CN106957807A - A kind of lichem bacillus strain TA65 and its application in compost maturity is promoted - Google Patents

A kind of lichem bacillus strain TA65 and its application in compost maturity is promoted Download PDF

Info

Publication number
CN106957807A
CN106957807A CN201710184427.5A CN201710184427A CN106957807A CN 106957807 A CN106957807 A CN 106957807A CN 201710184427 A CN201710184427 A CN 201710184427A CN 106957807 A CN106957807 A CN 106957807A
Authority
CN
China
Prior art keywords
bacillus strain
compost
lichem
strain
lichem bacillus
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
CN201710184427.5A
Other languages
Chinese (zh)
Other versions
CN106957807B (en
Inventor
李群良
陆彦宇
徐佳琦
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Guangxi University
Original Assignee
Guangxi University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Guangxi University filed Critical Guangxi University
Priority to CN201710184427.5A priority Critical patent/CN106957807B/en
Publication of CN106957807A publication Critical patent/CN106957807A/en
Application granted granted Critical
Publication of CN106957807B publication Critical patent/CN106957807B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • 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
    • C12R2001/10—Bacillus licheniformis
    • 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
    • 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
    • C—CHEMISTRY; METALLURGY
    • C05—FERTILISERS; MANUFACTURE THEREOF
    • C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
    • C05F17/00—Preparation of fertilisers characterised by biological or biochemical treatment steps, e.g. composting or fermentation
    • 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
    • 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
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/0004—Oxidoreductases (1.)
    • C12N9/0055—Oxidoreductases (1.) acting on diphenols and related substances as donors (1.10)
    • C12N9/0057—Oxidoreductases (1.) acting on diphenols and related substances as donors (1.10) with oxygen as acceptor (1.10.3)
    • C12N9/0061—Laccase (1.10.3.2)
    • 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
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/0004—Oxidoreductases (1.)
    • C12N9/0065—Oxidoreductases (1.) acting on hydrogen peroxide as acceptor (1.11)
    • 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
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
    • C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
    • C12N9/2405—Glucanases
    • C12N9/2434—Glucanases acting on beta-1,4-glucosidic bonds
    • C12N9/2437—Cellulases (3.2.1.4; 3.2.1.74; 3.2.1.91; 3.2.1.150)
    • 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
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
    • C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
    • C12N9/2477—Hemicellulases not provided in a preceding group
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y110/00—Oxidoreductases acting on diphenols and related substances as donors (1.10)
    • C12Y110/03—Oxidoreductases acting on diphenols and related substances as donors (1.10) with an oxygen as acceptor (1.10.3)
    • C12Y110/03002—Laccase (1.10.3.2)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y111/00—Oxidoreductases acting on a peroxide as acceptor (1.11)
    • C12Y111/01—Peroxidases (1.11.1)
    • C12Y111/01013—Manganese peroxidase (1.11.1.13)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y111/00—Oxidoreductases acting on a peroxide as acceptor (1.11)
    • C12Y111/01—Peroxidases (1.11.1)
    • C12Y111/01014—Lignin peroxidase (1.11.1.14)
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
    • Y02W30/00—Technologies for solid waste management
    • Y02W30/40—Bio-organic fraction processing; Production of fertilisers from the organic fraction of waste or refuse

Landscapes

  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Genetics & Genomics (AREA)
  • Zoology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Wood Science & Technology (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • Medicinal Chemistry (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Virology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

The invention belongs to plant growth-promoting bacteria development technique field, and in particular to be a kind of lichem bacillus strain TA65 and its promote compost maturity in application.A kind of lichem bacillus strain TA65, its taxology is named as bacillus licheniformis TA65 (Bacillus licheniformis), China Committee for Culture Collection of Microorganisms's common micro-organisms center was preserved on 01 05th, 2017, preserving number is CGMCC 13531.The lichem bacillus strain TA65 of the present invention is screens isolated from compost megathermal period sample first, the bacterial strain high temperature resistant, increases breeding soon, and can produce lignocellulolytic enzymes；The microbial inoculum of the TA65 containing lichem bacillus strain can improve compost temperature, accelerate organic matter degradation and water-soluble organic matter (DOM) degraded, improve the content of kjeldahl nitrogen, promote compost maturity；Also there is the function of biosurfactant production, therefore, the bacterial strain can be used for preparing surfactant.

