CN108794187A - A kind of microbial liquid microbial inoculum and its preparation method and application - Google Patents
A kind of microbial liquid microbial inoculum and its preparation method and application Download PDFInfo
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
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- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
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Abstract
The invention discloses a kind of microbial liquid microbial inoculums, include the component of following mass percent：Promoting bacteria bacterium solution 1-10%, polyglutamic acid 0.1-10%, biological organic matter 50-70%；Also disclose the preparation method of microbial liquid microbial inoculum；Further disclose application of the microbial liquid microbial inoculum in improving crop yield, promoting crops quality, enhancing crops resistance in any one or a few.When microbial inoculum is in packed state, Promoting bacteria gemma is inhibited to sprout by providing high organic environment and hyperosmosis environment so that Promoting bacteria is in gemma state；When microbial liquid microbial inoculum is diluted application, organic matter can provide nutrition for the sprouting of gemma again, promote Promoting bacteria fast breeding；Biological organic matter is used in combination with polyglutamic acid, and the vigor of Promoting bacteria in liquid bacterial agent is maintained by synergistic effect, substantially prolongs the shelf life of microbial liquid microbial inoculum.
Description
Technical field
The present invention relates to microbial bacterial agents, and in particular to a kind of microbial liquid microbial inoculum and its preparation method and application.
Background technology
China is world agriculture big country, and with the rapid development of China's agricultural, the usage amount of chemical fertilizer is also increasing, every year
Dosage reach 1,800,000 tons.Although this solves the problems, such as increases in grain production and supply, causes very serious environment and ask
Topic.The Use out of range of chemical fertilizer has aggravated the hardened of soil and pollution, leads to the decline and degeneration of soil quality, this makes soil have
Machine matter content is seriously less than the average level in the whole world.Once soil organic matter content is low, fixed to heavy metal by being greatly reduced
Ability increases the harmfulness of heavy metal-polluted soil, compromises the sustainable development of China's agricultural.
Microbial bacterial agent be it is a kind of can optimize soil physico-chemical property, improve absorption and utilization ratio of the crop to nutrient, into
And promoting the novel agricultural biological agent of plant growth, it has economic and environment-friendly, the improvement effectiveness such as soil and fertilizer synergistic.This
Outside, moreover it is possible to enhance the drought-resistant of crop, low-temperature resistance, the anti-adversity abilities such as high salt tolerance.In recent years, using microbial bacterial agent as bio-fertilizer
The research of material is more and more, and especially microbial liquid microbial inoculum has low production cost, easy to use, growth-promoting and degeneration-resistant effect bright
The advantages that aobvious.
But because state of the art limits, all there is effective bacterium number content in the microbial liquid microbial inoculum sold currently on the market
Low, the defects of effective bacterium easy in inactivation, especially commercially available microbial inoculum is Yi Dan sealed off, and effective bacterial content drops cliff of displacement formula, significantly
The effectiveness for reducing microbial inoculum increases the use cost of peasant.
Invention content
Goal of the invention：In order to solve the problems, such as that the existing effective bacterial content of microbial liquid microbial inoculum is low and easy in inactivation, the present invention
A kind of microbial liquid microbial inoculum is provided, invention further provides the preparation methods of microbial liquid microbial inoculum, and the present invention is more
Further provide the application of microbial liquid microbial inoculum.
Technical solution：A kind of microbial liquid microbial inoculum of the present invention, includes the component of following mass percent：Promoting bacteria
Bacterium solution 1-10%, polyglutamic acid 0.1-10%, biological organic matter 50-70%, surplus are water or aqueous solution.
Preferably, Promoting bacteria bacterium solution 5%-10%, polyglutamic acid 0.5%-8%, biological organic matter 55%-70%, surplus
For water or aqueous solution.
Most preferably, Promoting bacteria bacterium solution 8%-10%, polyglutamic acid 1%-5%, biological organic matter 55%-68%, surplus
For water or aqueous solution.
