CN110972806A - Cultivation method and artificial cultivation method of sulphur vermilion strain - Google Patents
Cultivation method and artificial cultivation method of sulphur vermilion strain Download PDFInfo
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Abstract
The invention provides a cultivation method and an artificial cultivation method of a sulfur vermilion strain, and the method comprises three key technical points: (1) the spawn running technology is innovative in that oak sawdust and eucommia branches are used as main raw materials in an artificial culture medium, so that the growth rate of hyphae is improved; (2) the formation of the primordium is stimulated by utilizing low-temperature induction, so that the primordium is under the growth advantage condition, and a foundation is provided for further developing a large number of buds and ensuring the yield of the bag cultivation; (3) the primordium is differentiated to obtain the sporocarp, the primordium can be smoothly transited to reproductive growth at the stage according to the fruiting condition disclosed by the invention, the management of the differentiation period of buds is very critical, and the method is an important condition for obtaining the high and stable yield of the sulfur red bacterium.
Description
Technical Field
The invention belongs to the technical field of edible fungus cultivation, relates to strain cultivation, and particularly relates to a cultivation method and an artificial cultivation method for sulfur cinnabar strains.
Background
Laetiporus cinnabarinus belongs to Basidiomycota, Agaricales, Polyporales, Fomitopsidae and Laetiporus, and is a large fungus for both food and medicine, and has high nutritive value and medicinal value. The Laetiporus cinnabarinus is mainly distributed in the areas of Hebei, Tibet, Xinjiang, Shanxi, Sichuan, Fujian and the like in China, belongs to middle and low temperature type fungi, and is grown on the base parts of coniferous trunks such as larch and the like and broad-leaf trunks such as oak and the like. The product is similar to salmon, has good taste and rich nutrition, contains various amino acids and medicinal components essential for human body, is cheese-shaped after aging, and is not suitable for eating, but can be used as medicine. The sulfur red mushroom is medicinal, warm in nature and sweet in taste, and has the effects of reducing blood fat, resisting tumor, delaying aging, improving human immunity and the like; in addition, the fruit body contains the odontic acid, can synthesize steroid medicament adrenocortical hormone, and is also an important medicament for preventing and treating various diseases. Therefore, the artificial cultivation of the sulfur cinnabarinus has wide application and development prospects.
At present, the latest bag cultivation mode for artificially cultivating the sulfur red bacteria has appeared in China, but tests show that the technology has the phenomena of low yield and unstable growth and cultivation quality of the sulfur red bacteria. At present, two common analysis reasons exist, namely that the quality of the sulfur cinnabarinus strain is not excessive, for example, the yield is low due to low activity, and the artificial cultivation technology of the sulfur cinnabarinus is still to be improved.
Disclosure of Invention
Aiming at the defects of the existing sulfur red fungus bag cultivation technology, the invention aims to firstly obtain a strain which is high in activity and beneficial to the yield and quality of sulfur red fungus, and on the basis, the invention also aims to provide a set of advanced artificial cultivation technology of sulfur red fungus based on the obtained strain, so that the problems of the existing sulfur red fungus bag cultivation technology are thoroughly solved from two sources, and theoretical reference basis is provided for further development and utilization of sulfur red fungus.
Therefore, the technical means adopted by the invention are as follows:
firstly, the invention provides a cultivation method of a sulphur vermilion strain, which specifically comprises the following steps:
step 1, inoculating a sulfur cinnabar strain stock into an artificial culture medium for spawn running; the spawn running conditions comprise: culturing at 15-18 deg.C, air relative humidity of 55-65%, carbon dioxide concentration of 0.8-1.0% in dark until hypha grows in the bag; the preparation of the artificial cultivation medium comprises the following steps: placing the artificial culture medium in a polyethylene material bag for sterilization for later use, wherein the formula of the artificial culture medium comprises the following components in percentage by mass: 48% of oak sawdust, 32% of eucommia branches, 15% of bran, 2% of sucrose, 1% of gypsum, 1% of humus soil and 1% of calcium carbonate;
step 2, performing low-temperature induction on the material bag full of hyphae to generate primordium;
step 3, differentiating the primordium to form a sulfur bacterium sporophore under the conditions that the temperature is 14-18 ℃, the relative air humidity is 80-90%, the carbon dioxide concentration is 1.5-2.0% and the illumination intensity is 200-;
and 4, separating and preserving the collected sulfur cinnabarinus sporocarp on an inclined plane to obtain the sulfur cinnabarinus bacterial strain.
