CN110846234B - Pleurotus cornucopiae culture medium - Google Patents
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Abstract
The invention discloses an pleurotus cornucopiae culture medium which comprises a solid culture medium and a liquid culture medium, wherein the solid culture medium comprises 200g of potatoes, 15g of agar, 10g of glucose and 10-20 g of mannitol, and the volume is fixed to 1L by adding distilled water, and the liquid culture medium comprises 200g of potatoes, 10g of glucose and 10-20 g of mannitol and the volume is fixed to 1L by adding distilled water.
Description
Technical Field
The invention belongs to the technical field of pleurotus cornucopiae cultivation, and relates to a pleurotus cornucopiae culture medium.
Background
Pleurotus cornucopiae (Paul. Pers.) Rolland) is called Pleurotus cornucopiae, Pleurotus cornucopiae. The pleurotus cornucopiae is rich in nutritional ingredients such as proteins, various vitamins and minerals, is delicious in taste, and has the effects of resisting tumors, reducing blood pressure, improving metabolism, enhancing immunity and the like. Chinese patent 201210023901 discloses a liquid fermentation culture method of Pleurotus cornucopiae strain, and studies the influence of different carbon sources on the growth of Pleurotus cornucopiae, and finds that Pleurotus cornucopiae grows best in a culture medium with mannitol as the carbon source, the rest is fructose, glucose, sucrose and soluble starch in turn, the utilization rate of two monosaccharides of mannitol and fructose is good, and the optimal culture conditions are 3% of mannitol, 1% of yeast extract and 0.2% of KH2PO4And 0.05% MgSO4·7H2And O. However, the culture medium is added with mannitol, nitrogen source yeast extract and corn flour, and is subjected to fermentation culture.
Disclosure of Invention
The invention aims to provide a simple pleurotus cornucopiae culture medium which can remarkably accelerate the growth speed of pleurotus cornucopiae hyphae and improve the biomass of the pleurotus cornucopiae hyphae and the laccase activity.
The technical scheme for realizing the purpose of the invention is as follows:
an pleurotus cornucopiae culture medium comprises a solid culture medium and a liquid culture medium, wherein the solid culture medium comprises 200g of potatoes, 15g of agar, 10g of glucose and 10-20 g of mannitol, distilled water is added for fixing the volume to 1L, and the liquid culture medium comprises 200g of potatoes, 10g of glucose and 10-20 g of mannitol, distilled water is added for fixing the volume to 1L.
The solid culture medium and the liquid culture medium are sterilized at 121 ℃ for 20min, and are cooled for inoculation.
The inoculation is carried out on an ultra-clean workbench which is subjected to ultraviolet sterilization for 30 min.
The inoculation amount of the liquid culture medium is 10%.
The invention also provides a culture method of the pleurotus cornucopiae, which comprises the following specific steps:
inoculating Pleurotus cornucopiae mycelium into the above solid culture medium or liquid culture medium, and culturing at 25 deg.C in dark.
Preferably, the culture is carried out in a constant temperature shaker at 25 ℃ at a rotation speed of 150rpm while being inoculated in a liquid medium.
Preferably, when inoculated in a solid medium, the culture is carried out in an incubator at a constant temperature of 25 ℃.
Compared with the prior art, the invention has the following advantages:
the culture medium can provide good nutrient substances and a space environment for growth of the pleurotus cornucopiae, the preparation method is simple, mannitol is only added into a conventional glucose culture medium, the addition amount of the glucose is controlled to be 1% and the addition amount of the mannitol is controlled to be 1-2%, and when the mass ratio of the glucose to the mannitol is 1:2, the growth speed of pleurotus cornucopiae mycelia can be remarkably accelerated, the biomass of the mycelia is increased, the laccase activity of the mycelia is improved, the cost is low, and the growth vigor of the pleurotus cornucopiae is excellent.