Wherein, the Promoting bacteria bacterium solution is bacillus subtilis (Bacillus subtilis), bacillusmusilaginosiengineering
It is any one in (Bacillus mucilaginosus) and bacillus amyloliquefaciens (Bacillus amyloliquefaciens)
The combination of kind zymotic fluid or several zymotic fluids, bacillus subtilis fermentation liquor, bacillusmusilaginosiengineering zymotic fluid or solution starch bud
Viable count is 1 × 10 in spore bacillus fermentation liquid9-1×1011CFU/mL, viable count >=5 of the Promoting bacteria in microbial liquid preparation
×109CFU/mL.The additive amount of Promoting bacteria bacterium solution is according to bacillus subtilis, gel-shaped bacillus, bacillus amyloliquefaciens
In the summation of any one or combination calculate, living bacteria count >=5 × 10 in microbial liquid microbial inoculum9CFU/mL.Withered grass gemma
Bacillus, gel-shaped bacillus, bacillus amyloliquefaciens make thalline enter gemma obtained by high density fermentation by refrigeration
Form, gemma state are the dormant states of bacillus, can be survived in the state of not sprouting several years and to decades.
Preferably, the bacillus subtilis (Bacillus subtilis) is in China typical culture collection center
Deposit number is CCTCC NO:M 2016264, bacillusmusilaginosiengineering (Bacillus mucilaginosus) is in Chinese Typical Representative
The deposit number of culture collection is CCTCC NO:M 2016265, bacillus amyloliquefaciens (Bacillus
Amyloliquefaciens it is) CCTCC NO in the deposit number of China typical culture collection center:M 2016266 is recorded
In patent 2016106658324.
Preferably, the Promoting bacteria bacterium solution is bacillus subtilis fermentation liquor, bacillusmusilaginosiengineering zymotic fluid reconciliation shallow lake
The combination of afnyloliquefaciens zymotic fluid, volume ratio 0.1-10:0.1-10:0.1-10.
Wherein, the polyglutamic acid weight average molecular weight is 5-3000kDa, preferred molecular weight 5-50kDa, the polyglutamic
Acid is polyglutamic acid sterling, polyglutamate sterling, polyglutamic acid zymotic fluid, polyglutamic acid zymotic fluid dried object and polyglutamic acid
Any one or the combination of several of them in wettable powder.The additive amount of polyglutamic acid is with polyglutamic acid sterling or polyglutamic acid
Gather contained in salt sterling or polyglutamic acid zymotic fluid or polyglutamic acid zymotic fluid dried object or polyglutamic acid wettable powder
Glutamic acid calculates；The Related product of above-mentioned polyglutamic acid is prepared by existing fermentation technique.When polyglutamic acid is sent out for polyglutamic acid
When zymotic fluid, for the quality of polyglutamic acid with polyglutamic acid content calculation in zymotic fluid, aqueous solution is poly- in microbial liquid microbial inoculum
Glutami acid fermentation liquor (not including polyglutamic acid therein).
Polyglutamic acid is a kind of special anionic polymer polymer, and side chain contains a large amount of carboxylic groups, these groups
Its special bioactivity is assigned, as a kind of substance of adhesiveness, polyglutamic acid can provide folder film protection for Promoting bacteria, promote
Promoting bacteria forms a kind of external wrapping biological membranous system, thus separates the mass exchange of Promoting bacteria intraor extracellular, reaches a kind of
Balance stable state.
Wherein, the biological organic matter is any one or a few group in biochemical fulvic acid, molasses and yeast extract
It closes.The additive amount of biological organic matter is according to any one in yeast extract, molasses, biochemical fulvic acid or the summation meter of combination
It calculates.Above-mentioned biological organic matter can be bought or according to prior art preparation from market.
Wherein, yeast extract main component is polypeptide, amino acid, trace element etc..
Biological organic matter can inhibit the bud of bacillus under use ratio of the present invention by forming hyperosmosis environment
Spore is sprouted, and the Promoting bacteria in microbial liquid microbial inoculum is maintained to be in gemma state.And it is applied to crops after microbial inoculum is diluted
When, biological organic matter concentration reduces, and changes its " identity " immediately, becomes the nutritional ingredient of gemma sprouting and microbial growth.This
Outside, biological organic matter can also improve soil texture, the problems such as reducing soil compaction rate, solve soil acidification.
The present invention further provides the preparation methods of the microbial liquid microbial inoculum, include the following steps：
(1) Promoting bacteria that inclined-plane preserves is inoculated on activation medium, 25-35 DEG C of activation 18-24h；Picking activation training
The strain on base is supported, is transferred in fermentation medium, zymotic fluid or fermentation concentrated liquor are placed in 4 by 30-35 DEG C of culture 12-24h
Low-temperature treatment 28-50h in DEG C freezer；
(2) Promoting bacteria bacterium solution, polyglutamic acid and the biological organic matter for obtaining step (1) be according to mass percent mixing,
Up to microbial liquid microbial inoculum.