Further, the conditions for low-temperature induction of primordia include, in particular: the relative humidity of air is 75-85%, the concentration of carbon dioxide is 1.5-2.0%, the illumination intensity is 100-.
Furthermore, the preparation method of the sulfur cinnabarinus protospecies particularly comprises the following steps:
step 1, inoculating activated wild sulfur red bacterium strains on a sulfur red bacterium liquid culture medium for culture to obtain sulfur red bacterium liquid strains; the liquid culture medium formula comprises: 15g of glucose, 15g of cane sugar, 5.0g of peptone, 2g of yeast extract, 0.5g of monopotassium phosphate, 1.0g of magnesium sulfate and vitamin B42mg, 1000mL of water;
step 2, inoculating the liquid strains to a stock culture medium for dark culture, and then carrying out 200-300lx scattered light treatment to obtain a sulfur cinnabarinus stock; the stock culture medium comprises the following components in percentage by mass: 50% of oak sawdust, 30% of eucommia branches, 5% of wheat, 10% of bran, 2% of sucrose, 0.8% of monopotassium phosphate, 0.6% of magnesium sulfate, 1% of gypsum and 0.6% of calcium phosphate.
Finally, preferably, the formula of the storage medium for slant storage comprises: 25g of wheat, 20g of bran and 80-150mL of nutrient solution, wherein the formula of the nutrient solution comprises the following components: 10g of glucose, 10g of yeast extract, 1.0g of monopotassium phosphate, 1.0g of magnesium sulfate and 1000mL of water.
The invention also aims to directly protect the sulfur cinnabar strain cultured by the sulfur cinnabar strain culture method and provide convenience for the industrial utilization of the sulfur cinnabar strain. Specifically, the sulfur vermilion strain is named as sulfur vermilion (Laetiporus Sulphureus) LS-AK405 with the collection number of CGMCC No.18155, and the strain is preserved in the China general microbiological culture Collection center (CGMCC for short, with the address of No. 3 Xilu No. 1. Beijing Kogyo facing the Yangtze district, Beijing) within 26 months and 8 months in 2019.
Based on the obtained bacterial strain, the third purpose of the invention is to provide an artificial cultivation method of sulfur cinnabarinus, which specifically comprises the following steps:
step 1, spawn running: inoculating Sulfur Laetiporus Sulphureus LS-AK405(CGMCC No.18155) into artificial culture medium for spawn running; the spawn running conditions comprise: culturing at 15-18 deg.C, air relative humidity of 55-65%, carbon dioxide concentration of 0.8-1.0% in dark until hypha grows in the bag; the preparation of the artificial cultivation medium comprises the following steps: placing the artificial culture medium in a polyethylene material bag for sterilization for later use, wherein the formula of the artificial culture medium comprises the following components in percentage by mass: 48% of oak sawdust, 32% of eucommia branches, 15% of bran, 2% of sucrose, 1% of gypsum, 1% of humus soil and 1% of calcium carbonate;
step 2, low-temperature induction of primordium formation: performing low-temperature induction on the material bag full of hypha to generate primordium;
step 3, fruiting: the primordium is differentiated to form a sulfur red mushroom fruiting body under the conditions that the temperature is 14-18 ℃, the relative air humidity is 80-90%, the carbon dioxide concentration is 1.5-2.0% and the illumination intensity is 200-;
and 4, harvesting: collecting until the growth of the daughter entities of the sulfur bacteria is stopped and the color is changed from light yellow to orange to obtain the fresh product of the sulfur bacteria.
Further, conditions for low temperature induction of primordia include, inter alia: the relative humidity of air is 75-85%, the concentration of carbon dioxide is 1.5-2.0%, the illumination intensity is 100-.
Compared with the prior art, the invention has the following beneficial effects:
1. the method for obtaining the sulphur vermilion strain comprises three key technical points: (1) the spawn running technology is innovative, oak sawdust and eucommia branches are used as main raw materials in an artificial culture medium, and the growth rate of hyphae is improved; (2) the formation of the primordium is stimulated by utilizing low-temperature induction, so that the primordium is under the growth advantage condition, and a foundation is provided for further developing a large number of buds and ensuring the yield of the bag cultivation; (3) the primordium is differentiated to obtain the sporocarp, the primordium can be smoothly transited to reproductive growth at the stage according to the fruiting conditions disclosed by the invention, the management of the differentiation period of buds is very critical, and the method is an important period for obtaining the high and stable yield of the sulfur red bacterium.