Drawings
FIG. 1 is a graph showing the growth of the hyphae of Pleurotus cornucopiae of examples 1 and 2 and comparative examples 1 and 2 during the culture of Pleurotus cornucopiae;
FIG. 2 is a graph showing the growth rate of the hyphae of Pleurotus cornucopiae of examples 1 and 2 and comparative examples 1 and 2 during the solid culture of Pleurotus cornucopiae;
FIG. 3 is a graph showing the dry weight of Pleurotus cornucopiae mycelia of examples 1 and 2 and comparative examples 1 and 2 during the liquid culture of Pleurotus cornucopiae;
FIG. 4 is a graph showing laccase activity of Agaricus blazei Murill mycelia of examples 1 and 2 and comparative examples 1 and 2 obtained by liquid culture of Pleurotus cornucopiae;
FIG. 5 is a protein electrophoresis chart of Agaricus blazei Murill mycelia in examples 1 and 2 and comparative examples 1 and 2 obtained by liquid culture of Agaricus blazei Murill.
Detailed Description
The invention is further illustrated below with reference to specific embodiments and the accompanying drawings. 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.
Comparative example 1
The method comprises the steps of preparing a common culture medium for pleurotus cornucopiae, wherein the common culture medium comprises a solid culture medium and a liquid culture medium, the solid culture medium comprises 200g of potatoes, 20g of glucose and 100M L of liquid culture medium per bottle, the distilled water is added to the solid culture medium to achieve a constant volume of 1L, the liquid culture medium comprises 200g of potatoes, 20g of glucose and 1 g of distilled water to achieve a constant volume of L, the pleurotus cornucopiae hyphae is inoculated on an ultraclean workbench after ultraviolet sterilization for 30min and then cultured under the condition of darkness at 25 ℃, the liquid culture medium is cultured in a constant temperature shaking table at 25 ℃ with the rotating speed of 150rpm for 20min, the solid culture medium is cultured in a constant temperature incubator at 25 ℃, the process of dry weight pleurotus cornucopiae culture is monitored in real time, the growth condition and colony morphology are observed, when the dry weight pleurotus cornucopiae grows to the 5d, the result is photographed, the result is shown as a comparative example in fig. 1 in the solid culture process, the growth speed of the constant weight of the pleurotus cornucopiae is evaluated, the growth rate of the constant weight of the dry weight of the pleurotus cornucopiae is 2, the growth of the mycelium, the growth of the pleurotus cornucopiae is observed, the growth of the mycelium, the colony morphology of the pleurotus cornucopiae is observed, the mycelium, the pleurotus cornucopiae is observed, the colony morphology of the mycelium, the colony growth of the pleurotus cornucopiae mycelium, the colony growth is obtained by the culture medium, the culture medium is obtained by the result is obtained by the average growth of the result is obtained by the culture medium, the result of the culture medium is obtained by the result of the.
Example 1
Example 1 the solid medium had a composition and content of 200g of potato, 15g of agar, 10g of glucose, 10g of mannitol, and a volume of 1.34% by adding distilled water, and the liquid medium had a composition and content of 200g of potato, 10g of glucose, 10g of mannitol, and a volume of 1.56% by adding distilled water, and a volume of 250M L Erlenmeyer flask was used, each flask was filled with 100M L of the liquid medium, all of which were sterilized at 121 ℃ for 20min, and after cooling, the mycelia of Pleurotus cornucopiae were inoculated on a super clean bench after 30min of UV sterilization, and then cultured under a dark condition at 25 ℃, the liquid medium was cultured on a 25 ℃ shaker at a rotation speed of 150rpm, the solid medium was cultured in a 25 ℃ incubator, the process of the mycelia of Pleurotus cornucopiae was monitored in real time, the growth and colony morphology were observed, and when the mycelia of Pleurotus cornucopiae grow to the 5d, the results were obtained as shown in example 1, the case where the solid medium was evaluated on the dry weight, the growth rate of the mycelia after inoculation, the mycelia of the mycelia was evaluated, the mycelia of the mycelia, the mycelia of the mycelia was measured by the medium, the medium was measured by the average of the culture medium was taken by the filtration of the culture medium, the average of the culture medium was measured by the average of the culture medium was taken by the culture medium, the average culture of the culture medium of the culture of.
