CN110771424B - Phellinus igniarius cultivation method for accumulating anti-tumor effective ingredients - Google Patents
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Abstract
The present disclosure relates to a phellinus igniarius cultivation method for accumulating antitumor active ingredients, which includes the steps of: in the process of the growth of Phellinus linteus fruiting body, the fruiting body surface is contacted with an aqueous solution containing zinc and calcium. By adopting the cultivation method disclosed by the invention, the phellinus igniarius sporocarp can effectively accumulate anti-tumor effective components.
Description
Technical Field
The present disclosure relates to a phellinus igniarius cultivation method for accumulating antitumor active ingredients.
Background
The information in this background section is only for enhancement of understanding of the general background of the disclosure and is not necessarily to be construed as an admission or any form of suggestion that this information forms the prior art that is already known to a person of ordinary skill in the art.
Phellinus genus is Basidiomycetes, Polyporales, Polyporaceae, Phellinus genus (Phellinus), and is also called Morriscus, Tree chicken, Hoffonia simplicifolia, Phellinus linteus, Phellinus igniarius, Phellinus linteus, and Prunus mume. Modern medical research finds that phellinus igniarius has various pharmacological actions and has obvious curative effects in the aspects of antibiosis, immunoregulation, sweat gland secretion inhibition, tumor resistance, fibrosis resistance, inflammation diminishing, pain relieving, oxidation resistance and the like. The phellinus igniarius polysaccharide is an effective active ingredient of phellinus igniarius, and related research reports show that the phellinus igniarius polysaccharide plays a main anti-tumor role, and the research finds that the phellinus igniarius polysaccharide can effectively inhibit the growth of tumor cells and can also prevent the metastasis of the tumor cells.
With the enhancement of economic level and health care consciousness of people, the demand of phellinus igniarius is large, and the wild phellinus igniarius resources are in short supply, so that the artificial cultivation technology of phellinus igniarius is produced at the same time.
In recent years, reports on artificial culture of Phellinus linteus mainly focus on the formation rate of fruiting bodies, uniformity of fruiting body size, contamination rate, etc., and a specific culture method for Phellinus linteus accumulating antitumor active ingredients has been rare.
Disclosure of Invention
Against the background art, the present disclosure provides a phellinus linteus cultivation method for accumulating antitumor active ingredients, which can rapidly and effectively cause fruiting bodies to accumulate antitumor active ingredients such as polysaccharides.
Specifically, the following technical scheme is adopted in the disclosure:
the present disclosure provides a phellinus igniarius cultivation method for accumulating antitumor active ingredients, comprising the steps of:
in the process of the growth of phellinus linteus fruiting body, the surface of the fruiting body is contacted with an aqueous solution containing zinc ions and calcium ions.
Furthermore, the anti-tumor effective component is phellinus igniarius polysaccharide.
Compared with the related technology known by the inventor, one technical scheme of the present disclosure has the following beneficial effects:
(1) by adopting the cultivation method disclosed by the invention, the phellinus igniarius sporocarp can effectively accumulate anti-tumor effective components.
(2) By adopting the cultivation method disclosed by the invention, the yield and the quality of phellinus igniarius are good.
(3) By adopting the cultivation method disclosed by the invention, the growth cycle of the phellinus igniarius sporocarp is relatively shortened compared with that of the conventional method.
(4) The cultivation method disclosed by the invention is convenient to operate, reliable and effective.
Drawings
The accompanying drawings, which are incorporated in and constitute a part of this disclosure, illustrate embodiments of the disclosure and, together with the description, serve to explain the disclosure and not to limit the disclosure.
FIG. 1 shows the stock species in example 1 of the present disclosure.
Fig. 2 is a phellinus linteus fruit body to be harvested in example 1 of the present disclosure.
FIG. 3 is a graph showing the variation of dry weight (per bag) and crude polysaccharide content of Phellinus linteus fruiting body at different growth stages in example 1 of the present disclosure.
FIG. 4 is a graph showing the variation of dry weight (per bag) of Phellinus linteus fruiting body with crude polysaccharide content at different growth stages in example 1 of the present disclosure.
FIG. 5 is a graph showing the variation of dry weight (per bag) and crude polysaccharide content of Phellinus linteus fruiting body at different growth stages in Experimental example 1 of the present disclosure.