2. The method for obtaining the sulfur red bacterial strains adopts sulfur red bacterial stock to carry out spawn running and adopts liquid bacterial to prepare the stock, thereby greatly shortening the culture period and further improving the hypha activity.
3. The artificial cultivation method of the sulfur red mushroom is carried out based on the obtained sulfur red mushroom strain, and mainly comprises three key technical control points of spawn running, low-temperature induction primordium formation and fruiting. Oak sawdust and eucommia branches are used as main raw materials in an artificial cultivation medium, so that the cultivation period is greatly shortened, and the hypha activity is improved; the low-temperature induction is utilized to stimulate the formation of primordium, the problem of low yield of artificial cultivation of the sulfur red mushroom is successfully solved, and a foundation is laid for the industrial cultivation of the sulfur red mushroom.
Drawings
FIG. 1 shows artificially cultivated Thiram cinnabarinum produced by the method of the present invention.
Detailed Description
The invention will be further illustrated with reference to the following specific examples and the accompanying drawings.
The oak sawdust in the invention is taken from crushed oak branches, and is required to be free of impurities and mildew; the eucommia branch is an eucommia branch, and is required to be free of mildew; the bran in the invention is taken from shells of wheat, rice and the like after grain threshing; the humus soil is a layer of mixture consisting of rotten plant substances and various organic wastes, or a product formed by rotting and fermenting dead branches and residual leaves of trees in surface soil layers in forests for a long time. In particular, the different sources of oak sawdust, bran and humus according to the present invention do not affect the technical reproducibility of the present invention, and any formulation satisfying the above requirements may be used in the technical means of the present invention.
Meanwhile, when the sulfur cinnabar strain is prepared, the spawn running is performed by using an original strain, wherein the original strain refers to a strain for expanding the mother strain of the sulfur cinnabar strain to a culture material. The stock seed is prepared by the following steps of firstly, preparing an activated wild sulphur red mushroom strain, performing conventional operation on the activated wild sulphur red mushroom strain, then, preparing a liquid sulphur red mushroom strain by using the activated wild sulphur red mushroom strain, and finally obtaining the stock seed, wherein the method can be referred to as the following method:
step 1, collecting wild sulphur mushroom sporocarp: collecting wild sulfur red fungus in Zhengxin county of Ankang city in 2013 at 9 months, performing tissue separation by using a tissue separation method, and performing purification culture on a composite P DA culture medium, wherein the strain is numbered as sulfur red fungus LS-AK405, and the strain is preserved by edible fungus institute of agricultural science research institute of Ankang city.
Step 2, activation of sulfur red mushroom strains: preparing a solid culture medium by using a basic culture medium formula, and inoculating the preserved sulfur cinnabarinus strain to the solid culture medium for activated culture. The basic culture medium formula is as follows: preparing a basic culture medium: 200g of potato, 10g of glucose, 10.0g of sucrose, 2.0g of peptone, 5g of yeast extract, 1.0g of monopotassium phosphate and magnesium sulfate0.5g, vitamin B46mg, agar 12g, water 1000 mL.
Step 3, preparing liquid strains of the sulfur red fungus: inoculating the activated sulfur cinnabar strain on a liquid culture medium, and culturing for 7d in a constant-temperature oscillation incubator at the temperature of 18-20 ℃, the relative air humidity of 55-65% and the shaking table rotating speed of 150r/min to prepare a liquid strain. The liquid culture medium has the following formula: 15g of glucose, 15g of cane sugar, 5.0g of peptone, 2g of yeast extract, 0.5g of monopotassium phosphate, 1.0g of magnesium sulfate and vitamin B42mg, 1000mL of water.
Step 4, preparing raw strains of sulfur cinnabar: preparing original seeds by adopting a round-mouth glass bottle with the specification of 110mm multiplied by 60 mm. Inoculating the prepared liquid strain to a stock culture medium, placing at the temperature of 16-19 ℃ and the relative air humidity of 55% -65%, culturing in the dark until hyphae grow over a glass bottle. After the hyphae overgrow, the hyphae are treated by scattered light of 200-300lx for 15d, and when the color of the hyphae in the fungus bag is changed into orange red, the fungus bag is refrigerated for standby. According to the mass percentage, the stock culture medium formula is as follows: 50% of oak sawdust, 30% of eucommia branch, 5% of wheat, 10% of bran, 2% of sucrose, 0.8% of monopotassium phosphate, 0.6% of magnesium sulfate, 1% of gypsum and 0.6% of calcium phosphate.
