CN117187083B - Photosensitive hericium erinaceus strain suitable for industrialization and cultivation method thereof - Google Patents
Photosensitive hericium erinaceus strain suitable for industrialization and cultivation method thereof Download PDFInfo
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Abstract
The invention discloses a photosensitive Hericium coralloides strain suitable for industrialization and a cultivation method thereof, relates to the field of biotechnology, and aims to solve the problems that a photosensitive Hericium coralloides variety is lacking in the prior art, and the fruiting uniformity is low, the fruiting time is long and the like. The invention provides a photosensitive coralloid Hericium erinaceus strain which is preserved in China general microbiological culture Collection center (CGMCC) with a preservation number of 40392 in 2023 and 03 and 17 days. The invention also provides a factory cultivation method of the photosensitive coralloid Hericium erinaceus fruiting body, which comprises the following steps: inoculating Hericium coralloides into liquid culture medium, and culturing to obtain liquid strain; inoculating the liquid strain into a bacterial tray filled with a culture medium, performing different light treatments to promote bacteria, primordium formation and fruiting body development, and collecting to obtain the final product of coralloid Hericium erinaceus fruiting body.
Description
Technical Field
The invention relates to the technical field of biology, in particular to a photosensitive hericium erinaceus strain suitable for industrialization and a cultivation method thereof.
Background
Hericium coralloides [ Hericium coralloides (chop.) Pers. ], belonging to the phylum Basidiomycetes (Basidiomycetas), the class Agaricales (Agariomycetes), the order Russules (Russules), the family Hericiaceae (Hericiaceae), the genus Hericium (Hericium), also known as Tricholoma matsutake (Changbai mountain area), yu Tou, and Hericium erinaceum [ Herieium erinaeeus (Bun.: fr.) Pers. ] are the same genus group. Hericium erinaceus is a well-known edible and medicinal fungus resource in China, and mycelium polysaccharide has the effects of improving immunity, resisting coagulation, reducing blood fat and the like. Coralloid Hericium erinaceus also has effects of benefiting five viscera, nourishing, and promoting digestion, and can be used for treating neurasthenia and gastric ulcer.
As the coralloid Hericium erinaceus are distributed sparsely, reports on the industrial cultivation of the coralloid Hericium erinaceus are less, most of the strains used in the production at present have the phenomena of low fruiting uniformity, poor photosensitivity and the like, and most of the strains in the production are cultivated in traditional facilities by using solid strains, so that the cultivation time is longer and the standardization degree is lower.
The existing coralloid Hericium erinaceus variety lacks photosensitive coralloid Hericium erinaceus, most of the coralloid Hericium erinaceus variety is obtained by wild domestication, and the problems of poor photosensitivity, low fruiting uniformity, long fruiting time, difficult harvesting, low standardization degree and the like exist. The existing coralloid Hericium erinaceus facility cultivation method lacks mature coralloid Hericium erinaceus industrial cultivation technology, and is difficult to form an industrial cultivation mode.
Disclosure of Invention
The invention aims to solve the technical problems that:
the prior art lacks photosensitive coralloid Hericium erinaceus variety, and has the problems of poor photosensitivity, low fruiting uniformity, long fruiting time, difficult harvesting, low standardization degree and the like.
The invention adopts the technical scheme for solving the technical problems:
the invention provides a photosensitive Hericium coralloides strain, which is Hericium coralloides ZSH-2 with a preservation number of CGMCC No.40392.
A fruiting body of Hericium coralloides is obtained by cultivating Hericium coralloides Hericium coralloidesZLs h-2 according to the above technical scheme.
An industrial cultivation method of coralloid Hericium erinaceus fruiting body comprises the following steps:
(1) Inoculating the coralloid Hericium erinaceus in the technical scheme into a liquid culture medium, and culturing to obtain liquid strains;
(2) Inoculating the liquid strain into a bacterial tray filled with a culture medium, performing different light treatments to promote bacteria, primordium formation and fruiting body development, and finally harvesting to obtain the coralloid Hericium erinaceus fruiting body product.
Further, the liquid medium in step (1) includes: 100g/L of potato, 50g/L of cotton seed hull, 15g/L of glucose, 20g/L of bovine bone animal protein powder, 2g/L of monopotassium phosphate, 0.5g/L of magnesium sulfate and 0.3ml/L of glycerin.
Further, in the step (2), the bacterial tray is square in shape and comprises a tray body and a tray cover, wherein a fruiting hole is formed in the tray cover, and the tray cover is a transparent tray cover.
Further, the size of the fungus disk is 60x60x10cm, the number of fruiting holes is 4, 4 fruiting holes are uniformly distributed on the disk cover, the distance between the fruiting holes is 15cm, and the diameter of each fruiting hole is 3-4 cm.