Description

A kind of lichem bacillus strain TA65 and its application in compost maturity is promoted
Technical field
The invention belongs to plant growth-promoting bacteria development technique field, and in particular to be a kind of lichem bacillus strain TA65 And its application in compost maturity is promoted.
Background technology
Chemical fertilizer plays great function in modern agricultural production.But with the increase of fertilizer application amount, its utilization rate Reduce year by year.The residual of agricultural chemicals and chemical fertilizer, the substantial amounts of harmful substance to Environment release, pollutes soil, water source and food, right Human health and living environment constitute great threat, it is desirable to preserve the ecological environment increasingly strong with the cry of production safety food It is strong.Therefore, the new source of manure for substituting chemical fertilizer is developed, to adapt to be to work as the need for developing green agriculture and pollution-free food Business is anxious.And microbial manure can make invalid nutrition validation in soil, prevention and control corps diseases, agricultural chemicals and change are reduced The use of fertilizer, is the basic by way of being generally considered a kind of environment-friendly, economy has of solution soil, water source and food pollution The method of the raising crop yield of effect.
Microbial manure is the particular product that a class contains living microorganisms, applied in agricultural production, results in spy Fixed fertilizer effect.Wherein viable microbial plays a crucial role in product.At present, that microorganism fertilizer material products typically are divided into two is big Class：One class is the microbial manure of narrow sense, refers to the vital movement by microorganism, adds the supply of plant nutrient, Include the total supply of plant nutrient in soil and production environment, cause the improvement of plant nutrient situation, and then increase production Amount.The representative of this quasi-microorganism fertilizer is rhizobia fertilizer；Another kind of is the microbial manure of broad sense, is referred to by wherein microorganism Vital movement, can not only improve the supply of plant nutrient, moreover it is possible to produce auxin, promote Plant To Nutrient The pathogenic effects for absorbing or having some pathogenic microorganisms of antagonism of element, mitigate diseases and pests of agronomic crop brief introduction raising and make produce Amount.Compared with chemical fertilizer, microbial manure has advantages below：Do not destroy soil texture；It is protecting ecology, free from environmental pollution, It is nontoxic to people and animals；Fertilizer efficiency is lasting；Crop yield is improved, improves crop quality；It is with low cost, economical and effective.
Plant growth-promoting rhizobacteria (Plant growth promoting rhizobacteria, PGPR) is that a class can be highly dense Degree colonizes the microorganism in plant rhizosphere, has suppression phytopathogen, rhizosphere harmful microorganism concurrently, and promote plant growth simultaneously Increase the effect of crop yield.As the valuable source storehouse of bio-feritlizer and biological pesticide, PGPR research and application have risen Very important effect is arrived.Microbial manure is researched and developed from the angle of resource regeneration then to comprehensive utilization of resources Realistic meaning is had more with environmental protection.
Bacillus licheniformis (Bacillus licheniformis), its cellular morphology and arrangement are in shaft-like, Dan Sheng, adjustable Whole flora imbalance reaches therapeutic purposes, and body can be promoted to produce antibacterial substance, kill pathogenic bacteria.Resistant activity thing can be produced Matter, and with unique biology take oxygen mechanism, the growth and breeding of pathogenic bacteria can be suppressed.Bacillus licheniformis mainly apply with Under several aspects：(1) effectively prevention aquatic livestock enteritis, the disease such as gill rot.(2) poisonous and harmful substance in culturing pool is decomposed, Purify water.(3) there is stronger protease, lipase, the activity of amylase, promote nutrient in feed to degrade, make aquatic products Class animal is more abundant to absorbing for feed.(4) development of aquatic livestock immune organ is stimulated, strengthens immunity of organisms. (5) promote the growth of normal physiological anaerobic bacteria in enteron aisle, adjust intestinal bacilli illness, recover function of intestinal canal；(6) it is thin to enteron aisle Bacterium infection has special efficacy, to light-duty or heavy chordapsus, light-duty and medium-sized acute bacillary dysentery etc., there is obvious curative effects；(7) Resistant activity material can be produced, and with unique biology take oxygen mechanism, the growth and breeding of pathogenic bacteria can be suppressed.
At present, bacillus licheniformis is mainly used as animal feed additive, and is promoting compost for bacillus licheniformis Middle application become thoroughly decomposed there is not yet relevant report.
The information for being disclosed in the background section is merely intended to understanding of the increase to the general background of the present invention, without answering When the prior art for being considered as recognizing or implying the information structure in any form well known to persons skilled in the art.
The content of the invention
Present invention solves the problem in that providing a kind of lichem bacillus strain TA65, the bacterial strain can be applied to prepare table Face activating agent and promotion compost maturity.
The purpose of the present invention is achieved through the following technical solutions：
The invention discloses a kind of lichem bacillus strain TA65, its taxology is named as bacillus licheniformis A65 (Bacillus licheniformis), was preserved in China Committee for Culture Collection of Microorganisms general on 01 05th, 2017 Logical microorganism center, preserving number is CGMCC 13531.
Present invention also offers purposes of the described lichem bacillus strain TA65 in lignocellulose degradation.
Present invention also offers purposes of the described lichem bacillus strain TA65 in surfactant is prepared.
Present invention also offers described lichem bacillus strain TA65 prepare laccase, lignin peroxidase, Purposes in manganese peroxidase, cellulase and hemicellulase.
Present invention also offers a kind of microbial inoculum, the lichem bacillus strain TA65 seed liquors described in claim 1 are pressed 1% (v/v) inoculum concentration, is inoculated into the LB nutrient solutions after 1L sterilizings, and the culture 24h in shaking table (50 DEG C, 120rmp) is collected Nutrient solution, produces microbial inoculum.
Present invention also offers purposes of the described microbial inoculum in compost maturity is promoted.
Compared with prior art, the present invention has following beneficial technique effect：