The selection of the collection places of the wild sulfur red mushrooms is only exemplified locally, the technical reproducibility of the wild sulfur red mushrooms collected in other places is not affected by the difference of the collection places, and the wild sulfur red mushrooms collected in other places can be used in the technical scheme of the invention.
Meanwhile, the primordium of the invention refers to the primordium or embryonic period of the dauriculus cinnabarinus sporocarp, and is generally in a granular or needle shape, and the primordium can be further developed into buds or young mushrooms. The formation of primordia, marks the mycelium has gone from vegetative to reproductive growth stages. When the mass-propagated vegetative hyphae meet the physical conditions of proper light, temperature, humidity and the like and mechanical stimulation, and the biochemical change of a culture medium and the like, primordia are formed, and only the primordia under the growth advantage condition can develop into mature sporocarp.
In summary, the method for artificially cultivating the sulfur cinnabarinus provided by the invention is detailed as follows:
1. collecting wild sulfur red mushroom fruiting body
Collecting wild sulfur red fungus in Zhengshan county of Ankang city in 2013 for 9 months, performing tissue separation by using a tissue separation method, and performing purification culture on a composite PDA culture medium, wherein the strain is numbered as sulfur red fungus LS-AK405, and the strain is preserved by edible fungus institute of agricultural science research institute of Ankang city.
2. Sulfur red bacterium strain activation
Preparing a solid culture medium by using a basic culture medium formula, and inoculating the preserved sulfur cinnabarinus strain to the solid culture medium for activated culture. The basic culture medium formula is as follows: preparing a basic culture medium: 200g of potato, 10g of glucose, 10.0g of sucrose, 2.0g of peptone, 5g of yeast extract, 1.0g of monopotassium phosphate, 0.5g of magnesium sulfate and vitamin B46mg, agar 12g, water 1000 mL.
3. Preparation of liquid spawn of sulfur red
Inoculating the activated sulfur cinnabar strain on a liquid culture medium, and culturing for 7d in a constant-temperature oscillation incubator at the temperature of 18-20 ℃, the relative air humidity of 55-65% and the shaking table rotating speed of 150r/min to prepare a liquid strain. The liquid culture medium has the following formula: 15g of glucose, 15g of cane sugar, 5.0g of peptone, 2g of yeast extract, 0.5g of monopotassium phosphate, 1.0g of magnesium sulfate and vitamin B42mg, 1000mL of water.
4. Preparation of raw seed of sulfur red
Preparing original seeds by adopting a round-mouth glass bottle with the specification of 110mm multiplied by 60 mm. Inoculating the prepared liquid strain to a stock culture medium, placing at the temperature of 16-19 ℃ and the relative air humidity of 55% -65%, culturing in the dark until hyphae grow over a glass bottle. After the hyphae overgrow, the hyphae are treated by scattered light of 200-300lx for 15d, and when the color of the hyphae in the fungus bag is changed into orange red, the fungus bag is refrigerated for standby. The formula of the stock culture medium is as follows: 50% of oak sawdust, 30% of eucommia branches, 5% of wheat, 10% of bran, 2% of sucrose, 0.8% of monopotassium phosphate, 0.6% of magnesium sulfate, 1% of gypsum and 0.6% of calcium phosphate.
5. Preparation of culture Medium
The method adopts a polyethylene angular folded plastic bag with the specification of 14cm multiplied by 28cm multiplied by 0.006cm, and each bag is filled with about 0.4 kg. The formula is as follows: 48% of oak sawdust, 32% of eucommia branches, 15% of bran, 2% of sucrose, 1% of gypsum, 1% of humus soil and 1% of calcium carbonate. Bagging, sterilizing under high pressure, cooling and inoculating.
6. Inoculation of
Under the aseptic environment, the cotton-free cover is opened to perform inoculation at one end, and the inoculation amount of each bag is about 20 g.
