CN106011018B - A kind of complex micro organism fungicide of degradable agricultural product castoff - Google Patents
A kind of complex micro organism fungicide of degradable agricultural product castoff Download PDFInfo
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- CN106011018B CN106011018B CN201610498520.9A CN201610498520A CN106011018B CN 106011018 B CN106011018 B CN 106011018B CN 201610498520 A CN201610498520 A CN 201610498520A CN 106011018 B CN106011018 B CN 106011018B
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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
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
Abstract
The invention discloses a kind of complex micro organism fungicides, its active constituent is the one or more of the following bacterial strain isolated from insect gut: sphingolipid bacillus (Sphingobacterium sp.) HP455, Fen Shi fiber germ (Cellulosimicrobium sp.) HP181, Pseudomonas alcaligenes (Pseudomonas sp.) HP207, acid-producing Klebsiella bacterium (Klebsiella oxytoca) BD177, Ke Shi citric acid bacillus (Citrobacter koseri) BD195.The composite bacteria agent can significantly promote crop growth, and raising crops seedling stage is disease-resistant, salt resistance, and freeze proof equal resistance environment-stress ability increases the survival rate of crop growth early stage.
Description
Technical field
The invention belongs to microbial bacterial agent fields, and in particular to a kind of composite microbial bacteria of degradable agricultural wastes
Agent.
Background technique
With the continuous transformation of China's rapid development of economy and the rural area mode of production and life, agricultural and the farming in rural area
Object waste is more and more, and the processing pressure of waste increasingly increases.On the one hand, the scientific processing farming of unreasonable shortage
Object waste will cause serious environmental pollution, for example burn crop material, discard the random discarding of vegetables and fruits, poultry waste
Improper processing etc..Especially crop straw burning is polluted caused by air, have become urban air pollution major reason it
One, haze, PM2.5 have become the focus of social concerns.On the other hand, China farmland organic matter lacks, and restricts arable land life
Force of labor is badly in need of organic substance circulation investment to the high dependency of chemical fertilizer, therefore agricultural waste material degradation treatment seems especially
It is important.
A large amount of microorganism is inhabited in insect gut, these microorganisms are suitable to the growth of host, development, pathogenic and environment
Aspect is answered to have important influence, they both participate in metabolism, provide nutriment for host, while also helping host defense
The invasion of external microorganism.At least following two aspect of the function of plant-feed insect enteric microorganism: first is that the life of nutriment
Object synthesis, first is that the depolymerization of biomass.Why termite can be food with timber, exactly because its enteric microorganism can degrade
Macromolecular cellulose, and it is converted to nutrition.Therefore existing in insect gut largely can use rich in cellulose, xylan etc.
Microorganism of the plant as energy source.
Summary of the invention
It is an object of the invention to provide a kind of degradable agricultural wastes using the microorganism for deriving from insect gut
Complex micro organism fungicide, to reduce environmental pollution.
The invention is realized by the following technical scheme:
Applicant isolates five kinds of microorganisms from the enteron aisle of plant-feed insect first, identified to be respectively designated as sphingolipid bar
Bacterium (Sphingobacterium sp.) HP455, Fen Shi fiber germ (Cellulosimicrobium sp.) HP181, alkali is produced
Pseudomonad (Pseudomonas sp.) HP207, acid-producing Klebsiella bacterium (Klebsiella oxytoca) BD177, Ke Shi lemon
Lemon acidfast bacilli (Citrobacter koseri) BD195, this five kinds of microorganisms have certain degradation capability to agricultural wastes.
Then, the complex micro organism fungicide of degradable agricultural product castoff is prepared using this five kinds of microorganisms, lives
Property ingredient be above-mentioned bacterial strains it is one or more.The complex micro organism fungicide further includes carrier, the carrier be wheat bran, rice husk,
One of wheat husk, vinasse, chitosan, sawdust, bacteria residue, plant ash, bagasse, peel residue, stalk, fowl and animal excrement are a variety of.
