CN104488557A - Method for purification and rejuvenation of edible fungus strain - Google Patents
Method for purification and rejuvenation of edible fungus strain Download PDFInfo
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- CN104488557A CN104488557A CN201410828485.3A CN201410828485A CN104488557A CN 104488557 A CN104488557 A CN 104488557A CN 201410828485 A CN201410828485 A CN 201410828485A CN 104488557 A CN104488557 A CN 104488557A
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- 238000000034 method Methods 0.000 title claims abstract description 21
- 238000000746 purification Methods 0.000 title claims abstract description 21
- 230000003716 rejuvenation Effects 0.000 title claims abstract description 21
- 241000233866 Fungi Species 0.000 title claims abstract description 20
- 238000012360 testing method Methods 0.000 claims abstract description 39
- 241000894006 Bacteria Species 0.000 claims abstract description 25
- 239000000203 mixture Substances 0.000 claims abstract description 11
- 230000001954 sterilising effect Effects 0.000 claims abstract description 11
- 239000007864 aqueous solution Substances 0.000 claims abstract description 7
- 239000012286 potassium permanganate Substances 0.000 claims abstract description 7
- YFNCATAIYKQPOO-UHFFFAOYSA-N thiophanate Chemical compound CCOC(=O)NC(=S)NC1=CC=CC=C1NC(=S)NC(=O)OCC YFNCATAIYKQPOO-UHFFFAOYSA-N 0.000 claims abstract description 7
- 239000002361 compost Substances 0.000 claims description 55
- 239000002023 wood Substances 0.000 claims description 10
- 239000000243 solution Substances 0.000 claims description 6
- 240000004922 Vigna radiata Species 0.000 claims description 5
- 235000010721 Vigna radiata var radiata Nutrition 0.000 claims description 5
- 235000011469 Vigna radiata var sublobata Nutrition 0.000 claims description 5
- 238000009835 boiling Methods 0.000 claims description 5
- 238000005538 encapsulation Methods 0.000 claims description 5
- 239000002994 raw material Substances 0.000 claims description 5
- 241000700605 Viruses Species 0.000 abstract description 7
- 239000001963 growth medium Substances 0.000 abstract 11
- 238000012258 culturing Methods 0.000 abstract 3
- 230000001105 regulatory effect Effects 0.000 abstract 2
- 229920000742 Cotton Polymers 0.000 abstract 1
- 244000013123 dwarf bean Species 0.000 abstract 1
- 235000021331 green beans Nutrition 0.000 abstract 1
- 238000004659 sterilization and disinfection Methods 0.000 abstract 1
- 235000001674 Agaricus brunnescens Nutrition 0.000 description 16
- 230000001580 bacterial effect Effects 0.000 description 9
- 241000196324 Embryophyta Species 0.000 description 7
- 238000004519 manufacturing process Methods 0.000 description 7
- 238000001784 detoxification Methods 0.000 description 6
- 241000234427 Asparagus Species 0.000 description 5
- 235000005340 Asparagus officinalis Nutrition 0.000 description 5
- 238000011109 contamination Methods 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 230000002538 fungal effect Effects 0.000 description 5
- 208000035143 Bacterial infection Diseases 0.000 description 2
- 240000007313 Tilia cordata Species 0.000 description 2
- 208000022362 bacterial infectious disease Diseases 0.000 description 2
- 238000003306 harvesting Methods 0.000 description 2
- 208000015181 infectious disease Diseases 0.000 description 2
- 238000012827 research and development Methods 0.000 description 2
- 230000001568 sexual effect Effects 0.000 description 2
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 231100000053 low toxicity Toxicity 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 210000000697 sensory organ Anatomy 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
Classifications
-
- 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
Abstract
The invention relates to a method for the purification and rejuvenation of an edible fungus strain. According to the method, a first culture medium and a second culture medium are adopted, wherein the first culture medium is obtained by mixing 95 to 97 weight percent of sawdust and 3 to 5 weight percent of river silt and regulating the moisture content of the mixture with a 1 percent potassium permanganate aqueous solution, and the second culture medium is obtained by mixing 95 to 97 weight percent of sawdust and 3 to 5 weight percent of river silt and regulating the moisture content with a 0.5 percent topsin aqueous solution. The method comprises the following steps: pushing 80 grams of the first culture medium and 80 grams of the second culture medium into the centers of bottomless test tubes respectively, and plugging cotton plugs into the two ends of the bottomless test tubes for sterilization; selecting fungus blocks sized as green beans in an aseptic technique manner, inoculating the fungus blocks into the first culture medium, performing culturing for 4 to 6 days at 25 DEG C, selecting mycelium at the uninoculated end to be inoculated to one end of the second culture medium when the mycelium in the first culture medium reaches the other end, performing culturing for 4 to 6 days at 25 DEG C, selecting the mycelium at the uninoculated end to be inoculated to a PDA culture medium when the mycelium penetrates through the second culture medium, and performing culturing for 4 days at 25 DEG C to obtain sterile mother strain. The method is short in cycle and high in accuracy, the sources are simple, the virus, bacterium and fungus pollution of the strain can be rapidly removed, and meanwhile, rejuvenation and strain purification functions are realized.