7. Spawn running
Transferring the inoculated fungus bags into a fungus growing chamber, keeping the temperature at 15-18 ℃ in the fungus growing stage, the relative humidity of air at 55% -65%, the concentration of carbon dioxide at 0.8% -1.0%, and culturing in the dark. And (5) turning over the pile to check the germination and pollution conditions of the strains after the hypha takes about 2cm without moving the fungus bags before the hypha grows. Culturing for 25-30 days under normal condition, and allowing mycelia to grow over the material bag.
8. Low temperature induced primordial formation
Transferring the fungus bag full of mycelia to a fruiting room, and removing the non-cotton cover and the lantern ring at the upper end of the fungus bag. The formation of primordium is promoted by low-temperature stimulation, the temperature in the daytime is controlled to be 18-20 ℃ in the low-temperature induction stage, the temperature at night is controlled to be 8-12 ℃, the relative humidity of air is 75-85%, the concentration of carbon dioxide is kept to be 1.5-2.0%, the illumination intensity is 100-200lx, the primordium is cultured for about 15 days, and the primordium is formed around and on the fungus bags.
9. Fruiting management
After fungus buds appear at the bag mouth, the fruiting stage is started, the temperature is kept at 14-18 ℃ during the fruiting period, the air relative humidity is 80% -90%, the carbon dioxide concentration is kept at 1.5% -2.0%, and the illumination intensity is 200-.
10. Harvesting
The optimal period for harvesting is when the sporocarp basically stops growing and the color changes from light yellow to orange.
11. Strain preservation
The excellent strain is a key factor of artificial cultivation. The prior common inclined plane preservation method has the defects of short preservation time, easy degradation and the like. The culture medium adopted by the invention can keep the excellent genetic performance of the strain for a long time. The formula of the preservation medium is as follows: 25g of wheat, 20g of bran, 10g of glucose, 10g of yeast extract, 1.0g of monopotassium phosphate, 1.0g of magnesium sulfate and 1000mL of water.
Example 1:
the embodiment provides a cultivation method of a sulfur vermilion strain (the strain number is CGMCC No.18155, the strain is preserved in China general microbiological culture Collection center (CGMCC) in 26 months and 8 months in 2019, the CGMCC is short, and the address is No. 3 of No.1 Hospital No. 3 of Xilu Beijing Kogyo of Chaoyang district in Beijing), which comprises the following steps:
step 1, inoculating a sulfur cinnabar strain stock into an artificial culture medium for spawn running; the spawn running conditions comprise: culturing at 15-18 deg.C, air relative humidity of 55-65%, carbon dioxide concentration of 0.8-1.0% in dark until hypha grows in the bag; the preparation of the artificial cultivation medium comprises the following steps: placing the artificial culture medium in a polyethylene material bag for sterilization for later use, wherein the formula of the artificial culture medium comprises the following components in percentage by mass: 48% of oak sawdust, 32% of eucommia branches, 15% of bran, 2% of sucrose, 1% of gypsum, 1% of humus soil and 1% of calcium carbonate;
step 2, performing low-temperature induction on the material bag full of hyphae to generate primordium;
step 3, differentiating the primordium to form a sulfur bacterium sporophore under the conditions that the temperature is 14-18 ℃, the relative air humidity is 80-90%, the carbon dioxide concentration is 1.5-2.0% and the illumination intensity is 200-;
and 4, separating and preserving the collected sulfur cinnabarinus sporocarp on an inclined plane to obtain the sulfur cinnabarinus bacterial strain.
Wherein, the conditions for generating the primordium by low-temperature induction comprise: the relative humidity of air is 75-85%, the concentration of carbon dioxide is 1.5-2.0%, the illumination intensity is 100-.
The preparation method of the sulfur cinnabar strain stock comprises the following steps:
step 1, inoculating activated wild sulfur red bacterium strains on a sulfur red bacterium liquid culture medium for culture to obtain sulfur red bacterium liquid culture mediumSeed growing; the liquid culture medium formula comprises: 15g of glucose, 15g of cane sugar, 5.0g of peptone, 2g of yeast extract, 0.5g of monopotassium phosphate, 1.0g of magnesium sulfate and vitamin B42mg, 1000mL of water;
step 2, inoculating the liquid strains to a stock culture medium for dark culture, and then carrying out 200-300lx scattered light treatment to obtain a sulfur cinnabarinus stock; the stock culture medium comprises the following components in percentage by mass: 50% of oak sawdust, 30% of eucommia branches, 5% of wheat, 10% of bran, 2% of sucrose, 0.8% of monopotassium phosphate, 0.6% of magnesium sulfate, 1% of gypsum and 0.6% of calcium phosphate.
