CN106978366A - A kind of mix bacterium agent and its application in compost maturity is promoted - Google Patents
A kind of mix bacterium agent and its application in compost maturity is promoted Download PDFInfo
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Abstract
The invention belongs to plant growth-promoting bacteria development technique field, and in particular to be a kind of mix bacterium agent and its promote compost maturity in application.A kind of mix bacterium agent, urea Bacillus strain TB42 seed liquors, lichem bacillus strain TA65 seed liquors and hot denitrification ground bacillus bacterial strain TB62 seed liquors are pressed to 1% (v/v) inoculum concentration respectively, it is inoculated into the LB nutrient solutions after 1L sterilizings, in (50 DEG C of shaking table, culture 24h in 120rmp), nutrient solution is collected, mix bacterium agent is produced.The mix bacterium agent of the present invention can improve compost temperature, accelerate organic matter degradation and DOM degradeds, improve the content of kjeldahl nitrogen, promote compost maturity.
Description
Technical field
The invention belongs to plant growth-promoting bacteria development technique field, and in particular to be a kind of mix bacterium agent and its promote
Application in compost maturity.
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,
Great threat is constituted to human health and living environment, it is desirable to preserve the ecological environment with the cry of production safety food increasingly
Strongly.Therefore, develop substitute chemical fertilizer the new source of manure, with adapt to development green agriculture and pollution-free food the need for
It is the task of top priority.And microbial manure can make invalid nutrition validation in soil, prevention and control corps diseases, agriculture is reduced
The use of medicine and chemical fertilizer, be solve soil, water source and food pollution it is basic by way of being generally considered a kind of environment friend
Good, the cost-effective method for improving crop yield.
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 micro- life
The vital movement of thing, can not only improve the supply of plant nutrient, moreover it is possible to produce auxin, promote plant pair
The pathogenic effects for absorbing or having some pathogenic microorganisms of antagonism of nutrient, mitigate diseases and pests of agronomic crop brief introduction and improve
Crop yield.Compared with chemical fertilizer, microbial manure has advantages below：Do not destroy soil texture；It is protecting ecology, not dirty
Environment is contaminated, 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 high
Density colonizes the microorganism in plant rhizosphere, has suppression phytopathogen, rhizosphere harmful microorganism concurrently, and promote plant life
Grow and increase the effect of crop yield.As the valuable source storehouse of bio-feritlizer and biological pesticide, PGPR research and application are
Through playing the important and pivotal role.Microbial manure is researched and developed from the angle of resource regeneration then comprehensive to resource
Close and have more realistic meaning using with environmental protection.
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
The existing skill that should be considered as recognizing or imply the information structure in any form well known to persons skilled in the art
Art.
The content of the invention
Present invention solves the problem in that providing a kind of mix bacterium agent, the mix bacterium agent can promote compost maturity.
The purpose of the present invention is achieved through the following technical solutions：
A kind of mix bacterium agent, by urea Bacillus strain TB42 seed liquors, lichem bacillus strain TA65 seed liquors and
Hot denitrification ground bacillus bacterial strain TB62 seed liquors press 1% (v/v) inoculum concentration respectively, are inoculated into the LB trainings after 1L sterilizings
In nutrient solution, the culture 24h in shaking table (50 DEG C, 120rmp) collects nutrient solution, produces mix bacterium agent.
Preferably, described urea Bacillus strain TB42, its taxology is named as urea bacillus TB42
(Ureibacillus suwonensis), China Committee for Culture Collection of Microorganisms was preserved on 01 05th, 2017
Common micro-organisms center, preserving number is CGMCC 13529.
Preferably, described lichem bacillus strain TA65, its taxology is named as bacillus licheniformis A65
(Bacillus licheniformis), China Committee for Culture Collection of Microorganisms was preserved on 01 05th, 2017
Common micro-organisms center, preserving number is CGMCC 13531.
Preferably, described hot denitrification ground bacillus bacterial strain TB42, its taxology with being named as hot denitrification bud
Spore bacillus TB62 (Geobacillus thermodenitrificans), China Microbiological was preserved on 01 05th, 2017
Culture presevation administration committee common micro-organisms center, preserving number is CGMCC 13530.
Present invention also offers purposes of the described mix bacterium agent in compost maturity is promoted.
Compared with prior art, the present invention has following beneficial technique effect：
The mix bacterium agent that the present invention is provided can improve compost temperature, accelerate organic matter degradation and DOM degradeds, improve triumphant
The content of family name's nitrogen, promotes compost maturity.
