CN109306334A - One plant of Bei Laisi bacillus L1-21 and its prevention and treatment Citrus Huanglongbing pathogen application - Google Patents

One plant of Bei Laisi bacillus L1-21 and its prevention and treatment Citrus Huanglongbing pathogen application Download PDF

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CN109306334A
CN109306334A CN201811266027.XA CN201811266027A CN109306334A CN 109306334 A CN109306334 A CN 109306334A CN 201811266027 A CN201811266027 A CN 201811266027A CN 109306334 A CN109306334 A CN 109306334A
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bei laisi
bacillus
laisi bacillus
grams per
per liter
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Inventor
何月秋
何鹏搏
赵正龙
亢波
吴毅歆
何鹏飞
李兴玉
李咏梅
崔文艳
何朋杰
王娟
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Yunnan Microbial Fermentation Engineering Research Center Co ltd
Yunnan Agricultural University
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Yunnan Microbial Fermentation Engineering Research Center Co ltd
Yunnan Agricultural University
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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
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/07—Bacillus
    • 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
    • A01G13/00—Protecting plants
    • 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
    • A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
    • A01N43/34—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one nitrogen atom as the only ring hetero atom
    • A01N43/46—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one nitrogen atom as the only ring hetero atom rings with more than six members
    • 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
    • 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
    • A01N63/10—Animals; Substances produced thereby or obtained therefrom
    • C—CHEMISTRY; METALLURGY
    • C05—FERTILISERS; MANUFACTURE THEREOF
    • C05D—INORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C; FERTILISERS PRODUCING CARBON DIOXIDE
    • C05D1/00—Fertilisers containing potassium
    • 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
    • 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

Abstract

The present invention relates to technical field of biological control, and in particular to one plant of Bei Laisi bacillus L1-21 and its prevention and treatment Citrus Huanglongbing pathogen application.One plant of Bei Laisi bacillus L1-21 is deposited in China Microbial Culture Preservation Commission's common micro-organisms center, deposit number are as follows: CGMCC NO.15726, classification naming are as follows: Bei Laisi bacillus L1-21 (Bacillus velezensis L1-21).Bei Laisi bacillus L1-21 of the invention has the advantages such as resistance is strong, nutrition is simple, reproduction speed is fast, and living bacteria count amount is high, performance is stablized, and Storage period is long.Bei Laisi bacillus L1-21 can controlling plant diseases can generate a variety of antibacterial materials derived from it, including multiple compounds such as lipopeptid class, polyketone classes, these antibacterial materials can inhibit the normal growth of disease fungus, bacterium, kill disease fungus and bacterium, play the effect of prevention and treatment yellow twig.

Description

One plant of Bei Laisi bacillus L1-21 and its prevention and treatment Citrus Huanglongbing pathogen application
Technical field
The present invention relates to technical field of plant disease biological control, and in particular to one plant of Bei Laisi bacillus L1-21 and It prevents and treats Citrus Huanglongbing pathogen application.
Background technique
Citrus Huanglongbing pathogen (Huanglongbing, HLB) is a kind of worldwide disease.Since 1919 find for the first time, Occur in a countries and regions in the world more than 40.In addition to Sichuan, Chongqing and Hubei, other producing regions are equal in south China citrus producing region There is generation, and in the gesture to expand.Its classical symptom be the mottled yellow of blade, disleave, " red nose fruit ", shedding, yield fall sharply, Disease fruit deformity becomes smaller, and pericarp softens, matt, sour, and fruit quality reduces etc., seriously affects the yield and quality of citrus, sends out The fruit tree of sick 5-6 loses commodity value or even garden is ruined in death, because of uncontrollable and high lethality rate, is known as " the cancer of citrus The title of disease ".Citrus Huanglongbing pathogen is, disease caused by Candidatus liberibacter category bacterium (CandidatusLiberibacter spp.) Pseudomonas gramnegative bacterium.It is distributed in citrus fruit trees (mandarin orange, tangerine, lemon, shaddock, fingered citron, citron) and wild host is (such as Fructus Aurantii) each tissue bast, and propagated by wood louse and infect host.
