CN108373978A - A kind of microorganism formulation and its application - Google Patents
A kind of microorganism formulation and its application Download PDFInfo
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- CN108373978A CN108373978A CN201810145996.3A CN201810145996A CN108373978A CN 108373978 A CN108373978 A CN 108373978A CN 201810145996 A CN201810145996 A CN 201810145996A CN 108373978 A CN108373978 A CN 108373978A
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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
- C12N1/14—Fungi; Culture media therefor
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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
- A01G22/00—Cultivation of specific crops or plants not otherwise provided for
- A01G22/05—Fruit crops, e.g. strawberries, tomatoes or cucumbers
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/30—Microbial fungi; Substances produced thereby or obtained therefrom
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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 microorganism formulation and its applications, belong to microorganisms technical field.The present invention is compounded using the agriculture microorganism such as photosynthetic bacteria, rhizobium, trichoderma pseudokiningii bacterium with that can be metabolized the lactobacillus plantarum for generating synergistic effect with growth, the microbial inoculum of safety non-pollution is prepared, microorganism formulation prepared by the present invention prepares in field of food and all has important application potential by the product of primary raw material or field of agricultural cultivation of capsicum.
Description
Technical field
The present invention relates to a kind of microorganism formulation and its applications, belong to microorganisms technical field.
Background technology
Capsicum is the main cultivated plant in China, but capsicum pest and disease damage is increasing on the influence of the yield and quality of capsicum,
Effective prevention of capsicum pest and disease damage is the basis of capsicum harvesting high-quality high-yield cultivation.With the continuous expansion of cultivated area, pest and disease damage is anti-
Controlling is particularly important for increasing yield, reducing economic loss.
Chemical method prevention at present is still the main method during the prevention and control of plant diseases, pest control.Chemical pesticide is because it is to disease height
The features such as imitating, is quick-acting, easy to operate is widely used in the prevention of tomato disease, but being excessively used for these medicaments can make to prevent
It controls object to develop immunity to drugs, and destroys soil microflora, also cause the problems such as agricultural product remain, is exceeded, seriously affect
People, animal safety, or even destroy the balance of the natural agriculture ecosystem.With the raising of people's environmental protection and nuisanceless consciousness,
Start to explore the use for reducing chemical pesticide using various methods, to reduce harm and environmental pollution to people, animal.Plant source
Fungicide as a kind of novel biological agent, have the characteristics that it is nontoxic, pollution-free, be easy to preserve transport, thus with wide
Application before
Invention content
The invention solves first technical problem be to provide it is a kind of reduce capsicum pest and disease damage microorganism formulation, it is described
Microbial inoculum is active biological product, and active constituent is conciliate by red pseudomonas, trichoderma pseudokiningii bacterium, rhizobium, lactobacillus plantarum
Bacillus amyloliquefaciens form.
In one embodiment of the invention, the microorganism formulation is to comprise the following steps：By red pseudomonas,
Trichoderma pseudokiningii bacterium, rhizobium, lactobacillus plantarum and bacillus amyloliquefaciens are respectively according to 1~9.9 × 105The inoculation of CFU/mL
Amount is inoculated in same fermentation medium, and 30 DEG C~35 DEG C are sealed by fermentation 5~7 days, until OD600It is 3.0~3.5, is cultivated
Ripe microbial inoculum
In one embodiment of the invention, the fermentation medium components are by weight percent：
KH2PO40.05%, K2HPO4·3H2O 0.05%, MgSO4·7H2O 0.05%, NaCl 0.1%, urea 1%~4%, second
Sour sodium 0.2%~0.5%, yeast extract 0.01%~0.06%, boils 15min, is cooled to 40 DEG C hereinafter, using NaHCO3It adjusts
PH 6.8~7.2.
