CN109105153B - Efficient cultivation method of tussah cordyceps militaris - Google Patents
Efficient cultivation method of tussah cordyceps militaris Download PDFInfo
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G18/00—Cultivation of mushrooms
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G18/00—Cultivation of mushrooms
- A01G18/20—Culture media, e.g. compost
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G18/00—Cultivation of mushrooms
- A01G18/40—Cultivation of spawn
Abstract
The invention discloses a high-efficiency cultivation method of tussah pupa cordyceps sinensis. The method comprises the steps of treating autumn tussah cocoons which are just cocooned and pupated at the diapause period for 6-8 days at the temperature of 19-28 ℃ and the humidity of 70-80%, storing for 17-23 days at the temperature of 0-4 ℃, taking disinfected tussah living pupae, inoculating 0.3-0.7ml of cordyceps militaris liquid strain from the 3 rd-5 th body node of the back of the tussah living pupae, placing the tussah living pupae in a plastic disc, and culturing the cordyceps militaris for 45-50 days under the conditions of proper temperature, humidity, illumination and the like to obtain mature cordyceps militaris. The invention ensures that the grass yield of the tussah cordyceps militaris reaches 89.2%, improves the space utilization rate by 56.7%, obviously improves the product quality, the production efficiency and the economic benefit, provides technical guarantee for the industrial production of the tussah cordyceps militaris, provides sufficient raw materials for the food and medicine health-care product industry, and has wide market application prospect.
Description
Technical Field
The invention belongs to the technical field of edible fungus cultivation, and particularly relates to a high-efficiency cultivation method of tussah silkworm chrysalis cordyceps sinensis.
Background
Cordyceps militaris (L.) Link, also known as Cordyceps militaris, is a model species of Cordyceps of Clavicipitaceae of Ascomycotina, is a species different from Cordyceps sinensis in the same genus, is a medical-edible fungus with high nutritive value and medicinal efficacy, and is classified as a new resource food in 2009 by the Ministry of health. Has effects in improving immunity, resisting tumor, and improving myocardial function.
The tussah cordyceps militaris is prepared by separating and domesticating wild cordyceps militaris, inoculating the wild cordyceps militaris to living tussah pupas, and culturing fruiting bodies, which are complexes of cordyceps fruiting bodies of the ergomycetaceae family and tussah pupas. Wherein the cordycepin is a specific nucleoside active substance in the cordyceps sinensis and has the functions of resisting cancer, bacteria and viruses; cordycepic acid (mannitol) can remarkably reduce intracranial pressure and promote metabolism, thereby relieving cerebral hemorrhage and cerebral thrombosis, and has diuretic and cough relieving effects; cordyceps sinensis polysaccharide is considered to be one of the excellent immune promoters in the world at present, and can enhance the immune capability of the organism. Therefore, the tussah cordyceps militaris can replace wild cordyceps sinensis, make up for the deficiency of natural cordyceps sinensis resources, provide raw materials for the fields of food and medical health-care products and have wide market application prospect. However, although the prior artificial cultivation technology of the tussah cordyceps militaris has been successful, the cultivation effect of the tussah cordyceps militaris is greatly influenced by the quality, cultivation mode, cultivation conditions and the like of the tussah silkworm chrysalis, so that the yield of the product in the current industrialized production of the tussah silkworm chrysalis cordyceps militaris is low. According to Chendan (edible fungi, 2006 (3): 33-34) and Marui (Chinese livestock and veterinary abstracts, 2013, 9 (2): 97-110), autumn commodity tussah pupa is taken as a raw material, a conventional glass bottle culture method is adopted, the grass yield of the tussah pupa cordyceps sinensis is respectively 64% and 65%, the grass yield and the production efficiency of the product are lower, and the healthy development of the tussah pupa cordyceps militaris industry is restricted.