And then, applicant additionally provides a kind of straw degradative complex micro organism fungicide, and active constituent is above-mentioned strain
In Fen Shi fiber germ HP181 and Pseudomonas alcaligenes HP207.Applicant has carried out straw degradative examination to the composite bacteria agent
It tests, the results showed that composite bacteria agent is much higher than single culture to the degradation capability of stalk.
Finally, applicant also is prepared for can promote the composite bacteria agent of plant growth using above five kinds of microorganisms, this is compound
Microbial inoculum is prepared as follows: by 100ml bacillus containing sphingolipid HP455, Fen Shi fiber germ HP181, Pseudomonas alcaligenes
HP207, acid-producing Klebsiella bacterium BD177 and Ke Shi citric acid bacillus BD195 are 107~109The bacterium solution of CFU/ml and 100g wheat
Bran is uniformly mixed, and is then uniformly sprinkling upon on the discarded citrus fruit of 1000g and is carried out stacking, ferments 30 days and be made.Test result shows:
The composite bacteria agent can significantly promote crop growth, and raising crops seedling stage is disease-resistant, salt resistance, freeze proof equal resistance environment-stress ability,
Increase the survival rate of crop growth early stage.
Specific embodiment
The present invention is described in detail below with reference to embodiment.
1 isolation and identification for strains of embodiment
(1) separation of insect gut microorganism: picking citrus fruit fly, dacus dorsalis, scarab beetle, longicorn, termite, Huang
The adult or larva of the plant-feed insects such as mealworm, stratiomyiid carry out surface sterilization processing to polypide, to exclude external microorganism
It influences, solution is put into the sterile homogenizer equipped with 300ul~500ul sterile water after cutting complete enteron aisle, careful dilute after grinding sufficiently
It releases, bacterium screening can be cultivated by applying the progress of NA plate.
(2) identification of insect gut microorganism: single colonie is chosen in NA plate culture, is inoculated in LB liquid medium, 28-37
DEG C constant temperature incubation 12-48h.Using the type of microbiology and Protocols in Molecular Biology identification bacterial strain, and preservation is spare.Pass through mirror
It is fixed to obtain five kinds of bacterial strains, respectively sphingolipid bacillus, Fen Shi fiber germ, production alkali vacation unit cell altogether out of plant-feed insect enteron aisle
Bacterium, acid-producing Klebsiella bacterium and Ke Shi citric acid bacillus, five kinds of bacterial strains are delivered on October 20th, 2015 positioned at Wuhan City
China typical culture collection center (CCTCC) in Wuhan University carries out preservation, and the name of each strain and deposit number are as follows:
Sphingolipid bacillus (Sphingobacterium sp.) HP455, deposit number are CCTCC NO:M2015618.
Fen Shi fiber germ (Cellulosimicrobium sp.) HP181, deposit number are CCTCC NO:
M2015619。
Pseudomonas alcaligenes (Pseudomonas sp.) HP207, CCTCC NO:M2015620.
Acid-producing Klebsiella bacterium (Klebsiella oxytoca) BD177, CCTCC NO:M2015621.
Ke Shi citric acid bacillus (Citrobacter koseri) BD195, CCTCC NO:M2015622.
Embodiment 2: the composite bacteria agent of degrading straw
By separated plant-feed insect enteric microorganism Fen Shi fiber germ HP181, the Pseudomonas alcaligenes identified
Both HP207 be inoculated in together enriched medium building mixed culture, 32 DEG C, 1100r/min shaking table culture for 24 hours.
The component of enriched medium are as follows: 0.1% yeast extract, 0.5% tryptone, 0.5%CaCO3, 0.5%NaCl,
KH2PO40.094g, K2HPO40.19g, distilled water 200ml, pH are natural (about 7).
Through detecting, OD value of the mixed bacteria liquid at 600nm after culture is 0.6, wherein the germ of fiber containing Fen Shi 0.8~
1.5×108CFU/ml contains Pseudomonas alcaligenes 0.9~1.7 × 108CFU/ml, by the mixed bacteria liquid in the ratio of 5% (v/v)
Straw degradative culture medium is accessed, 35 DEG C of stationary cultures 15d and 30d detect the quality of remaining stalk.