FIG. 6 shows the change in the content of crude polysaccharide in Experimental example 3 of the present disclosure.
Detailed Description
It should be noted that the following detailed description is exemplary and is intended to provide further explanation of the disclosure. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs.
It is noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of example embodiments according to the present disclosure. As used herein, the singular forms "a", "an" and "the" are intended to include the plural forms as well, and it should be understood that when the terms "comprises" and/or "comprising" are used in this specification, they specify the presence of the stated features, steps, operations, and/or combinations thereof, unless the context clearly indicates otherwise.
As described in the background art, there is currently a lack of a specific method for culturing phellinus linteus accumulating antitumor active ingredients, and in order to solve the above technical problems, in an exemplary embodiment of the present disclosure, there is provided a method for culturing phellinus linteus accumulating antitumor active ingredients, the method comprising the steps of:
in the process of the growth of Phellinus linteus fruiting body, the fruiting body surface is contacted with an aqueous solution containing zinc and calcium.
In one or more specific embodiments of the present disclosure, the specific step of contacting the fruiting body with an aqueous solution containing zinc and calcium is: an aqueous solution containing zinc and calcium is applied in the form of a spray to the surface of the fruit body.
The whole growth process of phellinus igniarius basically comprises the following steps: spawn running stage and sporophore development stage. The development stage of the fruiting body basically comprises: differentiation of primordium of mushroom bud, stem of Phellinus linteus, opening umbrella of Phellinus linteus, and mature fruiting body. The inventor finds that the polysaccharide content is highest in the early stage of opening the phellinus igniarius (20-25 days after opening), and the polysaccharide content is reduced rapidly along with the growth of fruit bodies through long-term practice and a large number of experiments. Although the polysaccharide content is highest at the initial stage of opening the umbrella, the fruiting body is smaller and the weight is lower at the stage, and if the fruiting body is directly harvested at the stage, the yield of the phellinus igniarius polysaccharide which is actually obtained is not high, and the economic benefit is lower. In view of this, the present inventors have started from the viewpoint of sugar metabolism and tried to stimulate sugar-metabolizing enzymes for polysaccharide synthesis by a specific means, so that the sugar-metabolizing enzymes in the fruit body part can maintain a high enzyme activity for a long period of time, thereby promoting the synthesis of phellinus linteus polysaccharides. After long-term diligent efforts and intensive studies by the inventors, it has been found that when the fruit body of phellinus linteus is brought into contact with an aqueous solution containing zinc ions and calcium ions at a specific growth period of the fruit body of phellinus linteus, the polysaccharide content in the fruit body can be maintained at a high level, so that the polysaccharide content is reduced slowly, the growth rate of the fruit body can be made relatively fast, the contamination rate is already low along with the growth of the fruit body, and thus more phellinus linteus polysaccharides can be obtained by obtaining the fruit body at the specific period, and the economic benefit is high.
In the course of the test, the present inventors also tested aqueous solutions of various ions, such as an aqueous solution containing only zinc ions, an aqueous solution containing only calcium ions, an aqueous solution containing iron ions, an aqueous solution containing magnesium ions, an aqueous solution containing molybdenum ions, an aqueous solution containing copper ions, and the like, but found that the effect is excellent and the synthesis of polysaccharide can be promoted more significantly when only zinc ions and calcium ions are present at the same time.
In one or more embodiments of the present disclosure, the anti-tumor effective ingredient is phellinus linteus polysaccharide.
Different types of phellinus igniarius have certain differences in effective components and growth conditions. In one or more embodiments of the present disclosure, the species of Phellinus linteus is Phellinus igniarius.
In one or more embodiments of the present disclosure, the phellinus linteus fruiting body growth process is performed at any time point or time period from 18 to 22 days of the opening of the fungus sack. Experiments prove that in the period before the opening of phellinus igniarius on day 18-22 of the opening of the fungus bag, the spraying of the aqueous solution containing zinc ions and calcium ions can keep the polysaccharide content in the fruiting body in the later period at a higher level, so that the polysaccharide content is reduced slowly, and the growth speed of the fruiting body is relatively high.