Description
Technical field
The present invention relates to edible mushroom technical field, the method for a kind of edible fungus species purification and rejuvenation specifically.
Technical background
Along with the fast development of edible fungus industrial cultivation in recent years, the control of strain quality is more shown in important.In continuous print production process, the original mother of edible mushroom plants and easily occurs that virus, bacterium, fungal contamination are with kind of a sexual involution phenomenon, after edible fungus species is polluted or degenerates, a large amount of underproduction of later stage fruiting can be caused even to have no harvest.Edible fungus species is degenerated and extremely difficultly to be judged from sense organ, edible mushroom strain pollution occurred in a lot of edible mushroom factory and to degenerate the event of the phenomenon causing the underproduction even to be had no harvest.Pollute and degenerate problem for edible fungus species at present, the method of the most advanced and sophisticated detoxification of many employings and bridge cut-off detoxification, find in actual production most advanced and sophisticated detoxification and bridge cut-off detoxification limited in one's ability, cannot ensure the safe and reliable of bacterial classification completely, most advanced and sophisticated detoxification simultaneously and bridge cut-off detoxification cannot play the effect of rejuvenation edible bacterium.
Summary of the invention
According to above-mentioned weak point, the object of the present invention is to provide a kind of method of purification, rejuvenation edible fungus species, the method cycle is short, accuracy is high, can slough fast bacterial classification effect with also playing rejuvenation bacterial classification while virus, bacterium, fungal contamination.
For achieving the above object, technical program of the present invention lies in: a kind of method of edible fungus species purification and rejuvenation, it adopts following composts or fertilisers of cultivating:
No. 1 composts or fertilisers of cultivating: the percentage of wood chip weight is 95-97%, the percentage of river silt weight is 3-5%, is used by said mixture 1% potassium permanganate solution to be adjusted between 50-58% by moisture content;
No. 2 composts or fertilisers of cultivating: the percentage of wood chip weight is 95-97%, the percentage of river silt weight is 3-5%, is used by said mixture 0.5% topsin aqueous solution moisture content to be adjusted between 50-58%;
Concrete operation step is as follows:
(1) composts or fertilisers of cultivating makes
(1) 2 bottomless test tubes are prepared;
(2) No. 1 composts or fertilisers of cultivating mixed and No. 2 each 80g of composts or fertilisers of cultivating are taken;
(3) load weighted raw material are used the test tube of little No., push Boiling tube central authorities respectively;
(4) bottomless test tube two ends are filled in tampon respectively and are carried out encapsulation process;
(5) to put into high-pressure sterilizing pot 121 DEG C of sterilizing 30min stand-by for bottomless test tube;
(2) purification and rejuvenation operation
(1) in aseptic operating platform, the large small bacteria block of picking mung bean grain is seeded in bacterium of having gone out and is equipped with in the test tube of No. 1 composts or fertilisers of cultivating;
(2) test tube inoculated is placed in 25 DEG C of incubators cultivates, cultivate 4-6 days, observed once every 12 hours;
(3) when the hypha of edible fungus of No. 1 composts or fertilisers of cultivating penetrate No. 1 medium arrive the other end time, the mycelia that picking does not inoculate end accesses one end of No. 2 composts or fertilisers of cultivating of bacterium of having gone out under sterile working;
(4) test tube of band No. 2 composts or fertilisers of cultivating inoculated, cultivates 4-6 days at placing 25 DEG C, observes once, provoke the mycelia not inoculating end, access common PDA medium when mycelia penetrates No. 2 composts or fertilisers of cultivating every 12 hours;
(5) access the common PDA medium of mycelia, cultivate 4 days at 25 DEG C, namely become aseptic mother's kind and can be used for producing or preserving.