1. the lichem bacillus strain TA65 that the present invention is provided separates to be screened first from compost megathermal period sample Arrive, the bacterial strain high temperature resistant, growth and breeding is fast, and can produce lignocellulolytic enzymes.
2. the microbial inoculum for the TA65 containing lichem bacillus strain that the present invention is provided can improve compost temperature, accelerate organic matter The degraded generation molten organic matter of micromolecular water (DOM), improves the content of kjeldahl nitrogen, promotes compost maturity.
3. the lichem bacillus strain TA65 that the present invention is provided also has the function of biosurfactant production, therefore, The bacterial strain can be used for preparing surfactant.
Preservation information explanation
Bacillus licheniformis A65 (Bacillus licheniformis), deposit number is CGMCC 13531, preservation day Phase is on 01 05th, 2017, and depositary institution is China Committee for Culture Collection of Microorganisms's common micro-organisms center, preservation Address is Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, Institute of Microorganism, Academia Sinica.
Brief description of the drawings
Fig. 1 is temperature with compost time changing curve.RT：Room temperature, blank group (CK), experimental group (T)
Fig. 2 is pH with compost time changing curve.Blank group (CK), experimental group (T)
Fig. 3 is moisture with compost time changing curve.Blank group (CK), experimental group (T)
Fig. 4 is the content of organic matter with compost time changing curve.Blank group (CK), experimental group (T)
Fig. 5 is kelvin nitrogen content with compost time changing curve.Blank group (CK), experimental group (T)
Fig. 6 is ammonia-nitrogen content with compost time changing curve.Blank group (CK), experimental group (T)
Fig. 7 is nitrate nitrogen content with compost time changing curve.Blank group (CK), experimental group (T)
Fig. 8 is DOM contents with compost time changing curve.Blank group (CK), experimental group (T)
Fig. 9 is Bacillus licheniformis TA65 bacterium colony figures.
Figure 10 is performances of the Bacillus licheniformis TA65 on blood agar plate.
Figure 11 is glucose Standard for Sugars curve.
Figure 12 is xylose standard curve.
Figure 13 is the gel electrophoresis spectrum for filtering out bacterial strain；
About the explanation of reference：
M-2000bpMake；1-TB21；2-TB22；3-TB23；4-TB24；5-TB25；6-TB41；7-TB42；8-TB43； 9-TB44；10-TB45；11-TB61；12-TB62；13-TB63；14-TB64；15-TB65；16-TA65；17-TB46.
Embodiment
With reference to specific embodiment, the present invention is described in further detail, it is described be explanation of the invention and It is not to limit.
Material and reagent used, unless otherwise specified, are commercially obtained in following embodiments.It is following to implement Experimental method used in example is conventional method unless otherwise specified.
Embodiment 1：Bacillus licheniformis A65 (Bacillus licheniformis) screening
1. experiment material and equipment
1.1 main agents
The main agents of table 1
1.2 key instruments and equipment
The key instrument of table 2 and equipment
Instrument title Model Manufacturer
Ultraviolet-visible spectrophotometer UV759 Upper Nereid section
Clean bench SW-CJ-1F SuZhou Antai Air Tech Co., Ltd.
Autobalance centrifuge PSZ4-1.2 Medical centrifuge factory of Beijing
Thunder such pH meter PHS-2S Shanghai Yi electricity instruments limited company
Constant-temperature table HQ45Z Wuhan Zhong Ke science and technology Development Co., Ltd
Biochemical cultivation case SPX-150B Shanghai leap medical apparatus and instruments factory
Adjust the temperature electronically universal electric furnace DK-98-II Tianjin Stettlen Instrument Ltd.
Refrigerator
Electronic balance YP6102 The upper positive Medical Instruments Co., Ltd of sea light
Electrically heated drying cabinet
Vertical pressure steam sterilizer YXQ-LS-50SII Shanghai Medical Equipment Plant of Bo Xun Industrial Co., Ltd.s
Electric-heated thermostatic water bath Medical Equipment Plant of Beijing
Precision electronic balance JJ500Y
1.3 culture mediums
Bacteria culture media：Nutrient agar 33g, deionized water 1000mL；
LB nutrient solutions：Peptone 10g, dusty yeast 5g, sodium chloride 10g, deionized water 1000mL.
2. the separation screening of lignin-degrading bacteria
2.1 samplings
The sample of sieve bacterium is respectively 2, the compost sample of 4,6 days, in heap temperature higher position multidraw (typically in heap body Lower face 10-20cm), it is fitted into sample sack after sample blending in being preserved in -4 DEG C of refrigerator.
The screening and purifying of 2.2 high temperature resistant bacterium
Sample 10g is weighed, is added in the triangular flask containing the NaCl solution (0.9%w/v) after 90mL sterilization treatments, puts In vibrating 30min on shaking table, it is therefore an objective to break up zoogloea, bacterium is scattered in unicellular in solution.By sterilization treatment NaCl solution afterwards is added separately in 7 test tubes (same sterilization treatment is crossed), every addition 9mL, takes 1mL points of bacterium solution after concussion Test tube is not added, i.e., be diluted to 10 respectively-1、10-2、10-3、10-4、10-5、10-6、10-7, then respectively by the bacterium solution under each dilution factor 0.1mL is taken to be coated on bacteria culture media flat board, 50 DEG C incubated.The flat board of every kind of dilution gradient make 5 it is parallel.Can be with It was observed that bacterial community is 10-5Growing way is preferable under dilution factor.The larger carry out separation training of bacterium colony is chosen in five parallel flats Support and repeatedly line purifying.
As shown in figs. 9-10, bacillus licheniformis A65 colonial morphologies are relatively regular, and bacterium colony is smaller, and rough surface is dried, edge Moistening, white, colony edge is more neat.
3. crude enzyme liquid is produced
LB nutrient solutions are prepared, sterilization treatment is poured into same sterilized serum bottle (25mL), isolated strain bacterium It is inoculated into serum bottle, is put into shaking table (50 DEG C, 120rmp) culture, prepares seed liquor.LB nutrient solutions are prepared, are fallen in conical flask Enter 100mL, sterilization treatment draws above-mentioned 1mL seed liquors and shaking table culture (50 DEG C, 120rmp) is put into conical flask, equally, makes Into bacterium solution.
Liquid producing enzyme culture：Bacterium solution centrifuges 5min through 5000r/mim, supernatant is taken as crude enzyme liquid, with heat inactivation enzyme liquid (taking 0.6mL crude enzyme liquids in centrifuge tube, to boil 10min) is used as control.
4. the assay method of enzyme activity
The measure of 4.1 laccases (Lac) activity
1st, reagent
(1) determined by oxydasis ABTS speed；
(2) 0.1moL/L citric acid-sodium citrate buffer solutions:2.941g sodium citrates and 2.1014g lemons are weighed respectively Acid, plus deionized water 100mL, the adding citric acid into sodium citrate adjust pH=5.0 with pH meter；
(3)0.5mL 0.5moL/L ABTS；
2nd, operating procedure
(1) at 25 DEG C, 2mL citric acid-sodium citrate buffers (0.1mmoL/L, pH=5) are added in test tube, With 0.5mL ABTS, mixed liquor is poured into cuvette；