Currently, the preventions for Citrus Huanglongbing pathogen mainly have following three aspect: first is that strict control vector elder brother Worm wood louse；Second is that cultivating disease-free nursery stock；Third is that the serious sick plant of felling.However, since wood louse is except citrus cultivation orchard of causing harm Outside plant, wild host of also causing harm.During practical prevention and control, only focus on the prevention and control of wood louse in orchard, it is also not possible to thoroughly disappear It goes out all wood louses in orchard, that is, is difficult to completely cut through the approach that wood louse passes disease.Disease-free nursery stock is exposed to wood louse for a long time after transplanting In the environment of causing harm, final also inevitable aggrieved final result, and there is also certain negative effects to environmental ecology for chemical insecticide. The serious sick plant of felling reduces germ source, but does not cut the potential plant of falling ill for not showing Disease symptoms, causes in citrus orchard Germ still largely exists.Therefore, Citrus Huanglongbing pathogen is still worldwide uncontrollable disease the most serious.
Summary of the invention
To solve the above-mentioned problems, the present invention provides one plant of Bei Laisi bacillus L1-21 and its prevention and treatment citrus Huanglong Disease application.
Specific technical solution are as follows: one plant of Bei Laisi bacillus L1-21, the Bei Laisi bacillus L1-21 preservation In China Microbial Culture Preservation Commission's common micro-organisms center, preservation address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3 Number, preservation date: on May 4th, 2018, deposit number are as follows: CGMCC NO.15726, classification naming are as follows: Bei Laisi bacillus L1-21(Bacillus velezensisL1-21)。
A kind of fermentation liquid, the fermentation liquid is Bei Laisi bacillus L1-21 fermentation liquid, by above-mentioned Bei Laisi gemma bar Bacterium L1-21 fermentation is made.
A kind of preparation method of fermentation liquid, specific steps include:
Step 1: Bei Laisi bacillus L1-21 bacterial strain is cultivated in the fermentation medium；
Step 2: culture medium sterilizes 30 minutes at 121 DEG C, cools to 37 DEG C, is inoculated into three by the strain amount of 1%-2% In the bottle of angle；
Step 3: it under 160 revs/min of revolving speed, is cultivated 24 hours in 37 DEG C of culture mediums, when Bei Laisi bacillus The gemma amount of L1-21 reaches 105When CFU/mL, stops fermentation, obtain Bei Laisi bacillus L1-21 seed fermentation liquid；
Step 4: seed fermentation liquid described in step 3 is transferred in fermentor by 1%-2%, at 100 revs/min It under revolving speed, is cultivated 48-54 hours in 37 DEG C of culture mediums, stops fermentation, obtain Bei Laisi bacillus L1-21 fermentation liquid；
Wherein, the component of the culture medium includes: 14 grams per liter of soybean powder, 4 grams per liter of corn flour, 3 grams per liter of sucrose, fish meal 2 grams per liters, 2 grams per liter of peptone, 6 grams per liter of calcium carbonate, 0.5 grams per liter of ammonium sulfate, 0.5 grams per liter of ammonium sulfate, 0.3 gram of magnesium sulfate/ Liter, 0.3 grams per liter of potassium dihydrogen phosphate, 0.3 grams per liter of dipotassium hydrogen phosphate, 0.2 gram of manganese sulfate, 0.25 grams per liter of sodium hydroxide.
Preferably, the concentration of Bei Laisi bacillus L1-21 bacterial strain is every milliliter 10 in the step 19To 1010Range Colony Forming Unit (CFU).
A kind of biological agent, the biological agent include above-mentioned Bei Laisi bacillus L1-21 fermentation liquid, the life Object preparation is Bei Laisi bacillus L1-21 aqua and Bei Laisi bacillus L1-21 biological organic fertilizer.