In step (1), when Promoting bacteria includes two or more, zymotic fluid or hair are respectively obtained according to step (1)
Then zymotic fluid concentrate mixes zymotic fluid according to volume ratio；The activation medium is LB culture mediums, and formula is as follows：
Peptone 10g/L, yeast powder 5g/L, sodium chloride 10g/L, pH6.0-8.0, the solvent are water.The fermentation medium is matched
Side is as follows：Carbon source 10-100g/L, nitrogen source 1-40g/L, inorganic salts 0.01-50g/L, solvent are water, pH5.0-9.0.
Preferably, the carbon source be glucose, sucrose, maltose, lactose, xylose, fructose, lactic acid, citric acid, glycerine,
Any one or the combination of several of them in starch and molasses；The nitrogen source is yeast powder, beef extract, peptone, yeast extract, corn
Slurry, beancake powder, cottonseed meal, urea, (NH4)2SO4、NH4Cl、(NH4)2HPO4And NH4NO3In any one or a few group
It closes；The inorganic salts are any one or a few in sulfate, phosphate, dihydric phosphate, phosphoric acid hydrogen disalt and hydrochloride
Combination.
The present invention further provide the microbial liquid microbial inoculum improve crop yield, promoted crops quality,
Enhance the application in any one or a few in crops resistance.
Wherein, the microbial liquid microbial inoculum can be used cooperatively with conventional chemical fertilizers, also be can be used as bacterial manure and directly applied；
Microbial liquid microbial inoculum is diluted 10-1000 times, by water planting, is sprayed or pouring root is applied to crops.
Preferably, the crops are cereal crops, greengrocery crop or melon or fruit type crops.
Advantageous effect：(1) polyglutamic acid provides pod membrane protection in microbial liquid microbial inoculum for Promoting bacteria, and makes Promoting bacteria
Biofilm system is formed, reaches a kind of stable equilibrium state, is at gemma state；Polyglutamic acid is produced by microbial fermentation
It is raw, it is formed by glutamy amine key homopolymerization by D-Glu and Pidolidone monomer, there is nontoxic, biodegradable and environment friend
The advantages that good, also has the absorption and other effects for improving plant to nutrient.(2) when microbial liquid microbial inoculum is in packed state,
Biological organic matter inhibits the sprouting of Promoting bacteria gemma by providing high organic environment and hyperosmosis environment, so that growth-promoting
Bacterium is in gemma state；When microbial liquid microbial inoculum is diluted application, organic matter can provide again for the sprouting of Promoting bacteria gemma
Nutrition promotes the fast breeding of Promoting bacteria；When biological organic matter and polyglutamic acid are used in combination, except with nutrition synergistic function
Outside, the vigor that Promoting bacteria in liquid bacterial agent is mainly maintained by synergistic effect, substantially prolongs the shelf of microbial liquid microbial inoculum
Phase makes the number of viable of the Promoting bacteria in microbial inoculum maintain >=5 × 10 in 18 months9CFU/mL.(3) microorganism liquid of the invention
Body microbial inoculum, crop root biological community structure can be improved by being applied in crops, increase quantity of useful microbe in soil, energy
Soil texture is enough improved, is kept soil from packing together, crop yield can be improved, enhancing crop drought resistance, salt resistance, disease-resistant, cold-resistant etc. are anti-
Inverse ability.(4) microbial liquid microbial inoculum of the invention can significantly improve the cereal crops such as rice, wheat, pakchoi, rape etc.
Leaf vegetables, the yield of the industrial crops such as melons and fruits class such as muskmelon, grape, can be improved 35-40%.In the feelings for ensureing the crop yield not underproduction
Under condition, it is possible to reduce apply the applying quantity of chemical fertilizer of 20-40% mass, improve crop quality, enhance the anti-adversity ability of plant, promote
Into the growth of plant.
Description of the drawings
Fig. 1 is the number of viable survival curve of different microbial inoculums.
Specific implementation mode
A kind of microbial liquid microbial inoculum of the present invention, includes the component of following mass percent：Promoting bacteria bacterium solution 1-
10%, polyglutamic acid 0.1-10%, biological organic matter 50-70%, surplus are water or aqueous solution.