Disclosure of Invention
Aiming at solving the technical problems, the invention systematically studies the aspects of tussah silkworm pupa raw material quality, inoculation position, inoculation dosage, culture conditions and the like aiming at the actual production, and prepares a set of tussah cordyceps militaris high-efficiency cultivation method, so that the quality, the product yield and the production efficiency of the tussah cordyceps militaris are obviously improved, the technical guarantee is provided for the industrial production of the tussah cordyceps militaris, the industrial economic benefit is greatly improved, the income increase of farmers and the industrial efficiency increase are realized, and a new way is opened up for the healthy sustainable development of the Liaoning characteristic tussah industry.
The invention relates to a high-efficiency cultivation method of tussah pupa cordyceps sinensis, which comprises the following specific steps:
(1) treating autumn tussah cocoon in diapause period after just cocooning and pupating at 19-28 deg.C and humidity of 70-80% for 6-8 days, and storing at 0-4 deg.C for 17-23 days;
(2) inoculating slant strain of Cordyceps militaris to Cordyceps militaris liquid culture medium, and culturing for 3-5 days at an inoculation ratio of: taking 0.2-0.7cm2Inoculating 5-7 pieces of slant strain of Cordyceps militaris into 200ml Cordyceps militaris liquid culture medium;
sieving with 30-50 mesh sieve under aseptic condition to obtain filtrate, transferring the filtrate into liquid culture medium at volume ratio of 2.5-7.5% to obtain Cordyceps militaris liquid strain;
(3) taking the tussah living pupae obtained in the step (1), sterilizing, puncturing a needle head into the direction of the head of the pupae for about 1cm from the 3 rd to 5 th body node of the back of the tussah living pupae, inoculating 0.3ml to 0.7ml of cordyceps militaris liquid strain prepared in the step (2), laying the needle opening of the inoculated tussah pupae upwards and laying the silkworm pupae on a culture plane, covering the silkworm pupae with a transparent film, culturing under the conditions of the temperature of 18 ℃ to 25 ℃, the humidity of 60% to 80% and the illumination intensity of 600 plus of 1000lux, illuminating 8 hours to 12 hours every day, ventilating 2 times every day, culturing for 45 days to 50 days, maturing sporocarps, and harvesting and carrying out deep processing.
For the technical scheme, the formula of the cordyceps militaris liquid strain culture medium is as follows:
potato 200g glucose 20g potassium dihydrogen phosphate 3g magnesium sulfate 1.5g vitamin B210mg peptone 3g beef extract 5g water 1000ml, pH 7.
In the above technical means, preferably, the culture temperature of the autumn tussah cocoon in the diapause period in the step (1) is 19-28 ℃, and the optimal culture temperature is 25 ℃; the best culture time is 7 days; the optimal temperature for standing is 2 ℃, and the product is stored for standby after 20 days.
For the above technical scheme, preferably, the cordyceps militaris slant strain is inoculated into the cordyceps militaris liquid culture medium in the step (2), and the culture time is 4 days;
preferably, in step (2), the slant strain size of Cordyceps militaris is preferably 0.3cm2The 6 blocks are inoculated, and the filtering of a screen is the best of a 40-mesh screen;
preferably, in the step (2), the volume ratio of the filtrate after the filtration by the screen to the liquid culture medium is 5%; for the above technical solution, preferably, the tussah pupae disinfection mode in step (3) generally adopts a mode of surface disinfection, cleaning and airing by 75% ethanol.
In the above technical solution, preferably, the culture method in which the inoculated tussah pupa needles are placed upward in a sterilized plastic tray, the plastic tray is placed in a culture chamber, and the plastic tray is covered with a transparent film is adopted in the step (3). The mode breaks through the traditional bottle type culture, fully utilizes the culture space and greatly improves the culture efficiency.
For the above technical scheme, preferably, the 4 th body node on the back of the tussah silkworm live pupae in the step (3) is the optimal injection position;
for the above technical scheme, preferably, the optimal amount of the cordyceps militaris liquid strain inoculated in the step (3) is 0.5 ml.
In the above technical solution, preferably, in the step (3), the inoculated tussah pupae are cultured in a plastic tray, the plastic tray has a length of 53cm, a width of 40cm and a height of 10cm, and about 200 tussah pupaes are flatly placed on each tray.
