CN107568248A - A kind of controlling plant diseases simultaneously strengthen microbial bacterial agent of plant stress-resistance and its preparation method and application - Google Patents
A kind of controlling plant diseases simultaneously strengthen microbial bacterial agent of plant stress-resistance and its preparation method and application Download PDFInfo
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Abstract
It is used to controlling disease and promote microbial bacterial agent of cotton growth and its preparation method and application the invention provides a kind of, belongs to agricultural microorganism bacterial manure technical field.The present invention embodies synergistic function by the way that Te Jila bacillus and Sphingobacterium are compounded, to the biocidal property aspect of plant disease, while strengthens plant stress-resistance ability, makes the function diversification of microbial bacterial agent, realizes the effect of one multi-purpose.The microbial bacterial agent is in preventing and treating cotton soil-borne disease bacterium and strengthens application of the Resistance Strain of Cotton in.
Description
Technical field
The invention belongs to agricultural microorganism bacterial manure technical field, and in particular to one kind is used for controlling disease and strengthens Genes For Plant Tolerance
Microbial bacterial agent of inverse ability and its preparation method and application.
Background technology
Cotton soil-borne disease is worldwide important cotton disease, has destructiveness, and feature is wide distribution, harm weight, host
Scope is wide, route of transmission is more, the time-to-live is long.China prevents and treats cotton disease mainly based on chemical pesticide at present, biological pesticide
Supplemented by, and chemical pesticide has the deficiencies of high poison, high residue and difficult degradation.Because people largely use for a long time so that the ecological balance
By destroying, injure natural enemy, drug resistance pest and disease damage substantial increase etc..
With society progress, emphasis of concern be increasingly turned to low toxicity, low-residual, it is pollution-free, do not hinder natural enemy,
Difficulty such as develops immunity to drugs at the harmless boilogical agricultural chemicals.Comprehensive measures for the prevention and control based on biological prevention and control because its have it is environment-friendly, sick
Opportunistic pathogen is not likely to produce the advantages such as resistance and is increasingly subject to people's attention, and achieves certain effect, and verticillium wilt is passed in cotton soil
Had very important significance in prevention and control.It is concentrated mainly on currently with biological method prevention and control cotton soil-borne disease by artificially adding
The biocontrol microorganisms of cotton germ can be suppressed by adding, and suppress the sprouting of pathogen spore in soil, reduce the number of pathogenic bacteria germ in soil
Amount.Therefore, applying has antagonism to cotton pathogen, and has the Biological agents of facilitation to cotton growth to suppress
The harmful microorganism such as germ bacterium grows in the soil of cotton root area, promotes beneficial bacterium growth and breeding, for effectively control Continuous Mono-cropping
Under cotton disease generation, realize Cotton Industry production sustainable development there is highly important theory significance and using valency
Value.Microbial pesticide is developed, the use of chemical pesticide is reduced or avoided, the highly effective and safe production for realizing Cotton Production is situation
Become.But because microorganism biocontrol agent is live body bacterium, there is also many up for solving the problems, such as in its exploitation.Example
Such as, microorganism biocontrol microorganisms are usually prepared using single bacterium at present, and effect is single, and antimicrobial spectrum is narrow, influences microorganism system
Standby application and development.
The content of the invention
In view of this, it is used for controlling plant diseases it is an object of the invention to provide a kind of and strengthens plant stress-resistance ability
Microbial bacterial agent and its preparation method and application, make the microbial bacterial agent that there are efficient long-acting controlling plant diseases to strengthen plant
Degeneration-resistant function.
In order to realize foregoing invention purpose, the present invention provides following technical scheme：
The invention provides a kind of microbial bacterial agent for controlling plant diseases, include the group of following weight/mass percentage composition
Point：Mixed fungus fermentation liquid 1%~50%, wetting agent 1%~5%, dispersant 1%~5%, binding agent 1%~5%, disintegrant 0~
15%th, protective agent 0.1%~2%, the filler of surplus；
Viable bacteria concentration >=5 × 10 of the mixed fungus fermentation liquid in microbial bacterial agent10CFU/g；The mixed fungus fermentation liquid bag
Include Te Jila bacillus and Sphingobacterium；The ratio of the thalline number of the Te Jila bacillus and Sphingobacterium is
5~9：4~1.
Preferably, the component of following weight/mass percentage composition is included：Mixed fungus fermentation liquid 10%~40%, wetting agent 2%~
4%th, dispersant 2%~4%, binding agent 2%~4%, disintegrant 1~10%, protective agent 0.5~1.5%, the filler of surplus.
