 2.5m, about 6000 hours, and the shortest being only 0.059m, which dies completely within a few days to more than ten days; (2) selecting new tissues or ascospores from fresh Morchella every year to produce Morchella, but due to spontaneous mutation of active cells, it is easy to increase the genetic instability of Morchella varieties; (3) cryopreservation method, which requires strict cryopreservation conditions and cryopreservation facilities, and has high cost; (4) sealing preservation of Morchella with liquid protective agents such as liquid paraffin, which not only has high cost of liquid protective agents, but also has poor preservation effect. Therefore, it is necessary to study more efficient and stable Morchella preservation medium and preservation method. SUMMARY

[0004] The present application relates to a Morchella preservation medium, which consists of 5-7 parts of soil, 1-2 parts of sawdust, 1 part of wheat, 1-2 parts of chaff and 6-6.5 parts of water by weight.

[0005] In a preferred technical solution of the present application, the preservation medium for Morchella consists of 7 parts of soil, 1 part of sawdust, 1 part of wheat, 1 part of chaff and 6 parts of water by weight.

[0006] In a preferred technical solution of the present application, the preservation medium for Morchella consists of 6 parts of soil, 2 parts of sawdust, 1 part of wheat, 1 part of chaff and 6 parts of water by weight.

[0007] In a preferred technical solution of the present application, the preservation medium for Morchella consists of 5 parts of soil, 2 parts of sawdust, 1 part of wheat, 2 parts of chaff and 6.5 parts of water by weight.

[0008] In a preferred technical solution of the present application, the soil is sandy loam.

[0009] In the preferred technical solution of the present application, the soil particle size is 0.25-1mm.

[0010] In the preferred technical solution of the present application, the sawdust is a broad-leaved tree of Quercus, preferably any one or combination of Quercus acutissima Carruth, Quercus aliena, Quercus acutissima Carruth and Quercus aliena.

[0011] In the preferred technical solution of the present application, the sawdust particle size is 30-50 mesh.

[0012] In the preferred technical solution of the present application, the chaff and wheat are fresh and non-moldy.

[0013] In the preferred technical solution of the present application, the preparation method of the Morchella preservation medium is to mix the required amount of soil, sawdust, wheat, and chaff with water, sterilize, and obtain.

[0014] The present application relates to a kind of Morchella preservation medium preparation method, by weight parts, the Morchella preservation medium is composed of 5-7 soil, 1-2 sawdust, 1 wheat, 1-2 chaff and 6-6.5 water, comprising the following steps: the required amount of soil, sawdust, wheat, chaff, mix with water uniformly, sterilize, obtain.

[0015] In the preferred technical solution of the present application, by weight parts, the Morchella preservation medium is composed of 7 soil, 1 sawdust, 1 wheat, 1 chaff and 6 water.

[0016] In the preferred technical solution of the present application, by weight parts, the Morchella preservation medium is composed of 6 soil, 2 sawdust, 1 wheat, 1 chaff and 6 water.

[0017] In the preferred technical solution of the present application, by weight parts, the Morchella preservation medium is composed of 5 soil, 2 sawdust, 1 wheat, 2 chaff and 6.5 water.

[0018] In the preferred technical solution of the present application, the soil is sandy loam.

[0019] In the preferred technical solution of the present application, the soil particle size is 0.25-1mm.

[0020] In the preferred technical solution of the present application, the sawdust is a broad-leaved tree of Quercus, preferably any one or combination of Quercus acutissima Carruth, Quercus aliena, Quercus acutissima Carruth and Quercus aliena.

[0021] In the preferred technical solution of the present application, the sawdust particle size is 30-50 mesh.

[0022] In the preferred technical solution of the present application, the chaff and wheat are fresh and non-moldy.

[0023] Another object of the present application is to provide the use of the Morel preservation medium of the present application in the preservation of Morel.

[0024] In the preferred technical solution of the present application, the use is to inoculate the mycelium culture into the preservation medium.

[0025] In the preferred technical solution of the present application, the use is to inoculate the mycelium culture into the preservation medium without sealing.