(2) extinction value changes and then addition 1mL enzyme liquids start reaction, are determined under 420nm.
3rd, computational methods
Enzyme activity is defined：ABTS for 1 μm of oL of oxidation per minute is an enzyme-activity unit,
Extinction coefficient epsilon=3.6*104[(moL/L)-1cm-1], enzyme-activity unit：U/L
Laccase (Lac) vigor=AOD*106/ΔT*VEnzyme liquid*∈
=(Δ OD/ Δ T) * 106/0.5*3.6*104
In formula：The value for the adjacent absorbance value difference maximum that Δ OD- is determined；The corresponding time difference of Δ T- Δ OD values；
The volume of V- enzyme liquids.
The measure of 4.2 lignin peroxidases (Lip) activity
1st, reagent
(1) 0.24moL/L sodium tartrates buffer solution：3.60216g tartaric acid and 5.52192g sodium tartrates are weighed respectively, Plus deionized water 100mL, tartaric acid is added into sodium tartrate, pH=3.0 is adjusted；
(2) 0.1mL 24mmoL/L veratryl alcohols；
(3) 0.05mL 6.0mmoL/L H2O2；
2nd, operating procedure
(1) at 37 DEG C, overall reaction system is 3mL, and 1.85mL 0.24moL/L sodium tartrates buffering is added into test tube Liquid (pH=3.0), and 0.1mL 24mmoL/L veratryl alcohols and 1.0mL enzyme liquids；
(2) 37 DEG C are preheated to solution is poured into cuvette, 0.05mL 6.0mmoL/L H is then added again2O2Start anti- Should, determine extinction value changes in the case where wavelength is 310nm.
3rd, computational methods
It is an enzyme-activity unit that enzyme activity, which is defined as 1 μm of oL of oxidation per minute veratryl alcohol,
Extinction coefficient epsilon=9.3*103[(moL/L)-1cm-1].Enzyme-activity unit：U/L
Peroxidase (Lip) vigor=Δ OD*106/ΔT*VEnzyme liquid*∈
=(Δ OD/ Δ T) * 106/1*9.3*103
In formula：The value for the adjacent absorbance value difference maximum that Δ OD- is determined；The corresponding time difference of Δ T- Δ OD values；
The volume of V- enzyme liquids.
The measure of 4.3 manganese peroxidases (Mnp) activity
1st, reagent
(1) 0.11moL/L sodium lactate buffer solution：1.10098g lactic acid and 2.05516g sodium lactates are weighed, adds go respectively Ionized water 100mL, lactic acid is added into sodium lactate, adjusts pH=4.5；
(2) 0.025M l40mmoL/L manganese sulfates；
(3) 0.025mL 1.6mmoL/L H2O2；
2nd, operating procedure
(1) according to enzyme in H2O2In the presence of Mn2+Aoxidize Mn3+Speed determine；
(2) total reaction volume is 1mL, and 0.85mL 0.11moL/L sodium lactate buffer solution (pH=is added into test tube 4.5) 0.025mL, 40mmoL/L MnSO are added4With 1mL enzyme liquid；
(3) preheat after 37 DEG C, solution is poured into cuvette, 0.025mL 1.6mmoL/L H is added in cuvette2O2Open Dynamic reaction, determines extinction value changes under 240nm.
3rd, computational methods
The definition of enzyme activity is 1 μm of oL of oxidation per minute Mn2+For Mn3+For an enzyme-activity unit,
Extinction coefficient epsilon=6.5*103[(moL/L)-1cm-1].Enzyme-activity unit：U/L
Manganese peroxidase (Mnp) vigor=Δ OD*106/ΔT*VEnzyme liquid*∈
=(Δ OD/ Δ T) * 106/1*6.5*103
In formula：The value for the adjacent absorbance value difference maximum that Δ OD- is determined；The corresponding time difference of Δ T- Δ OD values；
The volume of V- enzyme liquids.
The measure of 4.4 cellulase activities
1st, reagent
(1) 0.1moL/L sodium citrate-citric acids sodium buffer solution：2.941g sodium citrates and 2.1014g lemons are weighed respectively Lemon acid, plus deionized water 100mL, the adding citric acid into sodium citrate adjust pH=4.8 with pH meter；
(2)1.5mLDNS；
(3) glucose standard：The deionized water that 1.0000g glucose is dissolved in 1000ml is configured to 1mg/mL glucose Titer.
2nd, standard glucose curve plotting
1mg/mL standard glucose liquid glucoses each 0 are taken, 0.2,0.4,0.6,0.8,1.0,1.2mL in test tube, adds deionization Water adds 2.0mL DNS reagents to 2.0mL, and tool plug, boiling water bath 10min is settled to 15mL after cooling, uses spectrophotometer OD values are surveyed in the case where wavelength is 550nm, 3 repetitions are tested, and take average to chart, glucose Standard for Sugars curve is referring to Figure 11.By optical density Value invites glucose amount.
3rd, operating procedure
(1) 1cm*2cm Whatman NO1 quantitative test papers are added into test tube, 1.0cm sodium citrates-lemon is added Sour sodium buffer solution (0.1moL/L, pH=4.8) and 0.5mL enzyme liquids；
(2) 1h is incubated at 50 DEG C, then takes out flowing water after addition 1.5mL DNS terminating reactions, then waste water bath 5min cold But, it is settled to 25mL；
(3) its absorbance is determined under 540nm.
4th, computational methods
Cellulase activity=y*1000ug/VEnzyme liquid/ T, wherein y obtain y=0.6916x by mark song
In formula：The absorbance that X- is determined；The time of T- water-baths；VEnzyme liquidThe volume of-enzyme liquid.
4.5 hemicellulase activities are determined
1st, reagent
(1) 1.8mL 1% xylan solution：1% solution is made into pH=4.8 acetate buffer solutions with xylan；
(2) 1.8mL acetate buffer solutions：0.82g sodium acetates are weighed, 0.6mL acetic acid is measured, deionized water 100mL is added respectively, Add acetic acid into sodium acetate, adjust PH=4.8；
(3)2mL DNS；
(4) Xylose Standard：The deionized water that 1.0000g xyloses are dissolved in 1000mL is configured to 1mg/mL xylose standard Liquid.
2nd, xylose curve plotting
Take 1mg/ml standard Xyloses each 0,0.2,0.4,0.6,0.8,1.0,1.2mL in test tube, add deionized water To 2.0mL, 2.0mL DNS reagents are added, tool plug, boiling water bath 10min are settled to 15mL after cooling, existed with spectrophotometer Wavelength is survey OD values under 550nm, and 3 repetitions are tested, and take average to chart, xylose standard curve is referring to Figure 12.Drawn by OD value Obtain xylose amount.
3rd, operating procedure
(1) enzyme liquid 0.2mL, then each xylan solution for drawing 1.8mL 1% are added in 15mL scale test tubes；
(2) blank adds 1.8mL acetate buffer solutions with 0.2mL enzyme liquids, is not added with xylan solution, shakes up；
(3) 50 DEG C of water-bath 60min, after taking-up, then draw 2mL DNS reagents and shake up, tool plug, and boiling water bath reacts immediately 10min, after cooling, adds water constant volume to 15mL, gently shakes up up and down；
(4) blank zeroising is used, in surveying absorbance under wavelength 550nm.
4th, computational methods
Calculated according to formula：Hemicellulose enzyme activity=y*1000ug/VEnzyme liquid/ T, wherein y obtain y=1.1504x by mark song +0.008；In formula：The absorbance that X- is determined；The time of T- water-baths；VEnzyme liquidThe volume of-enzyme liquid.
5th, result
The Bacillus licheniformis TA65 strain enzyme-producing determinations of activity of table 3
As shown in table 3, the vigor highest of bacillus licheniformis A65 cellulase-producings, is 851U/L, next to that manganese peroxidating Thing enzyme, its vigor is 264U/L, and the vigor of laccase, lignin peroxidase and hemicellulase is respectively 23U/L, 44U/ L and 16U/L.
Embodiment 2：Bacillus licheniformis A65 (Bacillus licheniformis) identification
2.1 experiment key instruments
Table 4 tests instrument
2.2 experimental procedures
2.2.1 DNA is extracted
DNA uses raw work《Ezup pillar bacterial genomes DNA extraction agent box specifications》Extract.Concrete operation step is detailed See specification.
2.2.2 PCR is expanded