Further, the light transmittance of the disc cover is not less than 95%.
Further, the conditions of the trigger-promoting bacteria include: the illumination stimulation, the light quality adopts red light, the illumination intensity is 100Lux, the illumination time is 2 h/day, and the rest time is dark treatment; conditions for promoting bacteria also include: the temperature is 24-26 ℃, the relative humidity of air is 63-66%, and ventilation is kept good.
Further, the conditions for the formation of primordia include: light stimulation, wherein white light is adopted as light quality, the illumination intensity is 300Lux, the illumination time is 12 h/day, and the rest time is dark treatment; the conditions for pro-group formation also include: the temperature is controlled at 18-24 ℃, the humidity is controlled at 90-95%, and ventilation is kept good.
Further, the conditions for promoting the development of the fruiting body include: light stimulation, blue light is adopted as light, the illumination intensity is 300Lux, the illumination time is 3 h/day, white light is adopted to complement and irradiate to 12 h/day, and the rest time is dark treatment; conditions that promote the development of the fruiting body also include: the temperature is controlled at 20-25 ℃, the humidity is controlled at 85-90%, and ventilation is kept good.
Compared with the prior art, the invention has the beneficial effects that:
1. the coralloid hericium erinaceus strain ZSH-2 provided by the invention has the sensitivity characteristic of illumination with different colors in different growth periods, and according to the characteristic, different illumination stimulus is given to different periods in the factory cultivation process of the strain, so that the hypha growth speed can be effectively improved, the primordial differentiation time can be shortened, the fruiting body yield can be improved, and the accumulation of fruiting body nutrient substances can be promoted; the fruiting time of each tide of mushrooms is basically consistent, the form is stable, the mushrooms are fragrant, the color is white, the quality is higher, the fruiting rate is more than 95%, and the method is very suitable for industrial cultivation production.
2. The culture medium of the liquid strain of the coralloid Hericium erinaceus is optimized, so that the growth of the ZSH-2 hyphae of the coralloid Hericium erinaceus is further promoted, and the hyphae are thicker and stronger.
3. The industrial cultivation method also adopts the square fungus tray with the fruiting holes and the transparent fungus tray cover, so that the fruiting is more orderly, the harvesting is convenient, the pollution is reduced, the transparent fungus tray cover can enable the strains to absorb illumination stimulus more effectively, the optimal cultivation effect is achieved, the production efficiency is improved, and the production cost is saved.
Drawings
FIG. 1 is a diagram of a fruiting body of Hericium coralloides ZSH-2 in an embodiment of the invention;
FIG. 2 is a diagram of wild Hericium coralloides SHS-1 fruiting body in an embodiment of the invention;
FIG. 3 is a diagram showing antagonism of Hericium coralloides H002 and wild Hericium coralloides H001 in the example of the present invention, wherein the wild Hericium coralloides H001 is on the left side and the Hericium coralloides H002 is on the right side;
FIG. 4 is a diagram of a first level liquid spawn in an embodiment of the present invention;
FIG. 5 is a diagram showing fruiting bodies of Hericium coralloides ZSH-2 cultivated in the example of the invention.
Detailed Description
In the description of the present invention, it should be noted that the terms "first," "second," and "third" mentioned in the embodiments of the present invention are used for descriptive purposes only and are not to be construed as indicating or implying relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defining "a first", "a second", or a third "may explicitly or implicitly include one or more such feature.
In order that the above objects, features and advantages of the invention will be readily understood, a more particular description of the invention will be rendered by reference to specific embodiments thereof which are illustrated in the appended drawings.
Example 1
1. Strain isolation and mutagenesis purification
The strain is separated from a natural protection area of cold water in the Ischun city of Heilongjiang province, a wild strain is collected, the wild coralloid Hericium erinaceus SHS-1 is obtained by separation, and the photosensitive coralloid Hericium erinaceus strain ZSH-2 is obtained by further carrying out ultraviolet mutagenesis on the SHS-1, and the specific method is as follows:
1.1. preparation of culture medium
The formula of the culture medium comprises: 200g of potato, 20g of glucose, 5g of peptone, 3g of monopotassium phosphate, 1.5g of magnesium sulfate, 20g of agar and 1000ml of water; the pH value of the culture medium is 5-6.
Weighing 200g of peeled potatoes, cutting into slices, cleaning, adding 1000ml of water, boiling for 20min, filtering with gauze, taking filtrate, adding 1000ml of agar to make up for less than 1000ml of filtrate, adding 20g of agar, boiling to melt, adding 20g of glucose, 5g of peptone, 3g of potassium dihydrogen phosphate and 1.5g of magnesium sulfate, melting, packaging, sterilizing test tubes while the test tubes are hot, sealing the test tubes by using a cotton plug, sterilizing at 121 ℃ for 20min, and standing for inclined plane for standby after sterilizing.