Preferably, viable bacteria concentration >=8 × 10 of the mixed fungus fermentation liquid in microbial bacterial agent10CFU/g。
Preferably, the ratio of the thalline number of the Te Jila bacillus and Sphingobacterium is 9：1.
Preferably, the filler is diatomite.
Preferably, the protective agent is humic acid or xanthan rubber powder.
The invention provides the preparation for controlling plant diseases and the microbial bacterial agent for strengthening plant stress-resistance ability
Method, comprise the following steps：
1) Te Jila bacillus and Sphingobacterium seed liquor are seeded in PDB fluid nutrient mediums in 25~28 DEG C of temperature
Aerobic fermentation 24~96h of culture is carried out under the conditions of degree, obtains mixed fungus fermentation liquid；
2) the mixed fungus fermentation liquid for obtaining the step 1) and wetting agent, dispersant, suspending agent, protective agent, disintegrant and
Filler mixes, and obtains mixed material；
3) mixed material is mixed with binding agent, crushes, obtain microbial bacterial agent.
Preferably, the pH value of the PDB fluid nutrient mediums of fermented and cultured process is 7.0 in the step 1).
Preferably, the particle diameter of crushing is 40~50 mesh.
Present invention also offers microbial bacterial agent prepared by the microbial bacterial agent or methods described in preventing and treating cotton soil
Pass germ and promote the application in cotton growth.
Preferably, the cotton soil-borne disease bacterium bag includes that verticillium wilt, alternaria solani sorauer, eggplant phytophthora, eggplant Huang wither bacterium or tomato withers
Bacterium.
The invention provides a kind of microbial bacterial agent for controlling plant diseases and promoting growth of plants, by by Te Jila
Bacillus and Sphingobacterium compounding, synergistic function is embodied to the biocidal property aspect of plant disease, while strengthen plant
Thing anti-adversity ability, make the function diversification of microbial bacterial agent, realize the effect of one multi-purpose.Microbial bacterial agent provided by the invention
Increase substantially in the biocidal property method of cotton soil-borne disease bacterium, press down than single microbial inoculum (Te Jila bacillus or Sphingobacterium)
Bacterium rate improves 36.7%~131.3%.
Further, the present invention is aided with humic acid as protective agent, using diatomite as filler, same to dispersant, wetting agent
Acted on simultaneously Deng auxiliary agent, ensure that there is higher biocompatibility between each auxiliary agent and thalline, microbial cells can be effectively improved
Germination rate and survival rate, so as to providing certain guarantee for microorganism long-acting bacteriostatic, diatomite makes biocontrol microorganisms as filler
The equal ＞ 92% of gemma germination rate and survival rate of (Te Jila bacillus and Sphingobacterium), is carried compared with other conventional fillers
It is high by 10%~30%；Using humic acid as protective agent, the viable bacteria rate ratio of the microbial bacterial agent that is prepared after 150d is applied
Blank control group is compared and improves 15%.
Brief description of the drawings
Fig. 1 is the finished product aspect graph of microbial bacterial agent provided by the invention.
Embodiment
The invention provides a kind of microbial bacterial agent for being used for controlling plant diseases and promoting growth of plants, including following quality
The component of percentage composition：Mixed fungus fermentation liquid 1%~50%, wetting agent 1%~5%, dispersant 1%~5%, binding agent 1%~
5%th, disintegrant 0~15%, protective agent 0.1%~2%, the filler of surplus；
Viable bacteria concentration >=5 × 10 of the mixed fungus fermentation liquid in microbial bacterial agent10CFU/g；The mixed fungus fermentation liquid bag
Include Te Jila bacillus (Bacillus tequilensis) and Sphingobacterium；The Te Jila bacillus and sphingol
The ratio of the thalline number of bacillus is 5~9：4~1.