The component of straw degradative culture medium are as follows: 0.1% yeast extract, 0.5%CaCO3, 0.5%NaCl,
KH2PO40.094g, K2HPO40.19g, 1% rice straw (long 1cm), distilled water 200ml, pH are natural.
The measuring method of straw degradative rate: the supernatant of culture solution is carefully discarded, by remaining stalk dilute hydrochloric acid repeated flushing
For several times, then with sterile water repeated flushing, filtered through gauze is until neutral, and 60 DEG C drying to constant weight and claims its quality, calculates weight-loss ratio.
Single Fen Shi fiber germ, Pseudomonas alcaligenes are separately taken, is operated by method identical with mixed bacteria liquid, as control,
It the results are shown in Table 1.
1. straw degradative situation of table
The results showed that Fen Shi fiber germ, Pseudomonas alcaligenes and its mixed bacteria liquid energy obvious degradation stalk, and
The premature degradation ability of single bacterium solution is higher than Mixed Microbes, and the later stage degradation ability of mixed bacteria liquid is stronger, the degradation weight-loss ratio of stalk
It can reach 58%.
The composite bacteria agent of the promotion plant growth of embodiment 3
1) preparation of composite bacteria agent
By Fen Shi fiber germ HP181, Pseudomonas alcaligenes HP207, sphingolipid bacillus HP455, acid-producing Klebsiella bacterium
BD177, Ke Shi citric acid bacillus BD195 are inoculated in 100ml LB liquid medium with the access amount of in total 5% (v/v), 30 DEG C,
100r/min shaking table culture is 0.5 using OD value of the spectrophotometer measurement bacterium solution at 600nm, through detecting, containing sweet smell in bacterium solution
Family name's fiber germ 0.2 × 109CFU/ml, Pseudomonas alcaligenes 0.7 × 108CFU/ml, sphingolipid bacillus 1.5 × 108CFU/ml, production
Sour Klebsiella 5.6 × 108CFU/ml, Ke Shi citric acid bacillus 8.9 × 107CFU/ml。
Above-mentioned 100ml bacterium solution and 100g wheat bran are sufficiently mixed uniformly, are then uniformly sprinkling upon the wheat bran containing bacterium solution
1000g, which is discarded on citrus fruit, carries out stacking, and composite bacteria agent, living bacteria count >=0.5 hundred million of the microbial inoculum are made after fermentation 30 days.
2) quality testing of composite bacteria agent
Composite bacteria agent is detected according to the agricultural industry examination criteria NY525-2012 of organic fertilizer, the results are shown in Table 2.
The quality index of 2. composite bacteria agent of table
| The project indicator | Organic fertilizer technique standard | Composite bacteria agent |
| PH value | 5.5~8.5 | 6.0 |
| Organic matter (in terms of butt) | ≥40 | 88.50% |
| Full nitrogen (in terms of N) | 2.63% | |
| Available phosphorus is (with P2O5Meter) | 1.79% | |
| Effective potassium is (with K2O meter) | 1.31% | |
| Total nutrient (N+P2O5=K2O) (in terms of butt) | >=5% | 5.73% |
| Moisture, % | ≤ 30% | 27.39% |
| Excrement colibacillus group, a/g | ≤100 | 60 |
| Induced worm egg death rate, % | ≥95 | 98 |
| Limited period, the moon | ≥6 | 12 |
3) growth promoting function of the composite bacteria agent to crop:
The sandy soil 2kg for weighing composite bacteria agent 200g and drying, adds water and stirs and is sufficiently mixed, and adjusting soil moisture is 65%,
18-25 DEG C of temperature, and 30 days natural degradation products of discarded tangerine fruit that composite bacteria agent processing is not used is used to carry out potting as control
Test, kind be in double 11 Brassica campestris L seedling as test material, 3 basins of each processing, respectively measurement getting up early survival rate and strain in 30 days
Height, and it is for statistical analysis, it the results are shown in Table 3.