1.2. Tissue separation
Cutting off the base parts of the collected wild coralloid Hericium erinaceus fruiting bodies, washing with sterile water in a sterile room for several times, absorbing surface water with sterile paper, and soaking in 0.1% mercuric chloride for 5min. After taking out, washing with sterile water for several times (or directly coating the surface of fruiting body with 75% alcohol), and sterilizing the surface. Sterilizing, cutting to remove fungus thorn above flame of alcohol lamp with clean knife, breaking off with hands, and rapidly picking about 0.5cm of middle part of fruiting body of wild Hericium coralloides with inoculating needle 2 Inoculating the fruiting body tissue blocks on a test tube slant culture medium, and separating 4 test tubes from each fruiting body.
1.3. Cultivation and mutagenesis
Culturing the inoculated test tube in an incubator at 25deg.C, growing white mycelia around the tissue block after 2-3d, and growing slant with mycelia after 7-8d to obtain strain H001.
Washing activated tender mycelium fragments with physiological saline, filtering with absorbent cotton, shaking for 30min, and placing mycelium suspension under 15W ultraviolet lamp at 30cm position for 25s. The bacterial liquid after mutagenesis is diluted in gradient and then coated on a PDA plate, and the bacterial liquid is cultivated for 4d in a dark place.
Selecting a colony with large single colony diameter and luxuriant hypha growth after ultraviolet mutagenesis, transferring the colony into a PDA culture medium, culturing for 7d in a dark place, and continuously carrying out passage for 3 times to obtain a strain with more stable heredity; finally, culturing the strain in an incubator at 25 ℃, and obtaining the strain H002 after mycelium grows over the inclined plane.
2. Antagonism test
The strain H001 and the strain H002 are used as test materials, and the test method refers to the NY/T1845 edible fungus strain distinguishing identification antagonistic reaction.
The antagonistic test flat-plate culture medium comprises 200g of potato, 20g of glucose, 20g of agar, 5g of peptone, 3g of monopotassium phosphate, 1.5g of magnesium sulfate and 1000ml of water.
Inoculating Hericium coralloides H002 and Hericium coralloides H001 on antagonistic test plate medium; culture conditions: the cells were incubated at 25℃for 15 days, and the test results are shown in Table 1 and FIG. 3.
TABLE 1
Note that: "+" indicates antagonism.
Antagonism experiments show that: the strain H001 and the strain H002 are different strains, and the effectiveness of ultraviolet mutagenesis of the invention is further proved.
3. Authentication
3.1. Morphological identification
The mycelium of the H002 strain of the invention is white and dense and snowflake-shaped. The branches of the fruiting body are gathered together, short and compact spines are arranged at the periphery of the fruiting body, the spines are soft, the whole fruiting body is fluff-shaped, and the length of the fruiting body is 0.1-0.5cm. The base of the main branch is healed into blocks, and the width of a single fruiting body is 12-15cm and the height is 15cm.
The fruiting body has short and small branches, has faint scent, stable morphological characteristics, white color when fresh, and light yellow brown color after drying, and white fungus flesh (see figure 1). The classification characteristics of coral Hericium erinaceus in China (Dai Yucheng, et al, science Press, 2007) and the pictures of coral Hericium erinaceus in China (Mao Zhao Xiao lan, henan science and technology Press, 2000) are consistent.
3.2. Molecular biological identification
ITS sequencing is carried out on the H002 strain, and an ITS sequence is obtained through amplification and DNA sequencing:
the molecular biology identification of H002 strain belongs to basidiomycota, agaricales, rhodomycota, hericiaceae, hericium. Comprehensive morphological identification, the H002 strain belongs to Hericium coralloides Hericium coralloides.
4. Preserving
The wild domesticated Hericium coralloides strain is preserved in China general microbiological culture Collection center (China Committee for culture Collection) at a preservation address of China academy of sciences of China, including North Chen Silu No. 1, no. 3, of the area of Korean, beijing, named Hericium coralloides, strain named SHS-1, latin named Hericium coralloides, and preservation number of CGMCC No.40352 (fruiting body shown in FIG. 2), which is remarkably different from the fruiting body of ZLsh-2 obtained by separation mutagenesis of the present invention.
The strain obtained by separating and mutagenizing the invention is preserved in China general microbiological culture Collection center (China Committee for culture Collection) at the collection address of China academy of sciences of China, including national academy of sciences of China, no. 3, north Chen, west Lu, 1, beijing, and the region of the city, and the strain is named as Hericium coralloides, ZLsh-2, hericium coralloides, and CGMCC No.40392.
