CN107216201A - A kind of India's pyriform spore straw medium preparation method for cultivating mushroom - Google Patents
A kind of India's pyriform spore straw medium preparation method for cultivating mushroom Download PDFInfo
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- CN107216201A CN107216201A CN201710497457.1A CN201710497457A CN107216201A CN 107216201 A CN107216201 A CN 107216201A CN 201710497457 A CN201710497457 A CN 201710497457A CN 107216201 A CN107216201 A CN 107216201A
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- mushroom
- india
- pyriform spore
- straw medium
- culture
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
- C05G3/00—Mixtures of one or more fertilisers with additives not having a specially fertilising activity
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05D—INORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C; FERTILISERS PRODUCING CARBON DIOXIDE
- C05D3/00—Calcareous fertilisers
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F17/00—Preparation of fertilisers characterised by biological or biochemical treatment steps, e.g. composting or fermentation
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
- Y02W30/00—Technologies for solid waste management
- Y02W30/40—Bio-organic fraction processing; Production of fertilisers from the organic fraction of waste or refuse
Abstract
The invention provides a kind of India's pyriform spore straw medium preparation method for cultivating mushroom, step includes：(1) India's pyriform spore is inoculated in Kafer culture mediums, rotating speed 120~150r/min, 6~8d of shaken cultivation at 26~30 DEG C, culture terminates that viable count is measured by sampling, and culture, which is dissolved in distilled water, makes viable count in every milliliter of solution be 1 × 106It is individual；(2) bacterium number for obtaining step (1) is 1 × 106Individual/mL bacterium solution is sprayed onto in the ratio of inoculum concentration 1~15% produces the India's pyriform spore straw medium for cultivating mushroom after the 12~24h that fermented at straw medium surface, 25~37 DEG C.The present invention with the addition of India's pyriform spore, choose optimal breeding condition, can promote the silk growth of mushroom root, and then improve mushroom production.
Description
Technical field
The invention belongs to microbial technology field, and in particular to a kind of India's pyriform spore straw medium system of cultivation mushroom
Preparation Method.
Background technology
Cultivating champignon stock is in China, the existing history of more than 800 years so far.Most earlier than Song dynasty Zhejiang Qingyuan County Longyan village
Peasant Wu Sangong invented and cut colored cultivation, the rear diffusion whole nation.Incoming Japan is associated through Buddhist monk.The colored cultivation of cutting of mushroom comes from
China, existing Duan Muchun mycelium inoculations cultivation then comes from Japan.To 1989, Lentinus Edodes In China total yield exceeded Japan, one first
Jump turns into the big country of world's Lentnus edodes first.
India's pyriform spore is a kind of class mycorrhizal fungi for carrying out pure culture, the bacterium with mycelium, mycelia volume, branch or
The mode of round colonizes the intercellular and intracellular in host plant root.There are some researches show：The bacterium can promote 12 section, 24 kinds of plants
Growth, and plant generation system resistance can be induced and improved to the patience of environment stress, the formation for lateral root of accelerating to transplant and
Promote the hardening of micropropagation plant.
How by microbiological culture media apply in the cultivation of mushroom be widely studied the problem of, existing microculture
Base also has the weak point of oneself, and the long-term activity of such as microbiological culture media, engagement each other is studied not fine etc..
The content of the invention
In view of this, the invention provides a kind of India's pyriform spore straw medium preparation method for cultivating mushroom, the party
Method optimizes the handling process of India's pyriform spore, is adopted to improve the deficiency of existing mushroom culture medium, and further provide
The method for carrying out mushroom cultivation with India's pyriform spore straw medium.
First aspect present invention provides a kind of India's pyriform spore straw medium preparation method for cultivating mushroom, step bag
Include：
(1) India's pyriform spore is inoculated in Kafer culture mediums, rotating speed 120~150r/min, shaken cultivation at 26~30 DEG C
6~8d, culture terminates that viable count is measured by sampling, and culture is dissolved in distilled water make in every milliliter of solution viable count be 1 ×
106It is individual；
(2) bacterium number for obtaining step (1) is 1 × 106Individual/mL bacterium solution is sprayed onto in the ratio of inoculum concentration 1~15%
Straw medium surface, produces the India's pyriform spore straw medium for cultivating mushroom at 25~37 DEG C after 12~24h of fermentation.