Example 2
Example 2 solid medium was composed of potato 200g, agar 15g, glucose 10g, mannitol 20g, distilled water to volume 1L, liquid medium was composed of potato 200g, glucose 10g, mannitol 20g, distilled water to volume 1L, 250M L Erlenmeyer flask was used, 100M L liquid medium was filled in each flask, all the media were sterilized at 121 ℃ for 20min, and after cooling, they were inoculated, Agaricus blazei Murill mycelium inoculation was performed on a clean bench after 30min of UV sterilization, and then they were cultured under dark condition at 25 ℃, liquid medium was cultured on a 25 ℃ shaker at 150rpm, solid medium was cultured in a 25 ℃ incubator, the process of Agaricus blazei Murill mycelium culture was monitored in real time, the growth and colony morphology were observed, photographing was performed when the mycelium grows to the 5d, the results were as shown in example 2 in fig. 1, the speed of dry weight inoculation, the growth of Agaricus blazei Murill mycelium was evaluated after inoculation, the speed of the culture was measured by the average radius of Agaricus blazei Murill mycelium, the culture medium was measured by filtration under the three-medium, the average culture medium strain culture medium after the culture (SDS-PAGE) was analyzed by the average culture speed 2d, the culture was measured by the three times of the culture medium, the culture medium was analyzed by the average culture medium was the culture medium after the culture medium was analyzed by the culture medium was analyzed by the average culture medium was the culture medium was analyzed by the electrophoresis, the culture medium was analyzed by the average of the culture medium was the culture under the culture medium (SDS-submerged culture medium after the culture medium was inoculated at the average of the culture radius 3 mm, the average of the culture under the culture radius 3 mm.
Comparative example 2
Comparative example 2 solid medium was composed of potato 200g, agar 15g, mannitol 50g, distilled water to a constant volume of 1L, liquid medium was composed of potato 200g, mannitol 50g, distilled water to a constant volume of 1L, 250M L flasks each containing 100M L liquid medium, all the media were sterilized at 121 ℃ for 20min, cooled and ready to inoculate, pleurotus cornucopiae hyphae inoculation was performed on an ultraclean bench after uv sterilization for 30min, then cultured under dark conditions at 25 ℃, liquid medium was cultured on a constant temperature shaker at 150rpm, solid medium was cultured in a constant temperature incubator at 25 ℃, the process of pleurotus cornucopiae cultivation was monitored in real time, its growth and colony morphology were observed, when the hyphae grew to the 5d, the culture was performed as shown in comparative example 2 in fig. 1, the growth rate of pleurotus cornucopiae was evaluated, the 2d, 5d was inoculated with a constant dry weight, the radius of the hyphae was measured by cross-cell culture, the radius of the hyphae was measured by filtration under the filter paper, the average culture medium was measured by the procedure of the culture medium after the culture (abm-cell culture), the procedure, the culture medium was measured by the average cell culture medium after the procedure, the procedure was measured by the average cell culture medium was calculated as the average cell culture medium, the average cell culture medium was found by the procedure, the average cell culture of the procedure of the culture medium was found by the procedure of the culture medium was found by the culture medium, the procedure of the culture medium was found by the procedure of.
TABLE 1
TABLE 2
FIG. 1 is a graph showing the growth of the hyphae of Pleurotus cornucopiae of examples 1 and 2 and comparative examples 1 and 2 during the culture of Pleurotus cornucopiae. FIG. 1 shows that: the pleurotus cornucopiae mycelia of examples 1 and 2 had better growth vigor than the pleurotus cornucopiae mycelia of comparative example 1, and the pleurotus cornucopiae mycelia of example 2 had the most growing branches and had the best growth vigor, while the pleurotus cornucopiae mycelia of comparative example 2 had sparse mycelia and had similar growth vigor to that of comparative example 1.
FIG. 2 is a graph showing the growth rate of the mycelia of Pleurotus cornucopiae of examples 1 and 2 and comparative examples 1 and 2 during the solid culture of Pleurotus cornucopiae. FIG. 2 shows that: the growth rates of the pleurotus cornucopiae hyphae of the example 1 and the example 2 are very different from those of the comparative example 1, and are both significantly higher than those of the comparative example 2, and the culture medium of the example 2 has the most significant effect of promoting the growth of the pleurotus cornucopiae hyphae.