For the above technical solution, preferably, the plastic tray is placed in the culture chamber in the step (3), and the plastic tray is usually covered with plastic cloth;
for the technical scheme, preferably, the better culture temperature of the tussah pupae inoculated in the step (3) is 19-22 ℃; the optimal culture temperature is 22 ℃; the optimal illumination intensity is 800 lux; the optimal illumination time was 12 hours per day.
The application of the method can ensure that the yield of the tussah cordyceps militaris product reaches 89.2 percent, the space utilization rate is improved by 56.7 percent, the yield and the quality of the cordyceps militaris are improved, the production efficiency and the economic benefit are obviously improved, a foundation is laid for the industrial production of the tussah cordyceps militaris, and sufficient cordyceps militaris raw materials are provided for the food and medicine health-care product industry.
Advantageous effects
1. According to the morphological and structural characteristics of the tussah pupa, in the production process of the cordyceps militaris, different inoculation parts and different inoculation doses have large influence on the yield and quality of the cordyceps militaris, and through research and test, the effect of inoculating 0.5ml of liquid strain at the 4 th body node on the back of the tussah pupa is good.
2. At present, the production of the tussah cordyceps militaris generally adopts a glass bottle (diameter is 10cm multiplied by height is 10cm) cultivation method, 5-6 cordyceps militaris are cultivated in each bottle, and a bottle opening is sealed by a plastic film; and a plastic tray (53 cm in length, 40cm in width and 10cm in height) cultivation method is adopted, so that the technical operation is simple and convenient, the space utilization rate is high, the industrial production is easy, and the production efficiency can be obviously improved.
3. The method has the advantages of rich raw material resources, convenient process, high production efficiency and good product quality, and is suitable for industrial production; the method can remarkably improve the yield of the tussah pupa cordyceps militaris product, ensure the yield and quality of the cordyceps militaris, make up for the deficiency of natural cordyceps militaris resources, provide sufficient raw materials for the fields of foods and medical health-care products, and have wide market application prospect.
Detailed Description
The following non-limiting examples will allow one of ordinary skill in the art to more fully understand the present invention, but are not intended to limit the invention in any way.
Example 1
Research on influence of different treatment temperatures on tussah pupa quality
1.1 test methods
Selecting autumn tussah cocoons which are the same in variety and are just picked off in the diapause period under the same stocking condition, dividing the test into 4 groups, randomly selecting 1000 tussah cocoons in each group, respectively treating for 7 days under the conditions of 19, 22, 25 and 28 ℃ and 60-70% of humidity, then cutting cocoons, taking pupas, investigating the number of the diseased pupas and the number of normal pupas, and calculating the tussah pupa morbidity of each test group.
1.2 test results
The results of the tests carried out according to the test methods described above are shown in Table 1.
TABLE 1 influence of different treatment temperatures on tussah pupa disease
| Treatment temperature (. degree.C.) | Number of test cocoons/number | Number of pupae/number of disease | Normal pupa number/number | Incidence rate/%) |
| 19 | 1000 | 38 | 962 | 3.8 |
| 22 | 1000 | 54 | 946 | 5.4 |
| 25 | 1000 | 89 | 911 | 8.9 |
| 28 | 1000 | 92 | 908 | 9.2 |
As can be seen from the test results in Table 1, the incidence of tussah pupae increases gradually with the increase of temperature when the tussah pupae is acted for 7 days at the temperature of 19-28 ℃, the incidence of the tussah pupae is 8.9% at the temperature of 25 ℃, and the maximum incidence of the tussah pupae reaches 9.2% at the temperature of 28 ℃, but considering the influence of overhigh temperature on the quality of the tussah pupae and the problem of energy, and the incidence of the tussah pupae treated at the temperature of 25 ℃ and the temperature of 28 ℃ is low, so that the tussah cocoon treatment temperature is preferably selected to be 25 ℃.
