CN106929457B - Paenibacillus kribbensis granules and preparation method thereof - Google Patents

Paenibacillus kribbensis granules and preparation method thereof Download PDF

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CN106929457B
CN106929457B CN201710323746.XA CN201710323746A CN106929457B CN 106929457 B CN106929457 B CN 106929457B CN 201710323746 A CN201710323746 A CN 201710323746A CN 106929457 B CN106929457 B CN 106929457B
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parts
granule
granules
kribbensis
paenibacillus
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CN106929457A (en
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冯文文
刘熙文
梁作栯
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Jian College
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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
    • C12N1/20—Bacteria; Culture media therefor
    • C12N1/205—Bacterial isolates
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • 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
    • A01N25/00—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
    • A01N25/12—Powders or granules
    • 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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    • 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 relates to a Paenibacillus kribbensis granule and a preparation method thereof, belonging to the technical field of pesticides. The invention provides a Paenibacillus kribbensis granule which comprises the following components in parts by weight: 4 parts of Klibobacillus; 13-18 parts of an auxiliary agent; 55-68 parts of a filler; 18-20 parts of water. The paenibacillus kribbensis granules provided by the invention have high biological activity and good bacteriostatic effect, and are easy to transport and store.

Description

Paenibacillus kribbensis granules and preparation method thereof
Technical Field
The invention relates to the technical field of pesticides, and particularly relates to a Paenibacillus kribbensis granule and a preparation method thereof.
Background
The Klibobacillus belongs to the Bacillaceae, one of the Paenibacillus, the broad spectrum of the bacterium is good, the bacteriostatic activity is high, and the prevention and treatment effects are achieved on various plant diseases.
At present, the Paenibacillus kribbensis aqua is commonly used, is inconvenient to transport, has high packaging cost and is difficult to store, has short biological activity duration and influenced drug effect, and has certain regionality in popularization and use.
Disclosure of Invention
The invention aims to provide a Paenibacillus kribbensis granule. The paenibacillus kribbensis granules provided by the invention have high biological activity and good bacteriostatic effect, and are easy to transport and store.
The invention provides a Paenibacillus kribbensis granule which comprises the following components in parts by weight:
4 parts of Klibobacillus;
13-18 parts of an auxiliary agent;
55-68 parts of a filler;
18-20 parts of water.
Preferably, the biological preservation number of the bacillus kribbensis strain is CGMCC No. 7996.
Preferably, the auxiliary agent comprises white granulated sugar and corn starch, and the mass ratio of the white granulated sugar to the corn starch is 1: (4-7.5).
Preferably, the filler comprises talc and kaolin; the mass ratio of the talcum powder to the kaolin is (1-2.5): 1.
preferably, the content of the bacillus kribbensis in the granules is 190-210 IU/mg.
Preferably, the drying loss of the granules is less than or equal to 6 percent.
Preferably, the pH value of the granules is 6-8.
Preferably, the suspension rate of the granules is more than or equal to 80 percent.
The invention also provides a preparation method of the granules in the technical scheme, which comprises the following steps: mixing the Paenibacillus kribbensis, the auxiliary agent, the filler and water, stirring, and carrying out jet milling to obtain the granules.
The invention also provides the application of the granules in the technical scheme or the granules prepared by the preparation method in the technical scheme in preventing and treating rice sheath blight disease.
The invention provides a Paenibacillus kribbensis granule which comprises the following components in parts by weight: 4 parts of Klibobacillus; 13-18 parts of an auxiliary agent; 55-68 parts of a filler and water. The paenibacillus kribbensis granules provided by the invention have high biological activity and good bacteriostatic effect, and are easy to transport and store.
Biological preservation Instructions
The Paenibacillus kriibbens (Paenibacillus kribbensis) is preserved in China general microbiological culture Collection center at 13.08.2013 with the address of No. 3 Siro-1 of Beijing Kogyo-Yang district, No. 3 of China academy of sciences, and the biological preservation number is CGMCC No. 7996.
Drawings
FIG. 1 shows the bacterial inhibition profile of Paenibacillus kribbensis provided in example 3 of the present invention.
Detailed Description
The invention provides a Paenibacillus kribbensis granule which comprises the following components in parts by weight:
4 parts of Klibobacillus;
13-18 parts of an auxiliary agent;
55-68 parts of a filler;
18-20 parts of water.
In the invention, the bacillus kribbensis granules comprise 4 parts of bacillus kribbensis. The strain of the paenibacillus kribbensis is preserved in the common microorganism center of China Committee for culture Collection of microorganisms, and the biological preservation number is CGMCC No. 7996. The method for preparing the bacillus kribbensis is not particularly limited, and the method for preparing the bacillus kribbensis known to those skilled in the art can be adopted. In the invention, the content of the bacillus kribbensis in the granules is 190-210 IU/mg, and more preferably 200 IU/mg.
Based on 4 parts of the bacillus kribbensis, the bacillus kribbensis granule provided by the invention comprises 13-18 parts of an auxiliary agent, and preferably 15 parts. In the invention, the auxiliary agent preferably comprises white granulated sugar and corn starch, and the mass ratio of the white granulated sugar to the corn starch is preferably 1: (4-7.5), more preferably 1: 5. in the invention, the white granulated sugar plays a role of a binder, and the corn starch belongs to a disintegrating agent, so that the diluting speed in water is increased. The sources of the white granulated sugar and the corn starch are not particularly limited in the present invention, and commercial products of white granulated sugar and starch known to those skilled in the art can be used.
Based on 4 parts of the bacillus kribbensis, the bacillus kribbensis granule provided by the invention comprises 55-68 parts of a filler, and preferably 60 parts. In the present invention, the filler preferably comprises talc and kaolin; the mass ratio of the talcum powder to the kaolin is preferably (1-2.5): 1. in the invention, the talcum powder plays a role in lubrication, and the granulation speed is accelerated during granulation; the kaolin belongs to the filling material and plays a role in diluting. The sources of the talc and kaolin are not particularly limited in the present invention, and conventional commercial products of talc and kaolin known to those skilled in the art may be used.
In the invention, the drying decrement of the granules is less than or equal to 6 percent.
In the invention, the pH value of the granules is 6-8.
In the invention, the suspension rate of the granules is more than or equal to 80 percent.
In the invention, the particle size of the granules is 1.0-1.2 mm.