In one or more embodiments of the present disclosure, the phellinus linteus fruiting body is harvested 32 to 38 days after the opening of the fungus sack. Although the content of crude polysaccharides in Phellinus linteus fruiting body is highest around day 25, the fruiting body weight is small at this time, and the total polysaccharide content is not high. In addition, the weight of the fruiting body is increased rapidly in the period of 32 to 38 days, and the harvesting is preferably carried out in the period of 32 to 38 days in comprehensive consideration.
In one or more embodiments of the present disclosure, the mass concentration of the zinc ion or the calcium ion in the aqueous solution is 0.01 to 0.1 w/w%. Tests prove that the effect is better when the mass concentration of the zinc ions or the calcium ions is 0.02 w/w%. Furthermore, the zinc ions are added in the form of one or more of zinc sulfate, zinc chloride, zinc nitrate and the like, and the calcium ions are added in the form of one or more of calcium chloride, calcium nitrate, calcium dihydrogen phosphate and the like.
In one or more embodiments of the present disclosure, the method comprises the steps of:
(1) cultivation of cultivars: filling the culture medium of the cultivated species into a fungus bag, inoculating the stock species, carrying out dark culture, and ending fungus growth when the fungus bag is full of the strains;
(2) fruiting and culturing:
and (4) continuing dark culture after spawn running is finished, opening openings at two sides of the fungus bag after hyphae in the fungus bag are completely changed into dark yellow from light yellow, and controlling the fruiting environmental conditions.
In the step (1), the stock is prepared by the following method:
A. activating the parent strain:
inoculating the mother seeds to a PDA comprehensive culture medium for activation, and carrying out dark culture at a constant temperature of 25-28 ℃ for 8-12 days;
the PDA comprehensive culture medium comprises: 200g of potato (peeled), 20g of glucose, 3g of monopotassium phosphate, 1.5g of magnesium sulfate, 18g of vitamin B, 20g of agar and 1000mL of water, wherein the pH value is natural;
B. stock culture:
putting the stock culture medium into a strain bag, sealing, sterilizing at high pressure, inoculating the activated strain into the stock culture medium, performing dark culture at a constant temperature of 25-28 ℃, growing the strain in the strain bag after 25-40 days, and refrigerating the strain bag full of the strain (namely the stock) at 4 ℃ for standby or immediate use.
The method aims to screen and optimize the formula of the stock culture medium, and the stock culture medium obtained in a short time is thick in hypha, and is prepared from the following raw materials in percentage by mass through a large number of tests: 20-25% of soybean straw, 20-25% of corncob, 2-3% of gypsum, 4-6% of glucose, 0.4-0.6% of potassium hydrogen phosphate, 0.4-0.6% of magnesium sulfate and the balance of mulberry sawdust. Adding water to make the water content of the culture medium be 55-65 w/w%.
In the step (1), the culture medium for the cultivars is prepared from the following raw materials in percentage by mass: 20-25% of soybean straw, 20-25% of corncob, 3-5% of cane sugar, 2-3% of gypsum, 0.5-1% of cinnamon waste and the balance of mulberry sawdust. Adding water to make the water content of the culture medium be 60-70 w/w%.
Wherein the cinnamon waste refers to cinnamon residue, cinnamon leaves and other waste generated after cinnamon oil production. Tests prove that the pollution rate of fungus sacks and phellinus igniarius sporocarp can be effectively reduced by adding a small amount of cinnamon waste into the culture medium. The using amount of the phellinus igniarius is 0.5-1%, and experiments show that the phellinus igniarius contains aromatic oil substances and terpenoids, has a certain inhibiting effect on the growth of phellinus igniarius, and is not suitable for higher content.
The mulberry sawdust, the soybean straw and the corncob are matched for use, so that the requirement of rapid growth of the phellinus igniarius can be met. The granularity of solid matters (mulberry sawdust, soybean straws, corncobs and cinnamon waste) in the culture medium is 0.1-2 cm. Tests prove that the culture medium of the cultivar can obviously reduce the pollution rate of fungus bags and phellinus igniarius sporocarp under the condition of providing sufficient nutrient precursors for phellinus igniarius.
In the step (1), the dark culture temperature is 25-28 ℃, and the culture time is 28-35 days.