Second aspect of the present invention provides a kind of mushroom breeding method, and the cultivation that this method obtains above-mentioned preparation method is fragrant
India's pyriform spore straw medium of mushroom is used to cultivate mushroom, and step includes：Aseptically, mushroom mushroom body is cleaned, disappeared
After poison, the bacterial context of stem and cap intersection is taken to be seeded to 50~60d of culture in India's pyriform spore straw medium.
The beneficial effects of the invention are as follows：The present invention is to prepare microbiological culture media, improve the clinical application effect of microorganism
Technical basis is provided；The present invention with the addition of India's pyriform spore, can promote the silk growth of mushroom root, and then improve mushroom production；This hair
It is bright that India's pyriform spore medium culture mushroom is optimized by single factor experiment, optimal breeding condition has been obtained, has been work
The big production of industryization provides technical basis.
Embodiment
First aspect present invention provides a kind of India's pyriform spore straw medium preparation method for cultivating mushroom, step bag
Include：
(1) India's pyriform spore is inoculated in Kafer culture mediums, 120~150r/min of rotating speed, 26~30 DEG C of underlying swingings
6~8d of shaken cultivation in constant temperature speed governing shaking flask cabinet, culture terminates that viable count is measured by sampling, and culture is dissolved in distilled water made
Viable count is 1 × 10 in every milliliter of solution6It is individual；
(2) bacterium number for obtaining step (1) is 1 × 106Individual/mL bacterium solution is sprayed onto in the ratio of inoculum concentration 1~15%
Straw medium surface, produces the India's pyriform spore straw medium for cultivating mushroom at 25~37 DEG C after 12~24h of fermentation.
It is preferred that, the straw medium formula is：By weight percentage, corn stalk powder 62%, wood chip 26%, wheat
Bran 10%, sucrose 1%, gypsum 1%.It is well mixed that straw medium is made according to straw medium formula by raw material grating,
For subsequently using.
More preferred, step (2) described inoculum concentration is 5%.In one embodiment of the invention, solid-state fermentation bacterium
Kind, it is 1 × 10 by viable count6Individual/mL bacterium solution is inoculated into straw medium, inoculum concentration is respectively 1%, 5%, 10%,
15% (V/g), is placed in 34 DEG C of culture 24h.Aseptically, after mushroom body being cleaned, sterilized, stem is removed in cap intersection
Bacterial context is inoculated with, the optimal cultivation temperature culture 50-60d of selection mushroom.Optimal strain inoculation is selected according to mushroom growth situation
Amount.
More preferred, the time of step (2) described fermentation is 18h.In one embodiment of the invention, other are trained
Foster condition is fixed, and is 1 × 10 by viable count6Individual/mL bacterium solution is uniformly sprayed onto in straw medium surface, respectively cultivate 12h,
18h、24h.Aseptically, after mushroom body being cleaned, sterilized, go stem to be inoculated with the bacterial context of cap intersection, select mushroom
Optimal cultivation temperature culture 55d.Optimal fermentation time is selected according to mushroom growth situation.
It is preferred that, India's pyriform spore is inoculated in after Kafer culture mediums in step (1), rotating speed 120~150r/min, 28
Shaken cultivation 7d at DEG C.
Second aspect of the present invention provides a kind of mushroom breeding method, and the cultivation that this method obtains above-mentioned preparation method is fragrant
India's pyriform spore straw medium of mushroom is used to cultivate mushroom, and step includes：Aseptically, mushroom mushroom body is cleaned, disappeared
After poison, the bacterial context of stem and cap intersection is taken to be seeded to 50~60d of culture in India's pyriform spore straw medium.