Preservation information explanation
TB42 (Ureibacillus suwonensis), deposit number is CGMCC 13529, and preservation date is 2017
05 day 01 month, depositary institution was China Committee for Culture Collection of Microorganisms's common micro-organisms center, and preservation address is Beijing
The institute 3 of city Chaoyang District North Star West Road 1, Institute of Microorganism, Academia Sinica.
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.
Hot denitrification ground bacillus TB62 (Geobacillus thermodenitrificans), deposit number is
CGMCC 13530, preservation date is on 01 05th, 2017, and depositary institution is China Committee for Culture Collection of Microorganisms
Common micro-organisms center, preservation address is the institute 3 of BeiChen West Road, Chaoyang District, BeiJing City 1, Chinese Academy of Sciences's microbe research
Institute.
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 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, and described is explanation of the invention
Rather than limit.
Material and reagent used, unless otherwise specified, are commercially obtained in following embodiments.Following realities
Apply experimental method used in example unless otherwise specified, be conventional method.
Embodiment 1：Urea Bacillus strain TB42, lichem bacillus strain TA65 and hot denitrification ground bacillus
The screening of tri- kinds of bacterial strains of bacterial strain TB62
1. experiment material and equipment
1.1 main agents
The main agents of table 1
| Nomenclature of drug | Molecular formula | Specification |
| ABTS | C18H24N6O6S4 | AR |
| Hydrogen peroxide | H2O2 | AR |
| Citric acid | C6H8O7 | AR |
| Sodium citrate | Na3C6H5O7 | AR |
| Glacial acetic acid | CH3COOH | AR |
| Anhydrous sodium acetate | CH3COONa | AR |
| Xylose | C5H10O5 | AR |
| Xylan | (C5H8O4)n | AR |
| Sodium tartrate | C4H5Na2O6 | AR |
| Tartaric acid | C4H6O6 | AR |
| Veratryl alcohol | C8H12O3 | AR |
| Lactic acid | C3H6O3 | AR |
| Sodium lactate | C3H5O3Na | AR |
| Manganese sulfate | MnSO4 | AR |
| Sodium hydroxide | NaOH | AR |
| Hydrochloric acid | HCl | AR |
| DNS reagents |
1.2 key instruments and equipment
The key instrument of table 2 and equipment
1.3 culture medium
Bacteria culture media：Nutrient agar 33g, deionized water 1000mL；LB nutrient solutions：Peptone 10 g, dusty yeast 5g, chlorine
Change sodium 10g, deionized water 1000mL.
2. the separation screening of lignin-degrading bacteria
2.1 sampling
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,
It is placed on shaking table and vibrates 30min, it is therefore an objective to break up zoogloea, bacterium is scattered in unicellular in solution.At sterilizing
NaCl solution after reason is added separately in 7 test tubes (same sterilization treatment is crossed), every addition 9mL, takes the bacterium solution after concussion
1mL is separately added into test tube, i.e., be diluted to 10 respectively-1、10-2、10-3、10-4、10-5、10-6、10-7, then respectively by under each dilution factor
Bacterium solution take 0.1mL to be coated on bacteria culture media flat board, 50 DEG C are incubated.The flat board of every kind of dilution gradient is made 5 and put down
OK.It is observed that bacterial community is 10-5Growing way is preferable under dilution factor.Choose that bacterium colony is larger enters in five parallel flats
Row is separately cultured and repeatedly line purifying.
From the bacterium colony figure of 3 kinds of bacterial strains, bacillus licheniformis A65 colonial morphologies are relatively regular, and bacterium colony is smaller, and surface is thick
Rough drying, edge moistening, white, colony edge is more neat；TB42 colonial morphologies are irregular, and bacterium colony is larger, and surface is smooth wet
Profit, translucent, milky, colony edge is irregular；TB62 colonial morphologies are irregular, and bacterium colony is smaller, rough surface, relatively moisten,
Slightly transparent, faint yellow, colony edge is irregular.
3. crude enzyme liquid is produced
LB nutrient solutions are prepared, sterilization treatment is poured into same sterilized serum bottle (25mL), isolated strain
Bacterium is inoculated into serum bottle, is put into shaking table (50 DEG C, 120rmp) culture, is prepared seed liquor.LB nutrient solutions are prepared, in conical flask
In pour into 100mL, sterilization treatment, draw above-mentioned 1mL seed liquors be put into conical flask, equally shaking table culture (50 DEG C,
120rmp), bacterium solution is made.