The inventor researches the influence of environmental conditions such as temperature, humidity and illumination on the polysaccharide in the phellinus igniarius sporocarp, and tests and researches find that the specific day-night temperature difference is more favorable for the formation and accumulation of the polysaccharide in the phellinus igniarius sporocarp. Therefore, in the step (2), the environmental conditions for fruiting are selected as follows: the day temperature (6: 00-18: 00) is 28-32 ℃, the night temperature (18: 00-6: 00 in the next day) is 22-26 ℃, the temperature difference is 5-6 ℃, the humidity is 85-95%, the light is scattered and irradiated, and the ventilation is carried out twice a day for 0.5-1 h each time.
The phellinus igniarius culture vessel according to the present disclosure uses a fungus bag, and the material of the fungus bag may be polyethylene or polypropylene.
The inoculation operation according to the present disclosure should be performed strictly in a sterile manner to prevent contamination by undesired bacteria.
Regarding the inoculation amount related to the present disclosure, 1.0-1.2g of the seed is used per 100g of the culture material, and if the inoculation amount is small, the entering mixed bacteria can be rapidly propagated in the culture material.
In order to make the technical solutions of the present disclosure more clearly understood by those skilled in the art, the technical solutions of the present disclosure will be described in detail below with reference to specific embodiments.
Unless otherwise specified, the following test materials and test methods were used in the following examples.
Test materials:
(1) the strain source is as follows: phellinus linteus species used in the present disclosure is Phellinus igniarius, No. BNCC231138, stored on PDA slant medium, which is available through conventional commercial routes.
(2) Stock activation medium (PDA integrated medium): 200g of potato (peeled), 20g of glucose, 3g of monopotassium phosphate, 1.5g of magnesium sulfate, 18g of vitamin B, 20g of agar and 1000mL of water, and the pH value is natural. The culture medium is mainly used for strain activation.
(3) The stock culture medium is composed of the following raw materials in percentage by mass: 20% of soybean straw, 20% of corncob, 2% of gypsum, 4% of glucose, 0.6% of potassium hydrogen phosphate, 0.6% of magnesium sulfate and the balance of mulberry sawdust. Adding water to make the water content of the culture medium 60%.
(4) The culture medium for the cultivars comprises the following raw materials in percentage by mass: 20% of soybean straw, 20% of corncob, 4% of sucrose, 2% of gypsum, 0.5% of cinnamon waste and the balance of mulberry sawdust. Adding water to make the water content of the culture medium 60%.
(5) The determination of the content of the crude polysaccharide is referred to agricultural industry standard NY/T676-2008 of the people's republic of China.
Example 1
A Phellinus linteus cultivation method for accumulating antitumor effective components comprises the following steps:
(1) activation of the preserved strain:
inoculating the preserved slant strain (mother strain) BNCC231138 to a PDA comprehensive culture medium for activation in 5 months and 10 days in 2018, and performing dark culture at a constant temperature of 25 ℃ for 8 days;
(2) stock culture:
the strain bag is made of polyethylene bag of 15 × 40cm, the stock culture medium is filled into the strain bag in 2018, 5 month and 18 day, the strain bag is compacted while being filled, then the strain bag is sealed, the activated strain is inoculated into the stock culture medium after autoclaving, dark culture is carried out at constant temperature of 25 ℃, the strain is overgrown with the strain bag after 30 days, and the strain bag overgrown with the strain (namely the stock) is refrigerated at 4 ℃ for standby as shown in figure 1.
(3) Cultivation of cultivars:
a spawn running stage: the fungus bag is made of 15 × 40cm polyethylene bag, the culture medium of the cultivated species is filled into the fungus bag in 2018, 6 and 1 months, the fungus bag is compacted while being filled, then the fungus bag is sealed, the original species is inoculated into the culture medium of the cultivated species after autoclaving, dark culture is carried out at the constant temperature of 25 ℃, the humidity in the culture room is 65%, and the fungus bag is full of the strains in 2018, 6 and 30 months (after 30 days).
(4) Fruiting and culturing:
continuously culturing at constant temperature of 25 ℃ in the dark, observing color change of hyphae in the fungus bag, when the hyphae in the fungus bag is changed from light yellow to dark yellow basically, opening rectangular openings (the size is about 3-4 multiplied by 1-2 cm) at two sides of the fungus bag, controlling the environmental conditions of a fruiting culture room, wherein the day temperature (6: 00-18: 00) is 30 ℃, the night temperature (18: 00-6: 00 in the second day) is 25 ℃, the humidity is about 90%, irradiating by scattered light, ventilating twice a day, and ventilating for 0.5h each time. And regularly sterilizing the fruiting culture chamber to prevent infection.