It is preferred that, the incubation time is 55d.In one embodiment of the invention, according to the optimal fermentation temperature of determination
Degree, fixes other condition of culture, is 1 × 10 by viable count6Individual/mL bacterium solution is inoculated into straw medium, is cultivated respectively
50d, 55d, 60d, rational incubation time is selected according to mushroom growth situation.
It is preferred that, the mushroom choose mushroom type just, robust growth, 7 ripe Fresh Fruit Body of Lentinula Edodes.
A kind of India's pyriform spore straw medium of the cultivation mushroom provided below in conjunction with specific embodiment the present invention
Preparation method is further described.The embodiments described below is exemplary, is only used for explaining the present invention, without being understood that
For limitation of the present invention.
Experimental method in following embodiments, is conventional method unless otherwise specified.Reality used in following embodiments
Test material unless otherwise specified, be that market is commercially available.
Embodiment 1
A kind of India's pyriform spore straw medium preparation method for cultivating mushroom is present embodiments provided, and for cultivating perfume (or spice)
Mushroom, and fermentation time in preparation process is screened.
The composition of kafer culture mediums used uses this area conventional formulation in the present embodiment.
Microorganism used therefor India pyriform spore is provided by Laboratorios Biologicos Farmaceuticos (LABIOFAM) of Changjiang University, microbe-derived to be not limited in
This, market purchase or isolated India's pyriform spore can obtain propinquity effect.
Straw medium formula used is：Corn stalk powder 62%, wood chip 26%, wheat bran 10%, sucrose 1%, gypsum
1%.
A kind of India's pyriform spore straw medium preparation method for cultivating mushroom, step includes：
(1) India's pyriform spore is inoculated in Kafer culture mediums, puts back into shaken cultivation in rotary constant-temperature speed governing shaking flask cabinet, turned
Speed is 120r/min, cultivates 17h at 28 DEG C.Culture terminates that viable count is measured by sampling, and it is 1 × 10 to adjust viable count6Individual/mL.
The method of counting of viable count uses micro- direct counting method.Specifically operating method is：1g cultures are taken to be dissolved in 5ml
It is standby in distilled water.One piece of the blood counting chamber of cleaning is removed, in count block lid lastblock cover glass.The suspension of medicine is shaken
It is even, a small amount of suspension is drawn with pipettor, tally is instilled.Standing is counted in a moment.Count block is found under present low power lens, then
Conversion high power sem observation is simultaneously counted.During counting, if count block is made up of 16 block plaids, by diagonal orientation, number upper left, a left side
Under, upper right, the bacterium number of 4 block plaids in bottom right.If the count block of 25 block plaid compositions, above-mentioned 4 of divisor is generous especially,
Also need the bacterium number of central 1 block plaid of number.Thalline is located on the two-wire of block plaid, and then number is reached the standard grade and do not count offline during counting, and number is left
Line does not count right line, to reduce error.
Contain bacterium average (N) × coefficient (K=4 × 10 in bacterial population=each small lattice in per ml suspensions6) × mixed
Suspension extension rate (d).
(2) other condition of culture are fixed, the bacterium solution that (1) is obtained uniformly is sprayed onto in straw medium surface, is put respectively
In cultivating 12h, 18h, 24h at 28 DEG C, the corresponding India's pyriform spore straw medium for cultivating mushroom of different fermentations condition is produced.
India's pyriform spore straw medium of above-mentioned cultivation mushroom is respectively used to mushroom culture, step is as follows：Selection is fragrant
Mushroom mushroom type just, robust growth, 7 ripe Fresh Fruit Body of Lentinula Edodes be used as inoculation material.Aseptically, mushroom body is cleaned,
After sterilization, stem is gone to be inoculated with the bacterial context of cap intersection, the optimal cultivation temperature culture 55d of selection mushroom.After culture terminates,
Determine the optimized proportion of root filament length degree and yield compared to straw medium yield of India pyriform spore culture medium mushroom.As a result see
Table 1.
Mushroom root filament length degree and output optimization ratio under the different fermentations time of table 1
According to the result of table 1, it is determined that optimal fermentation time is 18h.