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=Δ OD*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, the mmoL/L of 0.025mL 1.6 H is added in cuvette2O2
Start reaction, extinction value changes are determined 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 meters；
(2)1.5mLDNS；
(3) glucose standard：The deionized water that 1.0000g glucose is dissolved in 1000ml is configured to 1mg/mL grape
Standard for Sugars liquid.
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, take average to chart, obtain glucose Standard for Sugars curve.Drawn by OD value
Obtain 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 is added respectively
100mL, acetic acid is added into sodium acetate, adjusts 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, take average to chart, obtain xylose standard curve.Wood is invited by OD value
Sugar 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=by mark song
1.1504x+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 inulinase-producing activity of table tri- kinds of bacterial strains of 3 TA65, TB42 and TB62 is determined
As shown in table 3, the vigor highest of bacillus licheniformis A65 cellulase-producings, is 851U/L, next to that manganese peroxide
Compound enzyme, its vigor be 264U/L, and the vigor of laccase, lignin peroxidase and hemicellulase be respectively 23U/L,
44U/L and 16U/L；Bacillus licheniformis TB42 produces the vigor highest of manganese peroxidase, is 207U/L, next to that cellulose
Enzyme, its vigor be 78U/L, and the vigor of laccase, lignin peroxidase and hemicellulase be respectively 7U/L, 0U/L and
16U/L；Hot denitrification ground bacillus TB62 produces the vigor highest of manganese peroxidase, is 136U/L, next to that hemicellulose
Plain enzyme, its vigor be 27U/L, and the vigor of laccase, lignin peroxidase and cellulase be respectively 9U/L, 4U/L and
5U/L。
Embodiment 2：The identification of three plants of bacterial strains
2.1 experiment key instruments
Table 4 tests key instrument
2.2 experimental procedure
2.2.1 DNA is extracted
DNA uses raw work《Ezup pillar bacterial genomes DNA extraction agent box specifications》Extract.Concrete operation step
Refer to 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
Use the agarose gel electrophoresis of 1% concentration, voltage 120V, electrophoresis 30min, every hole loading 1uL, electrophoresis pattern
See Fig. 9.
As shown in figure 9, PCR primer electrophoresis result shows the 16S rDNA fragments of Successful amplification bacterium.
2.2.4 sequencing and comparison result
Sequencing uses the two-way sequencings of primer 2 7F/1492R, urea Bacillus strain TB42, hot denitrification ground bacillus
Bacterial strain TB62 and lichem bacillus strain TA65 sequence are respectively as shown in SEQ.1-3.
BLASTN comparisons are carried out in NCBI gene pools, strain idenfication result (being shown in Table 5) is obtained.
Table tri- kinds of bacterial strain qualification results of 5 TB42, TA65 and TB62
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)；Bacterial strain TB42 and Ureibacillus
Suwonensis similarities reach 99%, therefore, are urea bacillus TB42 (Ureibacillus by Strain Designation
suwonensis)；Bacterial strain TB62 reaches 99% with Geobacillus thermodenitrificans similarities, therefore, will
Strain Designation is hot denitrification ground bacillus TB62 (Geobacillus thermodenitrificans).
Embodiment 3：Application of the mix bacterium agent in compost maturity is promoted
It is prepared by 3.1 microbial inoculums
TB42, TB62 and TA65 seed liquor are pressed respectively 1% (v/v) inoculum concentration, are inoculated into the LB cultures after 1L sterilizings
In liquid, the culture 24h in shaking table (50 DEG C, 120rmp), it is microbial inoculum to collect nutrient solution.It is experimental group (T) to add microbial inoculum group,
LB nutrient solutions after blank group (CK) addition 1L sterilizings.
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 result
Mix bacterium agent belongs to bacterium not of the same race by three bacillus and constituted, and this three plants of bacterium high temperature resistants increase breeding
It hurry up, and lignocellulolytic enzymes can be produced.
Compared with the compost without microbial inoculum (CK), the compost (T) of addition TB42 microbial inoculums has the advantage that：
(1) in terms of the temperature of composting process, the heap temperature of mix bacterium agent is added than the heap temperature without microbial inoculum
It is high.Be conducive to killing the high temperature degradation of pathogen and acceleration compost material in heap body, promote heap body to become thoroughly decomposed (Fig. 1).