Enter performing PCR using 2 × Es Taq MasterMix to expand, reaction system is as follows：
Response procedures：
2.2.3 PCR primer electrophoresis result
Using the agarose gel electrophoresis of 1% concentration, voltage 120V, electrophoresis 30min, per hole loading 1uL, electrophoresis pattern is shown in Figure 13.
As shown in figure 13, PCR primer electrophoresis result shows the 16S rDNA fragments of Successful amplification bacterium.
2.2.4 it is sequenced and comparison result
Sequencing uses the two-way sequencings of primer 2 7F/1492R, and lichem bacillus strain TA65 sequence is as shown in SEQ.1.
BLASTN comparisons are carried out in NCBI gene pools, strain idenfication result is obtained, the results are shown in Table 5.
The lichem bacillus strain TA65 appraising datums of table 5
As shown in Table 5, bacterial strain TA65 reaches 99% with Bacillus licheniformis similarities, therefore, by bacterial strain It is named as bacillus licheniformis A65 (Bacillus licheniformis).
Embodiment 3：Application of the microbial inoculum of the TA65 containing lichem bacillus strain in compost maturity is promoted
It is prepared by 3.1 microbial inoculums
TA65 seed liquors are pressed 1% (v/v) inoculum concentration, be inoculated into the LB nutrient solutions after 1L sterilizings, in shaking table (50 DEG C, 120rmp) middle culture 24h, it is microbial inoculum to collect nutrient solution.It is experimental group (T), blank group (CK) addition 1L to add microbial inoculum group LB nutrient solutions after sterilizing.
3.2 compost are tested
Composting material selects cow dung and sugarcane top, mass ratio 17:3, common 20kg.Compost carries out 45d, respectively the 0th, 5, 10,16,23,30,45d samplings, determine temperature, pH, moisture content, organic matter, kjeldahl nitrogen, inorganic nitrogen, the parameter such as DOM contents. Whole composting process is inoculated with microbial inoculum twice, respectively in 0d and 10d.Heap body turning three times, is 10d, 20d and 30d respectively.
3.3 experimental results
Bacillus licheniformis TA65 high temperature resistants, increase breeding soon, and can produce lignocellulolytic enzymes.
Compared with the compost without microbial inoculum (CK), the compost (T) of addition TA65 microbial inoculums has the advantage that：
(1) temperature of composting process is seen, the heap temperature for adding TA65 microbial inoculums is higher than the heap temperature without microbial inoculum.Have Beneficial to the high temperature degradation for killing pathogen and acceleration compost material in heap body, heap body is promoted to become thoroughly decomposed (Fig. 1).
(2) by Fig. 2, Fig. 3 is understood, the pH for adding the compost of TA65 microbial inoculums rises get Geng Gao, and moisture drops faster, that is, adds bacterium The heap precursor reactant of agent is more acutely so as to cause NH3A large amount of volatilization increase pH, heat production is more to take away substantial amounts of moisture.
(3) in terms of organic matter (OM) degradation rate, from 90.38% 78.67 are dropped to without the OM of microbial inoculum, degradation rate is 12.95%, the OM for adding TA65 microbial inoculums drops to 76.62% from 90.00%, and degradation rate is 14.87% (Fig. 4).
(4) from the point of view of heap body dissolved organic matter (DOM) changes of contents, without microbial inoculum DOM contents by 69.6mg/g drops to 9.8mg/g, have dropped 85.9%, the DOM contents of addition TA65 microbial inoculums are dropped to by 75.6mg/g 9.7mg/g, have dropped 87.2% (Fig. 8).
In summary, addition TA65 microbial inoculums can improve compost temperature, accelerate organic matter degradation and DOM degradeds, improve triumphant The content of family name's nitrogen, promotes compost maturity.
Embodiment 4：Lichem bacillus strain TA65 produces surfactant situation
By blood agar plate and surface tension instrument, three plants of bacterium of identification produce surfactant situations jointly.
Blood agar plate preparation method：Take after nutrient agar (Beijing overpass), sterilization treatment, treat that it is cooled to 50 DEG C or so, Gnotobasis adds the de- fiber Sheep Blood 5-10mL of sterilizing in every 100mL nutrient agars, gently shakes up, flat board is poured at once, make It is standby into inclined-plane.
Zymotic fluid surface tension test：Using around-France measure zymotic fluid surface tension, using full-automatic surface tension instrument, instrument Type number：BZY-1.
It is demonstrated experimentally that lichem bacillus strain TA65 can produce surfactant.
It is foregoing to the present invention specific illustrative embodiment description be in order to illustrate and illustration purpose.These descriptions It is not wishing to limit the invention to disclosed precise forms, and it will be apparent that according to above-mentioned teaching, can be much changed And change.The purpose of selecting and describing the exemplary embodiment is that explaining that the certain principles and its reality of the present invention should With so that those skilled in the art can realize and using the present invention a variety of exemplaries and A variety of selections and change.The scope of the present invention is intended to be limited by claims and its equivalents.
SEQUENCE LISTING
<110>Guangxi University
<120>A kind of lichem bacillus strain TA65 and its application in compost maturity is promoted
<130> ZYWS
<160> 1
<170> PatentIn version 3.3
<210> 1
<211> 1450
<212> DNA
<213>Artificial sequence
<400> 1
ccccgggcgc tcctataatg cagtcgagcg gaccgacggg agcttgctcc cttaggtcag 60
cggcggacgg gtgagtaaca cgtgggtaac ctgcctgtaa gactgggata actccgggaa 120
accggggcta ataccggatg cttgtttgaa ccgcatggtt caaacataaa aggtggcttt 180
tcgctaccac ttacagatgg acccgcggcg cattagctag ttggtggggt aacggctcac 240
caaggcgacg atgcgtagcc gacctgagag ggtgatcggc cacactggga ctgagacacg 300
gcccagactc ctacgggagg cagcagtagg gaatcttccg caatggacga aagtctgacg 360
gagcaacgcc gcgtgagtga tgaaggtttt cggatcgtaa aactctgttg ttagggaaga 420
acaagtaccg ttcgaacagg gcggtacctt gacggtacct aaccagaaag ccacggctaa 480
ctacgtgcca gcagccgcgg taatacgtag gtggcaagcg ttgtccggaa ttattgggcg 540
taaagcgcgc gcaggcggtt tcttaagtct gatgtgaaag cccccggctc aaccggggag 600
ggtcattgga aactggggaa cttgagtgca gaagaggaga gtggaattcc acgtgtagcg 660
gtgaaatgcg tagagatgtg gaggaacacc agtggcgaag gcgactctct ggtctgtaac 720
tgacgctgag gcgcgaaagc gtggggagcg aacaggatta gataccctgg tagtccacgc 780
cgtaaacgat gagtgctaag tgttagaggg tttccgccct ttagtgctgc agcaaacgca 840
ttaagcactc cgcctgggga gtacggtcgc aagactgaaa ctcaaaggaa ttgacggggg 900
cccgcacaag cggtggagca tgtggtttaa ttcgaagcaa cgcgaagaac cttaccaggt 960
cttgacatcc tctgacaacc ctagagatag ggcttcccct tcgggggcag agtgacaggt 1020
ggtgcatggt tgtcgtcagc tcgtgtcgtg agatgttggg ttaagtcccg caacgagcgc 1080
aacccttgat cttagttgcc agcattcagt tgggcactct aaggtgactg ccggtgacaa 1140
accggaggaa ggtggggatg acgtcaaatc atcatgcccc ttatgacctg ggctacacac 1200
gtgctacaat gggcagaaca aagggcagcg aagccgcgag gctaagccaa tcccacaaat 1260
ctgttctcag ttcggatcgc agtctgcaac tcgactgcgt gaagctggaa tcgctagtaa 1320
tcgcggatca gcatgccgcg gtgaatacgt tcccgggcct tgtacacacc gcccgtcaca 1380
ccacgagagt ttgtaacacc cgaagtcggt gaggtaacct ttggagccag ccgccgaagg 1440
tgatcagagt 1450