Embodiment 2
A kind of India's pyriform spore straw medium preparation method for cultivating mushroom is present embodiments provided, and for cultivating perfume (or spice)
Mushroom, and strain inoculum concentration optimal in preparation process is screened.
(1) according to the result of embodiment one, the solid-state fermentation time is 18h, and matching somebody with somebody for culture medium is changed by single factor experiment
Side, specific formula is shown in Table 2.
The formula of the India's pyriform spore straw medium of table 2
According to straw medium formula by raw material grating, straw medium culture is mixed and made into standby.
(2) change the formula of culture medium by single factor experiment, be 1 × 10 by viable count according to the testing program of table 26
Individual/mL bacterium solution is inoculated into straw medium, and inoculum concentration is respectively 1%, 5%, 10%, 15% (V/g), is placed in 34 DEG C of cultures
24h, produces the corresponding India's pyriform spore straw medium for cultivating mushroom of different fermentations condition；
India's pyriform spore straw medium of above-mentioned cultivation mushroom is respectively used to mushroom culture, step is as follows：Selection is fragrant
Mushroom mushroom type just, robust growth, 7 ripe Fresh Fruit Body of Lentinula Edodes be used as inoculation material.Aseptically, mushroom body is cleaned,
After sterilization, stem is gone to be inoculated with the bacterial context of cap intersection, the optimal cultivation temperature culture 50d of selection mushroom.The mushroom training
Foster temperature can be 28 DEG C.The root filament length degree and yield of India pyriform spore culture medium mushroom are determined compared to straw medium yield
Optimized proportion.It the results are shown in Table 3.
Mushroom root filament length degree and Unit Weight optimization ratio under the conditions of the different vaccination amount of table 3
According to the result of table 3, it is No. 2, i.e. India's pyriform spore to determine microbiological culture media：Straw medium is 5:95.
Embodiment 3
A kind of India's pyriform spore straw medium preparation method for cultivating mushroom is present embodiments provided, and for cultivating perfume (or spice)
Mushroom, and mushroom incubation time optimal in preparation process is screened.
A kind of India's pyriform spore straw medium preparation method for cultivating mushroom, step includes：
(1) India's pyriform spore is inoculated in Kafer culture mediums, rotating speed 150r/min, shaken cultivation 7d, culture knot at 28 DEG C
Beam sampler determines viable count, and culture, which is dissolved in distilled water, makes viable count in every milliliter of solution be 1 × 106It is individual；
(2) bacterium number for obtaining step (1) is 1 × 106Individual/mL bacterium solution is sprayed onto stalk in the ratio of inoculum concentration 5%
Media surface, produces the India's pyriform spore straw medium for cultivating mushroom at 34 DEG C after fermentation 24h.
India's pyriform spore straw medium of above-mentioned cultivation mushroom is respectively used to mushroom culture, step is as follows：
Selection mushroom mushroom type just, robust growth, 7 ripe Fresh Fruit Body of Lentinula Edodes be used as inoculation material.In aseptic condition
Under, after mushroom body is cleaned, sterilized, go stem to be inoculated with the bacterial context of cap intersection, the optimal cultivation temperature culture of selection mushroom
50d、55d、60d.The root filament length degree and yield for determining India pyriform spore culture medium mushroom are excellent compared to straw medium yield
Change ratio.It the results are shown in Table 4.
Mushroom root filament length degree and Weight-optimised ratio under each incubation time of table 4
According to the result of table 4, it is determined that the time of culture mushroom is 55d.
The foregoing is only presently preferred embodiments of the present invention, be not intended to limit the invention, it is all the present invention spirit and
Within principle, any modification, equivalent substitution and improvements made etc. should be included in the scope of the protection.
Claims (8)
1. a kind of India's pyriform spore straw medium preparation method for cultivating mushroom, step includes：
(1) India's pyriform spore is inoculated in Kafer culture mediums, rotating speed 120~150r/min, shaken cultivation 6 at 26~30 DEG C~
8d, culture terminates that viable count is measured by sampling, and culture, which is dissolved in distilled water, makes viable count in every milliliter of solution be 1 × 106
It is individual；
(2) bacterium number for obtaining step (1) is 1 × 106Individual/mL bacterium solution is sprayed onto stalk training in the ratio of inoculum concentration 1~15%
Support and produce the India's pyriform spore straw medium for cultivating mushroom after 12~24h of fermentation at primary surface, 25~37 DEG C.