Wherein, the formula of the preservation medium for slant preservation comprises: 25g of wheat, 20g of bran and 80-150mL of nutrient solution, wherein the formula of the nutrient solution comprises the following components: 10g of glucose, 10g of yeast extract, 1.0g of monopotassium phosphate, 1.0g of magnesium sulfate and 1000mL of water.
Example 2:
the embodiment provides an artificial cultivation method of sulfur cinnabarina, which specifically comprises the following steps:
step 1, spawn running: inoculating Sulfur Laetiporus Sulphureus LS-AK405(CGMCC No.18155) into artificial culture medium for spawn running; the spawn running conditions comprise: culturing at 15-18 deg.C, air relative humidity of 55-65%, carbon dioxide concentration of 0.8-1.0% in dark until hypha grows in the bag; the preparation of the artificial cultivation medium comprises the following steps: placing the artificial culture medium in a polyethylene material bag for sterilization for later use, wherein the formula of the artificial culture medium comprises the following components in percentage by mass: 48% of oak sawdust, 32% of eucommia branches, 15% of bran, 2% of sucrose, 1% of gypsum, 1% of humus soil and 1% of calcium carbonate;
step 2, low-temperature induction of primordium formation: performing low-temperature induction on the material bag full of hypha to generate primordium;
step 3, fruiting: the primordium is differentiated to form a sulfur red mushroom fruiting body under the conditions that the temperature is 14-18 ℃, the relative air humidity is 80-90%, the carbon dioxide concentration is 1.5-2.0% and the illumination intensity is 200-;
and 4, harvesting: collecting until the growth of the daughter entities of the sulfur bacteria is stopped and the color is changed from light yellow to orange to obtain the fresh product of the sulfur bacteria.
Further, conditions for low temperature induction of primordia include, inter alia: the relative humidity of air is 75-85%, the concentration of carbon dioxide is 1.5-2.0%, the illumination intensity is 100-.
The effect proves that:
in order to determine the optimal proportion of the oak sawdust to the eucommia ulmoides branches, the influence of different substrates on the growth and yield of the mycelium of the sulfur red fungi is researched, and the experimental design is shown in table 1.
TABLE 1 design of different substrate formulas for Laetiporus cinnabarinus
The results of the effects of different substrate formulations on the growth of the sulfur red hyphae are shown in table 2.
As can be seen from Table 2, different substrate formulations have a greater effect on the growth of the mycelium of the Thiram cinnabarinum. When the formula 2 is used as a culture medium, the growth rate of the sulfur red hyphae is the fastest and is 1.216cm/d, the bag filling time is the shortest and is 25d, and at the moment, the hyphae are dense, grow well, and the infectious rate of mixed bacteria is the lowest; secondly, formulation 2, with a hyphal growth rate of 1.109cm/d, formulation 7 performed the worst.
TABLE 2 influence of different substrate formulations on the growth of the mycelium of Thiram cinnabarina
Remarking: the + and + hyphae grow strongly and densely, the + hyphae grow more densely, and the + hyphae grow sparsely; different upper case letters represent a 1% significance level and different lower case letters represent a 5% significance level; the specification of the fungus bag is as follows: 14cm 28cm 0.006 cm.
The results of the effect of different substrate formulations on the yield of Laetiporus cinnabarinus are shown in Table 3.
As can be seen from Table 3, different substrate formulations have a great influence on the differentiation rate and yield of the sulfur cinnabarinus primordium. When formula 3 was used as the medium (FIG. 1), the differentiation rate of the primordia was high, the yield was 142.15 g/bag, and the yield was increased by 33.69 g/bag compared to CK. Indicating that proper formulation is a key factor in improving yield.
Table 3. Effect of different substrate formulations on Thiobacillus cinnabarinus yield and biological efficiency
Remarking: different upper case letters represent a 1% significance level and different lower case letters represent a 5% significance level.