Include mixed fungus fermentation liquid provided by the present invention for the microbial bacterial agent of controlling plant diseases and promoting growth of plants.With
The quality of microbial bacterial agent is 100% meter, and the mass percent of the mixed fungus fermentation liquid is 1%~50%, more preferably 10%
~40%, most preferably 25%.Viable bacteria concentration >=5 × 10 of the mixed fungus fermentation liquid in microbial bacterial agent10CFU/g, preferably
For >=8 × 1010CFU/g, more preferably (9~10) × 1010CFU/g.The mixed fungus fermentation liquid include Te Jila bacillus and
Sphingobacterium；The ratio of the thalline number of the Te Jila bacillus and Sphingobacterium is 5~9：4~1, preferably 7
~9：3~1, more preferably 9：1.In the mixed fungus fermentation liquid form of Te Jila bacillus and Sphingobacterium be thalline or
Spore.The growth phase of the thalline is preferably logarithmic phase.The Te Jila bacillus and Sphingobacterium can effectively press down
The growth of plant soil-borne germ processed, while effectively improve plant stress-resistance ability by promoting plant growth.The present invention is to institute
The bacterial strain for stating Te Jilatejila bacillus and Sphingobacterium is not particularly limited, and use is well known to those skilled in the art
Bacterial strain.In the embodiment of the present invention, the bacterial strain of the Te Jila bacillus is Te Jila bacillus C-9, the sheath
The bacterial strain of ammonia alcohol bacillus is that Sphingobacterium A1, the Te Jila bacillus C-9 and Sphingobacterium A1 are big by Shihezi
Biotechnology key lab provides, and promises to undertake and freely provide Sphingobacterium A1 and Te Jila bacillus to the public
C-9, time limit are from the applying date in 20 years.
Include wetting agent provided by the present invention for the microbial bacterial agent of controlling plant diseases and promoting growth of plants.With micro- life
The quality of thing microbial inoculum is 100% meter, and the weight/mass percentage composition of the wetting agent is 1%~5%, preferably 2%~4%, more excellent
Elect 3% as.The species of the wetting agent is preferably neopelex.The source of the wetting agent is not particularly limited,
Using wetting agent well-known to those skilled in the art.The wetting agent has as a kind of anion surfactant
Good surface-active, hydrophily are stronger.The tension force of oil-water interface is effectively reduced, reaches emulsification.
Include dispersant provided by the present invention for the microbial bacterial agent of controlling plant diseases and promoting growth of plants.With micro- life
The quality of thing microbial inoculum is 100% meter, and the mass percent of the dispersant is 1%~5%, preferably 2%~4%, more preferably
For 3%.The dispersant is preferably sodium lignin sulfonate (CMN) or sodium tripolyphosphate (STPP), more preferably lignin sulfonic acid
Sodium.The source of the dispersant is not particularly limited, using dispersant well-known to those skilled in the art.It is described scattered
Agent can greatly improve the suspensibility and wettability of wettable powder.
Include disintegrant provided by the present invention for the microbial bacterial agent of controlling plant diseases and promoting growth of plants.With micro- life
The quality of thing microbial inoculum is 100% meter, the mass percent of the disintegrant is 0%~15%, preferably 1%~10%, it is more excellent
Elect 5% as.The disintegrant is preferably ammonium sulfate or sodium chloride, more preferably ammonium sulfate.The source of the disintegrant is without spy
Different limitation, using disintegrant well-known to those skilled in the art.The disintegrant can guarantee that particle has and preferably collapse
Solution and dispersion effect.
Include binding agent provided by the present invention for the microbial bacterial agent of controlling plant diseases and promoting growth of plants.With micro- life
The quality of thing microbial inoculum is 100% meter, and the mass percent of the binding agent is 0~15%, preferably 1~10%, more preferably
5%.The species of the binding agent is preferably soluble starch or polyvinyl alcohol.The source of the binding agent is not particularly limited,
Using binding agent well-known to those skilled in the art.The binding agent can greatly improve granulating.
Include protecting provided by the present invention for preventing and treating plant soil-borne diseases and strengthening the microbial bacterial agent of plant stress-resistance ability
Protect agent.Counted using the quality of microbial bacterial agent as 100%, protectant mass percent is 0.1%~2%, preferably 0.5
~1.5%, more preferably 1%.Protectant species is preferably humic acid or xanthan rubber powder, more preferably humic acid.Institute
State protectant source to be not particularly limited, using protective agent well-known to those skilled in the art.The protective agent energy
A stable environment is provided for microbial bacterial agent, effectively viable bacteria is by poor environments such as ultraviolet, temperature effectively in reduction microbial inoculum
Influence, extend the storage time of microbial inoculum；On the other hand, humic acid can be effectively improved the situations such as soil hardening, there is enhancing plant
Degeneration-resistant ability.