Example 2
Comparison of grass production conditions of tussah pupa cordyceps militaris treated by different methods
1. Test method
The formula of the cordyceps militaris liquid strain culture medium comprises: potato 200g glucose 20g potassium dihydrogen phosphate 3g magnesium sulfate 1.5g vitamin B210mg peptone 3g beef extract 5g water 1000ml pH 7.
Sterilizing at 121 deg.C under high pressure of 0.1-0.11MPa for 30 min.
2. Preparation of cordyceps militaris liquid strain
Taking 0.3cm of Bingyu No. 1 Cordyceps militaris test tube mother strain selected and documented by Dalian biological technology research of Liaoning academy of agricultural sciences2Inoculating 6 pieces of the strain into 200ml of Cordyceps militaris liquid culture medium, performing static culture at 25 deg.C for 2d, performing shaking culture at 150rpm/min for 3-5d, filtering with 40 mesh screen under aseptic condition to obtain filtrate, and transferring the filtrate into liquid culture medium according to volume ratio of 5% for inoculating Antheraea pernyi pupa.
3. Test procedure
Treating autumn tussah cocoons which are just picked up in a diapause period at 25 ℃ for 7 days, storing the tussah cocoons in a refrigerator at 0-4 ℃ for 20 days, cutting the cocoons to obtain pupas, randomly taking 1000 tussah pupas, sterilizing the surfaces of the 1000 tussah pupas by using 75% ethanol, putting the cordyceps militaris bacterial liquid which is obtained by culture and filtration into a continuous injector under an aseptic condition, taking the sterilized tussah pupas, inoculating 0.5ml of cordyceps militaris liquid strain from the 4 th body node on the back of the tussah pupas, putting the cordyceps militaris liquid bacterial liquid in a sterilized plastic tray, covering the sterilized plastic tray with a plastic film, culturing the cordyceps militaris liquid bacterial liquid under the conditions of 19-22 ℃, humidity 60-80% and 600 plus 1000lux, illuminating for 12 hours every day, ventilating for 2 times every day, culturing for about 45 days, and investigating and counting the number of the tu. Meanwhile, 1000 tussah pupas are randomly cut from autumn tussah cocoons in the incubation period, which are not subjected to heating treatment and normally stored in a refrigerator at 0-4 ℃ for a grass-growing test, and the test method is the same as the above, and investigation and analysis are carried out.
4. Test results
The results of the tests carried out according to the test methods described above are shown in Table 1.
TABLE 2 influence of different treatments on tussah pupa emergence
| Processing method | Test pupa number/number | Number of fruiting body pupa/number | Grass yield/%) |
| Treatment at 25 deg.C | 1000 | 892 | 89.2 |
| Normal processing | 1000 | 674 | 67.4 |
As can be seen from the test results in Table 2, live autumn tussah pupae treated at 25 ℃ for 7 days are used as the material, each pupae is inoculated with 0.5ml of cordyceps militaris liquid strain, after the cultivation for 45 days according to the cordyceps militaris culture technology, 892 tussah pupaes with fruiting bodies exceeding 10 are grown, and the weed yield is 89.2%; 674 tussah pupas without heating treatment are preserved under normal condition, and the grass yield is 67.4%. The grass yield of the tussah pupa is improved by 21.8 percent by heating treatment compared with that of the tussah pupa preserved conventionally.
Example 3
Influence of different inoculation parts on growth of Chinese tussah cordyceps militaris
1. Test method
The cordyceps militaris liquid strain is prepared according to the method of example 1, live tussah pupae treated at 25 ℃ for 7 days are used as a material, the test is divided into 3 groups, 1000 tussah pupaes are randomly selected from each group, the inoculation part of the 1 st group is about 1cm from the head of the tussah pupae, the inoculation part of the 2 nd group is the 4 th body node (about 1cm is punctured into the head of the tussah pupae) from the back of the tussah pupae, the inoculation part of the 3 rd group is about 1cm from the tail of the tussah pupae, the tussah pupae is placed into a plastic tray to be cultured in a culture room after inoculation, the cordyceps militaris culture management method is the same as that of the 1.3 in the example 1, and the growth.