The invention also provides a preparation method of the granules in the technical scheme, which comprises the following steps: mixing the Paenibacillus kribbensis, the auxiliary agent, the filler and water, stirring, and carrying out jet milling to obtain the granules.
In the invention, the stirring time is preferably 0.8-2 h, and more preferably 1 h.
The jet mill of the present invention is not particularly limited, and a jet mill known to those skilled in the art may be used. Such as a QYF-60 jet mill. Specifically, in the specific jet milling process, the shaft sealing pressure is preferably 0.1-0.15 Mpa, and more preferably 0.12 Mpa; the preferable pressure of the discharging air seal is 0.15 Mpa; the pulse pressure is preferably 0.4-0.6 MPa, and more preferably 0.5 MPa; the feeding pressure is preferably 0.75-0.85 MPa, and more preferably 0.8 MPa; the crushing and beating speed is preferably 200-300 kg/h, and more preferably 240 kg/h; the preferable crushing material amount is 800-1000 kg per batch, and the more preferable crushing material amount is 850kg per batch; the kneading water amount is preferably controlled to be 20-25%, and more preferably controlled to be 23%; the granulation temperature is preferably controlled to be 50-60 ℃, and more preferably to be 55 ℃.
The invention also provides the application of the granules in the technical scheme or the granules prepared by the preparation method in the technical scheme in preventing and treating rice sheath blight disease.
The paenibacillus kribbensis granules and the preparation method thereof provided by the present invention are further described in detail with reference to the following specific examples, and the technical solutions of the present invention include, but are not limited to, the following examples.
Example 1
Preparing a first-order seed solution of the Paenibacillus kribbensis: inoculating the preserved Paenibacillus kribbensis strain into an eggplant-shaped bottle filled with an activation culture medium, and culturing at 28 ℃ for 12h for activation culture to complete strain activation; inoculating the activated Paenibacillus kribbensis strain into the first-stage liquid of Paenibacillus kribbensis according to the inoculation amount of 1%In the culture medium, the primary liquid culture medium comprises the following components in percentage by mass: 3% of sucrose, 0.15% of sodium nitrate, 0.1% of dipotassium phosphate, 0.05% of magnesium sulfate, 0.05% of potassium chloride, 0.002% of ferric sulfate and the balance of water. The first-stage liquid culture temperature of Paenibacillus kribbensis is 30 ℃, the initial pH value is 7.5, and the Paenibacillus kribbensis is cultured for 24 hours according to the length of 15m3Ventilating at a ventilation rate of 20m after liquid culture for 24h3Ventilating for culture at a ventilation rate of/h, and stopping culture after illumination culture for 30h to obtain a first-class seed solution of the Paenibacillus kribbensis.
Preparing a second-stage seed solution of the Paenibacillus kribbensis: inoculating the obtained first-stage seed liquid of the Paenibacillus kribbensis into a second-stage liquid culture medium of the Paenibacillus kribbensis according to the inoculation amount of 10%, wherein the second-stage liquid culture medium comprises the following components in percentage by mass: 1.5 percent of cane sugar, 1.5 percent of glucose, 1.0 percent of soybean meal, 0.05 percent of ammonium sulfate, 0.05 percent of dipotassium hydrogen phosphate, 0.1 percent of calcium chloride and the balance of water. The secondary liquid culture is carried out at 32 deg.C and pH 7.5 for 24 hr, and the aeration amount is 150m3H, after 24h of culture, the aeration is 250m3And h, stirring for 2 times per hour, wherein the stirring time is 15min each time, and stopping culturing after illumination culturing for 48 hours to obtain the second-stage seed liquid of the Paenibacillus kribbensis.
Preparing the Paenibacillus kribbensis: inoculating the obtained second-stage seed liquid of the Paenibacillus kribbensis into a third-stage liquid culture medium of the Paenibacillus kribbensis by using the inoculation amount of 10%, wherein the third-stage liquid culture medium comprises the following components in percentage by mass: 2.5% of glucose, 1% of corn flour, 0.05% of ammonium sulfate, 0.05% of dipotassium hydrogen phosphate, 0.1% of calcium chloride and the balance of water. The temperature of the third-stage liquid culture is 30 deg.C, pH is 7.5, and the aeration amount is 2000m3And h, after 24h of culture, the ventilation volume is 1000m3/h, stirring is carried out for 2 times per hour, the stirring time is 15min each time, after 65h of illumination culture, the culture is stopped, a third-stage culture solution of the Paenibacillus kriging is obtained, a butterfly centrifuge is adopted to centrifuge for 20min at 5000rpm, a supernatant and a concentrated solution are obtained, and the concentrated solution is dried by a spray drying tower, so that the Paenibacillus kriging is obtained.
Example 2
Mixing 4kg of the Bacillus kribbensis obtained in the embodiment 1 with 10kg of white granulated sugar, 5.5kg of corn starch, 10kg of talcum powder, 5kg of kaolin and 20kg of water, stirring for 1 hour, and carrying out jet milling with the shaft sealing pressure of 0.12 Mpa; the pressure of the discharged material gas seal is 0.15 Mpa; the pulse pressure is 0.6 Mpa; the feeding pressure is 0.85 Mpa; the crushing and material beating speed is 240 kg/h; the crushing material amount is 850 kg/batch; the kneading water amount is controlled to be 20 percent; the granulation temperature is controlled to be 55 ℃, and 200IU/mg kribber bacillus granules are prepared.
Example 3
The bacillus kribbensis granules prepared in example 2 were diluted 50 to 1000 times with water, and the inhibitory activities against sheath blight were compared with propiconazole registered in the ministry of agriculture.
1) Bacterial inhibition spectrum of Klibobacillus
The kriben fermentation stock solution is diluted by 50 times to measure the bacteriostatic activity of 22 plant pathogenic bacteria, and the experimental result is shown in figure 1. The results show that the kriben fermentation liquor shows different degrees of antibacterial activity on all 22 tested pathogenic bacteria, has a certain inhibition effect on common diseases of vegetables, fruits and tobacco, can be used for preventing and treating various fungal diseases of crops, and has a wide antibacterial spectrum.
2) Different dilution times of 200IU/mg kriben paenibacillus granules for bacteriostasis effect
In order to explore the optimal inhibition dilution multiple of the kriben paenibacillus granules, dilution multiple gradient is set to observe the inhibition effect, 0.5ppm propiconazole solution is used as a control, under the same condition, the inhibition rate of the 0.5ppm propiconazole solution to rice sheath blight is 80.06%, and the inhibition effects of the kriben paenibacillus granules diluted by different times are shown in table 1. It can be seen that the bacteriostatic effect of the bacillus kribbensis granules is in a descending trend along with the increase of the dilution times, the bacteriostatic effect when the bacillus kribbensis granules are diluted by 200 times is equivalent to the bacteriostatic effect of a propiconazole solution of 0.5ppm, and the bacillus kribbensis granules still have a certain inhibitory effect on rice sheath blight bacteria under the condition of dilution by 1000 times.
TABLE 1 dilution of Paenibacillus kribbensis granules with different times of bacteriostasis rate
Figure BDA0001290540440000051
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.