Include filler provided by the present invention for controlling plant diseases and the microbial bacterial agent that strengthens plant stress-resistance ability.With
The quality of microbial bacterial agent is 100% meter, and the mass percent of the filler is to be supplemented to 100%.The species of the filler is excellent
Elect diatomite as.The source of the filler is not particularly limited, using filler well-known to those skilled in the art.It is described
Filler can maintain microbial bacterial agent pH value neutral, nontoxic, and suspendability is good, and absorption property is strong, and unit weight is light, and oil absorbency is high, mixing
Uniformity is good, will not block agricultural machinery pipeline during use, and moisturizing, loose soil property can be played in soil, extends drug effect fertilizer efficiency time,
Encourage crop growth effect.
The invention provides the preparation method of microbial bacterial agent described in above-mentioned technical proposal, comprise the following steps：
1) Te Jila bacillus and Sphingobacterium seed liquor are seeded in PDB fluid nutrient mediums in 25~37 DEG C of temperature
Aerobic fermentation 24~96h of culture is carried out under the conditions of degree, obtains mixed fungus fermentation liquid；
2) the mixed fungus fermentation liquid for obtaining the step 1) and wetting agent, dispersant, suspending agent, protective agent, disintegrant and
Filler mixes, and obtains mixed material；
3) mixed material is mixed with binding agent, crushes, obtain microbial bacterial agent.
The present invention by the Te Jila bacillus of activation and Sphingobacterium in PDB fluid nutrient mediums in 25~37 DEG C of temperature
Aerobic fermentation 24~96h of culture is carried out under the conditions of degree, obtains mixed fungus fermentation liquid.
In the present invention, the Te Jila bacillus and Sphingobacterium are preferably activated respectively.The side of the activation
Method is not particularly limited, using activation method well-known to those skilled in the art.The activation is preferably with culture medium
PDA culture medium.The thalline quantity ratio of the Te Jila bacillus and Sphingobacterium is 5~9：4~1.
In the present invention, the method for the inoculation is not particularly limited, using inoculation side well-known to those skilled in the art
Method.The inoculum concentration of the inoculation is preferably 1~10%, more preferably 4~8%, most preferably 5%.
In the present invention, the pH value of the PDB fluid nutrient mediums of the fermented and cultured process is preferably 5.5~8.0, more preferably
7.0。
The temperature of the fermentation is preferably 28 DEG C.The time of the fermentation is preferably 40~80h, more preferably 50~70h,
Most preferably 60h.
It is of the invention by the mixed fungus fermentation liquid and wetting agent, dispersant, suspending agent, protection after obtained mixed fungus fermentation liquid
Agent, disintegrant and filler mixing, obtain mixed material.In the present invention, the method for the mixing is not particularly limited, using ability
Mixed method known to field technique personnel.
After obtaining mixed material, the present invention mixes the mixed material with binding agent, crushes, obtains microbial bacterial agent.
The present invention is not particularly limited to the method for crushing, using breaking method well-known to those skilled in the art.Crush
Particle diameter is preferably 30~60 mesh, more preferably 40 mesh.
In the present invention, after the crushing, preferably add water to be granulated, dry obtained comminuting matter and screening is packed, obtain
Microbial bacterial agent granule finished product.
In the present invention, the method that described plus water is granulated is not particularly limited, using granulation well known to those skilled in the art
Method.Described plus water adds the volume of water to be the 10~30% of microbial bacterial agent gross mass when being granulated.The size of the particle
For 1~3mm.
In the present invention, the method for the drying is not particularly limited, using drying side well-known to those skilled in the art
Method.The temperature of the drying is preferably 40~60 DEG C, more preferably 45 DEG C.
In the present invention, the method for the packaging is not particularly limited, using packaging side well-known to those skilled in the art
Method.The specification of the packaging is preferably 100~1000g/ bags.
In the present invention, after the microbial inoculum stores 150d respectively under the conditions of 4 DEG C, 25 DEG C, 45 DEG C, effective viable bacteria amount distinguishes energy
Reach 85%, 78%, more than 75%, this illustrates that microbial inoculum stability of the present invention is preferable.
According to the performance of mentioned microorganism microbial inoculum, present invention also offers the microbial bacterial agent or methods described to prepare
Microbial bacterial agent preventing and treating cotton soil-borne disease bacterium and promote cotton growth in application.
In the present invention, the cotton soil-borne disease bacterium preferably include verticillium wilt, alternaria solani sorauer, eggplant phytophthora, eggplant Huang wither bacterium or kind
Eggplant Fusarium oxysporum.
In the present invention, the amount of application of the microbial bacterial agent is 500~2000g/ mus, more preferably 1000g/ mus.It is described
The application process of microbial bacterial agent is drip irrigation or spreaded manuer in holes.The number of days of applying of the microbial bacterial agent is 3~5d, more preferably 5d.