Example 2
1. Preparation of first-order liquid strain
1.1. Preparation of culture Medium
The formula of the culture medium comprises: 100g of potato, 50g of cotton seed hulls, 15g of glucose, 20g of bovine bone animal protein powder, 2g of monopotassium phosphate, 0.5g of magnesium sulfate, 0.3ml of glycerol and 1000ml of water; the pH of the culture medium is 5-6.
Weighing 100g of peeled potatoes, cutting into slices, cleaning, adding 1000ml of water, boiling for 5min, adding 50g of cotton seed hulls, boiling for 15min, filtering with gauze, taking filtrate, adding 1000ml of the filtrate, adding 15g of glucose, 20g of bovine bone animal protein powder, 2g of potassium dihydrogen phosphate, 0.5g of magnesium sulfate and 0.3ml of glycerin, melting, subpackaging in triangular flasks when the mixture is hot, sterilizing at 121 ℃ under high pressure for 20min, cooling and standing for later use.
1.2. Inoculation and culture
Taking 5-8 blocks of 0.5cm with a puncher 2 Inoculating the mother strain into primary liquid strain culture medium, shake culturing at 20-25deg.C and 140-160r/min, as shown in figure 4, for 5-7d to obtainThe first-stage liquid strain can be seen to have higher density of liquid strain balls, white fungus balls and higher quality.
2. Preparation of secondary liquid strain
2.1. Preparation of culture Medium
The secondary liquid strain culture medium comprises: 100g/L of potato, 50g/L of cotton seed hull, 15g/L of glucose, 20g/L of bovine bone animal protein powder, 2g/L of monopotassium phosphate, 0.5g/L of magnesium sulfate and 0.3ml/L of glycerol; the pH of the culture medium is 5-6.
Decocting potato and cotton seed hull, filtering to obtain juice, adding filtered water according to culture medium formula, loading into fermentation tank, heating to 120deg.C, maintaining the pressure of fermentation tank at 0.1Mpa (gauge pressure), maintaining for 30min, and cooling.
2.2. Inoculation and culture
Inoculating the first-stage liquid strain into a fermentation tank, and culturing under stirring at 20-25deg.C and 140-160r/min for 5 days to obtain the second-stage liquid strain. If the mother strain is directly adopted for the culture period of the fermentation tank culture, the primary liquid strain is adopted for the fermentation tank fermentation culture to obtain the secondary liquid strain, so that the culture efficiency is improved.
3. Preparation of cultivars
3.1. Preparation of culture Medium
The culture medium of the cultivar medium comprises: 76% of tussah wood dust, 20% of wheat bran, 1% of bean powder, 1% of sucrose, 1% of gypsum powder and 1% of lime powder.
Fresh and mildew-free culture materials are selected, wheat bran, gypsum powder and bean powder are put together according to a proportion and are evenly dry-mixed, then tussah sawdust is added for dry-mixing, sucrose is put into water for melting, the mixed dry materials and sucrose water are evenly mixed, the prepared culture material (the water content is 60 wt%) is filled into square fungus trays with the size of 60x60x10cm, and a highly transparent fungus tray cover is covered for sterilization. 5kg of culture medium of cultivar is filled into each tray, sterilized for 12 hours under normal pressure, and stood until the temperature is reduced to 25 ℃. Wherein, be equipped with 4 fruiting holes on every transparent fungus dish lid, every fruiting hole is the circular hole that the diameter is 3cm, and fruiting hole interval is 15cm, and the luminousness of this fungus dish lid is not less than 95%. By adopting the fungus disk for cultivation, fruiting is tidier, harvesting is convenient, pollution is reduced, the fruiting hole spacing is reasonable in design, fruiting body growth is facilitated, and meanwhile, the design of the fungus disk cover which is highly transparent is convenient for the photosensitive hericium erinaceus to receive light stimulation in the cultivation process.
3.2. Inoculation and culture
Promoting bacteria: under aseptic condition, the fungus disk is moved to the fungus grafting equipment, the second-level liquid strain is inoculated through the fungus growing hole of the fungus disk cover by the fungus grafting gun, after inoculation, the fungus growing hole is sealed by using a cotton plug, and the fungus growing hole is moved to an aseptic culture room for unified culture. The bacteria are cultivated for 2 hours by irradiation of red light for each day, the illumination intensity is 100Lux, the rest time is dark, the temperature is kept at 24-26 ℃, the relative humidity of air is 63-66%, and ventilation is good.
4. Fruiting period management
Primordium formation: and 3.2, after the strain culture is completed, transferring the strain trays to a factory fruiting frame for fruiting management. Firstly, placing a fungus disk on a fruiting frame, and taking down a cotton plug in a fruiting hole to fruiting. And (3) in the period from fruiting to primordial differentiation in the fruiting chamber, white light irradiation is given for 12 hours every day, the illumination intensity is 300Lux, and the rest 12 hours are dark treatments. The temperature between fruiting is controlled at 18-24 deg.C, the humidity is controlled at 90-95%, and ventilation is ensured.