Preferably, the Bei Laisi bacillus L1-21 aqua includes Bei Laisi bacillus L1-21 fermentation liquid and dense Degree is 1 ‰~4 ‰ azone and 1 ‰~4 ‰ Tween-20；The Bei Laisi bacillus L1-21 biological organic fertilizer includes shellfish This bacillus L1-21 fermentation liquid of Lay and nitrogen content 5.1% contain available phosphorus 1.5%, contain potassium oxide 3.0%, contain organic matter 97.0% decomposed rape oil cake.
Preferably, Bei Laisi bacillus L1-21 biological organic fertilizer bacillus containing Bei Laisi L1-21 gemma amount exists 105-106CFU/ grams.
A kind of application method of biological agent, the method are as follows: in the foliage-spray Bei Laisi bacillus L1-21 of citrus Aqua, while Bei Laisi bacillus L1-21 biological organic fertilizer is applied in citrus root.
Preferably, the method specifically: spray Bei Laisi bacillus L1- before in non-dusk rainy day or the morning 8 21 aquas spray once, continuous spraying 6 months for every 7~10 days in blade tow sides；Trunk four is surround in the drip of citrus All wide 30cm of ditching × depth 25cm, mix with soil by every plant 5-10 kilograms of biological organic fertilizer of bacillus L1-21 of application Bei Laisi Afterwards, it is further filled in circular groove, earthing.
A kind of purposes of biological agent, the biological agent is for preventing and treating Citrus Huanglongbing pathogen.
The beneficial effects of the present invention are: the Bei Laisi bacillus L1-21 in the present invention has, resistance is strong, nutrition is simple The advantages such as list, reproduction speed are fast, and living bacteria count amount is high, performance is stablized, and Storage period is long.Bei Laisi bacillus L1-21 can Controlling plant diseases can generate the multiple compounds such as a variety of antibacterial materials, including lipopeptid class, polyketone class, these antibacterials derived from it Substance can inhibit the normal growth of disease fungus, bacterium, kill disease fungus and bacterium, play the effect of diseases prevention.
Specific embodiment
Below in conjunction with the specific embodiment of the invention, technical solution of the present invention is clearly and completely described, is shown So, described embodiments are only a part of the embodiments of the present invention, instead of all the embodiments.Based on the reality in the present invention Example is applied, every other embodiment obtained by those of ordinary skill in the art without making creative efforts all belongs to In the scope of protection of the invention.
Embodiment 1:
One plant of Bei Laisi bacillus L1-21, the Bei Laisi bacillus L1-21 are deposited in Chinese microorganism strain guarantor Hiding committee common micro-organisms center, deposit number are as follows: CGMCC NO.15726, classification naming are as follows: Bei Laisi bacillus L1-21(Bacillus velezensisL1-21)。
A kind of fermentation liquid, the fermentation liquid is Bei Laisi bacillus L1-21 fermentation liquid, by above-mentioned Bei Laisi gemma bar Bacterium L1-21 fermentation is made.
A kind of preparation method of fermentation liquid, specific steps include:
Step 1: Bei Laisi bacillus L1-21 bacterial strain is cultivated in the fermentation medium；
Step 2: culture medium sterilizes 30 minutes at 121 DEG C, cools to 37 DEG C, is inoculated into three by the strain amount of 1%-2% In the bottle of angle；
Step 3: it under 160 revs/min of revolving speed, is cultivated 24 hours in 37 DEG C of culture mediums, when Bei Laisi bacillus The gemma amount of L1-21 reaches 105When CFU/mL, stops fermentation, obtain Bei Laisi bacillus L1-21 seed fermentation liquid；
Step 4: seed fermentation liquid described in step 3 is transferred in fermentor by 1%-2%, at 100 revs/min It under revolving speed, is cultivated 48-54 hours in 37 DEG C of culture mediums, stops fermentation, obtain Bei Laisi bacillus L1-21 fermentation liquid；
Wherein, the component of the culture medium includes: 14 grams per liter of soybean powder, 4 grams per liter of corn flour, 3 grams per liter of sucrose, fish meal 2 grams per liters, 2 grams per liter of peptone, 6 grams per liter of calcium carbonate, 0.5 grams per liter of ammonium sulfate, 0.5 grams per liter of ammonium sulfate, 0.3 gram of magnesium sulfate/ Liter, 0.3 grams per liter of potassium dihydrogen phosphate, 0.3 grams per liter of dipotassium hydrogen phosphate, 0.2 gram of manganese sulfate, 0.25 grams per liter of sodium hydroxide.