The adjacent administered twice interval number of days of the microbial bacterial agent is 10~20d.
A kind of it be used for controlling plant diseases to provided by the invention with reference to embodiment and strengthen plant stress-resistance ability
Microbial bacterial agent and its preparation method and application is described in detail, but they can not be interpreted as protecting model to the present invention
The restriction enclosed.
Embodiment 1
The Te Jila bacillus and Sphingobacterium bacterium provided by biotechnology key lab of Shihezi Univ unit
Kind is activated in PDA culture medium respectively.Te Jila bacillus and Sphingobacterium are inoculated in PDB respectively after being activated
28 DEG C in culture medium, 160r/min culture 16h, obtain the seed liquor of Te Jila bacillus and Sphingobacterium.
It is 9 by volume by Te Jila bacillus and Sphingobacterium seed liquor:1st, inoculum concentration is seeded in PDB for 10%
Aerobic fermentation culture 96h is carried out in fluid nutrient medium under 28 DEG C, 160r/min temperature conditionss, obtains mixed fungus fermentation liquid.According to
Agar block diffusion method measure mixed fungus fermentation liquid Huang is withered compared with single bacterial strain and zymotic fluid bacterium antibacterial circle diameter influence be respectively-
20% and+41.3%
Obtained mixed fungus fermentation liquid is determined into OD values under UV detector device, according to standard curve, obtains mixed bacterium
The cell concentration of zymotic fluid is 5~9 × 1010CFU/ml。
Embodiment 2
The Te Jila bacillus and Sphingobacterium bacterium provided by biotechnology key lab of Shihezi Univ unit
Kind is activated in PDA culture medium respectively.Te Jila bacillus and Sphingobacterium after being activated are inoculated in respectively
28 DEG C in PDB culture mediums, 160r/min culture 16h, obtain the seed liquor of Te Jila bacillus and Sphingobacterium.
By Te Jila bacillus and Sphingobacterium seed liquor by volume 9:1st, inoculum concentration 5% is seeded in PDB liquid
Aerobic fermentation culture 96h is carried out in culture medium under 28 DEG C, 160r/min temperature conditionss, obtains mixed fungus fermentation liquid.According to agar
Block diffusion method measures mixed fungus fermentation liquid compared with the suppression of single bacterial strain Te Jila bacillus or Sphingobacterium zymotic fluid to the yellow bacterium that withers
The influence of bacterium loop diameter is respectively+36.7% and+116%
Obtained mixed fungus fermentation liquid is determined into OD values under UV detector device, according to standard curve, obtains mixed bacterium
The cell concentration of zymotic fluid is 5~10 × 1010CFU/ml。
Embodiment 3
The Te Jila bacillus and Sphingobacterium bacterium provided by biotechnology key lab of Shihezi Univ unit
Kind is activated on LB solid mediums respectively.Te Jila bacillus C-9 and Sphingobacterium connect respectively after being activated
Kind is 28 DEG C in LB fluid nutrient mediums, 160r/min culture 16h, obtains the seed of Te Jila bacillus and Sphingobacterium
Liquid.
It is 9 by volume by Te Jila bacillus and Sphingobacterium seed liquor:1st, inoculum concentration 5% is seeded in LB liquid
28 DEG C in culture medium, aerobic fermentation culture 96h is carried out under the conditions of 160r/min, obtains mixed fungus fermentation liquid.Spread according to agar block
It is straight to the inhibition zone of the yellow bacterium that withers compared with single bacterial strain Te Jila bacillus or Sphingobacterium zymotic fluid that method measures mixed fungus fermentation liquid
The influence in footpath is respectively -36.7% and -25%.
Obtained mixed fungus fermentation liquid is determined into OD values under UV detector device, according to standard curve, obtains mixed bacterium
The cell concentration of zymotic fluid is 5~9 × 1010CFU/ml。
Comparative example 1
Had phytopathogen compared with high inhibition effect by what biotechnology key lab of Shihezi Univ unit provided
Bacillus cercus X4 and N4 preservation of bacteria strain activated respectively on PDA solid mediums.Wax-like bud after being activated
Spore bacillus X4 and N4 are inoculated in PDB fluid nutrient mediums 28 DEG C, 160r/min culture 16h respectively, obtain Bacillus cercus X4
With N4 seed liquor.