(2) by Fig. 2, Fig. 3 is understood, the pH for adding the compost of mix bacterium agent rises get Geng Gao, and moisture drops faster, that is, added
The heap precursor reactant of microbial inoculum 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, degradation rate are dropped to without the OM of microbial inoculum
For 12.9%, the OM for adding mix bacterium agent drops to 75.32% from 90.91%, and degradation rate is 17.1% (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 mix bacterium agent are dropped to by 74.1mg/g
9.5mg/g, have dropped 87.2% (Fig. 8).
In summary, addition mix bacterium agent can improve compost temperature, accelerate organic matter degradation and DOM degradeds, improve triumphant
The content of family name's nitrogen, promotes compost maturity.
It is foregoing to the present invention specific illustrative embodiment description be in order to illustrate and illustration purpose.These are retouched
State and be not wishing to limit the invention to disclosed precise forms, and it will be apparent that according to above-mentioned teaching, can carry out a lot
Change and change.The purpose of selecting and describing the exemplary embodiment is that explaining the certain principles and in fact of the present invention
Border is applied, so that those skilled in the art can realize and utilize a variety of exemplary embodiment party of the present invention
Case 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 mix bacterium agent and its application in compost maturity is promoted
<130> ZYWS
<160> 3
<170> PatentIn version 3.3
<210> 1
<211> 1458
<212> DNA
<213>Artificial sequence
<400> 1
cgggggggga tgcctataca tgcaagtcga gcggaccaat tagaaagctt gctttttaat 60
tggttagcgg cggacgggtg agtaacacgt gggtaacctg ccctatagac cgggataact 120
cgcggaaacg cgtgctaata ccggataaca caccgaagcg catgcttcgg ggttgaaaga 180
tggttctgct atcactatag gatgggcccg cggcgcatta gctggttggt ggggtaacgg 240
cctaccaagg cgacgatgcg tagccgacct gagagggtga tcggccacac tgggactgag 300
acacggccca gactcctacg ggaggcagca gtagggaatc ttccacaatg ggcgaaagcc 360
tgatggagca acgccgcgtg agcgaagaag gtcttcggat cgtaaagctc tgttgtaagg 420
gaagaacaag cgcagcagtc actggctgcg ccctgacggt accttactag aaagccacgg 480
ctaactacgt gccagcagcc gcggtaatac gtaggtggca agcgttgtcc ggaattattg 540
ggcgtaaagc gcgcgcaggc ggtctcttaa gtctgatgtg aaagcccccg gctcaaccgg 600
ggagggtcat tggaaactgg gagacttgag tgcaggagag ggaagyggaa ttccatgtgt 660
agcggtgaaa tgcgtagaga tatggaggaa caccagtggc gaaggcggct tcctggcctg 720
taactgacgc tgaggcgcga aagcgtgggg agcaaacagg attagatacc ctggtagtcc 780
acgccgtaaa cgatgagtgc taggtgttag ggggtttccg ccccttagtg ctgcagctaa 840
cgcattaagc actccgcctg gggagtacgg tcgcaagact gaaactcaaa ggaattgacg 900
ggggcccgca caagcggtgg agcatgtggt ttaattcgaa gcaacgcgaa gaaccttacc 960
aggtcttgac atcccgctga ccgccatgga gacatggctt tcccttcggg gacagcggtg 1020
acaggtggtg catggttgtc gtcagctcgt gtcgtgagat gttgggttaa gtcccgcaac 1080
gagcgcaacc cttgtcctta gttgccatca ttcagttggg cactctaagg agactgccgt 1140
acaaatacgg aggaaggtgg ggatgacgtc aaatcatcat gccccttatg acctgggcta 1200
cacacgtgct acaatgggtg gtacaaaggg cggcaaaccc gcgaggggga gcgaatccca 1260
aaaagccact ctcagttcgg attgcaggct gcaactcgcc tgcatgaagc cggaatcgct 1320
agtaatcgcg gatcagcatg ccgcggtgaa tacgttcccg ggccttgtac acaccgcccg 1380
tcacaccacg agagtctgta acacccgaag tcggtgaggt aaccctccgg gagccagccg 1440
ccgaaaggtg acccgagt 1458
<210> 2
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<212> DNA
<213>Artificial sequence
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tggggggggt gctatacatg cagtcgagcg gaccgaacga gagcttgctc ttgttcggtc 60