Beneficial effect of the present invention is: the present invention adopts the medium of two kinds of low toxicities can slough virus, bacterium, fungal contamination through twice bottomless Tube propagation, purification, rejuvenation edible fungus species can be played simultaneously, solve the original mother of edible mushroom and plant the problem easily occurring virus, bacterium, fungal contamination so that plant sexual involution.The kind retention that the invention enables the original mother of edible mushroom to plant is good, and formula is simple, processing ease.This method source is simple, and the cycle is short, and accuracy is high, slough fast bacterial classification play the bacterial classification purification effect of rejuvenation effect with virus, bacterium, fungal contamination simultaneously.Utilize potassium permanganate strong oxidation, the strong bactericidal action of topsin solution in the present invention, alternately kill the virus in female kind, bacterium, mould at twice, and sawdust medium is the barren medium of nutrition, can play the effect of superseded weak tendency mycelia rejuvenation bacterial classification.
Embodiment
Below in conjunction with specific embodiment, the present invention will be further described.
Embodiment 1
Asparagus purification and rejuvenation example: in April, 2013, there is a large amount of underproduction in the Asparagus that Qingdao company produces, some needle mushroom bacterium bag is not sent out and is completely namely occurred putrefactive phenomenon, Asparagus bacterial infection is confirmed as through verification, the source of infection is that Asparagus mother plants, infect decision employing the inventive method purification and rejuvenation Needle mushroom strain for solving Asparagus, concrete operations are as follows:
(1) composts or fertilisers of cultivating is prepared
No. 1 composts or fertilisers of cultivating: the percentage of wood chip weight is 97%, the percentage of river silt weight is 3%, is used by said mixture 1% potassium permanganate solution that moisture content is adjusted to 58%;
No. 2 composts or fertilisers of cultivating: the percentage of wood chip weight is 97%, the percentage of river silt weight is 3%, is used by said mixture 0.5% topsin aqueous solution moisture content to be adjusted between 58%;
(2) composts or fertilisers of cultivating makes
(1) 2 bottomless test tubes are prepared;
(2) No. 1 composts or fertilisers of cultivating mixed and No. 2 each 80g of composts or fertilisers of cultivating are taken;
(3) load weighted raw material are used the test tube of little No., push Boiling tube central authorities respectively;
(4) bottomless test tube two ends are filled in tampon respectively and are carried out encapsulation process;
(5) to put into high-pressure sterilizing pot 121 DEG C of sterilizing 30min stand-by for bottomless test tube;
(3) purification and rejuvenation operation
(1) in aseptic operating platform, the large small bacteria block of picking mung bean grain is seeded in bacterium of having gone out and is equipped with in the test tube of No. 1 composts or fertilisers of cultivating;
(2) test tube inoculated is placed in 25 DEG C of incubators cultivates, cultivate 6 days, observed once every 12 hours;
(3) when the hypha of edible fungus of No. 1 composts or fertilisers of cultivating penetrate No. 1 medium arrive the other end time, the mycelia that picking does not inoculate end accesses one end of No. 2 composts or fertilisers of cultivating of bacterium of having gone out under sterile working;
(4) test tube of band No. 2 composts or fertilisers of cultivating inoculated, cultivates 6 days at placing 25 DEG C, observes once, provoke the mycelia not inoculating end, access common PDA medium when mycelia penetrates No. 2 composts or fertilisers of cultivating every 12 hours;
(5) access the common PDA medium of mycelia, cultivate 4 days at 25 DEG C, for the production of.
By the Needle mushroom strain after these purification and rejuvenation, be applied to production and do not find bacterial infection phenomenon.