[0026] In the preferred technical solution of the present application, the use is to inoculate the mycelium culture into the preservation medium without subculture during the preservation period.

[0027] In the preferred technical solution of the present application, the use is to inoculate the mycelium culture into the preservation medium, and the Morel can be preserved for not less than 5 years, preferably not less than 7 years, and more preferably not less than 10 years.

[0028] Another object of the present application is to provide a preservation method of Morel, comprising the following steps: inoculating the mycelium culture into the Morel preservation medium of the present application.

[0029] In the preferred technical solution of the present application, the preservation method does not need to be sealed.

[0030] In the preferred technical solution of the present application, the preservation period does not need to be subcultured.

[0031] In the preferred technical solution of the present application, the method can preserve the Morel for not less than 5 years, preferably not less than 7 years, and more preferably not less than 10 years.

[0032] Unless otherwise specified, when the present application involves the percentage between liquids, the percentage is volume / volume percentage; when the present application involves the percentage between liquids and solids, the percentage is volume / weight percentage; when the present application involves the percentage between solids and liquids, the percentage is weight / volume percentage; and the rest is weight / weight percentage.

[0033] Compared with the prior art, the present application has the beneficial technical effects including:

[0034] 1. The present application scientifically selects the composition and ratio of soil, sawdust, husk and wheat to prepare the Morel preservation medium, which is used for the preservation of Morel, can not only provide sufficient carbon source and matrix, but also improve the permeability of Morel growth, does not need subculture, does not need sealing, can be preserved for more than 7 years, improves the activity of enzymes (such as cellulase, multifunctional peroxidase and manganese peroxidase) in the Morel culture, and is used for field experiments to improve the yield, fruiting density and fruiting body maturation period of Morel, which is obviously superior to the traditional preservation method.

[0035] 2. The preservation method of the present invention is simple, easy to operate, and has low energy consumption. It does not require professional facilities and equipment for cryogenic preservation at -80°C or liquid nitrogen, and can be used for industrial production. Attached Figure Description

[0036] Figure 1a Yields from field experiments;

[0037] Figure 1b Mushroom density in field experiments;

[0038] Figure 1c The maturity period of fruiting bodies in field experiments. Detailed Implementation

[0039] The present invention will be described below with reference to the embodiments. However, the present invention is not limited to the embodiments.

[0040] In this example, the Sichuan Morel Mushroom No. 1 (Sichuan Approval No. 2013007) from the Sichuan Academy of Agricultural Sciences was used for the experiment.

[0041] Potato Dextrose Agar (PDA) medium: Contains 200 g potato, 20 g glucose, 20 g agar, and 1000 mL distilled water. Take 20 mL and place it into a test tube (20 mm × 200 mm diameter), sterilize (121℃, 30 min), and cool to obtain a test tube containing PDA solid medium. Take 20 mL of the sterilized medium and place it into a petri dish (90 mm diameter), cool to obtain a petri dish containing PDA solid medium.

[0042] Preparation of morel mycelial culture: The mycelium of morel No. 1 was placed in a petri dish (90 mm in diameter) containing PDA solid medium and cultured at 20℃ for 7 days to obtain the culture.

[0043] Cultivation bags (17 cm × 35 cm × 0.005 cm) containing 500 g of spawn culture medium: The spawn culture medium, by weight ratio, consists of 30 parts wheat, 18.5 parts soil, 20 parts rice husks, 30 parts sawdust, 1.5 parts lime, and 150 parts water. The cultivation bags are sterilized at 121℃ for 2.5 hours and then cooled before use.

[0044] Example 1: Preparation of the morel mushroom preservation culture medium of the present invention

[0045] (1) Select sandy loam soil with a particle size of 0.25~1mm as the soil;

[0046] (2) Select the sawdust from the oak tree and crush it. First, pass it through a 10-mesh sieve, then through a 30-50 mesh sieve. Take the sawdust from the 30-50 mesh sieve and set it aside.

[0047] (3) Select fresh, unmoldy rice husks and wheat from the current year;

[0048] (4) 7 parts of soil, 1 part of sawdust, 1 part of wheat, 1 part of chaff, and 6 parts of water are mixed uniformly, sterilized at 121 ℃ for 2.5 h, and then cooled to obtain the medium.