It is 9 by volume by Bacillus cercus X4 and N4 seed liquor:1st, inoculum concentration 5% is seeded in LB fluid nutrient mediums
28 DEG C, aerobic fermentation culture 96h is carried out under the conditions of 160r/min, obtain mixed fungus fermentation liquid.Measured according to agar block diffusion method mixed
The influence of the more single Bacillus cercus X4 of fermented liquid and N4 zymotic fluids to the antibacterial circle diameter of the yellow bacterium that withers is respectively+23.5%
With+7.7%.
Obtained mixed fungus fermentation liquid is determined into OD values under UV detector device, according to standard curve, obtains mixed bacterium
The cell concentration of zymotic fluid is 5~9 × 108CFU/ml。
Comparative example 1 test result indicates that, not any Bacillus species mixing can obtain the antibacterial effect of the present invention
Fruit.The high efficiency and optimality of two kinds of bacterial strains selected by the present invention in the antibacterial middle application of mixed fungus fermentation.
Embodiment 4
Mixed fungus fermentation liquid prepared by embodiment 1, with wetting agent neopelex, dispersant sodium lignin sulfonate
(CMN), protective agent humic acid (Humic acid), disintegrant ammonium sulfate, binding agent soluble starch and filler are made with diatomite
Carrier mixes, and specific quality proportioning is as follows：Mixed fungus fermentation liquid 50%, neopelex 1%, sodium lignin sulfonate
(CMN) 5%, humic acid (Humic acid) 0.1, ammonium sulfate 15%, soluble starch 1% and diatomite are supplemented to 100%.Institute
State viable bacteria concentration of the mixed fungus fermentation liquid in microbial bacterial agent and reach 5 × 1010CFU/g。
Embodiment 5
Mixed fungus fermentation liquid prepared by embodiment 2, with wetting agent neopelex, pentasodium triphosphate of dispersing agent
(STPP), protective agent xanthan rubber powder, disintegrant sodium chloride, binding agent polyvinyl alcohol and filler diatomite make carrier mixing.Specifically
Quality proportioning it is as follows：Mixed fungus fermentation liquid 20%, neopelex 5%, sodium tripolyphosphate (STPP) 1%, xanthans
Powder 1%, sodium chloride 10%, polyvinyl alcohol 2% and diatomite are supplemented to 100%.The mixed fungus fermentation liquid is in microbial bacterial agent
Viable bacteria concentration reach 8 × 1010CFU/g。
Embodiment 6
Mixed fungus fermentation liquid prepared by embodiment 3, with wetting agent neopelex, dispersant sodium lignin sulfonate
(CMN), protective agent humic acid, disintegrant sodium chloride, binding agent polyvinyl alcohol and diatomite make carrier mixing.Specific quality is matched somebody with somebody
Than as follows：Mixed fungus fermentation liquid 40%, neopelex 2%, sodium lignin sulfonate (CMN) 4%, humic acid 0.5%, chlorine
Change sodium 10%, polyvinyl alcohol 2%, diatomite complements to 100%.Viable bacteria concentration of the mixed fungus fermentation liquid in microbial bacterial agent
Reach 6 × 1010CFU/g。
Embodiment 7
Mixed fungus fermentation liquid prepared by embodiment 3, with wetting agent neopelex, dispersant sodium lignin sulfonate
(CMN), protective agent humic acid, disintegrant sodium chloride, binding agent polyvinyl alcohol and diatomite make carrier mixing.Specific quality is matched somebody with somebody
Than as follows：Mixed fungus fermentation liquid 10%, neopelex 4%, sodium lignin sulfonate (CMN) 2%, humic acid 1.5%, chlorine
Change sodium 1%, polyvinyl alcohol 4%, diatomite complements to 100%.Viable bacteria concentration of the mixed fungus fermentation liquid in microbial bacterial agent
Reach 5 × 1010CFU/g。
Embodiment 8
Mixed fungus fermentation liquid prepared by embodiment 2, with wetting agent neopelex, dispersant sodium lignin sulfonate
(CMN), protective agent humic acid (Humic acid), disintegrant ammonium sulfate, binding agent soluble starch and filler are made with diatomite
Carrier mixes, and specific quality proportioning is as follows：Mixed fungus fermentation liquid 25%, neopelex 3%, sodium lignin sulfonate
(CMN) 3%, humic acid (Humic acid) 1%, ammonium sulfate 5%, soluble starch 3% and diatomite are supplemented to 100%.Institute