Claims (6)

1. a kind of lichem bacillus strain TA65, its taxology is named as bacillus licheniformis A65 (Bacillus Licheniformis), it was preserved on 01 05th, 2017 in China Committee for Culture Collection of Microorganisms's common micro-organisms The heart, preserving number is CGMCC 13531.
2. purposes of the lichem bacillus strain TA65 according to claim 1 in lignocellulose degradation.
3. purposes of the lichem bacillus strain TA65 according to claim 1 in surfactant is prepared.
4. lichem bacillus strain TA65 according to claim 1 is preparing laccase, lignin peroxidase, manganese mistake Purposes in oxide enzyme, cellulase and hemicellulase.
5. a kind of microbial inoculum, it is characterised in that：Lichem bacillus strain TA65 seed liquors described in claim 1 are pressed into 1% (v/ V) inoculum concentration, is inoculated into the LB nutrient solutions after 1L sterilizings, and the culture 24h in shaking table (50 DEG C, 120rmp) collects culture Liquid, produces microbial inoculum.
6. purposes of the microbial inoculum according to claim 5 in compost maturity is promoted.
CN201710184427.5A 2017-03-24 2017-03-24 Bacillus licheniformis strain TA65 and application thereof in promoting compost maturity Active CN106957807B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201710184427.5A CN106957807B (en) 2017-03-24 2017-03-24 Bacillus licheniformis strain TA65 and application thereof in promoting compost maturity