2. Test results
The results of the tests carried out according to the test methods described above are shown in Table 3
TABLE 3 influence of different inoculation sites on the growth of Antheraea pernyi Cordyceps
| Inoculation site | Test pupa number/number | Average number of fruiting bodies/pupa | Average length/cm of fruiting body | Average fresh weight/g of single pupa |
| Pupa head | 1000 | 26.2 | 5.1 | 7.5 |
| Pupa back No. 4 body segment | 1000 | 29.0 | 5.5 | 8.1 |
| Pupa tail | 1000 | 22.4 | 5.4 | 7.6 |
As can be seen from the results in Table 3, the inoculation part has a large influence on the growth of the Antheraea pernyi Fritillaria. The inoculation test group of the 4 th body node on the back of the tussah pupa is higher than the inoculation test group of the head of the tussah pupa and the tail of the tussah pupa in terms of the average root number of the fruit body, the average length of the fruit body and the average fresh weight of single pupa, wherein the average root number of the fruit body is 1.1 times and 1.3 times of the inoculation test group of the head of the tussah pupa and the tail of the tussah pupa respectively, and the average fresh weight of the single pupa is 1.1 times of the inoculation test group of the head of the. Therefore, the quality and the yield of the cordyceps militaris are optimal when the 4 th section of the back of the tussah pupa is taken as an inoculation part.
Example 4
Influence of different inoculation doses on growth of cordyceps militaris of tussah
1. Test method
The cordyceps militaris liquid strain is prepared according to the method of the embodiment 1, live autumn tussah pupas treated at 25 ℃ for 7 days are used as materials, the experiment is divided into 3 groups, 1000 tussah pupas are randomly taken from each group, the inoculation positions are all the 4 th joints on the backs of the tussah pupas, the dosage of the inoculation bacterial liquid of the 1 st group is 0.3ml, the dosage of the inoculation bacterial liquid of the 2 nd group is 0.5ml, and the dosage of the inoculation bacterial liquid of the 3 rd group is 0.7ml, the inoculated tussah pupas are placed into a plastic disc to be cultured in a culture room, the cordyceps militaris culture management method is the same as that of the 1.3 in the embodiment 1, and after 45 days of culture, the growth conditions of.
2. Test results
The results of the tests carried out according to the test methods described above are shown in Table 3
TABLE 4 influence of different inoculation doses on the growth of Antheraea pernyi Cordyceps
| Inoculation dose (ml) | Test pupa number/number | Average number of fruiting bodies/pupa | Average length/cm of fruiting body | Average fresh weight/g of single pupa |
| 0.3 | 1000 | 23.5 | 4.5 | 6.2 |
| 0.5 | 1000 | 29.0 | 5.5 | 8.3 |
| 0.7 | 1000 | 27.8 | 4.3 | 6.4 |
As can be seen from the results in Table 4, different inoculation doses have a great influence on the growth of the cordyceps militaris of tussah. The inoculation dose of 0.5ml test group is higher than 0.3ml and 0.7ml dose groups in the aspects of the average number of the fruiting bodies, the average length of the fruiting bodies and the average fresh weight of single pupae. Wherein the average root number of the fruit body is 1.2 and 1.04 times of the dosage group of 0.3ml and 0.7ml respectively, the average length of the fruit body is 1.2 and 1.3 times of the dosage group of 0.3ml and 0.7ml respectively, and the average fresh weight of the single pupa is 1.3 times of the dosage group of 0.3ml and 0.7ml respectively. Therefore, the quality and the yield of the cordyceps militaris are optimal when the inoculation dose of the bacterial liquid is 0.5 ml.
Example 5
Influence of different cultivation temperatures on growth of Chinese tussah cordyceps militaris
1. Test method
The cordyceps militaris liquid strain is prepared according to the method of example 1, live tussah pupas treated at 25 ℃ for 7 days are taken as materials, the experiment is divided into 3 groups, 1000 tussah pupas are randomly taken from each group, the inoculation parts are all the 4 th body nodes on the backs of the tussah pupas, the dosage of the inoculation bacterial liquid is 0.5ml, and the inoculated tussah pupas are placed into a plastic disc to be cultured in a culture room. Wherein, the tussah pupa cultivation temperature of the 1 st group is 19 ℃, the tussah pupa cultivation temperature of the 2 nd group is 22 ℃, the tussah pupa cultivation temperature of the 3 rd group is 25 ℃, the cordyceps militaris cultivation management method is the same as that of 1.3 in the example 1, and after 45 days of cultivation, the growth condition of each test group of tussah cordyceps militaris is analyzed and researched.
2. Test results
The results of the tests carried out according to the test methods described above are shown in Table 5
TABLE 5 influence of different cultivation temperatures on the growth of Antheraea pernyi Cordyceps
| Culture temperature (. degree.C.) | Test pupa number/number | Average number of fruiting bodies/pupa | Average length/cm of fruiting body | Average fresh weight/g of single pupa |
| 19 | 1000 | 23.2 | 4.1 | 7.1 |
| 22 | 1000 | 29.0 | 5.5 | 8.0 |
| 25 | 1000 | 20.0 | 3.5 | 5.5 |
As can be seen from the results in Table 5, different cultivation temperatures have a great influence on the growth of the Antheraea pernyi Fr. The test groups with the cultivation temperature of 22 ℃ are higher than the test groups with the cultivation temperatures of 19 ℃ and 23 ℃ in the aspects of the average root number of the fruit bodies, the average length of the fruit bodies and the average fresh weight of single pupae. Wherein the average root number of the fruiting body is 1.3 and 1.5 times of the test group at 19 ℃ and 23 ℃, the average length of the fruiting body is 1.3 and 1.6 times of the test group at 19 ℃ and 23 ℃, and the average fresh weight of single pupa is 1.1 and 1.5 times of the test group at 19 ℃ and 23 ℃. Therefore, the quality and yield of the cordyceps militaris are optimal when the cultivation temperature is 22 ℃.
Example 6
Comparative study on space utilization rate and grass yield of tussah cordyceps militaris in different cultivation containers
1. Test method
The cordyceps militaris liquid strain is prepared according to the method of example 1, live tussah pupas treated at 25 ℃ for 7 days are taken as materials, the test is divided into 2 groups, 1000 tussah pupas are randomly taken from each group, the inoculation positions are all the 4 th body nodes on the backs of the tussah pupas, the inoculation bacterial liquid dosage is 0.5ml, the 1 st group is inoculated and then placed into a plastic tray (with the length of 53cm, the width of 40cm and the height of 10cm) for cultivation, about 200 tussah pupas are placed in each tray, the 2 nd group is inoculated and then placed into a glass bottle (with the diameter of 10cm, the height of 10cm) for cultivation, 6 tussah pupas are placed in each bottle, the cordyceps militaris cultivation management method is the same as that of the 1.3 in example 1, and after 45 days of cultivation, the space utilization rate and the grass discharge condition of.
2. Test results
The results of the tests carried out according to the test methods described above are shown in Table 6
TABLE 6 comparison of space utilization and yield of Antheraea pernyi Cordyceps in different cultivation containers
| Cultivation container | Test pupa number/number | Tussah pupa count | Culturing Cordyceps militaris number/m2 | Percentage of weed yielded (%) |
| Plastic dish | 1000 | 200 pieces/disc | 940 | 91.7 |
| Glass bottle | 1000 | 6 pieces/bottle | 600 | 89.4 |
As can be seen from the test results in Table 6, by using plastic trays (53 cm long, 40cm wide, 10cm high) as cultivation containers, 200 Antheraea pernyi pupa were cultivated per tray, 940 Cordyceps militaris were cultivated per square meter, and the grass yield was 91.7/%; and by using glass bottles (diameter is 10cm multiplied by 10cm high) as cultivation containers, 6 cordyceps militaris can be cultivated in each bottle, 600 cordyceps militaris can be cultivated in each square meter, the grass yield is 89.4/%, the space utilization rate of the plastic disc cultivation method is improved by 56.7% compared with the glass bottle cultivation method, and the grass yield is also improved by 2.3% compared with the glass bottle cultivation method. Therefore, the plastic disc cultivation method greatly improves the space utilization rate of large-scale production, and has convenient technical operation and high production efficiency.