State viable bacteria concentration of the mixed fungus fermentation liquid in microbial bacterial agent and reach 9 × 1010CFU/g。
Comparative example 2
Mixed fungus fermentation liquid prepared by embodiment 2, with wetting agent neopelex, dispersant sodium lignin sulfonate
(CMN), protective agent carboxymethyl cellulose, disintegrant ammonium sulfate, binding agent soluble starch and filler are mixed with diatomite as carrier
Close, specific quality proportioning is as follows：Mixed fungus fermentation liquid 25%, neopelex 3%, sodium lignin sulfonate (CMN)
3%th, carboxymethyl cellulose (Humic acid) 1%, ammonium sulfate 5%, soluble starch 3% and diatomite are supplemented to 100%.Institute
State viable bacteria concentration of the mixed fungus fermentation liquid in microbial bacterial agent and reach 6 × 1010CFU/g。
Comparative example 3
Mixed fungus fermentation liquid prepared by embodiment 3, with wetting agent neopelex, dispersant sodium lignin sulfonate
(CMN), protective agent humic acid, disintegrant ammonium sulfate, binding agent soluble starch and filler make carrier mixing with kaolin, specifically
Quality proportioning it is as follows：Mixed fungus fermentation liquid 25%, neopelex 3%, sodium lignin sulfonate (CMN) 3%, humic acid
(Humic acid) 1%, ammonium sulfate 5%, soluble starch 3% and kaolin are supplemented to 100%.The mixed fungus fermentation liquid is micro-
Viable bacteria concentration in bacteria agent reaches 6 × 1010CFU/g。
Embodiment 9
The microbial bacterial agent that embodiment 4~8 and comparative example 2~3 are prepared is applied to cotton seed premise germination process
In, the germination rate and the speed of growth of survey calculation cotton seed, (the n of germination percentage=nd germinations sum/seed sum × 100%
Refer to germination number of days)；The length of cotton plants epicotyl and hypocotyl growth in the speed of growth=unit interval.(set in the present invention
It is a unit interval to put 2d) 3 parallel laboratory tests of each group setting.The germination rate and the speed of growth of cotton seed the results are shown in Table 1.
The each group microbial bacterial agent of table 1 is to the germination rate of cotton seed and the influence of the speed of growth
| Treatment group | The germination rate of cotton seed | The speed of growth |
| Embodiment 4 | 78.56±4.8 | 2.61±0.31 |
| Embodiment 5 | 79.02±4.3 | 2.91±0.22 |
| Embodiment 6 | 79.22±3.4 | 2.82±0.09 |
| Embodiment 7 | 80.56±4.8 | 2.97±0.11 |
| Embodiment 8 | 83.42±3.7 | 3.28±0.19 |
| Comparative example 2 | 83.54±3.8 | 3.43±0.12 |
| Comparative example 3 | 80.32±2.6 | 2.50±0.13 |
Embodiment 10
The microbial bacterial agent that embodiment 4~8 and comparative example 2~3 are prepared is used for verticillium dahliae and tomato is withered
Wither in bacterium bacteriostatic experiment, count bacteriostasis rate.Each group bacteriostasis rate is as shown in table 2.
The each group microbial bacterial agent of table 2 is to verticillium dahliae and tomato Fusarium oxysporum bacteriostasis rate
| Treatment group | Verticillium dahliae | Tomato Fusarium oxysporum |
| Embodiment 4 | 67.31% | 36.25% |
| Embodiment 5 | 65.16% | 40.52% |
| Embodiment 6 | 35.42% | 31.02% |
| Embodiment 7 | 36.87% | 33.62% |
| Embodiment 8 | 76.13% | 59.68% |
| Comparative example 2 | 66.14% | 49.36% |
| Comparative example 3 | 31.26% | 30.26% |
Embodiment 11
Microbial bacterial agent prepared by embodiment 4 is used for the Inhibition test of variety classes pathology fungi, specific experiment operation ginseng
See embodiment 10.As a result it is as shown in table 3.
The microbial bacterial agent of table 3 is suppressing the inhibiting rate of variety classes pathology fungi
Note：+：Weak suppression, bacteriostasis rate<50%；++：Moderate suppresses；50%<Bacteriostasis rate<75%；+++：High inhibition, it is antibacterial
Rate>75%
Embodiment 12
In order to examine the long-acting stable performance drug effect of microbial bacterial agent, embodiment 8 and comparative example 2~3 are applied to cotton kind
In plant, using the microbial bacterial agent group for not being applied in cotton planting as control, after 150d, the viable bacteria of microbial bacterial agent is counted
Number.