[0049] Example 2 Preparation of the Morchella culture medium of the application

[0050] (1) The sandy soil with a particle size of 0.25-1 mm is selected as the soil;

[0051] (2) The sawdust of the green bamboo tree is crushed, passed through a 10-mesh sieve, and then passed through a 30-50-mesh sieve, and the sawdust on the 30-50-mesh sieve is taken for use;

[0052] (3) Fresh and non-molded chaff and wheat of the current year are selected;

[0053] (4) 6 parts of soil, 2 parts of sawdust, 1 part of wheat, and 1 part of chaff are mixed uniformly, sterilized at 121 ℃ for 2.5 h, and then cooled to obtain the medium.

[0054] Example 3 Preparation of the Morchella culture medium of the application

[0055] (1) The sandy soil with a particle size of 0.25-1 mm is selected as the soil;

[0056] (2) The sawdust of the green bamboo tree is crushed, passed through a 10-mesh sieve, and then passed through a 30-50-mesh sieve, and the sawdust on the 30-50-mesh sieve is taken for use;

[0057] (3) Fresh and non-molded chaff and wheat of the current year are selected;

[0058] (4) 7 parts of soil, 2 parts of sawdust, 1 part of wheat, and 2 parts of chaff are mixed uniformly, sterilized at 121 ℃ for 2.5 h, and then cooled to obtain the medium.

[0059] Test Example 1 Research on the Morchella culture medium of the application for the preservation of Morchella

[0060] 1. Morchella preservation experiment

[0061] 18 Morchella mycelium cultures with a diameter of 5 mm are taken with a sterile puncher with an inner diameter of 5 mm, divided into 3 groups, 6 parallel repeats in each group, inoculated into different culture media, and preserved at 4 ℃ for 7 years from February 2015 to February 2022:

[0062] CK group: inoculate the Morel mycelium culture to the test tube (test tube diameter 20mm x 200mm) containing PDA solid medium, and seal with rubber plug. The test tube strain is the starting test tube, which is subcultured to PDA solid medium twice a year for 14 times in 7 years. The specific subculture method is as follows: take 5mm diameter inoculation block from the starting test tube, and transfer to the new test tube containing PDA solid medium.

[0063] T1 group: inoculate the Morel mycelium culture to the test tube (test tube diameter 20mm x 200mm) containing the preservation medium of Example 1, and seal with rubber plug. No subculture is performed during the 7-year experiment.

[0064] T2 group: inoculate the Morel mycelium culture to the test tube (test tube diameter 20mm x 200mm) containing the preservation medium of Example 1, and seal with rubber plug. The test tube strain is the starting test tube, which is subcultured to the preservation medium of Example 1 twice a year for 14 times in 7 years. The specific subculture method is as follows: take 5mm diameter inoculation block from the starting test tube, and transfer to the new test tube containing the preservation medium of Example 1.

[0065] After 7 years, the mycelium cultures of each group are inoculated to the Petri dish (diameter 90mm) containing PDA solid medium, and activated at 20℃ in dark for 7 days, to obtain the activated Morel mycelium culture.

[0066] 2. Field experiment

[0067] Prepare the cultivation bag containing the Morel of GK group, T1 group and T2 group respectively, by the following method: take 1 piece of activated Morel mycelium culture with 5mm inner diameter sterile punch, inoculate to the center of the Petri dish (diameter 90mm) containing new PDA solid medium, and culture at 20℃ for 10 days. Then take 3 pieces of 5mm inoculation block, inoculate to the cultivation bag containing 500g cultivation medium, and culture at 20℃ in dark for 20 days.

[0068] The field experiment soil is located in the field cultivation test field of Sichuan Academy of Agricultural Sciences Modern Agricultural Science and Technology Innovation Demonstration Garden in Taixing Town, Xindu District, Chengdu City (soil pH 6.5-7.5), which is divided into 3 groups, each group has 6 plots of 5m x 1m, corresponding to 6 parallel repeats during preservation.