Further, the concentration of Bei Laisi bacillus L1-21 bacterial strain is every milliliter 10 in the step 19To 1010Model The Colony Forming Unit (CFU) enclosed.
A kind of biological agent, the biological agent include above-mentioned Bei Laisi bacillus L1-21 fermentation liquid, the life Object preparation is Bei Laisi bacillus L1-21 aqua and Bei Laisi bacillus L1-21 biological organic fertilizer.
Further, the Bei Laisi bacillus L1-21 aqua include Bei Laisi bacillus L1-21 fermentation liquid and Concentration is 1 ‰~4 ‰ azone and 1 ‰~4 ‰ Tween-20；The Bei Laisi bacillus L1-21 biological organic fertilizer includes Bei Laisi bacillus L1-21 fermentation liquid and nitrogen content 5.1% contain available phosphorus 1.5%, contain potassium oxide 3.0%, contain organic matter 97.0% decomposed rape oil cake.
Further, Bei Laisi bacillus L1-21 biological organic fertilizer bacillus containing Bei Laisi L1-21 gemma amount 105-106CFU/ grams.
Embodiment 2
(1) Shi Beilaisi bacillus L1-21 fermentation liquid is filled in the foliage-spray of citrus and root, sprays and uses shellfish with pouring root This bacillus L1-21 bacterial strain concentration of Lay is 1010The fermentation liquid of CFU/mL.
(2) Bei Laisi bacillus L1-21 aqua is sprayed at dusk in blade tow sides in the non-rainy day, spray within every 7~10 days It applies once, continuous spraying 6 months；The wide 30cm of trunk surrounding ditching × depth 25cm, every plant of application shellfish are surround in the drip of citrus This 5 kilograms of bacillus L1-21 biological organic fertilizer of Lay is further filled in circular groove, earthing after mixing with soil.
(3) Bei Laisi bacillus L1-21 aqua is diluted 1000 times -10000 times, at dusk or when the morning 8 before spray In blade tow sides, spray within Bei Laisi bacillus L1-21 aqua every 7~10 days once, continuous spraying 6 months；
In the present embodiment, kind is fertile mandarin orange, and the age of tree 2 years, control efficiency was shown in Table 1.
1 Bei Laisi bacillus L1-21 of table prevents and treats 2 years age of trees and irrigates mandarin orange yellow twig effect
Embodiment 3
(1) Shi Beilaisi bacillus L1-21 fermentation liquid is filled in the foliage-spray of citrus and root, sprays and uses shellfish with pouring root This bacillus L1-21 bacterial strain concentration of Lay is 109The fermentation liquid of CFU/mL.
(2) Bei Laisi bacillus L1-21 aqua is sprayed at dusk in blade face tow sides in the non-rainy day, spray within every 7~10 days It applies once, continuous spraying 6 months；The wide 30cm of trunk surrounding ditching × depth 25cm, every plant of application shellfish are surround in the drip of citrus This 7 kilograms of bacillus L1-21 biological organic fertilizer of Lay is further filled in circular groove, earthing after mixing with soil.