Wherein, on the 20 th day after opening, adopt the atomizer to spray the aqueous solution that contains zinc sulfate and calcium nitrate to the fruit body of fruiting department, the mass concentration of zinc sulfate and calcium nitrate is 0.02 w/w%, requires that the spraying is even unanimous, the spraying volume: the water mist-shaped droplets are only required to be formed on the surface of the fruit body, and are not sprayed more, otherwise, the fruit body is easy to mildew and rot.
On the 35 th day after the opening of the fungus sack, fruiting bodies can be harvested, as shown in fig. 2.
Example 2
A Phellinus linteus cultivation method for accumulating antitumor effective components comprises the following steps:
the difference from the example 1 is that on the 22 nd day after the opening, the water solution containing zinc sulfate and calcium nitrate is sprayed to the fruiting body at the fruiting part by a sprayer, the mass concentration of the zinc sulfate and the calcium nitrate is 0.04 w/w%, and the spraying is required to be uniform.
And (4) harvesting the fruiting bodies on the 38 th day after the opening of the fungus bag.
Other methods and conditions are the same.
Example 3
A Phellinus linteus cultivation method for accumulating antitumor effective components comprises the following steps:
the difference from the example 1 is that on the 20 th day after the opening, the water solution containing zinc sulfate and calcium nitrate is sprayed to the fruiting body at the fruiting part by a sprayer, the mass concentration of the zinc sulfate and the calcium nitrate is 0.04 w/w%, and the spraying is required to be uniform.
And on the 36 th day after the opening of the fungus bag, harvesting the fruiting bodies.
Other methods and conditions are the same.
Example 4
A Phellinus linteus cultivation method for accumulating antitumor effective components comprises the following steps:
(1) activation of the preserved strain:
inoculating the preserved slant strain (mother strain) BNCC231138 to a PDA comprehensive culture medium for activation in 5 months and 10 days in 2018, and performing dark culture at a constant temperature of 25 ℃ for 8 days;
(2) stock culture:
the strain bag is made of polyethylene bag of 15 × 40cm, the stock culture medium is filled into the strain bag in 2018, 5 and 18 months, the strain bag is sealed, the strain bag is autoclaved, the activated strain is inoculated to the stock culture medium, dark culture is carried out at a constant temperature of 25 ℃, the strain bag is full of strain after 30 days, and the strain bag full of strain (namely stock) is refrigerated at 4 ℃ for standby as shown in figure 1.
(3) Cultivation of cultivars:
a spawn running stage: the strain bag is made of polyethylene bag of 15 × 40cm, the culture medium of the cultivated species is filled into the strain bag, the bag is sealed and autoclaved, the stock is inoculated into the culture medium of the cultivated species, dark culture is carried out at the constant temperature of 28 ℃, and the strain grows over the strain bag after 30 days.
(4) Fruiting and culturing:
continuously carrying out constant-temperature dark culture at 28 ℃, observing the color change of the hyphae in the fungus bag, when the hyphae in the fungus bag are basically completely changed into dark yellow from the previous light yellow, forming rectangular openings (the size is about 2 multiplied by 1cm) at two sides of the fungus bag, controlling the environmental conditions of a fruiting culture room, wherein the day temperature (6: 00-18: 00) is 32 ℃, the night temperature (18: 00-6: 00) is 28 ℃, the humidity is about 95 percent, irradiating by scattered light, and ventilating twice a day for 0.5 hour each time. And regularly disinfect the fruiting culture room to prevent infection.
On the 20 th day after opening, a sprayer is adopted to spray an aqueous solution containing zinc sulfate and calcium nitrate to fruiting bodies at fruiting positions, the mass concentration of the zinc sulfate and the mass concentration of the calcium nitrate are both 0.02 w/w%, and the spraying is required to be uniform.
And harvesting fruiting bodies 35 days after the opening of the fungus bag.
Experimental example 1
The differences from example 1 are: on day 20 after opening, only the water spray control was applied and no aqueous solution containing zinc sulfate and calcium nitrate was applied.
Other methods and conditions were the same as in example 1.