Claims (7)

1. The Paenibacillus kribbensis granules are characterized by comprising the following components in parts by weight:
4 parts of Klibobacillus; the content of the Paenibacillus kribbensis in the granules is 200 IU/mg;
10 parts of white granulated sugar, 5.5 parts of corn starch, 10 parts of talcum powder, 5 parts of kaolin and 20 parts of water.
2. The granule as claimed in claim 1, wherein the biological preservation number of the Paenibacillus kribbensis strain is CGMCC No. 7996.
3. The granule formulation according to claim 1, wherein the granule formulation has a drying loss of 6% or less.
4. The granule according to claim 1, wherein the pH of the granule is 6 to 8.
5. The granule according to claim 1, wherein the suspension rate of the granule is 80% or more.
6. A method of preparing the granule of claim 1, comprising the steps of: mixing the Paenibacillus kribbensis, the auxiliary agent, the filler and water, stirring, and carrying out jet milling to obtain the granules.
7. The granule according to any one of claims 1 to 5 or the granule obtained by the preparation method according to claim 6, for use in controlling rice sheath blight disease.
CN201710323746.XA 2017-05-10 2017-05-10 Paenibacillus kribbensis granules and preparation method thereof Expired - Fee Related CN106929457B (en)

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CN108728380B (en) * 2018-06-06 2020-07-31 浙江省农业科学院 Strain with antagonistic action on Chinese cabbage sclerotinia sclerotiorum and culture method and application thereof
CN109438087A (en) * 2018-12-27 2019-03-08 天津环微生物科技有限公司 A kind of instant granular type agricultural microbial agent and preparation method and application method

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