In one embodiment of the invention, the step of microbial inoculum is also cultivated by expanding, the expansion culture
It is as follows：The ripe microbial inoculum of culture is directly inoculated in fermentation medium, inoculum concentration is 15%~25%, 30 in transparent vessel
DEG C~35 DEG C be sealed by fermentation 5~7 days, until OD600It is 3.0~3.5, microbial inoculum is cultivated ripe；The fermentation medium components press quality
Percentage is：KH2PO40.05%, K2HPO4·3H2O 0.05%, MgSO4·7H2O 0.05%, NaCl 0.1%, urea 1%
~4%, sodium acetate 0.2%~0.5%, yeast extract 0.01%~0.06% boils 15min, is cooled to 40 DEG C hereinafter, using
NaHCO3Adjust pH 6.8~7.2.
In one embodiment of the invention, the microorganism formulation, by the bacterium solution after inoculation in incubation
It is placed under outdoor solar light, cloudy day and night, intensity of illumination was 1500~2000lx, is cultivated using incandescent lamp supplementary light culture
Container is shaken in journey for every eight hours 2-3 times, illumination is obtained to help the bacterium precipitated to float.
Second technical problem that the present invention solves is to provide a kind of preparation of the microorganism formulation of prevention capsicum pest and disease damage
Method, the method are by red pseudomonas, trichoderma pseudokiningii bacterium, rhizobium, lactobacillus plantarum and bacillus amyloliquefaciens point
Not with 1~9.9 × 105The inoculum concentration of CFU/mL is inoculated in same fermentation medium, is placed under outdoor solar light, 30 DEG C~35 DEG C
It is sealed by fermentation 5~7 days, cloudy day and night use incandescent lamp supplementary light culture, intensity of illumination to be cultivated in 1500~2000lx
Timing daily shakes container 2-3 times in journey；To OD600It is 3.0~3.5, bacterium solution is cultivated ripe.
In one embodiment of the invention, the fermentation medium components are as follows：KH2PO40.05%, K2HPO4·
3H2O 0.05%, MgSO4·7H2O 0.05%, NaCl 0.1%, urea 1%~4%, sodium acetate 0.2%~0.5%, yeast
Cream 0.01%~0.06%.
The third technical problem that the present invention solves is to provide application of institute's microorganism formulation in capsicum production, cultivation.
In one embodiment of the invention, the application refers to using difference dilute respectively in capsicum cultivating process
Release microbial inoculum seed soaking, root dipping transplanting, pouring root or the sprinkling liquid level of multiple.
In one embodiment of the invention, the application is that microbial inoculum is diluted 200~300 times, 2~3d sprinklings one
Secondary solution.
Beneficial effects of the present invention：The present invention using the agriculture microorganism such as photosynthetic bacteria, rhizobium, trichoderma pseudokiningii bacterium with
It can be metabolized the lactobacillus plantarum compounding for generating synergistic effect with growth, prepare the microbial inoculum of safety non-pollution, can significantly carry
High capsicum emergence rate, height of seedling, overground part fresh weight, main root length etc. improve product quality and biological yield, the shortening training of bud capsicum
Educate the period；And have obvious action to the prevention of powdery mildew, anthracnose, gray mold, aphid, make gray mold incidence 2.32~
2.65%, anthracnose incidence 2.3~2.62%, aphid incidence 3.71~4.23%, powdery mildew incidence 2.48~
2.83%, 2~7 percentage points are reduced compared with the control.The microbial inoculum, which makes, simultaneously simply easily implements, is safe and reliable；It cultivated
Journey does not need shaking table, large fermentation tank, and summer can ferment under normal temperature environment, remaining season only needs to provide simple heat preservation
Measure.
Specific implementation mode
Below in conjunction with specific embodiment, the operating method that the present invention is further explained.But these embodiments are only used in detail
It describes the bright present invention in detail, rather than limits the scope of the invention.
