CN107502557B

CN107502557B - Hypsizygus marmoreus culture medium

Info

Publication number: CN107502557B
Application number: CN201710863705.XA
Authority: CN
Inventors: 杨焕玲; 赵妍; 陈明杰; 查磊; 黄建春; 余昌霞; 王晨光
Original assignee: SHANGHAI BAIXIN BIO-TECH CO LTD; Shanghai Academy of Agricultural Sciences
Current assignee: SHANGHAI BAIXIN BIO-TECH CO LTD; Shanghai Academy of Agricultural Sciences
Priority date: 2017-09-22
Filing date: 2017-09-22
Publication date: 2021-07-09
Anticipated expiration: 2037-09-22
Also published as: CN107502557A

Abstract

The invention relates to a Hypsizygus marmoreus culture medium, which comprises a solid culture medium and a liquid culture medium, wherein the solid culture medium comprises the following components in percentage by weight: 100g-200g of potato, 10g-20g of agar, 5g-10g of glucose and 5g-10g of trehalose, distilled water is added to the mixture to be constant volume of 0.5L-1L, and the composition and the content of a liquid culture medium are as follows: 100-200 g of potato, 5-10 g of glucose and 5-10 g of trehalose, and adding distilled water to a constant volume of 0.5L-1L. The culture medium can provide good nutrient substances and a space environment for the growth of the Hypsizygus marmoreus, the preparation method is simple, the growth speed of Hypsizygus marmoreus hyphae can be accelerated, the hypha biomass of the Hypsizygus marmoreus hyphae is obviously improved, the economic benefit is increased, and the cultured Hypsizygus marmoreus hyphae has good growth vigor.

Description

Hypsizygus marmoreus culture medium

Technical Field

The invention belongs to the field of Hypsizygus marmoreus cultivation, and particularly relates to a Hypsizygus marmoreus culture medium.

Background

Hypsizigus marmoreus (Hypsizigus marmoreus), also known as Hypsizigus marmoreus, is belonging to Basidiomycota (Basidiomycota), Hymenomycetes (Hymenomycetes), Agaricales (Agaricales), Tricholomataceae (Tricholomataceae), Hypsizigus (Hypsizigus). Hypsizygus marmoreus has tender meat and delicious taste, and is reputed to be fragrant in tricholoma matsutake and fragrant in Hypsizygus marmoreus. The mushroom is rich in various active ingredients such as protein, polysaccharide, amino acid, vitamins, etc., has effects of resisting oxidation, resisting virus, reducing blood sugar, reducing blood lipid, etc., and has edible and medicinal values. Hypsizygus marmoreus has been industrially cultivated in Shanghai, Jiangsu, Hebei, Shandong, Fujian, Guangdong provinces (city) in China, and the cultivation of robust Hypsizygus marmoreus mycelia is a precondition for realizing large-scale fruiting production.

Disclosure of Invention

The invention aims to solve the technical problem of providing a Hypsizygus marmoreus culture medium which is simple in preparation method and easy to popularize, and can accelerate the growth speed of Hypsizygus marmoreus hyphae and obviously improve the hypha biomass.

The Hypsizigus marmoreus culture medium comprises a solid culture medium and a liquid culture medium, wherein the solid culture medium comprises the following components in percentage by weight: 100g-200g of potato, 10g-20g of agar, 5g-10g of glucose and 5g-10g of trehalose, distilled water is added to the mixture to be constant volume of 0.5L-1L, and the composition and the content of a liquid culture medium are as follows: 100-200 g of potato, 5-10 g of glucose and 5-10 g of trehalose, and adding distilled water to a constant volume of 0.5L-1L.

And sterilizing the solid culture medium and the liquid culture medium at 121 ℃ for 20min, and cooling to be inoculated.

The inoculation is carried out on a clean bench with ultraviolet sterilization for 30 min.

The inoculum size of the liquid medium was 10%.

The solid culture medium and the liquid culture medium are both cultured under the dark condition of 25 ℃, the liquid culture medium is cultured in a constant temperature shaking table of 150rpm and 25 ℃, and the solid culture medium is cultured in a constant temperature incubator of 25 ℃.

The liquid culture medium is filled in a 250mL triangular flask with 100mL each.