Preferably, Promoting bacteria bacterium solution 5%-10%, polyglutamic acid 0.5%-8%, biological organic matter 55%-70%, surplus
For water or aqueous solution.
Most preferably, Promoting bacteria bacterium solution 8%-10%, polyglutamic acid 1%-5%, biological organic matter 55%-68%, surplus
For water or aqueous solution.
Wherein, the Promoting bacteria bacterium solution is bacillus subtilis (Bacillus subtilis), bacillusmusilaginosiengineering
It is any one in (Bacillus mucilaginosus) and bacillus amyloliquefaciens (Bacillus amyloliquefaciens)
The combination of kind zymotic fluid or several zymotic fluids, bacillus subtilis fermentation liquor, bacillusmusilaginosiengineering zymotic fluid or solution starch bud
Viable count is 1 × 10 in spore bacillus fermentation liquid9-1×1011CFU/mL, viable count >=5 of the Promoting bacteria in microbial liquid preparation
×109CFU/mL.The additive amount of Promoting bacteria bacterium solution is according to bacillus subtilis, gel-shaped bacillus, bacillus amyloliquefaciens
In the summation of any one or combination calculate, living bacteria count >=5 × 10 in microbial liquid microbial inoculum9CFU/mL.Withered grass gemma
Bacillus, gel-shaped bacillus, bacillus amyloliquefaciens make thalline enter gemma obtained by high density fermentation by refrigeration
Form, gemma state are the dormant states of bacillus, can be survived in the state of not sprouting several years and to decades.
Preferably, the bacillus subtilis (Bacillus subtilis) is in China typical culture collection center
Deposit number is CCTCC NO:M 2016264, bacillusmusilaginosiengineering (Bacillus mucilaginosus) is in Chinese Typical Representative
The deposit number of culture collection is CCTCC NO:M 2016265, bacillus amyloliquefaciens (Bacillus
Amyloliquefaciens it is) CCTCC NO in the deposit number of China typical culture collection center:M 2016266 is recorded
In patent 2016106658324.
Preferably, the Promoting bacteria bacterium solution is bacillus subtilis fermentation liquor, bacillusmusilaginosiengineering zymotic fluid reconciliation shallow lake
The combination of afnyloliquefaciens zymotic fluid, volume ratio 0.1-10:0.1-10:0.1-10.
Wherein, the polyglutamic acid weight average molecular weight is 5-3000kDa, preferred molecular weight 5-50kDa, the polyglutamic
Acid is polyglutamic acid sterling, polyglutamate sterling, polyglutamic acid zymotic fluid, polyglutamic acid zymotic fluid dried object and polyglutamic acid
Any one or the combination of several of them in wettable powder.The additive amount of polyglutamic acid is with polyglutamic acid sterling or polyglutamic acid
Gather contained in salt sterling or polyglutamic acid zymotic fluid or polyglutamic acid zymotic fluid dried object or polyglutamic acid wettable powder
Glutamic acid calculates；The Related product of above-mentioned polyglutamic acid is prepared by existing fermentation technique.When polyglutamic acid is sent out for polyglutamic acid
When zymotic fluid, for the quality of polyglutamic acid with polyglutamic acid content calculation in zymotic fluid, aqueous solution is poly- in microbial liquid microbial inoculum
Glutami acid fermentation liquor (not including polyglutamic acid therein).
Polyglutamic acid is a kind of special anionic polymer polymer, and side chain contains a large amount of carboxylic groups, these groups
Its special bioactivity is assigned, as a kind of substance of adhesiveness, polyglutamic acid can provide folder film protection for Promoting bacteria, promote
Promoting bacteria forms a kind of external wrapping biological membranous system, thus separates the mass exchange of Promoting bacteria intraor extracellular, reaches a kind of
Balance stable state.
Wherein, the biological organic matter is any one or a few group in biochemical fulvic acid, molasses and yeast extract
It closes.The additive amount of biological organic matter is according to any one in yeast extract, molasses, biochemical fulvic acid or the summation meter of combination
It calculates.Above-mentioned biological organic matter can be bought or according to prior art preparation from market.
Wherein, yeast extract main component is polypeptide, amino acid, trace element etc..