2. India's pyriform spore straw medium preparation method of mushroom is cultivated as claimed in claim 1, it is characterised in that：It is described
Straw medium formula is：By weight percentage, corn stalk powder 62%, wood chip 26%, wheat bran 10%, sucrose 1%, gypsum
1%.
3. India's pyriform spore straw medium preparation method of mushroom is cultivated as claimed in claim 1, it is characterised in that：Step
(1) India's pyriform spore is inoculated in after Kafer culture mediums in, rotating speed 120~150r/min, shaken cultivation 7d at 28 DEG C.
4. India's pyriform spore straw medium preparation method of mushroom is cultivated as claimed in claim 2, it is characterised in that：Step
(2) inoculum concentration is 5%.
5. India's pyriform spore straw medium preparation method of mushroom is cultivated as claimed in claim 2, it is characterised in that：Step
(2) time of the fermentation is 18h.
6. a kind of mushroom breeding method, it is characterised in that：The print for the cultivation mushroom that preparation method described in claim 1 is obtained
Degree pyriform spore straw medium is used to cultivate mushroom, and step includes：Aseptically, after mushroom mushroom body being cleaned, sterilized, take
The bacterial context of stem and cap intersection is seeded to 50~60d of culture in India's pyriform spore straw medium.
7. mushroom breeding method as claimed in claim 6, it is characterised in that：The incubation time is 55d.
8. mushroom breeding method as claimed in claim 6, it is characterised in that：The mushroom choose mushroom type just, robust growth, 7
Ripe Fresh Fruit Body of Lentinula Edodes.
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| CN201710497457.1A CN107216201B (en) | 2017-06-27 | 2017-06-27 | Preparation method of medium for cultivating sorrel straw of lentinus edodes |
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| CN201710497457.1A CN107216201B (en) | 2017-06-27 | 2017-06-27 | Preparation method of medium for cultivating sorrel straw of lentinus edodes |
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| WO2015001575A1 (en) * | 2013-07-05 | 2015-01-08 | Amity University | Plant growth promoting formulation of piriformospora indica and azotobacter chroococcum with talcum powder |
| CN105794455A (en) * | 2016-03-18 | 2016-07-27 | 浙江大学 | Method for utilizing piriformospora indica and Zhongshengmycin to jointly prevent and control bacterial wilt of tobaccos |
| US20170112137A1 (en) * | 2015-10-26 | 2017-04-27 | Ut-Battelle, Llc | Complex of mutualistic microbes designed to increase plant productivity |
-
2017
- 2017-06-27 CN CN201710497457.1A patent/CN107216201B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2015001575A1 (en) * | 2013-07-05 | 2015-01-08 | Amity University | Plant growth promoting formulation of piriformospora indica and azotobacter chroococcum with talcum powder |
| CN103858719A (en) * | 2014-03-11 | 2014-06-18 | 浙江大学 | Method for reducing cadmium content of overground part of paddy rice |
| US20170112137A1 (en) * | 2015-10-26 | 2017-04-27 | Ut-Battelle, Llc | Complex of mutualistic microbes designed to increase plant productivity |
| CN105794455A (en) * | 2016-03-18 | 2016-07-27 | 浙江大学 | Method for utilizing piriformospora indica and Zhongshengmycin to jointly prevent and control bacterial wilt of tobaccos |
Non-Patent Citations (2)
| Title |
|---|
| 刘晓曦: "印度梨形孢培养条件优化和剂型研制及在油菜上的应用研究", 《中国优秀硕士学位论文全文数据库 农业科学辑》 * |
| 朱志炎等: "印度梨形孢（Piriformospora indica）的生物学特性及对植物生长的互作效应研究进展", 《长江大学学报（自然科学版）》 * |
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