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201710184427.5A CN106957807B (en) 2017-03-24 2017-03-24 Bacillus licheniformis strain TA65 and application thereof in promoting compost maturity

Publications (2)

Publication Number Publication Date
CN106957807A true CN106957807A (en) 2017-07-18
CN106957807B CN106957807B (en) 2020-07-28

Family

ID=59471073

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201710184427.5A Active CN106957807B (en) 2017-03-24 2017-03-24 Bacillus licheniformis strain TA65 and application thereof in promoting compost maturity

Country Status (1)

Country Link
CN (1) CN106957807B (en)

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110093296A (en) * 2019-05-15 2019-08-06 广东省生物工程研究所（广州甘蔗糖业研究所） A kind of filter mud During High-Temperature Composting microbial inoculum LC and its application
WO2020030763A1 (en) * 2018-08-08 2020-02-13 Monaghan Mushrooms Ireland A bacterial composition capable of degrading complex carbohydrates
CN111621432A (en) * 2020-04-10 2020-09-04 四川轻化工大学 Bacillus licheniformis, screening method and application
CN114410536A (en) * 2022-01-28 2022-04-29 北京四良科技有限公司 Method for releasing intracellular enzyme by bacterial culture
CN114717125A (en) * 2021-01-04 2022-07-08 山东农业大学 Thermophilic bacillus licheniformis AMCC101380 and application thereof in high-temperature composting of tailstocks
CN115011509A (en) * 2022-05-05 2022-09-06 北京嘉博文生物科技有限公司 Bacterial strain for degrading cellulose in kitchen waste at high temperature and screening and application thereof
CN115786163A (en) * 2022-08-02 2023-03-14 四川农业大学 Bacillus licheniformis SCAU1602 and application thereof

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102174423A (en) * 2010-12-06 2011-09-07 中国农业科学院农业资源与农业区划研究所 Bacillus licheniformis CH15 for degrading straws and bacterial agent thereof
CN103820361A (en) * 2014-01-15 2014-05-28 深圳市铁汉生态环境股份有限公司 High-temperature cellulose degradation bacterium and application thereof
CN103667128B (en) * 2013-11-27 2015-09-16 北京昕大洋科技发展有限公司 A kind of Bacillus licheniformis and uses thereof

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102174423A (en) * 2010-12-06 2011-09-07 中国农业科学院农业资源与农业区划研究所 Bacillus licheniformis CH15 for degrading straws and bacterial agent thereof
CN103667128B (en) * 2013-11-27 2015-09-16 北京昕大洋科技发展有限公司 A kind of Bacillus licheniformis and uses thereof
CN103820361A (en) * 2014-01-15 2014-05-28 深圳市铁汉生态环境股份有限公司 High-temperature cellulose degradation bacterium and application thereof