Biological organic matter can inhibit the bud of bacillus under use ratio of the present invention by forming hyperosmosis environment
Spore is sprouted, and the Promoting bacteria in microbial liquid microbial inoculum is maintained to be in gemma state.And it is applied to crops after microbial inoculum is diluted
When, biological organic matter concentration reduces, and changes its " identity " immediately, becomes the nutritional ingredient of gemma sprouting and microbial growth.This
Outside, biological organic matter can also improve soil texture, the problems such as reducing soil compaction rate, solve soil acidification.
The fermentation of 1 Promoting bacteria of embodiment
Promoting bacteria bacterium solution is prepared as follows：
The bacillus subtilis CCTCC NO that 4 DEG C of inclined-planes are preserved:M 2016264, gel-shaped bacillus CCTCC NO:
M 2016265, bacillus amyloliquefaciens CCTCC NO:M 2016266 is activated, and is inoculated in eggplant shape culture bottle culture medium respectively (i.e.
Activation medium) on, 32 DEG C of culture 18h.The strain of activation is transferred in fermentation medium respectively, 30 DEG C of cultures for 24 hours, will be sent out
Zymotic fluid is placed in low-temperature treatment 48h in Cool Room 4 DEG C, is coated with and measures through tablet, and the number of viable in each microbial fermentation solution is reachable
To 8 × 109CFU/mL or more, by three kinds of microbial fermentation solutions according to volume ratio 1:1:1 ratio mixing compounds up to Promoting bacteria
Bacterium solution.
Culture medium in the eggplant shape culture bottle is LB culture mediums, and fermentation medium group is divided into：Glucose 50g/L, albumen
Peptone 10g/L, (NH4)2SO40.05g/L, NaCl 1g/L, MnSO4·4H2O 0.5g/L, KH2PO41.0g/L, pH 6.0-
8.0。
The ventilatory capacity of filtrated air is 0.6vvm, mixing speed 220rpm during the fermented and cultured of the above production tank.
The preparation of 2 complex microorganism liquid bacterial agent of embodiment
It is prepared as follows the complex microorganism liquid bacterial agent containing polyglutamic acid, biological organic matter, Promoting bacteria bacterium solution：
(1) polyglutamic acid prepares (bibliography Xu Z, Feng X, Dan Z, et al.Enhanced using fermentation method
poly(γ-glutamic acid)fermentation by Bacillus subtilis,NX-2immobilized in an
aerobic plant fibrous-bed bioreactor[J].Bioresour Technol,2014,155:8-14), it ferments
Polyglutamic acid content is 15% after liquid is concentrated；
(2) molasses are purchased by sugar refinery；
(3) it is according to mass ratio with polyglutamic acid Fermented Condensed liquid, molasses by the compound Promoting bacteria bacterium solution that embodiment 1 obtains
1:2.5:6.5 ratio is uniformly mixed and obtains complex microorganism liquid bacterial agent.After testing, polyglutamic acid content mass percent is
3.5%, content of organic matter mass percent is 67%, and living bacteria count is 7.3 × 109CFU/mL。
3 complex microorganism liquid bacterial agent of embodiment can open preservation and be up to 18 months
Microbial inoculum used in the present embodiment is as follows：
(1) it is 7.3 × 10 that the mix bacterium agent that embodiment 1 is prepared, which is diluted to living bacteria count,9CFU/mL, as microbial inoculum
1。
(2) mix bacterium agent that embodiment 1 is prepared is mixed in proportion with polyglutamic acid Fermented Condensed liquid, makes polyglutamic
Acid content is 3.5%, and living bacteria count is 7.3 × 109CFU/mL, as microbial inoculum 2.
(3) mix bacterium agent that embodiment 1 is prepared is mixed in proportion with molasses, makes the content of organic matter be
67%, living bacteria count is 7.3 × 109CFU/mL, as microbial inoculum 3.
(4) mix bacterium agent that embodiment 1 is prepared is mixed in proportion with polyglutamic acid, molasses, makes polyglutamic
Acid content is 3.5%, the content of organic matter 15%, and living bacteria count is 7.3 × 109CFU/mL, as microbial inoculum 4.
(5) the complex microorganism liquid bacterial agent containing polyglutamic acid, biological organic matter and Promoting bacteria for obtaining embodiment 2,
As microbial inoculum 5.
Above-mentioned 5 kinds of microbial inoculums are placed in 25 DEG C, is stored in the environment of relative humidity 40%, 1mL microbial inoculums was respectively taken every 2 months,
Dilution 108Times, it in applying solid LB culture mediums, is placed under 32 DEG C of environment, cultivates 18-24h, obtained using colony counting method effective
Number of viable, continuous sampling 20 months.If three groups of parallel laboratory tests, living bacteria count is changed over time such as Fig. 1 institutes in different microbial inoculums
Show.