It will be apparent to those skilled in the art from this disclosure that many changes and modifications can be made, or equivalents modified, in the embodiments of the invention without departing from the scope of the invention. Therefore, any simple modification, equivalent change and modification made to the above embodiments according to the technical essence of the present invention shall still fall within the protection scope of the technical solution of the present invention, unless the contents of the technical solution of the present invention are departed.
Claims (5)
1. An efficient cultivation method of tussah cordyceps militaris is characterized by comprising the following steps: the method comprises the following operation steps:
(1) treating autumn tussah cocoon in diapause period after just cocooning and pupating at 19-28 deg.C and humidity of 70-80% for 6-8 days, and storing at 0-4 deg.C for 17-23 days;
(2) inoculating slant strain of Cordyceps militaris to Cordyceps militaris liquid culture medium, and culturing for 3-5 days at the following inoculation ratio: taking 0.2-0.7cm2Inoculating 5-7 pieces of slant strains with size into 200ml Cordyceps militaris liquid culture medium; filtering with 30-50 mesh sieve under aseptic condition to obtain filtrate, and transferring the filtrate into liquid culture medium according to volume ratio of 2.5-7.5% for use;
(3) taking the tussah silkworm live pupae obtained in the step (1), sterilizing, puncturing a needle head to the direction of the head of the pupae for about 1cm from the 4 th body node on the back of the tussah silkworm live pupae, inoculating 0.3ml-0.7ml of cordyceps militaris liquid strain prepared in the step (2), laying the needle mouth of the inoculated tussah silkworm pupae upwards on a culture plane, covering the culture plane with a transparent film, culturing under the conditions of the temperature of 18-25 ℃, the humidity of 60-80% and the illumination intensity of 600 plus materials of 1000lux, illuminating 8-12 hours every day, ventilating 2 times every day, and culturing for 45-50 days to obtain the tussah silkworm live pupae.
2. The cultivation method as claimed in claim 1, wherein: the culture temperature of the autumn tussah cocoons in the diapause period in the step (1) is 19-28 ℃, and the culture temperature is 25 ℃; the culture time is 7 days; the mixture was stored at a temperature of 2 ℃ for 20 days.
3. The cultivation method as claimed in claim 1, wherein: and (3) inoculating the cordyceps militaris slant strains into the cordyceps militaris liquid culture medium in the step (2), wherein the culture time is 4 days.
4. The cultivation method as claimed in claim 1, wherein: the culture temperature of the tussah pupae inoculated in the step (3) is 19-22 ℃.
5. The cultivation method as claimed in claim 1, wherein: the illumination intensity of the tussah pupae inoculated in the step (3) is 800 lux; the optimal illumination time was 12 hours per day.
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| CN112391295B (en) * | 2020-11-17 | 2022-06-10 | 辽宁省海洋水产科学研究院 | Preparation method of wild cordyceps militaris strain |
| CN112586513B (en) * | 2020-12-11 | 2021-11-19 | 辽宁省海洋水产科学研究院 | Pure natural cordyceps militaris growth regulator |
| CN114467625B (en) * | 2022-01-20 | 2024-02-02 | 辽宁省海洋水产科学研究院 | Cultivation method of high-quality tussah cordyceps militaris |
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| KR100610983B1 (en) * | 2004-07-26 | 2006-08-10 | 한국식품연구원 | Method for a cultivation of Cordyceps militaris by using Ginsaeng Culture medium |
| CN102342216A (en) * | 2010-07-29 | 2012-02-08 | 于兴军 | Method by combining physical method and chemical method to improve quality of cordyceps militaris |
| CN102907256B (en) * | 2012-10-19 | 2014-03-19 | 湖北省农业科学院农产品加工与核农技术研究所 | Box-type cultivation method of tussah pupa cordyceps militaris |
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