[0069] Control group: inoculate the cultivation bag containing the Morel of GK group to the soil at the seeding amount of 300g / m2.

[0070] Test group 1: inoculate the cultivation bag containing the Morel of T1 group to the soil at the seeding amount of 300g / m2.

[0071] Experimental Group 2: Cultivation bags containing morel mushrooms from Group T2 were sown into the soil at a sowing rate of 300g / ㎡.

[0072] Morel mushrooms are cultivated and harvested using conventional methods. Calculate the yield (g / m³). 2 (dry weight), fruiting density (per m) 2 The number of ascocarps), the fruiting body maturity period (days from sowing to fruiting) (see...) Figure 1a , Figure 1b , Figure 1c ). Where * indicates a significant difference compared to the control group (P < 0.05), and △ indicates a significant difference compared to the experimental group 2 (P < 0.05).

[0073] The above description of specific embodiments of the present invention does not limit the present invention. Those skilled in the art can make various changes or modifications based on the present invention, and as long as they do not depart from the spirit of the present invention, they should all fall within the scope of protection of the claims of the present invention.

Claims

1. A Morei preservation medium consisting of 5-7 parts of sandy loam soil, 1-2 parts of oak broadleaf tree wood chips, 1 part of wheat, 1-2 parts of husk and 6-6.5 parts of water by weight, wherein, The Morchella is Morchella delavayii No.1, the sandy loam soil has a particle size of 0.25-1 mm, and the Quercus broadleaf wood chip has a particle size of 30-50 mesh.

2. The medium of claim 1, wherein the Morchella preservation medium is composed of 7 parts of sandy loam soil, 1 part of Quercus broadleaf wood chip, 1 part of wheat, 1 part of chaff, and 6 parts of water.

3. The medium of claim 1, wherein the Morchella preservation medium is composed of 6 parts of sandy loam soil, 2 parts of Quercus broadleaf wood chip, 1 part of wheat, 1 part of chaff, and 6 parts of water.

4. The medium of claim 1, wherein the Morchella preservation medium is composed of 5 parts of sandy loam soil, 2 parts of Quercus broadleaf wood chip, 1 part of wheat, 2 parts of chaff, and 6.5 parts of water.

5. The medium of any one of claims 1-4, wherein the Quercus broadleaf wood chip is any one of Quercus variabilis, Quercus aquifolioides, Quercus acutissima, Quercus aliena, or a combination thereof.

6. The medium of any one of claims 1-4, wherein the chaff and wheat are fresh and free of mildew.

7. The medium of any one of claims 1-4, wherein the Morchella preservation medium is prepared by mixing the required amounts of sandy loam soil, Quercus broadleaf wood chip, wheat, chaff, and water, and sterilizing the mixture.

8. The method for preparing the Morchella preservation medium of any one of claims 1-7, comprising the following steps: mixing the required amounts of sandy loam soil, Quercus broadleaf wood chip, wheat, chaff, and water, and sterilizing the mixture.

9. The use of the Morchella preservation medium of any one of claims 1-7 in the preservation of Morchella delavayii No.

1.

10. The use of claim 9, wherein the use is inoculating mycelial culture onto the preservation medium.

11. The use of claim 9, wherein the use is inoculating mycelial culture onto the preservation medium without subculture during the preservation period.

12. The use of any one of claims 9-11, wherein the use is inoculating mycelial culture onto the preservation medium, and Morchella delavayii No. 1 can be preserved for no less than 7 years.

13. The use of claim 12, wherein the use is inoculating mycelial culture onto the preservation medium, and Morchella delavayii No. 1 can be preserved for no less than 10 years.

14. A method of preserving Morchella comprising the steps of: Inoculating mycelial culture of Morchella delavayii No. 1 onto the Morchella preservation medium of any one of claims 1-7.

15. The preservation method of claim 14, wherein the preservation period is without subculture.

16. The preservation method of any one of claims 14-15, wherein the method can preserve Morchella delavayii No. 1 for no less than 7 years.

17. The preservation method of claim 16, wherein the method can preserve Morchella delavayii No. 1 for no less than 10 years.

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