(3) in the dusk of non-rainy day or the morning 8 before be sprayed on blade tow sides, Bei Laisi bacillus L1-21 It sprays within aqua every 7~10 days once, continuous spraying 6 months；In the present embodiment, kind is to irrigate mandarin orange, the age of tree 3 years, control efficiency It is shown in Table 2；
2 Bei Laisi bacillus L1-21 of table prevents and treats 3 years age of trees and irrigates mandarin orange yellow twig effect
Processing Diseased plant number Healthy tree number Preventive effect %
(1) 250 182 72.8
(2) 250 201 80.4
(3) 250 167 66.8
Untreated control 250 0
Embodiment 4
(1) Shi Beilaisi bacillus L1-21 fermentation liquid is filled in the foliage-spray of citrus and root, sprays and uses shellfish with pouring root This bacillus L1-21 bacterial strain concentration of Lay is 109The fermentation liquid of CFU/mL.
(2) Bei Laisi bacillus L1-21 aqua is sprayed at dusk in blade face tow sides in the non-rainy day, spray within every 7~10 days It applies once, continuous spraying 6 months；The wide 30cm of trunk surrounding ditching × depth 25cm, every plant of application shellfish are surround in the drip of citrus This 8 kilograms of bacillus L1-21 biological organic fertilizer of Lay is further filled in circular groove, earthing after mixing with soil.
(3) in the dusk of non-rainy day or the morning 8 before be sprayed on blade face tow sides, Bei Laisi bacillus L1-21 It sprays within aqua every 7~10 days once, continuous spraying 6 months；
In the present embodiment, kind is fertile mandarin orange, and the age of tree 4 years, control efficiency was shown in Table 3；
3 Bei Laisi bacillus L1-21 of table prevents and treats 4 years age of trees and irrigates mandarin orange yellow twig effect
Processing Diseased plant number Healthy tree number Preventive effect %
(1) 250 194 77.6
(2) 250 202 80.8
(3) 250 176 70.4
Untreated control 250 0
It is obtained by embodiment 2-4, compared with prior art, Bei Laisi bacillus L1-21 fermentation liquid, Bei Laisi gemma bar Bacterium L1-21 aqua and Bei Laisi bacillus L1-21 aqua and Bei Laisi bacillus L1-21 biological organic fertilizer are used in combination The effect of prevention and treatment will be played, in this application Bei Laisi bacillus L1-21 aqua and Bei Laisi bacillus L1-21 biology It is best that effect is used in combination in organic fertilizer.
Bei Laisi bacillus L1-21 has that resistance is strong, nutrition is simple, reproduction speed is fast, and living bacteria count amount is high, property It can stablize, the advantages such as Storage period length.Bei Laisi bacillus L1-21 can controlling plant diseases derived from its can generate it is a variety of The multiple compounds such as antibacterial material, including lipopeptid class, polyketone class, these antibacterial materials can inhibit disease fungus, bacterium it is normal Growth kills disease fungus and bacterium, plays the effect of anti-yellow twig.
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the scope of the present invention.It is all Any modification, equivalent replacement, improvement and so within the spirit and principles in the present invention, are all contained in protection scope of the present invention It is interior.

Claims (10)

1. one plant of Bei Laisi bacillus L1-21, which is characterized in that it is micro- that the Bei Laisi bacillus L1-21 is deposited in China Biological inoculum preservation committee common micro-organisms center, deposit number are as follows: CGMCC NO.15726, classification naming are as follows: Bei Laisi Bacillus L1-21 (Bacillus velezensis L1-21).
2. a kind of fermentation liquid, which is characterized in that the fermentation liquid is Bei Laisi bacillus L1-21 fermentation liquid, by claim 1 The Bei Laisi bacillus L1-21 fermentation is made.