Embodiment 1
(1) preparation of fermentation medium：Medium component is (by weight percent)：KH2PO40.05%, K2HPO4·
3H2O 0.05%, MgSO4·7H2O 0.05%, NaCl 0.1%, urea 1%~4%, sodium acetate 0.2%~0.5%, yeast
Cream 0.01%~0.06%, and continue to boil 15min, pH uses NaHCO after cooling3PH 6.8~7.2 is adjusted, by culture medium point
It is filled in sealable transparent round；
(2) preparation of microbial inoculum：The microbial inoculum is made of red pseudomonas, trichoderma pseudokiningii bacterium, rhizobium, by 3 kinds of microorganisms
Respectively according to 1~9.9 × 105The inoculum concentration of CFU/mL is inoculated with, and is inoculated in transparent vessel, is placed under outdoor solar light, 30 DEG C~
35 DEG C are sealed by fermentation 5~7 days, and cloudy day and night use incandescent lamp supplementary light culture, intensity of illumination to maintain 1500~2000lx,
Timing daily shakes container 2-3 times in cultivating process, and illumination is obtained to help the bacterium precipitated to float.It cultivates to OD600It is 2.0
~2.5, bacterium solution color is peony at this time.
(3) microbial inoculum simply expands culture：In order to which daily production is cultivated, microbial inoculum is simple and efficient, can be by culture in step (2)
Bacterium solution is inoculated in the culture medium in step (1) according to 20%~30% inoculum concentration.30 DEG C~35 DEG C are sealed by fermentation 5~7 days,
Cloudy day and night use incandescent lamp supplementary light culture, intensity of illumination to maintain 1500~2000lx, timing daily in cultivating process
It shakes container 2-3 times, illumination is obtained to help the bacterium precipitated to float.Cultivate ripe bacterium solution OD600It is 2.0~2.5.
In above-mentioned steps (2) and step (3), viable bacteria content can reach 150~20,000,000,000/mL in bacterium solution.
Embodiment 2
(1) preparation of fermentation medium：Medium component is (by weight percent) KH2PO40.05%, K2HPO4·3H2O
0.05%, MgSO4·7H2O 0.05%, NaCl 0.1%, urea 1%~4%, sodium acetate 0.2%~0.5%, yeast extract
0.01%~0.06%, and continue to boil 15min, pH uses NaHCO after cooling3PH 6.8~7.2 is adjusted, culture medium is dispensed
To in sealable transparent round；
(2) preparation of microbial inoculum：The microbial inoculum is formed sediment by red pseudomonas, trichoderma pseudokiningii bacterium, rhizobium, lactobacillus plantarum reconciliation
Afnyloliquefaciens form, by 5 kinds of microorganisms respectively according to 1~9.9 × 104The inoculum concentration of CFU/mL is inoculated in transparent sealing container
In, it is placed under outdoor solar light, 30 DEG C~35 DEG C are sealed by fermentation 5~7 days, and cloudy day and night use incandescent lamp supplementary light culture,
Intensity of illumination is between maintaining 1500~2000lx, and the container of timing shake daily 2-3 times in cultivating process is thin to help to precipitate
Bacterium, which floats, obtains illumination.It cultivates to OD600It is 2.7~3.2, bacterium solution color is peony at this time.
(3) microbial inoculum simply expands culture：In order to which daily production is cultivated, microbial inoculum is simple and efficient, can be by culture in step (2)
Bacterium solution is inoculated in the culture medium in step (1) according to 20%~30% inoculum concentration, and 30 DEG C~35 DEG C are sealed by fermentation 5~7 days,
Between cloudy day and night use incandescent lamp supplementary light culture, intensity of illumination to maintain 1500~2000lx, in cultivating process daily
Timing shakes container 2-3 times, and illumination is obtained to help the bacterium precipitated to float.Cultivate ripe bacterium solution OD600It is 2.7~3.2.
