CN103262751B - A kind of method utilizing Cordyceps militaris spawn infected silkworm larva to cultivate silkworm Chinese caterpillar fungus - Google Patents
A kind of method utilizing Cordyceps militaris spawn infected silkworm larva to cultivate silkworm Chinese caterpillar fungus Download PDFInfo
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Abstract
A breeding method for silkworm Chinese caterpillar fungus, the selection of it comprises the steps: (1) silkworm: select five age 3rd ~ 5d silkworm as host; (2) bacterial classification preparation: be inoculated in by Cordyceps militaris spawn in liquid nutrient medium, after concussion is cultivated, obtains liquid spawn after sterile gauze filtration sterilization ball; (3) preparation of Nutrition Soil: the soil of preparation water content 5% ~ 20%, as Nutrition Soil after autoclave sterilization; (4) inoculate: silkworm is put into sterilizing 20 ~ 50min under ultraviolet, then inject the liquid spawn of 0.2 ~ 0.4ml in Silkworm, Bombyx mori; (5) training orientation: postvaccinal silkworm is put into Nutrition Soil, is placed on lucifuge under 5 DEG C ~ 12 DEG C environment and cultivates 5 ~ 12d, after to proceed to light intensity be 50 ~ 200lx, temperature is 21 DEG C ~ 23 DEG C, and humidity is cultivate 35 ~ 60d under the condition of 86% ~ 89%, obtains silkworm Chinese caterpillar fungus.The present invention have simple to operate, cost is low, infection rate and go out the advantages such as careless high, good stability, be applicable to large-scale production, there is vast potential for future development.
Description
Technical field
The present invention relates to a kind of method utilizing Cordyceps militaris spawn to infect live body silkworm larva cultivation silkworm Chinese caterpillar fungus.
Background technology
Chinese caterpillar fungus is the medicine-food two-purpose fungi that a class has remarkable health care, and wherein foremost is Cordyceps sinensis, and its Related product relates to several large classes tens kinds such as health products, food, Chinese patent drug, all enjoys high reputation in the whole world.But host of Cordyceps sinensis is single-minded, living environment is harsh, artificial culture is difficult to success completely, and due to its natural host bat moth life cycle long, natural enemy is many, disease is many, the larva output wretched insufficiency of natural propagation, in addition in these years the environmental pollution of Cordyceps sinensis producing region is serious, too early artificial, excessively taste is adopted taste and is dug and grassy marshland desertification, the reasons such as the chance causing immature Cordyceps Militaris to infect host reduces in a large number, thus cause cordyceps yield and quality significantly to decline, the source of goods is rare, expensive, the growing demand in market can not be met far away.
Silkworm be a kind of can the lepidopterous insects of artificial feeding on a large scale, the polypide of its form and Cordyceps sinensis is the most close, and the silkworm Chinese caterpillar fungus cultivated using it as host is the substitute of wild cordyceps the best.What particularly Chinese medicine was particular about is the comprehensive of nutrient component; i.e. all herbal medicine; silkworm Chinese caterpillar fungus is the most close with Cordyceps sinensis in form and active ingredient, utilizes silkworm Chinese caterpillar fungus to substitute the ecological environmental protection of Cordyceps sinensis to the comprehensive development and utilization of silkworm resource and Qinghai-Tibet Platean all significant.
Pan Zhonghua (2001) with 5 ages silkworm larvae cultivate silkworm Chinese caterpillar fungus, cultivate in temperature 22 DEG C, humidity more than 95% after inoculation, its rigid rate is 48% ~ 80%, but spawn degeneration is obvious; Wen Lu (2004) and Zhu Fendi (2005) have studied cordyceps and infects the infection conditions playing silkworm 4 ages, play the silkworm of silkworm 5 ages, and find that the infectious effect in 2 ~ 4 days 5 ages is relatively good, rigid rate reaches about 80%.It is fruiting bodies of cordyceps militaris and the breeding method thereof of host that Chinese invention patent application 201110381394.6 discloses with silkworm larva, after have employed injecting fluid dilution bacterial classification, silkworm is inserted in the mixture of perlite and vermiculite, then cultivate under being 70% ~ 90% condition in temperature 18 ~ 24 DEG C, humidity, but do not mention the infection rate of its silkworm.Chinese invention patent application 201110306713.7 discloses a kind of method utilizing silkworm larvae to cultivate Cordyceps militaris, have employed 5 age matured silkworm, Cordyceps militaris hyphal suspension is injected after hypothermic anesthesia, then after in earlier stage constantly carrying out the temperature difference and cultivating a period of time, cultivate after 3 ~ 5 days under proceeding to temperature 19 ~ 20 DEG C, humidity 85% ~ 95% condition, cultivate to light.The method operation more complicated, technology is difficult to control.