Claims (7)
1. A cultivation method of a sulphur vermilion strain is characterized by comprising the following steps:
step 1, inoculating a sulfur cinnabar strain stock into an artificial culture medium for spawn running; the spawn running conditions comprise: culturing at 15-18 deg.C, air relative humidity of 55-65%, carbon dioxide concentration of 0.8-1.0% in dark until hypha grows in the bag; the preparation of the artificial cultivation medium comprises the following steps: placing the artificial culture medium in a polyethylene material bag for sterilization for later use, wherein the formula of the artificial culture medium comprises the following components in percentage by mass: 45% of oak sawdust, 40% of eucommia branches, 10% of bran, 2% of sucrose, 1% of gypsum, 1% of humus soil and 1% of calcium carbonate;
step 2, performing low-temperature induction on the material bag full of hyphae to generate primordium;
step 3, differentiating the primordium to form a sulfur bacterium sporophore under the conditions that the temperature is 14-18 ℃, the relative air humidity is 80-90%, the carbon dioxide concentration is 1.5-2.0% and the illumination intensity is 200-;
and 4, separating and preserving the collected sulfur cinnabarinus sporocarp on an inclined plane to obtain the sulfur cinnabarinus bacterial strain.
2. The method for cultivating a sulfur cinnabar strain according to claim 1, wherein the conditions for inducing the production of primordia at low temperature comprise: the relative humidity of air is 75-85%, the concentration of carbon dioxide is 1.5-2.0%, the illumination intensity is 100-.
3. The method for culturing a Laetiporus cinnabarinus strain according to claim 1, wherein the method for preparing a stock species of Laetiporus cinnabarinus comprises:
step 1, inoculating activated wild sulfur red bacterium strains on a sulfur red bacterium liquid culture medium for culture to obtain sulfur red bacterium liquid strains; the liquid culture medium formula comprises: 15g of glucose, 15g of cane sugar, 5.0g of peptone, 2g of yeast extract, 0.5g of monopotassium phosphate, 1.0g of magnesium sulfate and vitamin B42mg, 1000mL of water;
step 2, inoculating the liquid strains to a stock culture medium for dark culture, and then carrying out 200-300lx scattered light treatment to obtain a sulfur cinnabarinus stock; the stock culture medium comprises the following components in percentage by mass: 50% of oak sawdust, 30% of eucommia branch, 5% of wheat, 10% of bran, 2% of sucrose, 0.8% of monopotassium phosphate, 0.6% of magnesium sulfate, 1% of gypsum and 0.6% of calcium phosphate.
4. The method for cultivating a strain of sulfur cinnabar as claimed in claim 1, wherein the formula of the preservation medium for slant preservation comprises: 25g of wheat, 20g of bran and 80-150mL of nutrient solution, wherein the formula of the nutrient solution comprises the following components: 10g of glucose, 10g of yeast extract, 1.0g of monopotassium phosphate, 1.0g of magnesium sulfate and 1000mL of water.
5. The sulfur cinnabarinus strain obtained in claim 1, which is designated as sulfur cinnabarinus (Laetiporus Sulphureus) LS-AK405 with a preservation number of CGMCC No. 18155.
6. An artificial cultivation method of sulfur cinnabarina is characterized by comprising the following steps:
step 1, spawn running: inoculating Sulfur Laetiporus Sulphureus LS-AK405(CGMCC No.18155) into artificial culture medium for spawn running; the spawn running conditions comprise: culturing at 15-18 deg.C, air relative humidity of 55-65%, carbon dioxide concentration of 0.8-1.0% in dark until hypha grows in the bag; the preparation of the artificial cultivation medium comprises the following steps: placing the artificial culture medium in a polyethylene material bag for sterilization for later use, wherein the formula of the artificial culture medium comprises the following components in percentage by mass: 48% of oak sawdust, 32% of eucommia branches, 15% of bran, 2% of sucrose, 1% of gypsum, 1% of humus soil and 1% of calcium carbonate;
step 2, low-temperature induction of primordium formation: performing low-temperature induction on the material bag full of hypha to generate primordium;
step 3, fruiting: the primordium is differentiated to form a sulfur red mushroom fruiting body under the conditions that the temperature is 14-18 ℃, the relative air humidity is 80-90%, the carbon dioxide concentration is 1.5-2.0% and the illumination intensity is 200-;
and 4, harvesting: collecting until the growth of the sporophores of the sulfur bacteria is stopped and the color is changed from light yellow to orange yellow to obtain the sulfur bacteria.
7. The artificial cultivation method of Laetiporus cinnabarinus as claimed in claim 6, wherein the conditions for inducing the production of primordium at low temperature include: the relative humidity of air is 75-85%, the concentration of carbon dioxide is 1.5-2.0%, the illumination intensity is 100-.
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