3. based on a kind of preparation method of fermentation liquid as claimed in claim 2, which is characterized in that specific steps include:
Step 1: Bei Laisi bacillus L1-21 bacterial strain is cultivated in the fermentation medium；
Step 2: culture medium sterilizes 30 minutes at 121 DEG C, cools to 37 DEG C, is inoculated into triangular flask by the strain amount of 1%-2% In；
Step 3: it under 160 revs/min of revolving speed, is cultivated 24 hours in 37 DEG C of culture mediums, as Bei Laisi bacillus L1- 21 gemma amount reaches 105When CFU/mL, stops fermentation, obtain Bei Laisi bacillus L1-21 seed fermentation liquid；
Step 4: seed fermentation liquid described in step 3 is transferred in fermentor by 1%-2%, in 100 revs/min of revolving speed Under, it is cultivated 48-54 hours in 37 DEG C of culture mediums, stops fermentation, obtain Bei Laisi bacillus L1-21 fermentation liquid；
Wherein, the component of the culture medium include: 14 grams per liter of soybean powder, 4 grams per liter of corn flour, 3 grams per liter of sucrose, 2 grams of fish meal/ Liter, 2 grams per liter of peptone, 6 grams per liter of calcium carbonate, 0.5 grams per liter of ammonium sulfate, 0.5 grams per liter of ammonium sulfate, 0.3 grams per liter of magnesium sulfate, phosphorus 0.3 grams per liter of acid dihydride potassium, 0.3 grams per liter of dipotassium hydrogen phosphate, 0.2 gram of manganese sulfate, 0.25 grams per liter of sodium hydroxide.
4. a kind of preparation method of fermentation liquid according to claim 3, which is characterized in that Bei Laisi bud in the step 1 The concentration of spore bacillus L1-21 bacterial strain is every milliliter 109To 1010The Colony Forming Unit (CFU) of range.
5. a kind of biological agent, which is characterized in that the biological agent includes Bei Laisi bacillus as claimed in claim 2 L1-21 fermentation liquid, the biological agent are that Bei Laisi bacillus L1-21 aqua and Bei Laisi bacillus L1-21 biology have Machine fertilizer.
6. a kind of biological agent according to claim 5, which is characterized in that the Bei Laisi bacillus L1-21 aqua It is 1 ‰~4 ‰ azone and 1 ‰~4 ‰ Tween-20 including Bei Laisi bacillus L1-21 fermentation liquid and concentration；The shellfish This bacillus L1-21 biological organic fertilizer of Lay includes Bei Laisi bacillus L1-21 fermentation liquid and nitrogen content 5.1%, containing effective Phosphorus 1.5%, the decomposed rape oil cake containing potassium oxide 3.0%, containing organic matter 97.0%.
7. a kind of biological agent according to claim 6, which is characterized in that the Bei Laisi bacillus L1-21 biology The organic fertilizer L1-21 gemma amount of bacillus containing Bei Laisi is 105-106CFU/ grams.
8. the application method based on a kind of any biological agent of claim 5-7, which is characterized in that the method are as follows: Bei Laisi bacillus L1-21 is applied in the foliage-spray Bei Laisi bacillus L1-21 aqua of citrus, while in citrus root Biological organic fertilizer.
9. a kind of application method of biological agent according to claim 8, which is characterized in that the method specifically: Bei Laisi bacillus L1-21 aqua is sprayed before when non-dusk rainy day or the morning 8 in blade tow sides, is sprayed within every 7~10 days It applies once, continuous spraying 6 months；The wide 30cm of trunk surrounding ditching × depth 25cm, every plant of application shellfish are surround in the drip of citrus It 5-10 kilograms of biological organic fertilizer of this bacillus L1-21 of Lay, after being mixed with soil, is further filled in circular groove, earthing.
10. the purposes based on a kind of any biological agent of claim 6-7, which is characterized in that the biological agent is used In prevention and treatment Citrus Huanglongbing pathogen.