Advantageous effects

The culture medium can provide good nutritional value and space environment for the growth of hypsizigus marmoreus, the preparation method is simple, the growth speed of hypsizigus marmoreus hyphae can be accelerated, the hypha biomass of the hypsizigus marmoreus is obviously improved, the economic benefit is increased, and the cultured hypsizigus marmoreus hyphae has good growth vigor.

Drawings

FIG. 1 is a graph showing the growth of Hypsizygus marmoreus hyphae of example 1 (example) and Hypsizygus marmoreus hyphae of comparative example 1 (control group) during Hypsizygus marmoreus cultivation;

FIG. 2 is a graph showing the growth rate of Hypsizygus marmoreus hyphae of example 1 (example) and Hypsizygus marmoreus hyphae of comparative example 1 (control) during solid culture of Hypsizygus marmoreus;

FIG. 3 is a dry weight graph of Hypsizygus marmoreus hyphae of example 1 (example) and Hypsizygus marmoreus hyphae of comparative example 1 (control) during Hypsizygus marmoreus liquid culture;

FIG. 4 is a protein electrophoresis chart of Hypsizygus marmoreus mycelia of example 1 (example) and Hypsizygus marmoreus mycelia of comparative example 1 (control group) obtained by liquid culture of Hypsizygus marmoreus.

Detailed Description

The invention will be further illustrated with reference to the following specific examples. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. Further, it should be understood that various changes or modifications of the present invention may be made by those skilled in the art after reading the teaching of the present invention, and such equivalents may fall within the scope of the present invention as defined in the appended claims.

Example 1

A Hypsizygus marmoreus culture medium comprises a solid culture medium and a liquid culture medium, wherein the solid culture medium comprises the following components in percentage by weight: 200g of potato, 15g of agar, 10g of glucose and 10g of trehalose, distilled water is added to the mixture to be constant volume to 1L, and the composition and the content of a liquid culture medium are as follows: 200g of potato, 10g of glucose and 10g of trehalose, adding distilled water to a constant volume of 1L, and using a 250mL triangular flask, wherein each flask contains 100mL of liquid culture medium. All media were sterilized at 121 ℃ for 20min, cooled and ready for inoculation. Inoculating Hypsizygus marmoreus mycelium on an ultra-clean workbench sterilized by ultraviolet for 30min, culturing at 25 deg.C in dark, culturing in a liquid culture medium at 150rpm in a constant temperature shaking table at 25 deg.C, and culturing in a constant temperature incubator at 25 deg.C with a solid culture medium. The cultivation process of Hypsizygus marmoreus is monitored in real time, the growth and colony morphology are observed, and pictures are taken when hyphae grow to 7 days, and the results are shown in the example in FIG. 1. In the solid culture process, the growth rate of hypsizigus marmoreus hyphae is evaluated, the colony radius is measured by a cross method at 4d and 8d after inoculation, the ratio of the difference (mm) of the colony radius to the growth days (d) is the hypha growth rate (mm/d), the three times of the measurement are repeated, and the average measurement result is shown in an example in a table 1. The inoculation amount of the liquid culture medium is 10%, and in the liquid culture process, when the hypsizygus marmoreus mycelium pellets are cultured for 10 days under dark conditions, the hypsizygus marmoreus mycelium pellets basically grow in a triangular flask and are in a vigorous growth stage. The mycelium pellets were filtered through a filter paper dried to a constant weight (M1), the mycelium pellets and the filter paper were dried together to a constant weight (M2), and the dry weight M of the hypsizygus marmoreus mycelium pellets was calculated as (M2-M1) and repeated three times, and the average dry weight of the mycelium pellets was measured as shown in the examples in table 2. Collecting Hypsizygus marmoreus mycelium from liquid culture medium, extracting protein by TCA-acetone method, and performing SDS-PAGE electrophoretic analysis.