Embodiment 2
Mushroom strain is tamed: bacterial classification research and development centre of Qingdao company, collect the linden mushroom that a strain kind is excellent, obtain 5 original mothers by organizing separation and plant test tube, 5 test tubes are found while mushroom strain growth with bacterium, mycotic infection when Tube propagation, and now the excellent linden mushroom of kind property without, unfinishedly obtain this strain bacterial classification, determine to adopt the inventive method purification bacterial classification, concrete operations are as follows:
(1) composts or fertilisers of cultivating is prepared
No. 1 composts or fertilisers of cultivating: the percentage of wood chip weight is 95%, the percentage of river silt weight is 5%, is used by said mixture 1% potassium permanganate solution that moisture content is adjusted to 50%;
No. 2 composts or fertilisers of cultivating: the percentage of wood chip weight is 95%, the percentage of river silt weight is 5%, is used by said mixture 0.5% topsin aqueous solution moisture content to be adjusted between 50%;
(2) composts or fertilisers of cultivating makes
(1) 2 bottomless test tubes are prepared;
(2) No. 1 composts or fertilisers of cultivating mixed and No. 2 each 80g of composts or fertilisers of cultivating are taken;
(3) load weighted raw material are used the test tube of little No., push Boiling tube central authorities respectively;
(4) bottomless test tube two ends are filled in tampon respectively and are carried out encapsulation process;
(5) to put into high-pressure sterilizing pot 121 DEG C of sterilizing 30min stand-by for bottomless test tube;
(3) purification and rejuvenation operation
(1) in aseptic operating platform, the large small bacteria block of picking mung bean grain is seeded in bacterium of having gone out and is equipped with in the test tube of No. 1 composts or fertilisers of cultivating;
(2) test tube inoculated is placed in 25 DEG C of incubators cultivates, cultivate 4 days, observed once every 12 hours;
(3) when the hypha of edible fungus of No. 1 composts or fertilisers of cultivating penetrate No. 1 medium arrive the other end time, the mycelia that picking does not inoculate end accesses one end of No. 2 composts or fertilisers of cultivating of bacterium of having gone out under sterile working;
(4) test tube of band No. 2 composts or fertilisers of cultivating inoculated, cultivates 4 days at placing 25 DEG C, observes once, provoke the mycelia not inoculating end, access common PDA medium when mycelia penetrates No. 2 composts or fertilisers of cultivating every 12 hours;
(5) access the common PDA medium of mycelia, cultivate 4 days at 25 DEG C, for the production of.
Take turns through one the mushroom mycelium that purification is inoculated in PDA medium again and do not find any pollution again, for the production of respond well.
Embodiment 3
The purification of Xingbao mushroom is preserved: the Xingbao mushroom quality of Qingdao company research and development is very good, but after repeatedly going down to posterity, find that quality declines serious, and therefore need mother to plant to purify and preserve, in order to the use of production, concrete operations are as follows:
(1) composts or fertilisers of cultivating is prepared
No. 1 composts or fertilisers of cultivating: the percentage of wood chip weight is 96%, the percentage of river silt weight is 45%, is used by said mixture 1% potassium permanganate solution that moisture content is adjusted to 55%;
No. 2 composts or fertilisers of cultivating: the percentage of wood chip weight is 96%, the percentage of river silt weight is 4%, is used by said mixture 0.5% topsin aqueous solution moisture content to be adjusted between 55%;
(2) composts or fertilisers of cultivating makes
(1) 2 bottomless test tubes are prepared;
(2) No. 1 composts or fertilisers of cultivating mixed and No. 2 each 80g of composts or fertilisers of cultivating are taken;
(3) load weighted raw material are used the test tube of little No., push Boiling tube central authorities respectively;
(4) bottomless test tube two ends are filled in tampon respectively and are carried out encapsulation process;
(5) to put into high-pressure sterilizing pot 121 DEG C of sterilizing 30min stand-by for bottomless test tube;
(3) purification and rejuvenation operation
(1) in aseptic operating platform, the large small bacteria block of picking mung bean grain is seeded in bacterium of having gone out and is equipped with in the test tube of No. 1 composts or fertilisers of cultivating;
(2) test tube inoculated is placed in 25 DEG C of incubators cultivates, cultivate 5 days, observed once every 12 hours;
(3) when the hypha of edible fungus of No. 1 composts or fertilisers of cultivating penetrate No. 1 medium arrive the other end time, the mycelia that picking does not inoculate end accesses one end of No. 2 composts or fertilisers of cultivating of bacterium of having gone out under sterile working;
(4) test tube of band No. 2 composts or fertilisers of cultivating inoculated, cultivates 5 days at placing 25 DEG C, observes once, provoke the mycelia not inoculating end, access common PDA medium when mycelia penetrates No. 2 composts or fertilisers of cultivating every 12 hours;
(5) access the common PDA medium of mycelia, cultivate 4 days at 25 DEG C, for preserving.