agcggcggac gggtgagtaa cacgtgggca acctgcccgc aagaccggga taactccggg 120
aaaccggagc taataccgga taacaccaaa gaccgcatgg tctttggttg aaaggcggct 180
tcggctgtca cttgcggatg ggcccgcggc gcattagcta gttggtgagg taacggctca 240
ccaaggcgac gatgcgtagc cggcctgaga gggtgaccgg ccacactggg actgagacac 300
ggcccagact cctacgggag gcagcagtag ggaatcttcc gcaatggacg aaagtctgac 360
ggagcgacgc cgcgtgagcg aagaaggcct tcgggtcgta aagctctgtt gtgagggacg 420
aaggagcgcc gtttgaataa ggcggcgcgg tgacggtacc tcacgagaaa gccccggcta 480
actacgtgcc agcagccgcg gtaatacgta gggggcgagc gttgtccgga attattgggc 540
gtaaagcgcg cgcaggcggt cctttaagtc tgatgtgaaa gcccacggct caaccgtgga 600
gggtcattgg aaactggggg acttgagtgc aggagaggag agcggaattc cacgtgtagc 660
ggtgaaatgc gtagagatgt ggaggaacac cagtggcgaa ggcggctctc tggcctgtaa 720
ctgacgctga ggcgcgaaag cgtggggagc aaacaggatt agataccctg gtagtccacg 780
ccgtaaacga tgagtgctaa gtgttagagg ggtcacaccc tttagtgctg yagctaacgc 840
gataagcact ccgcctgggg agtacggccg caaggctgaa actcaaagga attgacgggg 900
gcccgcacaa gcggtggagc atgtggttta attcgaagca acgcgaagaa ccttaccagg 960
tcttgacatc ccctgacaac ccaagagatt gggcgttccc ccttcggggg gacagggtga 1020
caggtggtgc atggttgtcg tcagctcgtg tcgtgagatg ttgggttaag tcccgcaacg 1080
agcgcaaccc ttgcctctag ttgccagcat tcagttgggc actctagagg gactgccggc 1140
taaaagtcgg aggaaggtgg ggatgacgtc aaatcatcat gccccttatg acctgggcta 1200
cacacgtgct acaatgggcg gtacaaaggg ctgcgaaccc gcgaggggga gcgaatccca 1260
aaaagccgct ctcagttcgg attgcaggct gcaactcgcc tgcatgaagc cggaatcgct 1320
agtaatcgcg gatcagcatg ccgcggtgaa tacgttcccg ggccttgtac acaccgcccg 1380
tcacaccacg agagcttgca acacccgaag tcggtgaggt aacccttacg gagccagccg 1440
ccgaaaggtg ggcaatgt 1458
<210> 3
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<212> DNA
<213>Artificial sequence
<400> 3
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 (5)
1. a kind of mix bacterium agent, it is characterised in that：By urea Bacillus strain TB42 seed liquors, lichem bacillus strain TA65
Seed liquor and hot denitrification ground bacillus bacterial strain TB62 seed liquors press 1% (v/v) inoculum concentration respectively, are inoculated into after 1L sterilizings
LB nutrient solutions in, the culture 24h in the shaking table (50 DEG C, 120rmp) collects nutrient solution, produces mix bacterium agent.
2. mix bacterium agent according to claim 1, it is characterised in that：Described urea Bacillus strain TB42, it is classified
Be named as urea bacillus TB42 (Bacillus licheniformis), and Chinese micro- life was preserved on 01 05th, 2017
Thing culture presevation administration committee common micro-organisms center, preserving number is CGMCC 13529.
3. mix bacterium agent according to claim 1, it is characterised in that：Described lichem bacillus strain TA65, its point
Class is named as bacillus licheniformis A65 (Bacillus licheniformis), and China was preserved on 01 05th, 2017
Microbiological Culture Collection administration committee common micro-organisms center, preserving number is CGMCC 13531.
4. mix bacterium agent according to claim 1, it is characterised in that：Described hot denitrification ground bacillus bacterial strain
TB42, its taxology is named as hot denitrification ground bacillus TB62 (Geobacillus thermodenitrificans), in
It is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center within 01 05th, 2017, preserving number is CGMCC
13530。
5. according to purposes of any described mix bacterium agents of claim 1-4 in compost maturity is promoted.
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