Promoting the development of fruiting bodies: during the period from the completion of primordial differentiation to the harvesting of fruiting bodies, blue light irradiation is carried out for 3 hours every day, the illumination intensity is 300Lux, white light irradiation is carried out for 9 hours, the illumination intensity is 300Lux, and the rest time is dark treatment; the temperature is controlled at 20-25 deg.C, the humidity is controlled at 85-90%, and ventilation is ensured.
5. Harvesting
When the coralloid Hericium erinaceus thorns are fully stretched and start sagging, fruiting bodies (first tide mushrooms) are harvested for the first time; after the first tide mushroom is picked, the culture is carried out in a dark place, after 10d, mycelium grows up to the culture material, the culture is carried out again according to the conditions, the fruiting body (second tide mushroom) is picked up for the second time, and the coralloid Hericium erinaceus fruiting body is picked up continuously. As shown in FIG. 5, the fruiting body obtained is white in color, regular in fruiting and good in growth vigor.
Example 3
The coralloid Hericium erinaceus ZLsh-2, wild coralloid Hericium erinaceus SHS-1 and coralloid Hericium erinaceus ZLsh-1 (classified as Hericium coralloides ZLsh-1, strain name ZLsh-1, latin name Hericium coralloides, preservation number of CGMCC No.40391, and the collection of the same at the China general microbiological culture Collection of the China Committee at the month 17 of 2023, the collection address of the same being the national academy of sciences of China, including Beijing, kogyo, no. 1, no. 3 of the same), xueyu pine mushroom (Li Jie, peng Yanfang, yu Wenqing. Xueyu pine mushroom dry product nutrition and flavor component analysis and evaluation [ J ]. Food research and development, 2014, 35 (20): 8-12.) Hericium coralloides RT24 (Zhang Peng, wang Yanfeng, shi Lei, etc. straw sensitivity test materials were developed by using China coral fungus [ J ]. Edible fungus report, 2021,28 (02): 55-59.DOI: 10.88/j.cnki. 9873.2021.02.008).
The specific test method is as follows:
1. inoculating test tube stock of Hericium coralloides ZLsh-2, wild Hericium coralloides SHS-1, hericium coralloides ZLsh-1, hericium erinaceus Tricholoma matsutake, and Hericium coralloides RT24 into liquid strain culture medium containing potato 100g, cotton seed hull 50g, glucose 15g, bovine bone animal protein powder 20g, potassium dihydrogen phosphate 2g, magnesium sulfate 0.5g, glycerol 0.3ml and water 1000ml, and shake culturing at 20-25deg.C and 140-160r/min for 5-7d.
The method for respectively inoculating five coralloid Hericium erinaceus liquid strains into the culture materials comprises the following steps of: 76% of tussah wood chips, 20% of wheat bran, 1% of bean flour, 1% of sucrose, 1% of gypsum powder and 1% of lime powder (the water content is 60 wt%) and placing the inoculated five cultivars in the same culture environment for culture, wherein each cultivar is provided with 50 discs.
As the red light plays a role in promoting the growth of hyphae, the embodiment gives red light irradiation for 2 hours every day in the growth period of hyphae, the temperature is controlled at 24-26 ℃, the relative humidity of air is about 65%, and good ventilation is ensured. The sensitivity of the growth period of the mycelia of different Hericium coralloides to red light was analyzed by measuring the growth rate, concentration and color of the mycelia of different strains after red light treatment, and the results are shown in Table 2.
TABLE 2
| Hypha growth rate | Concentration of hypha | Hypha color | |
| Coralloides Hericium erinaceus ZSH-2 | 0.62cm/d | ++++ | Whitening device |
| Wild coralloid Hericium erinaceus SHS-1 | 0.46cm/d | ++ | White color |
| Hericium coralloides ZSH-1 | 0.49cm/d | ++++ | Whitening device |
| Radix Et rhizoma Nardostachyos | 0.47cm/d | +++ | White color |
| Hericium coralloides RT24 | 0.48cm/d | ++ | White color |
Wherein "+" indicates sparse and weak hyphae, "++" indicates sparse hyphae and good growth vigor, "+++" indicates dense hyphae the growth vigor is strong, growth vigor of is strong in the strength and the strength.
2. The white light irradiation is helpful for shortening primordial differentiation time of fruiting bodies and promoting fruiting uniformity; therefore, in the embodiment, the coralloid Hericium erinaceus after spawning are placed in the same culture environment, white light irradiation treatment is carried out for 12 hours every day from the beginning of fruiting to the primordial differentiation, the temperature is controlled at 18-24 ℃, the humidity is controlled at 90-95%, and good ventilation is ensured. The differentiation time and fruiting uniformity of the different coralloid Hericium erinaceus primordia were observed and counted, and the results are shown in Table 3.
TABLE 3 Table 3
Wherein "+" indicates irregular fruiting, "++" indicates that the fruiting is relatively clean, "+++" means is very neat
3. Because blue light treatment can remarkably improve the yield of fruiting bodies of Hericium coralloides, in the embodiment, blue light is applied to each Hericium coralloides every day for 3 hours in the period from the completion of primordium differentiation to fruiting body harvesting, the temperature is controlled at 20-25 ℃, the humidity is controlled at 85-90%, ventilation is ensured to be good, and fruiting bodies are harvested when the thorns of Hericium coralloides are fully stretched and start to droop. Observing and counting fruiting body yield, fruiting rate and crude protein content of different coralloid Hericium erinaceus; wherein, the yield is the total fresh weight of the multi-tide Hericium erinaceus fruiting body produced by each fruiting hole, the crude protein content is tested according to national standard test, and the crude protein GB 5009.5-2016, and the specific results are shown in Table 4.
TABLE 4 Table 4
| Yield (g) | Fruiting rate | Crude protein content | |
| Coralloides Hericium erinaceus ZSH-2 | 1275.4 | 94％ | 36.1％ |
| Wild coralloid Hericium erinaceus SHS-1 | 798.3 | 83％ | 18.3％ |
| Hericium coralloides ZSH-1 | 1117.2 | 92％ | 32.8％ |
| Radix Et rhizoma Nardostachyos | 499.8 | 86％ | 21.4％ |
| Hericium coralloides RT24 | 464.3 | 87％ | 17.5％ |
The results of the various comparative tests are combined to see: compared with other coralloid Hericium erinaceus in the prior art, the coralloid Hericium erinaceus ZSH-2 provided by the invention has higher sensitivity to red light in the mycelium growth stage, and has the advantages of high mycelium growth speed, thick mycelium and white color; in the stage from fruiting to primordial differentiation, the white light is more sensitive, the primordial differentiation time is shorter, and the fruiting uniformity is higher; the method has the advantages that the sensitivity to blue light is higher in the stage from the completion of primordium differentiation to fruiting body harvesting, the fruiting body yield and fruiting rate are higher, the crude protein content is obviously improved, and the nutritional value of the fruiting body is improved.
Example 4
In order to explore the most suitable growth illumination condition of the coralloid Hericium erinaceus ZSH-2, an illumination test is carried out, and the specific test method comprises the following steps:
1. red light treatment is carried out on the coralloid Hericium erinaceus ZSH-2 in the mycelium growth period, 7 groups of different illumination intensities and times are set, the rest time is dark treatment except red light treatment every day in each group of experiments, the temperature is controlled at 24-26 ℃, the relative air humidity is about 63-66%, and good ventilation is ensured. The specific illumination parameters for each set of experiments are shown in table 5.
TABLE 5
The growth rate, concentration and color of the coralloid Hericium erinaceus ZSH-2 mycelium treated with red light at different illumination intensities and different illumination times were measured, and specific test results are shown in Table 6.
TABLE 6
| Treatment group | Hypha growth rate | Concentration of hypha | Hypha color |
| A | 0.51cm/d | ++++ | Whitening device |
| B | 0.47cm/d | ++ | White color |
| C | 0.55cm/d | ++++ | Whitening device |
| D | 0.45cm/d | ++ | White color |
| E | 0.50cm/d | +++ | Whitening device |
| F | 0.47cm/d | ++ | White color |
| G | 0.49cm/d | +++ | White color |
Wherein "+" indicates sparse and weak hyphae, "++" indicates sparse hyphae and good growth vigor, "+++" indicates dense hyphae the growth vigor is strong, growth vigor of is strong in the strength and the strength.
2. Placing the coralloid Hericium erinaceus ZSH-2 after spawning in the same culture environment, and performing white light irradiation treatment every day during primordium differentiation period, wherein the rest time is dark treatment, the temperature between fruiting is controlled at 18-24deg.C, and the humidity is controlled at 90-95%, and ventilation is ensured to be good. The tests were divided into 5 groups, and specific illumination parameters of each group are shown in table 7.
TABLE 7
The differentiation time and fruiting uniformity of the coralloid Hericium erinaceus ZLsh-2 primordium subjected to white light treatment with different illumination intensities and different illumination times are observed and counted, and specific results are shown in Table 8.
TABLE 8
| Treatment group | Primordial differentiation time | Uniformity of fruiting |
| A | 120h | + |
| B | 84h | ++ |
| C | 72h | +++ |
| D | 96h | ++ |
| E | 108h | + |
Wherein "+" indicates irregular fruiting, "++" indicates that the fruiting is relatively clean, "+++" means is very neat
3. And (3) in the period from the completion of primordium differentiation to fruiting body harvesting, blue light irradiation treatment is carried out every day, white light is used for complement irradiation for 12 hours, the rest 12 hours are dark treatment, the temperature is controlled at 20-25 ℃, the humidity is controlled at 85-90%, and good ventilation is ensured. The tests were divided into 7 groups, and specific illumination parameters of each group are shown in table 9.
TABLE 9
| Treatment group | Blue light illumination intensity | Blue light illumination time | White light illumination time | Dark treatment time |
| A | 100Lux | 3h | 9h | 12h |
| B | 100Lux | 9h | 3h | 12h |
| C | 300Lux | 3h | 9h | 12h |
| D | 300Lux | 9h | 3h | 12h |
| E | 500Lux | 3h | 9h | 12h |
| F | 500Lux | 9h | 3h | 12h |
| G | 0Lux | 0h | 12h | 12h |
When the coralloid Hericium erinaceus thorns fully stretch and start sagging, fruit bodies are collected, and the output, fruiting rate and crude protein content of the coralloid Hericium erinaceus ZLsh-2 fruit bodies subjected to blue light treatment with different light intensities and different light times are observed and counted. Wherein, the yield is the total fresh weight of the multi-tide Hericium erinaceus fruiting body produced by each fruiting hole; the crude protein content was measured according to national standards, and the results are shown in Table 10 for crude proteins GB 5009.5-2016.
Table 10
According to the invention, as can be seen from the comprehensive illumination test results, when the coralloid Hericium erinaceus ZSH-2 is given to 100Lux red light irradiation for 2h/d in the mycelium growth period, the mycelium growth can be effectively promoted, the mycelium density can be increased, and the white color of the mycelium can be promoted; in the primordium differentiation period, the irradiation of 300Lux white light is given for 12 hours/d, so that primordium differentiation can be accelerated, and fruiting can be promoted more orderly; during the period from primordium differentiation to fruiting body harvesting, blue light irradiation of 300Lux is given for 3h/d, so that the increase of crude protein content of fruiting bodies can be promoted, the fruiting rate can be improved, and the yield can be increased.
Example 5
The formula of the culture medium for the coralloid Hericium erinaceus ZSH-2 liquid strain is improved, so that the growth speed of the coralloid Hericium erinaceus ZSH-2 hypha is faster, and the hypha is thicker and stronger; the culture effect of the strain medium of the present invention is compared with that of the conventional liquid medium.
Liquid strain medium a: 100g/L of potato, 50g/L of wood dust, 45g/L of wheat bran, 20g/L of corn meal, 3g/L of yeast extract powder, 1g/L of monopotassium phosphate, 0.5g/L of magnesium sulfate, 0.1g/L of vitamin B and 0.3ml/L of glycerin; the pH of the culture medium is 5-6.
Boiling filtrate of potato, wheat bran and wood chip (boiling potato for 20min, boiling wheat bran and wood chip for 15 min), adding other ingredients, stirring, packaging into triangular flask, sealing, sterilizing at 121deg.C for 20min, cooling to 25deg.C, inoculating Hericium coralloides ZSH-2 mother strain under aseptic condition, and shake culturing at 20-25deg.C and 140-160 r/min.
Liquid strain medium B: 100g/L of potato, 50g/L of wheat bran, 50g/L of brown sugar, 10g/L of glucose, 2.5g/L of peptone, 2g/L of monopotassium phosphate, 1g/L of magnesium sulfate, 12 tablets of vitamin B and 0.3ml/L of glycerin; the pH of the culture medium is 5-6.
Boiling filtrate of potato and wheat bran (boiling potato for 20min, boiling wheat bran for 15 min), adding other ingredients, stirring, packaging into triangular flask with liquid content not exceeding two fifths of total volume of triangular flask, sterilizing after sealing, sterilizing at 121deg.C under high pressure for 20min, cooling to 25deg.C, inoculating Hericium coralloides ZLsh-2 mother strain under aseptic condition, and shake culturing at 20-25deg.C and 140-160 r/min.
The liquid strain culture medium C comprises 100g/L of potato, 50g/L of cotton seed hull, 15g/L of glucose, 20g/L of bovine bone animal protein powder, 2g/L of monopotassium phosphate, 0.5g/L of magnesium sulfate and 0.3ml/L of glycerin; the pH of the culture medium is 5-6.
Boiling filtrate of potato and cotton seed hull (after boiling potato for 20min, boiling cotton seed hull for 15 min), adding other ingredients, stirring, packaging into triangular flask, packaging to obtain liquid strain culture medium C with liquid content not exceeding two fifths of total volume of triangular flask, sealing, sterilizing, autoclaving at 121deg.C for 20min, cooling to 25deg.C, inoculating Hericium coralloides ZLsh-2 mother strain under aseptic condition, and shake culturing at 20-25deg.C and 140-160 r/min.
Observing the growth condition of coralloid Hericium erinaceus ZSH-2 mycelia of three different liquid strain culture mediums, the mycelia using the liquid strain culture medium C are whiter than the mycelia of other two liquid strain culture mediums, the growth speed is high, and the density of mycelia is improved by approximately 7% compared with that of the liquid strain culture medium A and is improved by approximately 5% compared with that of the liquid strain culture medium B.
The invention adopts bovine bone animal protein powder with higher protein content and cotton seed hulls for promoting the health of hyphae to prepare the coral Hericium erinaceus liquid strain culture medium, which is beneficial to the production and cultivation of the coral Hericium erinaceus ZSH-2.
Although the present disclosure is disclosed above, the scope of the present disclosure is not limited thereto. Various changes and modifications may be made by one skilled in the art without departing from the spirit and scope of the disclosure, and such changes and modifications would be within the scope of the disclosure.
Claims (10)
1. A photosensitive Hericium coralloides strain is characterized in that the strain is Hericium coralloides strainHericiumcoralloides) ZSH-2 with a preservation number of CGMCC No.40392.
2. A fruit body of Hericium coralloides, characterized in that it is prepared by cultivating Hericium coralloides according to claim 1Hericium coralloides) Fruiting body of ZSH-2.
3. A method of industrially cultivating the fruit body of a coral-shaped Hericium erinaceus as defined in claim 2, comprising the steps of:
(1) Inoculating the coralloid Hericium erinaceus of claim 1 into a liquid culture medium, and culturing to obtain liquid strain;
(2) Inoculating the liquid strain into a bacterial tray filled with a culture medium, performing different light treatments to promote bacteria, primordium formation and fruiting body development, and finally harvesting to obtain the coralloid Hericium erinaceus fruiting body product.
4. A method of industrially culturing a fruit body of a coral-shaped Hericium erinaceus according to claim 3, wherein the liquid medium in step (1) comprises: 100g/L of potato, 50g/L of cotton seed hull, 15g/L of glucose, 20g/L of bovine bone animal protein powder, 2g/L of monopotassium phosphate, 0.5g/L of magnesium sulfate and 0.3ml/L of glycerin.
5. The method for industrially culturing the fruit bodies of coralloid Hericium erinaceus according to claim 3, wherein the fungus tray in the step (2) has a square shape and comprises a tray body and a tray cover, wherein the tray cover is provided with fruiting holes, and the tray cover is a transparent tray cover.
6. The method for industrially culturing the fruit bodies of Hericium coralloides according to claim 5, wherein the tray has a size of 60x60x10cm, the number of fruiting holes is 4, 4 fruiting holes are uniformly distributed on the tray cover, the interval between fruiting holes is 15cm, and the diameter of each fruiting hole is 3-4 cm.
7. An industrial cultivation method of coral Hericium erinaceus fruiting body as claimed in claim 6, wherein the light transmittance of the tray cover is not less than 95%.
8. A method of industrially culturing a fruit body of a coral-shaped Hericium erinaceus as defined in claim 3, wherein said conditions for promoting bacteria include: the illumination stimulation, the light quality adopts red light, the illumination intensity is 100Lux, the illumination time is 2 h/day, and the rest time is dark treatment; conditions for promoting bacteria also include: the temperature is 24-26 ℃, the relative humidity of air is 63-66%, and ventilation is kept good.
9. A method for the industrial cultivation of a fruit body of a coral-shaped hericium erinaceus as defined in claim 3, wherein the primordium-forming conditions include: light stimulation, wherein white light is adopted as light quality, the illumination intensity is 300Lux, the illumination time is 12 h/day, and the rest time is dark treatment; the conditions for pro-group formation also include: the temperature is controlled at 18-24 ℃, the humidity is controlled at 90-95%, and ventilation is kept good.
10. A method of industrially culturing a fruit body of a coral-shaped hericium erinaceus as defined in claim 3, wherein the conditions for promoting the development of the fruit body include: light stimulation, blue light is adopted as light, the illumination intensity is 300Lux, the illumination time is 3 h/day, white light is adopted to complement and irradiate to 12 h/day, and the rest time is dark treatment; conditions that promote the development of the fruiting body also include: the temperature is controlled at 20-25 ℃, the humidity is controlled at 85-90%, and ventilation is kept good.
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| CN106856984A (en) * | 2017-02-24 | 2017-06-20 | 武景江 | A kind of Hydnum tree and its cultural method |
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| CN106856984A (en) * | 2017-02-24 | 2017-06-20 | 武景江 | A kind of Hydnum tree and its cultural method |
| CN116515641A (en) * | 2023-04-26 | 2023-08-01 | 吉林农业大学 | Hericium coralloides and application thereof |
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