As shown in Figure 1, its living bacteria count amount of microbial inoculum 1, microbial inoculum 2, microbial inoculum 3, microbial inoculum 4 only maintains 2 months, 6 months, 6
The moon and 6 months, fail to reach 18 months standards.Microbial inoculum 5, i.e., the living bacteria count amount of complex microorganism liquid bacterial agent is 18
Reached >=5 × 10 before a month9The level of cfu/mL.It can be seen that the complex microorganism liquid bacterial agent that the present invention protects has
The effect phase is 18 months.
4 complex microorganism liquid bacterial agent of embodiment can reduce the application of chemical fertilizer
Complex microorganism liquid bacterial agent is prepared according to the method for embodiment 2.Using potting pakchoi as experimental subjects, examination
It tests and sets 3 processing, each handle 3 repetitions, wherein 1 (T of processing1) it is experimental control common fertilizer, handle 2 (T2) it is common fertilizer
Material+complex microorganism liquid bacterial agent handles 3 (T3) it is to subtract the common fertilizer+complex microorganism liquid bacterial agent for applying 20% mass, it is real
It tests and the results are shown in Table 1.The fertilizer applied is every kilogram of soil 150mg, and the dosage of complex microorganism liquid bacterial agent is every kilogram of soil
0.2mL。
Influence of the 1 complex microorganism liquid bacterial agent of table to potting Growth of Cabbage characteristic and yield
As shown in Table 1, complex microorganism liquid bacterial agent is mixed with fertilizer and is applied, and can be promoted the development of pakchoi root, be made root
Thickening, increase have apparent facilitation to pakchoi plant plant height, underground part fresh weight, overground part fresh weight, can increase production
49.5%.When subtracting the fertilizer for applying 20% mass, can still it illustrate complex microorganism liquid to Chinese cabbage output increased 23.4%
Microbial inoculum can effectively reduce the usage amount of chemical fertilizer.
5 complex microorganism liquid bacterial agent of embodiment significantly improves the quality of crops
Complex microorganism liquid bacterial agent is prepared according to the method for embodiment 2.Using muskmelon as experimental subjects, experiment is set altogether
3 processing, each handle 3 repetitions.Handle 1 (T1) it is to apply Common compound fertilizer (450kg/ha2), handle 2 (T2) it is that application is general
Logical composite fertilizer (450kg/ha2)+complex microorganism liquid bacterial agent (6L/ha2), handle 3 (T3) it is to reduce using the general of 30% mass
Logical composite fertilizer+complex microorganism liquid bacterial agent (315kg/ha2), muskmelon management is daily management, and experimental result is shown in Table 2.
Influence of the 2 complex microorganism liquid bacterial agent of table to muskmelon Vc and nitrate content
As shown in Table 2, compared with processing 1, complex microorganism liquid bacterial agent can carry high Vc content 47.5%, reduce nitric acid
Salt content 22.3%.When reducing the composite fertilizer using 30% mass, complex microorganism liquid bacterial agent can carry high Vc content
35.0%, nitrate content 35.3% is reduced, the quality of muskmelon is improved.Test result shows the complex microorganism in the present invention
Liquid bacterial agent can not only reduce the usage amount of chemical fertilizer, and can also significantly improve the quality of muskmelon.
6 complex microorganism liquid bacterial agent of embodiment significantly improves the anti-adversity ability of crops
Complex microorganism liquid bacterial agent is prepared according to the method for embodiment 2.Using Xiaoqinling Nature Reserve as experimental subjects, basin is carried out
It plants, experiment sets 3 processing altogether, each handles 3 repetitions.Water control method is drawn using Hsiao (1973) mesad gradation of moisture
Point-score handles 1 (T1) it is control 75%-80% (percentage for accounting for maximum water holding capacity), handle 2 (T2) it is that moderate is coerced
40%-45% handles 3 (T3) it is severe water stress 30%-35%.Normal management is carried out to potting Xiaoqinling Nature Reserve, is controlled before test observation
Moisture processed, when reaching 3 gradations of moisture, in processing 2,3 addition complex microorganism liquid bacterial agent of processing, applied amount is every kilogram of soil
0.2mL.Period measures volumetric water content of soil daily, and the moisture that the supplement same day loses makes each processing level reach containing for setting
Water, experimental result are shown in Table 3.