Growth-promoting growth of 3 composite bacteria agent of table to rape
| Treatment outcome (%) | 30 days plant heights (cm) | |
| Composite bacteria agent processing | 98 | 34.10±1.47 |
| Control treatment | 86 | 30.46±1.09 |
The results showed that composite bacteria agent can significantly promote crop growth, raising crops seedling stage is disease-resistant, and salt resistance is freeze proof
Deng resistance environment-stress ability, increase the survival rate of crop growth early stage.
Claims (3)
1. a kind of complex micro organism fungicide for degrading straw, it is characterised in that: active constituent is Fen Shi fiber germ HP181
With Pseudomonas alcaligenes HP207, the Fen Shi fiber germ HP181, it is deposited in China typical culture collection center, preservation is compiled
Number be CCTCC NO:M2015619；The Pseudomonas alcaligenes HP207, is deposited in China typical culture collection center, preservation
Number is CCTCC NO:M2015620.
2. a kind of for promoting the complex micro organism fungicide of plant growth, it is characterised in that: active constituent is sphingolipid bacillus
HP455, Fen Shi fiber germ HP181, Pseudomonas alcaligenes HP207, acid-producing Klebsiella bacterium (Klebsiella oxytoca)
BD177 and Ke Shi citric acid bacillus (Citrobacter koseri) BD195, the sphingolipid bacillus HP455 is deposited in Chinese allusion quotation
Type culture collection, deposit number CCTCCNO:M2015618；The Fen Shi fiber germ HP181, is deposited in China
Type Tissue Collection, deposit number are CCTCC NO:M2015619；The Pseudomonas alcaligenes HP207, is deposited in
State's Type Tissue Collection, deposit number are CCTCC NO:M2015620；The acid-producing Klebsiella bacterium BD177, preservation
In China typical culture collection center, deposit number is CCTCC NO:M2015621；The Ke Shi citric acid bacillus BD195,
It is deposited in China typical culture collection center, deposit number is CCTCC NO:M2015622.
3. a kind of method for preparing complex micro organism fungicide described in claim 2, it is characterised in that: by 100ml bacillus containing sphingolipid
HP455, Fen Shi fiber germ HP181, Pseudomonas alcaligenes HP207, acid-producing Klebsiella bacterium BD177 and Ke Shi citric acid bacillus
BD195 is 107~109The bacterium solution of CFU/ml is uniformly mixed with 100g wheat bran, is then uniformly sprinkling upon 1000g and is discarded citrus fruit
Upper carry out stacking is fermented 30 days and is made.
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| CN201610498520.9A CN106011018B (en) | 2016-06-29 | 2016-06-29 | A kind of complex micro organism fungicide of degradable agricultural product castoff |
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| CN106011018B true CN106011018B (en) | 2019-10-15 |
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Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104560810A (en) * | 2014-12-29 | 2015-04-29 | 河南城建学院 | Compound microbial fertilizer and application thereof |
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104560810A (en) * | 2014-12-29 | 2015-04-29 | 河南城建学院 | Compound microbial fertilizer and application thereof |
Non-Patent Citations (5)
| Title |
|---|
| Screening of a composite microbial system and its characteristics of wheat straw degradation.;Li Pei-pei et al.;《Agricultural sciences in China》;20111021;第10卷(第10期);第1586-1594页 * |
| 农林剩余物综合利用的研究现状;刘飞等;《农机化研究》;20150228(第2期);第232页第2栏第3段 * |
| 复合微生物菌剂在有机堆肥中的应用研究;孙干等;《北方园艺》;20150815(第15期);第166-170页 * |
| 暗黑鳃金龟不同发育阶段肠道微生物及糖苷水解酶多样性研究;圣平;《中国博士学位论文全文数据库农业科技辑》;20140915(第9期);第79页最后1段，摘要，第97页最后1段 * |
| 纤维素降解细菌DBJ的筛选鉴定及其特性研究;吴翔等;《西南农业学报》;20160131;第29卷(第1期);摘要，第81页第1栏第2段 * |
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