Claims (1)
1. a method for edible fungus species purification and rejuvenation, is characterized in that: it adopts following composts or fertilisers of cultivating:
No. 1 composts or fertilisers of cultivating: the percentage of wood chip weight is 95-97%, the percentage of river silt weight is 3-5%, is used by said mixture 1% potassium permanganate solution to be adjusted between 50-58% by moisture content;
No. 2 composts or fertilisers of cultivating: the percentage of wood chip weight is 95-97%, the percentage of river silt weight is 3-5%, is used by said mixture 0.5% topsin aqueous solution moisture content to be adjusted between 50-58%;
Concrete operation step is as follows:
(1) composts or fertilisers of cultivating makes
(1) 2 bottomless test tubes are prepared;
(2) No. 1 composts or fertilisers of cultivating mixed and No. 2 each 80g of composts or fertilisers of cultivating are taken;
(3) load weighted raw material are used the test tube of little No., push Boiling tube central authorities respectively;
(4) bottomless test tube two ends are filled in tampon respectively and are carried out encapsulation process;
(5) to put into high-pressure sterilizing pot 121 DEG C of sterilizing 30min stand-by for bottomless test tube;
(2) purification and rejuvenation operation
(1) in aseptic operating platform, the large small bacteria block of picking mung bean grain is seeded in bacterium of having gone out and is equipped with in the test tube of No. 1 composts or fertilisers of cultivating;
(2) test tube inoculated is placed in 25 DEG C of incubators cultivates, cultivate 4-6 days, observed once every 12 hours;
(3) when the hypha of edible fungus of No. 1 composts or fertilisers of cultivating penetrate No. 1 medium arrive the other end time, the mycelia that picking does not inoculate end accesses one end of No. 2 composts or fertilisers of cultivating of bacterium of having gone out under sterile working;
(4) test tube of band No. 2 composts or fertilisers of cultivating inoculated, cultivates 4-6 days at placing 25 DEG C, observes once, provoke the mycelia not inoculating end, access common PDA medium when mycelia penetrates No. 2 composts or fertilisers of cultivating every 12 hours;
(5) access the common PDA medium of mycelia, cultivate 4 days at 25 DEG C, namely become aseptic mother's kind and can be used for producing or preserving.
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| CN201410828485.3A CN104488557B (en) | 2014-12-26 | 2014-12-26 | A kind of method of edible fungus species purification and rejuvenation |
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| CN201410828485.3A CN104488557B (en) | 2014-12-26 | 2014-12-26 | A kind of method of edible fungus species purification and rejuvenation |
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| CN104488557A true CN104488557A (en) | 2015-04-08 |
| CN104488557B CN104488557B (en) | 2016-09-14 |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107384805A (en) * | 2017-08-25 | 2017-11-24 | 山西农业大学 | A kind of production method of edible mushroom rejuvenation type bacterial strain |
| CN107619792A (en) * | 2017-10-25 | 2018-01-23 | 翔天农业开发集团股份有限公司 | A kind of edible fungus species purification and rejuvenation technology |
| CN110637680A (en) * | 2018-06-27 | 2020-01-03 | 贵州金蟾大山生物科技有限责任公司 | Production method of Dictyophora rubrovalvata mildew-removing high-germination-rate cultivation stock |
| CN111466255A (en) * | 2020-03-27 | 2020-07-31 | 江苏华绿生物科技股份有限公司 | Method for rejuvenating flammulina velutipes strains by using wood chip culture medium |
| CN112280690A (en) * | 2020-10-20 | 2021-01-29 | 江苏华绿生物科技股份有限公司 | Method for rejuvenating and screening needle mushroom strains |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107384805A (en) * | 2017-08-25 | 2017-11-24 | 山西农业大学 | A kind of production method of edible mushroom rejuvenation type bacterial strain |
| CN107384805B (en) * | 2017-08-25 | 2020-12-29 | 山西农业大学 | Production method of rejuvenation type strain of edible fungi |
| CN107619792A (en) * | 2017-10-25 | 2018-01-23 | 翔天农业开发集团股份有限公司 | A kind of edible fungus species purification and rejuvenation technology |
| CN110637680A (en) * | 2018-06-27 | 2020-01-03 | 贵州金蟾大山生物科技有限责任公司 | Production method of Dictyophora rubrovalvata mildew-removing high-germination-rate cultivation stock |
| CN110637680B (en) * | 2018-06-27 | 2023-08-18 | 贵州金蟾大山生物科技有限责任公司 | Production method of Dictyophora rubrovalvata mildew-removed and high-germination-rate cultivation stock |
| CN111466255A (en) * | 2020-03-27 | 2020-07-31 | 江苏华绿生物科技股份有限公司 | Method for rejuvenating flammulina velutipes strains by using wood chip culture medium |
| CN112280690A (en) * | 2020-10-20 | 2021-01-29 | 江苏华绿生物科技股份有限公司 | Method for rejuvenating and screening needle mushroom strains |
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| Publication number | Publication date |
|---|---|
| CN104488557B (en) | 2016-09-14 |
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