Influence of the 3 complex microorganism liquid bacterial agent of table to Xiaoqinling Nature Reserve chlorophyll and fresh weight
As shown in Table 3, under the environmental condition of mild drought, by applying complex microorganism liquid bacterial agent, with control group
It compares, Xiaoqinling Nature Reserve can increase production 23.0%；It is small by applying complex microorganism liquid bacterial agent under the environmental condition of Severe drought
Yields of vegetables is compared with the control group without significant difference.The experimental results showed that complex microorganism liquid bacterial agent can improve the anti-of plant
Non-irrigated ability, to promote the growth of plant.
Claims (10)
1. a kind of microbial liquid microbial inoculum, which is characterized in that include the component of following mass percent：Promoting bacteria bacterium solution 1-
10%, polyglutamic acid 0.1-10%, biological organic matter 50-70%, surplus are water or aqueous solution.
2. microbial liquid microbial inoculum according to claim 1, which is characterized in that the Promoting bacteria bacterium solution is bacillus subtilis
(Bacillus subtilis), bacillusmusilaginosiengineering (Bacillus mucilaginosus) and bacillus amyloliquefaciens
The combination of any one zymotic fluid or several zymotic fluids in (Bacillus amyloliquefaciens), bacillus subtilis hair
Viable count is 1 × 10 in zymotic fluid, bacillusmusilaginosiengineering zymotic fluid or bacillus amyloliquefaciens zymotic fluid9-1×1011CFU/mL,
Viable count >=5 × 10 of the Promoting bacteria in microbial liquid preparation9CFU/mL。
3. microbial liquid microbial inoculum according to claim 2, which is characterized in that the bacillus subtilis (Bacillus
Subtilis it is) CCTCC NO in the deposit number of China typical culture collection center:M 2016264, gel-shaped gemma bar
Bacterium (Bacillus mucilaginosus) is CCTCC NO in the deposit number of China typical culture collection center:M
2016265, bacillus amyloliquefaciens (Bacillus amyloliquefaciens) are in China typical culture collection center
Deposit number is CCTCC NO:M 2016266.
4. microbial liquid microbial inoculum according to claim 3, which is characterized in that the Promoting bacteria bacterium solution is bacillus subtilis
The combination of zymotic fluid, bacillusmusilaginosiengineering zymotic fluid and bacillus amyloliquefaciens zymotic fluid, volume ratio 0.1-10:0.1-
10:0.1-10。
5. microbial liquid microbial inoculum according to claim 1, which is characterized in that the polyglutamic acid weight average molecular weight is 5-
3000kDa, the polyglutamic acid are polyglutamic acid sterling, polyglutamate sterling, polyglutamic acid zymotic fluid, polyglutamic acid fermentation
Any one or the combination of several of them in liquid dried object and polyglutamic acid wettable powder.
6. microbial liquid microbial inoculum according to claim 1, which is characterized in that the biological organic matter be biochemical fulvic acid,
Any one or the combination of several of them in molasses and yeast extract.
7. the preparation method of microbial liquid microbial inoculum described in claim 1-6 any one, which is characterized in that including following step
Suddenly：
(1) Promoting bacteria that inclined-plane preserves is inoculated on activation medium, 25-35 DEG C of activation 18-24h；Picking activation medium
On strain, transfer in fermentation medium, 30-35 DEG C culture 12-24h, by zymotic fluid or fermentation concentrated liquor be placed in 4 DEG C it is cold
Low-temperature treatment 28-50h in library；
(2) Promoting bacteria bacterium solution, polyglutamic acid and the biological organic matter for obtaining step (1) according to mass percent mixing to get
Microbial liquid microbial inoculum.
8. preparation method according to claim 7, which is characterized in that the formula of fermentation medium described in step (1) is such as
Under：Carbon source 10-100g/L, nitrogen source 1-40g/L, inorganic salts 0.01-50g/L, solvent are water, pH5.0-9.0.
9. microbial liquid microbial inoculum described in claim 1-6 any one is improving crop yield, is promoting crops quality, increasing
Application in any one or a few in strong crops resistance.
10. application according to claim 9, which is characterized in that the microbial liquid microbial inoculum by water planting, spray or fill
Root application is used for crops.
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Application publication date: 20181113 |