Summary of the invention
The object of the present invention is to provide a kind of infection rate and go out careless high silkworm Chinese caterpillar fungus artificial culturing method, the method have simple to operate, cost is low, infection rate and go out careless high, good stability, be easy to the advantages such as large-scale planting, has vast potential for future development.
The object of the invention is to be achieved through the following technical solutions.
It comprises the steps: 1, the breeding method of a kind of silkworm Chinese caterpillar fungus, the selection of it comprises the steps: (1) silkworm: select the silkworm in the 3rd day to the 5th day five ages as host; (2) preparation of Cordyceps militaris spawn: the Cordyceps militaris slant strains after rejuvenation is divided into soya bean size, every 2 ~ 3 pieces are inoculated in one bottle of liquid nutrient medium, 23 DEG C, 130 r/min concussion cultivate 5d obtain zymotic fluid, cross sterile gauze obtain liquid spawn; (3) preparation of Nutrition Soil: the soil choosing water content 5% ~ 20%, 121 DEG C, 0.1 Mpa sterilizing 30 min; (4) inoculate: above-mentioned silkworm is put into ultraviolet sterilization 20 ~ 50min on superclean bench, be then expelled in Silkworm, Bombyx mori with the liquid spawn of disposable syringe absorption 0.2 ~ 0.4ml; (5) early stage training orientation: postvaccinal silkworm is put into above-mentioned Nutrition Soil, is placed on lucifuge under 5 DEG C ~ 12 DEG C environment and cultivates 5 ~ 12d; (6) later stage training orientation: the silkworm of the cultivation in step (5) is placed on intensity of illumination 50 ~ 200lx, temperature 21 DEG C ~ 23 DEG C, cultivates 35 ~ 60d under humidity 86% ~ 89% condition, obtains silkworm Chinese caterpillar fungus.
Advantage of the present invention is: simple to operate, cost is low, technology is easily promoted, infection rate and go out careless high, good stability, be easy to large-scale planting.
Accompanying drawing explanation
Fig. 1 is operating process schematic diagram of the present invention.
Fig. 2 is the silkworm worm grass product figure that the present invention cultivates.
Embodiment
Embodiment 1
(1) selection of silkworm: select the silkworm in the 3rd day five ages as host;
(2) preparation of Cordyceps militaris spawn: the Cordyceps militaris slant strains after rejuvenation is divided into soya bean size, every 2 ~ 3 pieces are inoculated in one bottle of liquid nutrient medium, in 23 DEG C, 130r/min concussion cultivate 5d obtain zymotic fluid, cross sterile gauze obtain liquid spawn;
(3) preparation of Nutrition Soil: the soil of preparation water content 10%, 121 DEG C, 0.1 Mpa sterilizing 30 min;
(4) inoculate: above-mentioned silkworm is put into ultraviolet sterilization 30min on superclean bench, be then expelled in Silkworm, Bombyx mori with the liquid spawn of disposable syringe absorption 0.2ml;
(5) early stage training orientation: postvaccinal silkworm is put into above-mentioned Nutrition Soil, and under being placed on 9 DEG C of environment, lucifuge cultivates 10 d;
(6) later stage training orientation: the silkworm of the cultivation in step (5) is placed on intensity of illumination 50 lx, temperature 21 DEG C, cultivates 45 d under humidity 86% condition;
After cultivation terminates, its silkworm body infection rate 90%, going out carelessness is 89%, the long 3 ~ 6cm of fruit body, and after measured, the content of gained silkworm cordyceps militaris sporocarp cordycepin is 0.189%, and polysaccharide is 0.839%, and cordycepic acid is 1.238%; The content of polypide cordycepin is 0.077%, and polysaccharide is 0.281%, and cordycepic acid is 0.227%.Wherein in silkworm Chinese caterpillar fungus entirety, cordycepin content is 0.155%, and polysaccharide is 0.495%, and cordycepic acid is 0.873%.
Embodiment 2
(1) selection of silkworm: select the silkworm in the 5th day five ages as host;
(2) preparation of Cordyceps militaris spawn: the Cordyceps militaris slant strains after rejuvenation is divided into soya bean size, every 2 ~ 3 pieces are inoculated in one bottle of liquid nutrient medium, in 23 DEG C, 130r/min concussion cultivate 5d obtain zymotic fluid, cross sterile gauze obtain liquid spawn;
(3) preparation of Nutrition Soil: the soil of preparation water content 15%, 121 DEG C, 0.1 Mpa sterilizing 30 min;
(4) inoculate: above-mentioned silkworm is put into ultraviolet sterilization 50min on superclean bench, be then expelled in Silkworm, Bombyx mori with the liquid spawn of disposable syringe absorption 0.3ml;
(5) early stage training orientation: postvaccinal silkworm is put into above-mentioned Nutrition Soil, is placed on lucifuge under 7 DEG C of environment and cultivates 12d;
(6) later stage training orientation: it is 150lx that the silkworm of the cultivation in step (5) is placed on intensity of illumination, temperature 23 DEG C cultivates 60d under humidity 89% condition;
After cultivation terminates, its silkworm body infection rate 95%, going out carelessness is 92%, and the long 3 ~ 6cm of fruit body, in its sub-entities, the content of cordycepin is 0.191%, and polysaccharide is 0.911%, and cordycepic acid is 1.476%.Wherein in polypide, the content of cordycepin is 0.062%, and polysaccharide is 0.237%, and cordycepic acid is 0.309%.Wherein in silkworm Chinese caterpillar fungus entirety, cordycepin content is 0.169%, and polysaccharide is 0.523%, and cordycepic acid is 0.987%.
The silkworm Chinese caterpillar fungus more than obtained is tested through the research special topic " its mouse oral gavages silkworm Chinese caterpillar fungus powder acute toxicity experience (HNSE2013 (3) 005ATSI) " of non-clinical study mechanism Hunan Province's Experimental Animal Center and Drug Safety Evaluation Center of Hunan Province, result shows: its mouse oral gavage silkworm Chinese caterpillar fungus powder Cmax 0.48g/ml, maximum volume 40ml/kg, the same day 1 time, mouse autonomic activities is as usual, overt toxicity symptom is had no upon administration in observation period of 14 days, have no animal dead, observation period terminates rear anesthesia and puts to death all animals, visually observe each internal organs after gross anatomy and also do not find obvious exception.
Claims (2)
1. a breeding method for silkworm Chinese caterpillar fungus, the selection of it comprises the steps: (1) silkworm: select the silkworm in the 3rd day five ages as host; (2) preparation of Cordyceps militaris spawn: the Cordyceps militaris slant strains after rejuvenation is divided into soya bean size, every 2 ~ 3 pieces are inoculated in one bottle of liquid nutrient medium, in 23 DEG C, 130r/min concussion cultivate 5d obtain zymotic fluid, cross sterile gauze obtain liquid spawn; (3) preparation of Nutrition Soil: the soil of preparation water content 10%, 121 DEG C, 0.1Mpa sterilizing 30min; (4) inoculate: above-mentioned silkworm is put into ultraviolet sterilization 30min on superclean bench, be then expelled in Silkworm, Bombyx mori with the liquid spawn of disposable syringe absorption 0.2ml; (5) early stage training orientation: postvaccinal silkworm is put into above-mentioned Nutrition Soil, is placed on lucifuge under 9 DEG C of environment and cultivates 10d; (6) later stage training orientation: the silkworm of the cultivation in step (5) is placed on intensity of illumination 50lx, temperature 21 DEG C, cultivates 45d under humidity 86% condition.
2. a breeding method for silkworm Chinese caterpillar fungus, the selection of it comprises the steps: (1) silkworm: select the silkworm in the 5th day five ages as host; (2) preparation of Cordyceps militaris spawn: the Cordyceps militaris slant strains after rejuvenation is divided into soya bean size, every 2 ~ 3 pieces are inoculated in one bottle of liquid nutrient medium, in 23 DEG C, 130r/min concussion cultivate 5d obtain zymotic fluid, cross sterile gauze obtain liquid spawn; (3) preparation of Nutrition Soil: the soil of preparation water content 15%, 121 DEG C, 0.1Mpa sterilizing 30min; (4) inoculate: above-mentioned silkworm is put into ultraviolet sterilization 50min on superclean bench, be then expelled in Silkworm, Bombyx mori with the liquid spawn of disposable syringe absorption 0.3ml; (5) early stage training orientation: postvaccinal silkworm is put into above-mentioned Nutrition Soil, is placed on lucifuge under 7 DEG C of environment and cultivates 12d; (6) later stage training orientation: it is 150lx that the silkworm of the cultivation in step (5) is placed on intensity of illumination, temperature 23 DEG C cultivates 60d under humidity 89% condition.
Priority Applications (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310179341.5A CN103262751B (en) | 2013-05-15 | 2013-05-15 | A kind of method utilizing Cordyceps militaris spawn infected silkworm larva to cultivate silkworm Chinese caterpillar fungus |
| CN201510740832.1A CN105359825A (en) | 2013-05-15 | 2013-05-15 | Silkworm insect grass cultivating method of infecting bombyxmori larva by cordyceps fungus strain |
| HK16110372.9A HK1222092A1 (en) | 2013-05-15 | 2016-08-31 | A method of cultivating silkworm-based cordyceps militaris by infecting bombyx mori larva with cordyceps militaris |
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| CN201310179341.5A CN103262751B (en) | 2013-05-15 | 2013-05-15 | A kind of method utilizing Cordyceps militaris spawn infected silkworm larva to cultivate silkworm Chinese caterpillar fungus |
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| CN201510740832.1A Division CN105359825A (en) | 2013-05-15 | 2013-05-15 | Silkworm insect grass cultivating method of infecting bombyxmori larva by cordyceps fungus strain |
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| CN201510740832.1A Pending CN105359825A (en) | 2013-05-15 | 2013-05-15 | Silkworm insect grass cultivating method of infecting bombyxmori larva by cordyceps fungus strain |
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| CN103960055A (en) * | 2014-05-15 | 2014-08-06 | 高青和润丝绸有限公司 | Method for cultivating silkworm cordyceps |
| TWI580780B (en) * | 2015-05-29 | 2017-05-01 | 東海大學 | The cultivation method of Cordyceps sinensis |
| CN105940956B (en) * | 2016-06-06 | 2017-06-20 | 江苏科技大学 | A kind of method with tussah silkworm chrysalis as host artificial culture silkworm chrysalis cicada fungus |
| CN106034738B (en) * | 2016-06-06 | 2017-06-16 | 江苏科技大学 | A kind of method with silkworm pupa as host artificial culture silkworm chrysalis cicada fungus |
| CN106613314A (en) * | 2016-10-10 | 2017-05-10 | 贵州高山六芝园种植有限公司 | Artificial cordyceps stroma ripening method |
| CN106818206A (en) * | 2016-12-20 | 2017-06-13 | 湖南省中医药研究院 | A kind of artificial culturing method of snowy peak Chinese caterpillar fungus |
| CN108293606A (en) * | 2017-10-16 | 2018-07-20 | 常德炎帝生物科技有限公司 | A kind of soil cultivation method of Cordyceps militaris potted landscape |
| CN108112423A (en) * | 2017-12-27 | 2018-06-05 | 湖南省蚕桑科学研究所 | A kind of method that emulated ecological environment earthing cultivates silkworm grass |
| CN108184540B (en) * | 2018-01-29 | 2020-04-03 | 泰安市农业科学研究院 | Method for cultivating cordyceps militaris by using living tenebrio molitor larvae as hosts |
| CN109122027A (en) * | 2018-09-29 | 2019-01-04 | 杭州中非生物科技有限公司 | The inoculation method of silkworm grass |
| CN109644779B (en) * | 2019-01-31 | 2021-07-13 | 湖南省蚕桑科学研究所 | Method for cultivating cordyceps militaris by inoculating 5-instar larvae of castor silkworms with cordyceps militaris |
| CN110616154A (en) * | 2019-09-27 | 2019-12-27 | 江西省农业科学院农产品质量安全与标准研究所 | Cordyceps militaris cultured by taking hermetia illucens pupae as host and culturing method thereof |
| CN115053750A (en) * | 2022-06-10 | 2022-09-16 | 扬州大学 | Method for artificially cultivating Clanis bilineata tsingtauica cordyceps militaris by inoculating live bean worms |
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| CN103262751A (en) | 2013-08-28 |
| CN105359825A (en) | 2016-03-02 |
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