In above-mentioned steps (2) and step (3), in bacterium solution viable bacteria content can reach respectively 100~18,000,000,000/mL and 150~20,000,000,000/
mL。
Embodiment 3
(1) preparation of fermentation medium：Medium component (by weight percent) KH2PO40.05%, K2HPO4·3H2O
0.05%, MgSO4·7H2O 0.05%, NaCl 0.1%, urea 1%~4%, sodium acetate 0.2%~0.5%, yeast extract
0.01%~0.06%, and continue to boil 15min, pH uses NaHCO after cooling3PH 6.8~7.2 is adjusted, culture medium is dispensed
To in sealable transparent round；
(2) preparation of microbial inoculum：The microbial inoculum is formed sediment by red pseudomonas, trichoderma pseudokiningii bacterium, rhizobium, lactobacillus plantarum reconciliation
Afnyloliquefaciens form, by 5 kinds of microorganisms according to 1~9.9 × 105The inoculum concentration of CFU/mL is inoculated in transparent sealing container respectively
In, it is placed under outdoor solar light, 30 DEG C~35 DEG C are sealed by fermentation 5~7 days, and cloudy day and night use incandescent lamp supplementary light culture,
Between intensity of illumination maintains 1500~2000lx, the container of timing shake daily 2-3 times in cultivating process, until OD600For 3.0~
3.5。
(3) microbial inoculum simply expands culture：In order to which daily production is cultivated, microbial inoculum is simple and efficient, can be by culture in step (2)
Bacterium solution is inoculated in the culture medium in step (1) according to 20%~30% inoculum concentration, and 30 DEG C~35 DEG C are sealed by fermentation 5~7 days,
Between cloudy day and night use incandescent lamp supplementary light culture, intensity of illumination to maintain 1500~2000lx, in cultivating process daily
Timing shakes container 2-3 times, and illumination cultivation is obtained to OD to help the bacterium precipitated to float600It is 3.0~3.5, bacterium solution face at this time
Color is peony.
In above-mentioned steps (2) and step (3), viable bacteria content can reach 200~30,000,000,000/mL in bacterium solution.
Embodiment 4
The cultivation microbial inoculum for cultivating ripe in embodiment 1, embodiment 2 and embodiment 3 is diluted 200 times respectively, is with blank
Control, is divided into tetra- regions A, B, C, D by cultivated area, respectively with being broadcasted sowing after above-mentioned 4 kinds of solution treatments seed soaking 2h, covers
Shallow-layer；After about 20~30d, tomato is carried out at pouring root, root irrigation concentration corresponds to seed soaking concentration.
The pouring root same day takes 5 plants at random in above-mentioned 4 groups often processing, measures the height of seedling, remove sandy soil, claim its single plant small
The fresh weight of seedling, the fresh weight of seedling root, overground part fresh weight survey main root length, root cross growth carries out length, while calculating tomato
Emergence rate.The results are shown in Table 1.
Table 1
The result shows that conciliating starch gemma bar using red pseudomonas, trichoderma pseudokiningii bacterium, rhizobium, lactobacillus plantarum
The Hot Pepper Seedling of bacterium mix bacterium agent processing, after using three kinds of red pseudomonas, trichoderma pseudokiningii bacterium, rhizobium microorganism compoundings
Hot Pepper Seedling emergence rate is high, illustrate that 5 kinds of microorganisms can play synergistic effect, provide the promotion multiple-microorganism that Hot Pepper Seedling is grown,
Antiviral substance and somatomedin, make the Hot Pepper Seedling average height of seedling of cultivation be higher by 6.31cm, and emergence rate is improved than control
32.26%.
Chilli seedling investigates pest and disease damage, and each region records the bacterium of different embodiments 3 using 20m × 20m as unit
Agent different extension rates treated pest and disease damages such as pepper powdery mildew, anthracnose, gray mold, aphid Occurrence degree,
The results are shown in Table 2.
As can be seen from Table 2, the microbial inoculum of 200~300 times of dilution is for inhibiting powdery mildew, anthracnose, gray mold, aphid to have significantly
Effect, gray mold incidence 2.32~2.65%, anthracnose incidence 2.3~2.62%, aphid incidence 3.71~
4.23%, powdery mildew incidence 2.48~2.83%.2~7 percentage points are reduced compared with the control.
Table 2
Although the present invention has been described by way of example and in terms of the preferred embodiments, it is not limited to the present invention, any to be familiar with this skill
The people of art can do various change and modification, therefore the protection model of the present invention without departing from the spirit and scope of the present invention
Enclosing be subject to what claims were defined.
Claims (10)
1. a kind of microorganism formulation, which is characterized in that the microbial inoculum is active biological product, and active constituent is by red false unit cell
Bacterium, trichoderma pseudokiningii bacterium, rhizobium, lactobacillus plantarum and bacillus amyloliquefaciens composition.