The effective viable bacteria rate of microbial bacterial agent prepared using embodiment after 150d 8 is 69%, is not applied in cotton planting
The viable bacteria rate of microbial bacterial agent group is 73%, it can be seen that microbial bacterial agent and be not applied in cotton planting prepared by embodiment 8
Microbial bacterial agent group microbial cells survival rate variation it is little, illustrate microbial bacterial agent provided by the invention have length
Imitate bacteriostasis.
The viable bacteria rate of the microbial bacterial agent prepared using comparative example after 150d 2 is 54%, is made than embodiment 8 after applying 150d
The viable bacteria rate of standby microbial bacterial agent reduces 11%.Protective agent is humic acid in the microbial bacterial agent of this explanation the application protection
Being compared than carboxymethyl cellulose has preferably protection biological and ecological methods to prevent plant disease, pests, and erosion thalline survival, and have good biocompatibility with biocontrol microorganisms.
The viable bacteria rate of the microbial bacterial agent prepared using comparative example after 150d 3 is 52%, is made than embodiment 8 after applying 150d
The viable bacteria rate of standby microbial bacterial agent reduces 17%.Protective agent is diatomite in the microbial bacterial agent of this explanation the application protection
The carrier for being more suitable for doing biocontrol microorganisms is compared than kaolin, and there is good biocompatibility with biocontrol microorganisms.
Described above is only the preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, under the premise without departing from the principles of the invention, some improvements and modifications can also be made, these improvements and modifications also should
It is considered as protection scope of the present invention.
Claims (10)
1. a kind of microbial bacterial agent for being used for controlling plant diseases and promoting growth of plants, include the group of following weight/mass percentage composition
Point：Mixed fungus fermentation liquid 1%~50%, wetting agent 1%~5%, dispersant 1%~5%, binding agent 1%~5%, disintegrant 0~
15%th, protective agent 0.1%~2%, the filler of surplus；
Viable bacteria concentration >=5 × 10 of the mixed fungus fermentation liquid in microbial bacterial agent10CFU/g；The mixed fungus fermentation liquid includes spy
Base draws bacillus and Sphingobacterium；The ratio of the thalline number of the Te Jila bacillus and Sphingobacterium is 5~9:4
~1.
2. microbial bacterial agent according to claim 1, it is characterised in that include the component of following weight/mass percentage composition：It is mixed
Fermented liquid 10%~40%, wetting agent 2%~4%, dispersant 2%~4%, binding agent 2%~4%, disintegrant 1~
10%th, protective agent 0.5~1.5%, the filler of surplus.
3. microbial bacterial agent according to claim 1, it is characterised in that the mixed fungus fermentation liquid is in microbial bacterial agent
Viable bacteria concentration >=8 × 1010CFU/g。
4. microbial bacterial agent according to claim 1, it is characterised in that the Te Jila bacillus and Sphingobacterium
Thalline number ratio be 9:1.
5. according to the microbial bacterial agent described in claims 1 to 3 any one, it is characterised in that the filler is diatomite.
6. according to the microbial bacterial agent described in claims 1 to 3 any one, it is characterised in that the protective agent is humic acid
Or xanthan rubber powder.
7. it is used for the preparation method of the microbial bacterial agent of controlling plant diseases described in claim 1~6 any one, including it is following
Step：
1) Te Jila bacillus and Sphingobacterium are seeded in PDB fluid nutrient mediums under 25~37 DEG C of temperature conditionss to enter
Row aerobic fermentation 24~96h of culture, obtains mixed fungus fermentation liquid；
2) the mixed fungus fermentation liquid for obtaining the step 1) and wetting agent, dispersant, suspending agent, protective agent, disintegrant and filler
Mixing, obtains mixed material；
3) mixed material is mixed with binding agent, crushes, obtain microbial bacterial agent.
8. preparation method according to claim 7, it is characterised in that the PDB liquid of fermented and cultured process in the step 1)
The pH value of body culture medium is 5.5~8.0.
9. the preparation method of microbial bacterial agent according to claim 7, it is characterised in that the particle diameter of crushing is 30~60
Mesh.
10. prepared by microbial bacterial agent described in claim 1~6 any one or claim 7~9 any one methods described
Microbial bacterial agent preventing and treating cotton soil-borne disease bacterium application；The soil-borne disease bacterium bag includes cotton verticillium wilt, alternaria solani sorauer, eggplant
Phytophthora, eggplant Huang wither bacterium or tomato Fusarium oxysporum.
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