CN201811266027.XA 2018-10-29 2018-10-29 One plant of Bei Laisi bacillus L1-21 and its prevention and treatment Citrus Huanglongbing pathogen application Pending CN109306334A (en)

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CN111690576A (en) * 2020-07-24 2020-09-22 江苏师范大学 Bacillus belgii MMB-51 for producing biosurfactant and application thereof
CN111837766A (en) * 2020-07-27 2020-10-30 华中农业大学 Method for controlling pathogen of seedling phlobacterium phlogistii by composite microorganism treatment and application
CN113667620A (en) * 2021-09-15 2021-11-19 南宁汉和生物科技股份有限公司 Bacillus subtilis Bs1218m and its application
CN114058549A (en) * 2021-11-29 2022-02-18 中国农业科学院农产品加工研究所 Bacillus belgii Shui-6, production method and application
CN114369542A (en) * 2020-10-15 2022-04-19 南京理工大学 Alkane degrading bacterium and application thereof
CN114381403A (en) * 2022-01-21 2022-04-22 中山大学 Bacillus belgii LOH112 and application thereof
CN114540249A (en) * 2022-03-23 2022-05-27 云南农业大学 Antagonistic strain 1X1Y and application thereof
CN115399338A (en) * 2022-09-19 2022-11-29 华中农业大学 Application of Bacillus velezensis Bv-6 in insect prevention
CN116496941A (en) * 2023-04-04 2023-07-28 华中农业大学 Microbial fermentation compound and application thereof in preventing and treating citrus yellow dragon disease

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CN110117556A (en) * 2019-03-27 2019-08-13 海南大学 A kind of efficient biocontrol microorganisms HN-2 and zymotechnique for producing lipopeptid substance
CN111690576A (en) * 2020-07-24 2020-09-22 江苏师范大学 Bacillus belgii MMB-51 for producing biosurfactant and application thereof
CN111837766A (en) * 2020-07-27 2020-10-30 华中农业大学 Method for controlling pathogen of seedling phlobacterium phlogistii by composite microorganism treatment and application
CN111837766B (en) * 2020-07-27 2021-09-28 华中农业大学 Method for controlling pathogen of seedling phlobacterium phlogistii by composite microorganism treatment and application
CN114369542B (en) * 2020-10-15 2023-06-09 南京理工大学 Alkane degrading bacteria and application thereof
CN114369542A (en) * 2020-10-15 2022-04-19 南京理工大学 Alkane degrading bacterium and application thereof
CN113667620A (en) * 2021-09-15 2021-11-19 南宁汉和生物科技股份有限公司 Bacillus subtilis Bs1218m and its application
CN113667620B (en) * 2021-09-15 2023-09-22 云南农业大学 Bacillus subtilis Bs1218m and application thereof
CN114058549A (en) * 2021-11-29 2022-02-18 中国农业科学院农产品加工研究所 Bacillus belgii Shui-6, production method and application
CN114058549B (en) * 2021-11-29 2023-07-11 中国农业科学院农产品加工研究所 Bacillus bailii Shu-6, production method and application
CN114381403B (en) * 2022-01-21 2023-06-30 中山大学 Bacillus bailii LOH112 and application thereof
CN114381403A (en) * 2022-01-21 2022-04-22 中山大学 Bacillus belgii LOH112 and application thereof
CN114540249A (en) * 2022-03-23 2022-05-27 云南农业大学 Antagonistic strain 1X1Y and application thereof
CN115399338B (en) * 2022-09-19 2023-01-31 华中农业大学 Application of Bacillus velezensis Bv-6 in insect prevention
CN115399338A (en) * 2022-09-19 2022-11-29 华中农业大学 Application of Bacillus velezensis Bv-6 in insect prevention
CN116496941A (en) * 2023-04-04 2023-07-28 华中农业大学 Microbial fermentation compound and application thereof in preventing and treating citrus yellow dragon disease
CN116496941B (en) * 2023-04-04 2024-01-16 华中农业大学 Microbial fermentation compound and application thereof in preventing and treating citrus yellow dragon disease

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