2. microorganism formulation according to claim 1, which is characterized in that comprise the following steps：By red pseudomonas, intend
Koning trichoderma bacterium, rhizobium, lactobacillus plantarum and bacillus amyloliquefaciens are respectively according to 1~9.9 × 105The inoculum concentration of CFU/mL
It is inoculated in same fermentation medium, 30 DEG C~35 DEG C are sealed by fermentation 5~7 days, until OD600Be 3.0~3.5, obtain cultivate at
Ripe microbial inoculum.
3. microorganism formulation according to claim 2, which is characterized in that the fermentation medium components are by weight percent
For：KH2PO40.05%, K2HPO4·3H2O 0.05%, MgSO4·7H2O 0.05%, NaCl 0.1%, urea 1%~4%,
Sodium acetate 0.2%~0.5%, yeast extract 0.01%~0.06%, boils 15min, is cooled to 40 DEG C hereinafter, using NaHCO3It adjusts
Save pH 6.8~7.2.
4. microorganism formulation according to claim 2, which is characterized in that the microbial inoculum is described also by expanding culture
The step of expanding culture is as follows：The ripe microbial inoculum of culture is directly inoculated in fermentation medium, inoculum concentration is 15%~25%,
It is sealed by fermentation 5~7 days for 30 DEG C~35 DEG C in transparent vessel, until OD600It is 3.0~3.5, microbial inoculum is cultivated ripe；The fermentation training
Foster based component is by weight percent：KH2PO40.05%, K2HPO4·3H2O 0.05%, MgSO4·7H2O 0.05%, NaCl
0.1%, urea 1%~4%, sodium acetate 0.2%~0.5%, yeast extract 0.01%~0.06% boils 15min, is cooled to 40
DEG C hereinafter, using NaHCO3Adjust pH 6.8~7.2.
5. microorganism formulation according to claim 1, which is characterized in that be placed in the bacterium solution after inoculation in incubation
Under outdoor solar light, cloudy day and night use incandescent lamp supplementary light culture, and intensity of illumination is 1500~2000lx, in cultivating process
Container is shaken for every eight hours 2-3 times, illumination is obtained to help the bacterium precipitated to float.
6. a kind of method preparing microorganism formulation described in claim 1, which is characterized in that the method is by red false unit cell
Bacterium, trichoderma pseudokiningii bacterium, rhizobium, lactobacillus plantarum and bacillus amyloliquefaciens are respectively with 1~9.9 × 105CFU/mL's connects
Kind amount is inoculated in same fermentation medium, is placed under outdoor solar light, and 30 DEG C~35 DEG C are sealed by fermentation 5~7 days, cloudy day and night
Using incandescent lamp supplementary light culture, container 2-3 is shaken in timing to intensity of illumination daily in 1500~2000lx, cultivating process
It is secondary；To bacterium solution OD600It is 3.0~3.5, bacterium solution is cultivated ripe.
7. microorganism formulation according to claim 6, which is characterized in that the fermentation medium components are as follows：KH2PO4
0.05%, K2HPO4·3H2O 0.05%, MgSO4·7H2O 0.05%, NaCl 0.1%, urea 1%~4%, sodium acetate
0.2%~0.5%, yeast extract 0.01%~0.06%.
8. a kind of method of prevention capsicum pest and disease damage, which is characterized in that respectively using different dilutions times in capsicum cultivating process
Several microbial inoculum seed soaking, root dipping transplanting, pouring root or sprinkling liquid levels.
9. according to the method described in claim 8, it is characterized in that, microbial inoculum is diluted 200~300 times, 2~3d sprinklings are primary molten
Liquid.
10. microorganism formulation prepared by any the methods of claim 1-5 preparing using capsicum as the food of primary raw material or
The application of field of agricultural cultivation.
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| CN110819573B (en) * | 2019-12-05 | 2021-02-12 | 青岛市农业科学研究院 | Microbial agent for improving cold and moisture resistance of peas and application thereof |
| CN112358347A (en) * | 2020-11-10 | 2021-02-12 | 湖南金叶众望科技股份有限公司 | Special organic-inorganic compound fertilizer for ginseng fruit and preparation method thereof |
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