Comparative example 1

A Hypsizygus marmoreus common culture medium comprises a solid culture medium and a liquid culture medium, wherein the solid culture medium comprises the following components in percentage by weight: 200g of potato, 15g of agar and 20g of glucose, distilled water is added to the mixture to be constant volume to 1L, and the composition and the content of a liquid culture medium are as follows: 200g of potato and 20g of glucose, adding distilled water to a constant volume of 1L, and using a 250mL triangular flask, wherein each flask contains 100mL of liquid culture medium. All media were sterilized at 121 ℃ for 20min, cooled and ready for inoculation. Hypsizygus marmoreus hyphae in the same growth state as in example 1 were inoculated on an ultraclean bench after ultraviolet sterilization for 30min, and then cultured under dark conditions at 25 deg.C, a liquid medium was cultured on a constant temperature shaker at 25 deg.C at a rotation speed of 150rpm, and a solid medium was cultured in a constant temperature incubator at 25 deg.C. The culture process of Hypsizygus marmoreus was monitored in real time, the growth and colony morphology were observed, and photographs were taken when the hyphae grew to 7 days, with the results shown in FIG. 1 for the control group. In the solid culture process, the growth rate of hypsizigus marmoreus hyphae is evaluated, the colony radius is measured by a cross method at 4d and 8d after inoculation, the ratio of the colony radius difference (mm) to the growth days (d) is the hypha growth rate (mm/d), the three times of the measurement are repeated, and the average measurement result is shown in a control group in a table 1. The inoculation amount of the liquid culture medium is 10%, and in the liquid culture process, when the hypsizygus marmoreus mycelium pellets are cultured for 10 days under dark conditions, the hypsizygus marmoreus mycelium pellets basically grow in a triangular flask and are in a vigorous growth stage. The mycelium pellets were filtered through a filter paper dried to a constant weight (M1), the mycelium pellets and the filter paper were dried together to a constant weight (M2), and the dry weight M of the hypsizygus marmoreus mycelium pellets was calculated as (M2-M1) and repeated three times, and the average dry weight of the mycelium pellets was measured as shown in the control group in table 2. Collecting Hypsizygus marmoreus mycelium from liquid culture medium, extracting protein by TCA-acetone method, and performing SDS-PAGE electrophoretic analysis.

FIG. 1 shows that: the Hypsizygus marmoreus mycelia of example 1 (example) were white and strong, dense colonies, regular edges, and better in growth vigor, compared to the Hypsizygus marmoreus mycelia of comparative example 1 (control).

FIG. 2 shows that: the growth rate of Hypsizygus marmoreus hyphae of example 1 (example) was significantly faster than that of Hypsizygus marmoreus hyphae of comparative example 1 (control), indicating that the medium of example 1 has a promoting effect on the growth of Hypsizygus marmoreus hyphae.

FIG. 3 shows: the dry weight of Hypsizygus marmoreus hyphae of example 1 (example) was significantly different from that of Hypsizygus marmoreus hyphae of comparative example 1 (control), and the dry weight of Hypsizygus marmoreus hyphae of example was significantly higher than that of control, indicating that the culture medium of example 1 was capable of significantly increasing the biomass of Hypsizygus marmoreus hyphae.

FIG. 4 shows that: the Hypsizygus marmoreus hypha protein band composition of example 1 (example) was indistinguishable from the Hypsizygus marmoreus hypha protein band composition of this comparative example 1 (control), indicating that the culture medium in example 1 did not alter the protein expression of Hypsizygus marmoreus hyphae.

TABLE 1

TABLE 2

Claims

1. A Hypsizygus marmoreus culture medium is characterized by comprising a solid culture medium and a liquid culture medium, wherein the solid culture medium comprises the following components in percentage by weight: 100g-200g of potato, 10g-20g of agar, 5g-10g of glucose and 5g-10g of trehalose, distilled water is added to the mixture to be constant volume of 0.5L-1L, and the composition and the content of a liquid culture medium are as follows: 100-200 g of potato, 5-10 g of glucose and 5-10 g of trehalose, and adding distilled water to a constant volume of 0.5L-1L; the inoculum size of the liquid medium was 10%.

2. The Hypsizygus marmoreus culture medium according to claim 1, wherein the solid culture medium and the liquid culture medium are sterilized at 121 ℃ for 20min, cooled and then inoculated.

3. The Hypsizygus marmoreus culture medium according to claim 2, wherein said inoculation is carried out on a super clean bench sterilized by ultraviolet light for 30 min.

4. The Hypsizygus marmoreus culture medium according to claim 1, wherein the solid culture medium and the liquid culture medium are both cultured in dark at 25 ℃, the liquid culture medium is cultured on a 25 ℃ incubator with a rotation speed of 150rpm, and the solid culture medium is cultured in a 25 ℃ incubator.

5. The Hypsizygus marmoreus culture medium according to claim 1, wherein the liquid culture medium is contained in a 250mL triangular flask in a volume of 100 mL.