Experimental example 2
The total polysaccharide content of the sporocarp in different growth periods is researched:
all fruit bodies were collected on the 5 th, 10 th, 15 th, 25 th, 35 th, 45 th, 55 th and 65 th days after opening of the fungus sacks of example 1 and experimental example 1, respectively, and the dry weight and the crude polysaccharide content of the fruit bodies were measured.
As shown in FIG. 3, the dry weight of the fruiting body cultured by the method of example 1 increased with the increase of the culture time, while the content of crude polysaccharide in the fruiting body increased and decreased with the increase of the culture time, the content of crude polysaccharide reached the maximum in about 25 days (at the initial stage of opening the umbrella), decreased relatively gradually in about 25 to 35 days, and decreased sharply after 35 days. The inventors chose to harvest for 35 days, which resulted in the maximum amount of crude polysaccharide, as shown in fig. 4.
As shown in FIG. 5, the dry weight of the fruiting body cultured by the method of Experimental example 1 and the content of crude polysaccharide were lower than those of example 1 in the same day. And the content of the crude polysaccharide has no flat period when the content is reduced, and the reduction speed of the crude polysaccharide is very high.
Experimental example 3
The differences from example 1 are: on the 20 th day after opening, spraying an aqueous solution containing magnesium sulfate and calcium nitrate to fruiting bodies at fruiting positions by using a sprayer, wherein the mass concentrations of the magnesium sulfate and the calcium nitrate are both 0.02 w/w%, and the spraying is required to be uniform.
Other methods and conditions were the same as in example 1.
Collecting all fruiting bodies on 5 th, 10 th, 15 th, 25 th, 35 th, 45 th, 55 th and 65 th days after opening the bag, and determining crude polysaccharide content of the fruiting body.
As shown in fig. 6, the content of crude polysaccharide in the phellinus linteus fruit body was only 9.2% on the 25 th day of opening of the fungus sack, which was lower than that of example 1, and the rate of decrease of crude polysaccharide content was faster after 25 days.
The above embodiments are preferred embodiments of the present disclosure, but the embodiments of the present disclosure are not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present disclosure should be regarded as equivalent replacements within the scope of the present disclosure.
Claims (5)
1. A Phellinus igniarius cultivation method for accumulating anti-tumor effective components is characterized by comprising the following steps:
(1) cultivation of cultivars: filling the culture medium of the cultivated species into a fungus bag, inoculating the stock species, carrying out dark culture, and ending fungus growth when the fungus bag is full of the strains;
the culture medium for the cultivars comprises the following raw materials in percentage by mass: 20-25% of soybean straw, 20-25% of corncob, 3-5% of cane sugar, 2-3% of gypsum, 0.5-1% of cinnamon waste and the balance of mulberry sawdust; adding water to make the water content of the culture medium be 60-70 w/w%;
(2) fruiting and culturing:
continuing dark culture after spawn running is finished, opening openings at two sides of the fungus bag after hyphae in the fungus bag are completely changed into dark yellow from light yellow, and controlling the environment condition of fruiting;
wherein in the process of the growth of phellinus igniarius sporocarp, the surface of the sporocarp is contacted with an aqueous solution containing zinc and calcium;
the growth process of the phellinus igniarius sporocarp refers to any time point or time period in 18-22 days of opening of the fungus sack;
harvesting phellinus igniarius sporocarp on the 32 th to 38 th days after the opening of the fungus bag;
the anti-tumor active ingredient is phellinus igniarius polysaccharide.
2. The method according to claim 1, wherein the Phellinus linteus is Phellinus linteusPhellinus igniarius。
3. The method according to claim 1, wherein the mass concentration of zinc ions or calcium ions in the aqueous solution is 0.01 to 0.1 w/w%; the addition form of the zinc ions is one or more of zinc sulfate, zinc chloride and zinc nitrate, and the addition form of the calcium ions is one or more of calcium chloride, calcium nitrate and calcium dihydrogen phosphate.
4. The method according to claim 1, wherein in the step (1), the dark culture temperature is 25 to 28 ℃ and the culture time is 28 to 35 days.
5. The method as claimed in claim 1, wherein in the step (2), the environmental conditions for fruiting are as follows: the day temperature is 6: 00-18: 00 and 28-32 ℃, the night temperature is 18: 00-6: 00 and 22-26 ℃ on the next day, the humidity is 80-90%, the scattered light is irradiated, and the sun is ventilated twice every day for 0.5-1 h.
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