FIG. 3 is a dry weight chart of the mycelia of Pleurotus cornucopiae of examples 1 and 2 and comparative examples 1 and 2 during the liquid culture of Pleurotus cornucopiae. FIG. 3 shows: the culture media of example 2 and comparative example 2 were able to increase the biomass of pleurotus cornucopiae hyphae, with significant differences between the dry weight of pleurotus cornucopiae hyphae of example 2 and comparative example 2, and no significant difference between the dry weight of pleurotus cornucopiae hyphae of example 1 and comparative example 1.
FIG. 4 is a graph showing laccase activity of Agaricus blazei Murill mycelia of examples 1 and 2 and comparative examples 1 and 2 obtained by liquid culture of Agaricus blazei Murill. FIG. 4 shows that: the laccase activity of the pleurotus cornucopiae mycelia in the examples 1 and 2 is obviously higher than that of the pleurotus cornucopiae mycelia in the comparative example 1, while the laccase activity of the pleurotus cornucopiae mycelia in the comparative example 2 is not obviously different from that of the comparative example 1, and the culture mediums in the examples 1 and 2 can improve the lignin degradation capability of the pleurotus cornucopiae mycelia.
FIG. 5 is a protein electrophoresis chart of Agaricus blazei Murill mycelia in examples 1 and 2 and comparative examples 1 and 2 obtained by liquid culture of Agaricus blazei Murill. FIG. 5 shows that: the pleurotus cornucopiae hypha protein band compositions of examples 1 and 2 were not different from the pleurotus cornucopiae hypha protein band composition of comparative example 1, indicating that the culture media of examples 1 and 2 did not change the protein expression of pleurotus cornucopiae hyphae; the composition of the pleurotus cornucopiae hypha protein band of comparative example 2 is significantly different from that of the pleurotus cornucopiae hypha protein band of comparative example 1, indicating that the culture medium of comparative example 2 changes the protein expression of pleurotus cornucopiae hyphae.
In conclusion, the culture medium in example 2, compared to comparative example 1, while increasing growth rate and hyphal biomass, promoted secretion of hyphal laccase activity and did not affect hyphal protein expression. Example 1 is slightly inferior to example 2 in the hyphal growth state and growth rate; comparative example 2 is significantly inferior to example 2 in the hyphal growth state, growth rate and laccase activity. The above data indicate that the medium formulation in example 2 is the optimal formulation. The formula optimizes the composition of a carbon source in the formula of a basic culture medium (comparative example 1), and the culture effect of the formula with 1% of the addition amount of glucose and 2% of the addition amount of mannitol is superior to that of the formula with 1% of the addition amount of glucose and 1% of the addition amount of mannitol, and is obviously superior to that of the formula with 5% of the addition amount of mannitol.
Claims (5)
1. The pleurotus cornucopiae culture medium comprises a solid culture medium and a liquid culture medium, and is characterized in that the solid culture medium comprises 200g of potatoes, 15g of agar, 10g of glucose and 10-20 g of mannitol, and distilled water is added to the mixture to achieve a constant volume of 1L, and the liquid culture medium comprises 200g of potatoes, 10g of glucose and 10-20 g of mannitol, and distilled water is added to achieve a constant volume of 1L.
2. The culture medium of Pleurotus cornucopiae according to claim 1, wherein the solid medium and the liquid medium are sterilized at 121 ℃ for 20min and cooled to be inoculated, the inoculation is performed on a super clean bench sterilized by UV for 30min, and the inoculation amount of the liquid medium is 10%.
3. A method for culturing Pleurotus cornucopiae based on the Pleurotus cornucopiae culture medium according to any one of claims 1 to 2, comprising:
a solid medium or a liquid medium according to any one of claims 1 to 2, wherein the mycelia of Pleurotus cornucopiae are inoculated and cultured in the dark at 25 ℃.
4. The method according to claim 3, wherein the culture is carried out in a shaker at 25 ℃ and a rotation speed of 150rpm while being inoculated into a liquid medium.
5. The culture method according to claim 3, wherein the culture is carried out in an incubator at 25 ℃ while inoculating the culture medium.
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