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
JIAQI XU等: "A compost-derived thermophilic microbial consortium enhances the humification process and alters the microbial diversity during composting，Journal of environment management", 《JOURNAL OF ENVIRONMENTAL MANAGEMENT》 *
徐庆贤等: "几株猪粪堆肥发酵菌对堆肥发酵的促进作用", 《生态与农村环境学报》 *
李红亚等: "解淀粉芽孢杆菌MN-8对玉米秸秆木质纤维素的降解", 《应用生态学报》 *

Cited By (13)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2020030763A1 (en) * 2018-08-08 2020-02-13 Monaghan Mushrooms Ireland A bacterial composition capable of degrading complex carbohydrates
CN110093296A (en) * 2019-05-15 2019-08-06 广东省生物工程研究所（广州甘蔗糖业研究所） A kind of filter mud During High-Temperature Composting microbial inoculum LC and its application
CN110093296B (en) * 2019-05-15 2022-03-15 广东省生物工程研究所（广州甘蔗糖业研究所） Filter mud high-temperature composting microbial inoculum LC and application thereof
CN111621432A (en) * 2020-04-10 2020-09-04 四川轻化工大学 Bacillus licheniformis, screening method and application
CN111621432B (en) * 2020-04-10 2022-04-12 四川轻化工大学 Bacillus licheniformis, screening method and application
CN114717125A (en) * 2021-01-04 2022-07-08 山东农业大学 Thermophilic bacillus licheniformis AMCC101380 and application thereof in high-temperature composting of tailstocks
CN114717125B (en) * 2021-01-04 2023-06-20 山东农业大学 Thermophilic bacillus licheniformis AMCC101380 and application thereof in high-temperature composting of tail vegetables
CN114410536A (en) * 2022-01-28 2022-04-29 北京四良科技有限公司 Method for releasing intracellular enzyme by bacterial culture
CN114410536B (en) * 2022-01-28 2023-12-26 北京四良科技有限公司 Method for releasing intracellular enzyme by bacterial culture
CN115011509A (en) * 2022-05-05 2022-09-06 北京嘉博文生物科技有限公司 Bacterial strain for degrading cellulose in kitchen waste at high temperature and screening and application thereof
CN115011509B (en) * 2022-05-05 2023-12-22 北京嘉博文生物科技有限公司 Bacterial strain for degrading kitchen waste cellulose at high temperature and screening and application thereof
CN115786163A (en) * 2022-08-02 2023-03-14 四川农业大学 Bacillus licheniformis SCAU1602 and application thereof
CN115786163B (en) * 2022-08-02 2023-09-01 四川农业大学 Bacillus licheniformis SCAU1602 and application thereof

Also Published As

Publication number Publication date
CN106957807B (en) 2020-07-28

Similar Documents

Publication Publication Date Title
CN106957807A (en) A kind of lichem bacillus strain TA65 and its application in compost maturity is promoted
CN102433282B (en) Bacillus subtilis NB12, as well as culture method and application thereof
CN102952768B (en) Bacillus, bacterial agent, preparation method and applications thereof
CN107058177A (en) A kind of hot denitrification ground bacillus bacterial strain TB62 and its application in compost maturity is promoted
CN104263684B (en) A kind of product siderophore series bacillus and application thereof
CN106978366A (en) A kind of mix bacterium agent and its application in compost maturity is promoted
Ballardo et al. Valorization of soy waste through SSF for the production of compost enriched with Bacillus thuringiensis with biopesticide properties
CN105368747B (en) One bacillus amyloliquefaciens bacterial strain and its application
CN104130958B (en) One bacillus subtilis and the application in preventing and treating root knot nematode on tree peony disease thereof
CN107736379A (en) Application of the bacillus amyloliquefaciens in fungal diseases of plants is prevented and treated
CN109370939A (en) One plant of Bei Laisi bacillus and its separation method and application
CN106978367A (en) A kind of urea Bacillus strain TB42 and its application in compost maturity is promoted
CN109161506A (en) One bacillus subtilis and its application
CN106906172A (en) One plant of Streptomycesalbidoflhaving and its application in terms of apple tree canker preventing and treating
CN105670964B (en) Bacillus atrophaeus BsR05 strain and application thereof
CN106011005A (en) Bacillus amyloliquefaciens T600 and preparation method and application of microbial agent
CN107629978A (en) A kind of Pseudomonas nitroreducens and its application in the colony induction signaling molecule DSF that degrades
CN104726378B (en) The method for improving salt stress turfgrass defence enzyme activity using Salt-tolerant microbial agent is strengthened
CN106148232A (en) The bacterial isolates of one strain antagonism plant pathogenetic bacteria and application thereof
CN106190892B (en) One bacillus subtilis strain and its application
CN101519644B (en) Sinorhizobium sp. and application thereof
CN104263682A (en) Plant-growth-promoting endophytic bacterium having polycyclic aromatic hydrocarbons degrading function and application thereof
CN106011002A (en) Bacillus megatherium T317, microbial agent and preparation method of microbial agent
CN107699526A (en) One plant of actinomycetes strain for preventing and treating gray mold and its application
CN109321500A (en) One bacillus amyloliquefaciens bacterial strain and its application in prevention and treatment Oil Tea Anthracnose evil

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant