CN108424856A - A method of producing agricultural microbial agent using maize germ grease - Google Patents
A method of producing agricultural microbial agent using maize germ grease Download PDFInfo
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- CN108424856A CN108424856A CN201810239229.9A CN201810239229A CN108424856A CN 108424856 A CN108424856 A CN 108424856A CN 201810239229 A CN201810239229 A CN 201810239229A CN 108424856 A CN108424856 A CN 108424856A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
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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
- C05F11/00—Other organic fertilisers
- C05F11/08—Organic fertilisers containing added bacterial cultures, mycelia or the like
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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Abstract
The invention discloses a kind of methods producing agricultural microbial agent using maize germ grease, include the following steps：First, the pretreatment of maize germ；Secondly, continue wet steaming by water vapour to soften, control the rotating speed and temperature of bombing bore, and the grease of acquisition is filtered purifying；Furthermore above-mentioned maize germ grease is sterilized, and culture medium is added wherein；Finally, it moves up plant quarantine in mixed culture medium and surveys strain, until there is bacterium colony on mixed culture medium, carry out inactivation operation；Strain on above-mentioned mixed culture medium is subjected to constant temperature selectively culture, until all growing strain on culture medium, culture medium crushing is mixed into organic fertilizer and carries out constant temperature and humidity fermentation acquisition microbial inoculum；By directly preparing maize germ grease, convenient for the cultivation and detection of follow-up strain, gel solidification is carried out on the basis of culture medium, it is with strong applicability convenient for observation in real time, selective culture can be carried out, in a short time the specified microbial inoculum of mass propagation.
Description
Technical field
The present invention relates to technical field of microbial detection, specially a kind of to produce agricultural microorganism using maize germ grease
The method of microbial inoculum.
Background technology
Microbial inoculum can play the role of very important in agricultural production, wherein most significant effect is exactly to improve soil
Quality improves the yield of crops, and improves the quality of crops.Microbial inoculum can reach above-mentioned advantageous effect, and reason exists
In：
First, nutrient is more comprehensive, based on organic fertilizer, while being also configured with corresponding microorganism fungus kind, has
The characteristics of multifunctional fertilizers；
Secondly, organic principle and element in microbial inoculum can be quickly dissolved in water so that nutritional ingredient therein etc. with
Dampening and enter soil, reach better effect；
Third is a large amount of strains contained by inside, and a large amount of breeding in the soil releases protease and organic acid etc., has
There are dissolving phosphorus, potassium decomposing and fixed nitrogen and other effects, while the residual of pesticide can be reduced.
Although the using effect of microbial inoculum is clearly, the preparation of microbial inoculum is very difficult.As long as this due to
Under the conditions of the prior art, specified strain can not be quickly obtained, and then corresponding microbial inoculum is obtained by the mass propagation of strain.
And in configuration process, need to be detected strain, and the method for inspection of national standard is still very traditional, is primarily present
Following defect：It is cumbersome, it sterilizes from the preparation of culture medium to practice examining, last sterilizing and washing is in micro- inspection conventional method
Essential step, these operations are high to technical staff's professional skill requirement, and prepare to expend with aftermath a large amount of
Time seriously affects working efficiency；Sensitivity is low.
Therefore conventional method is unfavorable for quick detection of the enterprise to production line, and most importantly, specific to prepare
Process cost is higher, it is difficult to fast and effectively prepare the product for reaching requirement, and its product quality reliability for preparing
Difference.
Invention content
In order to overcome the shortcomings of prior art, present invention offer is a kind of to produce agricultural micro- life using maize germ grease
The method of object microbial inoculum is convenient for the cultivation and detection of follow-up strain, on the basis of culture medium by directly preparing maize germ grease
Upper carry out gel solidification, it is with strong applicability convenient for observation in real time, it can according to circumstances be improved, examined in real time at the scene
Fertile environment is rapidly achieved when survey, mass propagation obtains visible testing result in a short time, ensure that strain
The rapidity and reliability of detection can effectively solve the problem that the problem of background technology proposes.
The technical solution adopted by the present invention to solve the technical problems is：
A method of agricultural microbial agent being produced using maize germ grease, is included the following steps：
The pretreatment of step 100, maize germ, corn is crushed, and is screened by double-deck screen shale shaker；
The softening and squeezing of step 200, maize germ continue wet steamings by water vapour and soften, and adjusting temperature and vapor contain
Treated plumule is sent into press chamber, controls the rotating speed and temperature of bombing bore, and the grease of acquisition is filtered pure by amount
Change；
Step 300 sterilizes above-mentioned maize germ grease, and wherein according to mass fraction（8~10）：（2~4）It is added
Culture medium；
Step 400 moves up plant quarantine survey strain in mixed culture medium, and carries out constant temperature incubation, until gelatinous mixed culture
Occur bacterium colony on base upper surface, and directly inactivated by high pressure steam process, direct aseptic packaging is spare later；
Strain on above-mentioned mixed culture medium is carried out constant temperature selectively culture by step 500, until all growing bacterium on culture medium
Kind, culture medium crushing is mixed into organic fertilizer and carries out constant temperature and humidity fermentation acquisition microbial inoculum.
As a kind of preferred technical solution of the present invention, in step 100, then by being centrifuged caused by cyclone separator
Power isolates pure plumule, and the maize germ isolated, which is dried to moisture content, is less than 10%.
As a kind of preferred technical solution of the present invention, 1.5 х of top mesh, 1.5 mesh of double-deck screen shale shaker/centimetre, under
The mesh in portion be 4 х, 4 mesh/centimetre, 5 х, 5 mesh/centimetre or 7 х, 7 mesh/centimetre in any one.
As a kind of preferred technical solution of the present invention, in step 200, the wet temperature for steaming softening is controlled in 80-100
DEG C, humidity increases temperature to 120-140 DEG C after softening in 40-60% according to the rate of 5-8 DEG C/min, and humidity reduces
To 2-4%, in 8-10r/min, temperature is controlled at 140-180 DEG C for the rotating speed control of bombing bore.
As a kind of preferred technical solution of the present invention, the moisture content of plumule is reduced to 3%-4% after wet steaming softening.
As a kind of preferred technical solution of the present invention, in step 300, the ingredient of culture medium is specific according to parts by weight
For：
8-10 parts of Yeast protein, 20-25 parts of corn starch, 2-5 parts of peptone, 0.3-0.5 parts of sodium chloride, dipotassium hydrogen phosphate 1-3
Part, 2-5 parts of glucose, 70-90 parts of sterile purified water.
As a kind of preferred technical solution of the present invention, after culture medium is added, add wherein after adding water to stir evenly
Enter gel, until mixed culture medium becomes gel after cooling.
As a kind of preferred technical solution of the present invention, the detection strain is in lactic acid bacteria, saccharomycete or bacillus
Any one or a variety of combinations.
The area of bacterium colony is first appeared in 0.1- in step 400 as a kind of preferred technical solution of the present invention
0.5mm or clump count carry out high-temperature inactivation when being more than 3.
As a kind of preferred technical solution of the present invention, in step 500, selectively cultivate the specific steps are：
Mixed culture medium is carried out constant temperature incubation 1-2h by step 501, until the strain of media surface, which all activates, bacterium occurs
It falls；
Step 502 crushes mixed culture medium, and selective bactericidal agent is sprinkled on mixed culture medium, constant temperature incubation 2-3h；
Nutrient solution is sprinkled in step 503, the mixed culture medium after above-mentioned crushing persistently to cultivate, until strain all activates.
Compared with prior art, the beneficial effects of the invention are as follows：
（1）The present invention is by directly preparing maize germ grease, the case where capable of adjusting maize germ grease according to demand, and
And can also ensure that the actual product quality of maize germ grease, it is convenient for the cultivation and detection of follow-up strain；
（2）In the preparation of maize germ grease, the utilization rate of maize germ grease is improved, reduces original maize germ
Usage amount；
（3）Combine maize germ oil to form mixed culture medium with culture medium, and is coagulated on the basis of culture medium
Adhesive curing, the concrete condition convenient for the upper bacterium colony to culture medium are observed in real time, and in specific culture, Ke Yigen
According to needing the agricultural microorganism that detects to select different microorganism fungus kind types, and be not limited to specifically to limit in present embodiment
Strain, it is with strong applicability and during improved and uncomplicated, technology is required it is not high, can basis at the scene
Situation is improved in real time；
（4）The present invention by carrying out the culture of strain in advance so that strain has formd stable growing environment, can train
It is rapidly achieved fertile environment when foster, in a short time mass propagation, and visible selectivity can be quickly obtained
Cultivation results ensure that the rapidity and reliability of identification so that result more has convincingness, is selectively cultivating it
The strain for being suitble to culture can be gone out with mass propagation afterwards, obtain specified microbial inoculum.
Description of the drawings
Fig. 1 is the flow diagram of the present invention.
Specific implementation mode
Following will be combined with the drawings in the embodiments of the present invention, and technical solution in the embodiment of the present invention carries out clear, complete
Site preparation describes, it is clear that described embodiments are only a part of the embodiments of the present invention, instead of all the embodiments.It is based on
Embodiment in the present invention, it is obtained by those of ordinary skill in the art without making creative efforts every other
Embodiment shall fall within the protection scope of the present invention.
As shown in Figure 1, the present invention provides a kind of method producing agricultural microbial agent using maize germ grease, packet
Include following steps：
The pretreatment of step 100, maize germ, corn is crushed, and is screened by double-deck screen shale shaker.
Preferably, 1.5 х of top mesh, 1.5 mesh of double-deck screen shale shaker/centimetre, the mesh of its underpart be 4 х, 4 mesh/li
Rice, 5 х, 5 mesh/centimetre or 7 х, 7 mesh/centimetre in any one.
It is big miscellaneous among the above, being removed by the screening net on top, it is small miscellaneous by the screening net removing of bottom, at this point, by root
The screening net using which kind of specification is determined according to the number of screenings kind impurity, to reduce the loss of maize germ, in screening
After need timely by tray or chute using clear water rinsing several times, until surface is no longer contaminated with clast impurity, carry
The pure rate of high maize germ, avoids the pollution of impurity.
In step 100, go out pure plumule by centrifugal force separate caused by cyclone separator again later, will detach
The maize germ gone out is dried to moisture content and is less than 10%, reduces the moisture content of maize germ, is on the one hand to prevent maize germ
In the case that moisture content it is higher due to the effect heat production of the general breathings such as accumulation causes temperature to increase and the case where germinateing, occurs, separately
Outer one side is prevented during storage and transport, and maize germ itself goes mouldy.
The softening and squeezing of step 200, maize germ continue wet steaming by water vapour and soften, adjust temperature and vapor
Content, it is wet steam softening after the moisture content of plumule be reduced to 3%-4%, treated plumule is sent into press chamber, control
The rotating speed and temperature of bombing bore, and the grease of acquisition is filtered purifying.
The effect of steam humidifying is so that the plasticity of maize germ changes, and then in subsequent processing procedure not
It is easy to happen direct mechanical crushing, convenient for oil expression, and the impurity of oil expression can be prevented excessive.But in the process of softening
In, it needs to control specific temperature, prevents temperature is excessively high from leading to protein deformation etc. so that dispensing follows the string and influences follow-up
Steaming fry and oil expression etc..
In above-mentioned steps, the wet temperature control for steaming softening is at 80-100 DEG C, and humidity is in 40-60%, and after softening
Temperature is increased to 120-140 DEG C according to the rate of 5-8 DEG C/min, and humidity is reduced to 2-4%, the rotating speed of bombing bore to control in 8-10r/
Min, temperature are controlled at 140-180 DEG C.
In the steaming for reducing moisture content is fried, material embryo color transition can smell fragrance and burnt odor cannot occur to brownish red
It is advisable.
Step 300 sterilizes above-mentioned maize germ grease, and wherein according to mass fraction（8~10）：（2~4）
Culture medium is added.
Wherein, in step 300, the ingredient of culture medium is specially according to parts by weight：
8-10 parts of Yeast protein, 20-25 parts of corn starch, 2-5 parts of peptone, 0.3-0.5 parts of sodium chloride, dipotassium hydrogen phosphate 1-3
Part, 2-5 parts of glucose, 70-90 parts of sterile purified water.
After culture medium is added, be added gel after adding water to stir evenly wherein, after cooling until mixed culture medium at
For gel.
By the way that gel is added after stirring evenly, the culture medium for being originally used for liquid can be made to become solid-state, convenient for specific
Culture, and the practical growing state of bacterium colony can be more clearly seen on solid culture medium, be convenient for judging.
Step 400 moves up plant quarantine survey strain in mixed culture medium, and carries out constant temperature incubation, until gelatinous mixing
Occur bacterium colony on culture medium upper surface, and inactivation operation is directly carried out by high pressure steam process, direct aseptic packaging is spare later.
Further illustrate, it is described detection strain be lactic acid bacteria, saccharomycete or bacillus in any one or
A variety of combinations.
It is inoculated with bacterium colony on above-mentioned mixed culture medium, is so that bacterium colony forms stable growth ring on culture medium in advance
Border, and inactivated at once once after bacterium colony is formed, retain growing environment of the bacterium colony on gel, be convenient for the detection of follow-up strain,
It avoids bacterium colony during detection and needs to form specific environment again and can breed, improve the sensitive of identification
Property and reliability, can detect extremely micro strain, while can greatly shorten specific detection time.
In addition, it is desirable to illustrate, in step 400, the area of bacterium colony is first appeared in 0.1-0.5mm or bacterium
It falls when number is more than 3 and carries out high-temperature inactivation.
To ensure stability existing for bacterium colony, in the presence of existing bacterium colony, and can be so that bacterium colony forms one
The mark of comparison, so that identification is more for convincingness.
Strain on above-mentioned mixed culture medium is carried out constant temperature selectively culture by step 500, until all being grown on culture medium
Culture medium crushing is mixed into organic fertilizer and carries out constant temperature and humidity fermentation acquisition microbial inoculum by strain.
In above-mentioned steps, selectively cultivate the specific steps are：
Mixed culture medium is carried out constant temperature incubation 1-2h by step 501, until the strain of media surface, which all activates, bacterium occurs
It falls；
Step 502 crushes mixed culture medium, and selective bactericidal agent is sprinkled on mixed culture medium, constant temperature incubation 2-3h；
Nutrient solution is sprinkled in step 503, the mixed culture medium after above-mentioned crushing persistently to cultivate, until strain all activates.
Visible selective cultivation results can be quickly obtained, the rapidity and reliability of identification is ensure that, is selecting
The strain for being suitble to culture can be gone out after the culture of selecting property with mass propagation, obtain specified microbial inoculum.
It is obvious to a person skilled in the art that invention is not limited to the details of the above exemplary embodiments, Er Qie
In the case of without departing substantially from spirit or essential attributes of the invention, the present invention can be realized in other specific forms.Therefore, no matter
From the point of view of which point, the present embodiments are to be considered as illustrative and not restrictive, and the scope of the present invention is by appended power
Profit requires rather than above description limits, it is intended that all by what is fallen within the meaning and scope of the equivalent requirements of the claims
Variation is included within the present invention.Any reference signs in the claims should not be construed as limiting the involved claims.
Claims (10)
1. a kind of method producing agricultural microbial agent using maize germ grease, which is characterized in that include the following steps：
The pretreatment of step 100, maize germ, corn is crushed, and is screened by double-deck screen shale shaker；
The softening and squeezing of step 200, maize germ continue wet steamings by water vapour and soften, and adjusting temperature and vapor contain
Treated plumule is sent into press chamber, controls the rotating speed and temperature of bombing bore, and the grease of acquisition is filtered pure by amount
Change；
Step 300 sterilizes above-mentioned maize germ grease, and wherein according to mass fraction（8~10）：（2~4）It is added
Culture medium；
Step 400 moves up plant quarantine survey strain in mixed culture medium, and carries out constant temperature incubation, until gelatinous mixed culture
Occur bacterium colony on base upper surface, and directly inactivated by high pressure steam process, direct aseptic packaging is spare later；
Strain on above-mentioned mixed culture medium is carried out constant temperature selectively culture by step 500, until all growing bacterium on culture medium
Kind, culture medium crushing is mixed into organic fertilizer and carries out constant temperature and humidity fermentation acquisition microbial inoculum.
2. a kind of method producing agricultural microbial agent using maize germ grease according to claim 1, feature
It is, in step 100, then goes out pure plumule by centrifugal force separate caused by cyclone separator, the jade that will be isolated
Rice plumule is dried to moisture content and is less than 10%.
3. a kind of method producing agricultural microbial agent using maize germ grease according to claim 1, feature
Be, 1.5 х of top mesh, 1.5 mesh of double-deck screen shale shaker/centimetre, the mesh of its underpart be 4 х, 4 mesh/centimetre, 5 х, 5 mesh/li
Rice or 7 х, 7 mesh/centimetre in any one.
4. a kind of method producing agricultural microbial agent using maize germ grease according to claim 1, feature
Be, in step 200, the wet temperature control for steaming softening at 80-100 DEG C, humidity in 40-60%, and after softening according to
The rate of 5-8 DEG C/min increases temperature to 120-140 DEG C, and humidity is reduced to 2-4%, the rotating speed of bombing bore to control in 8-10r/min,
Temperature is controlled at 140-180 DEG C.
5. a kind of method producing agricultural microbial agent using maize germ grease according to claim 1, feature
It is, the moisture content of plumule is reduced to 3%-4% after wet steaming softening.
6. a kind of method producing agricultural microbial agent using maize germ grease according to claim 1, feature
It is, in step 300, the ingredient of culture medium is specially according to parts by weight：
8-10 parts of Yeast protein, 20-25 parts of corn starch, 2-5 parts of peptone, 0.3-0.5 parts of sodium chloride, dipotassium hydrogen phosphate 1-3
Part, 2-5 parts of glucose, 70-90 parts of sterile purified water.
7. a kind of method producing agricultural microbial agent using maize germ grease according to claim 1, feature
It is, after culture medium is added, gel is added after adding water to stir evenly wherein, until mixed culture medium becomes solidifying after cooling
It is gluey.
8. a kind of method producing agricultural microbial agent using maize germ grease according to claim 1, feature
It is, the detection strain is any one or a variety of combinations in lactic acid bacteria, saccharomycete or bacillus.
9. a kind of method producing agricultural microbial agent using maize germ grease according to claim 1, feature
It is, in step 400, the area for first appearing bacterium colony carries out high-temperature inactivation when 0.1-0.5mm or clump count are more than 3.
10. a kind of method producing agricultural microbial agent using maize germ grease according to claim 1, feature
Be, in step 500, selectively cultivate the specific steps are：
Mixed culture medium is carried out constant temperature incubation 1-2h by step 501, until the strain of media surface, which all activates, bacterium occurs
It falls；
Step 502 crushes mixed culture medium, and selective bactericidal agent is sprinkled on mixed culture medium, constant temperature incubation 2-3h；
Nutrient solution is sprinkled in step 503, the mixed culture medium after above-mentioned crushing persistently to cultivate, until strain all activates.
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| CN201810239229.9A CN108424856A (en) | 2018-03-22 | 2018-03-22 | A method of producing agricultural microbial agent using maize germ grease |
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Application publication date: 20180821 |