CN106495951A - A kind of high concentration microorganism microbial inoculum containing polyglutamic acid and preparation method and application - Google Patents

A kind of high concentration microorganism microbial inoculum containing polyglutamic acid and preparation method and application Download PDF

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Publication number
CN106495951A
CN106495951A CN201610954700.3A CN201610954700A CN106495951A CN 106495951 A CN106495951 A CN 106495951A CN 201610954700 A CN201610954700 A CN 201610954700A CN 106495951 A CN106495951 A CN 106495951A
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liquid
fermentation
bacillus
culture
polyglutamic acid
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展彬
许鹏飞
梁东明
李环环
杜绍国
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Dezhou Yuanhe Agricultural Technology Development Co Ltd
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Dezhou Yuanhe Agricultural Technology Development Co Ltd
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    • CCHEMISTRY; METALLURGY
    • C05FERTILISERS; MANUFACTURE THEREOF
    • C05DINORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C; FERTILISERS PRODUCING CARBON DIOXIDE
    • C05D3/00Calcareous fertilisers
    • C05D3/02Calcareous fertilisers from limestone, calcium carbonate, calcium hydrate, slaked lime, calcium oxide, waste calcium products
    • CCHEMISTRY; METALLURGY
    • C05FERTILISERS; MANUFACTURE THEREOF
    • C05GMIXTURES 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
    • C05G3/00Mixtures of one or more fertilisers with additives not having a specially fertilising activity
    • C05G3/80Soil conditioners
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00Microorganisms, 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
    • C12N1/20Bacteria; Culture media therefor

Abstract

The present invention relates to a kind of high concentration microorganism microbial inoculum containing polyglutamic acid and preparation method and application.High concentration microorganism microbial inoculum containing polyglutamic acid, component are as follows:Mixed microorganism yeast powder, precipitated calcium carbonate, polyglutamic acid, magnesium sulfate, boric acid and zinc sulfate.The present invention with the addition of polyglutamic acid in microbial bacterial agent, due to containing substantial amounts of superpower hydrophilic groups, can fully keep soil moisture in polyglutamic acid, improves porosity and the bulkiness of soil, improves the retain water and nutrients ability of soil;Strengthen phosphorus decomposing, ability of dissolving potassium;The addition of polyglutamic acid can be greatly improved the field planting rate of microorganism simultaneously.

Description

A kind of high concentration microorganism microbial inoculum containing polyglutamic acid and preparation method and application
Technical field
The present invention relates to a kind of high concentration microorganism microbial inoculum containing polyglutamic acid and preparation method and application, belongs to micro- Bacteria agent technical field.
Technical background
China is traditional large agricultural country, and agricultural is the basic industry of China, occupies very important in national economy Status.Since reform and opening-up, China's grain yield achieves significant growth, and modern agricultural technology has promoted the fast of agricultural industry The a large amount of use of speed development, wherein chemical fertilizer is the key factor for supporting China's increases in grain production.Using for long-term excess is chemical Fertilizer brings environmental pollution, soil compaction, Soil degradation, ecological degeneration and quality of agricultural product and increasingly serious the asking such as declines Topic.To account for the chemical fertilizer that the arable land in the world 8% consumes the world 32%, the chemical fertilizer utilization ratio of current China is only 30% for China, and European and American developed countries can reach more than 60%.Chemical fertilizer utilization ratio is low, increases agricultural cost, and productivity effect reduces.Actively seek Look for solution, realize that the sustainable development of agricultural is our urgent problems in agricultural production.
In recent years, with Environmental safety and the continuous attention of food safety, microbial bacterial agent has environment because of which The features such as close friend, improved soil, increase yield, improve quality and green safety, is in ecological environmental protection and safe and high quality agricultural product Receive much concern in production.Microorganisms with specific functions in microbial bacterial agent can promote thing in soil by the vital movement of itself The conversion of matter, improves crop nutrition level, promotes and/or assists alimentation, stimulates regulation and control plant growth, preventing and treating to be harmful to micro- life Thing etc., so that reach high yield, high-quality, efficient, ecological, safe purpose.
Polyglutamic acid has good water solublity, superpower adsorptivity and biodegradability, and catabolite is glutamic acid, Nutrient substance supply plant absorption is can act also as, is a kind of excellent environment-friendly type macromolecule material, can be used as water-retaining agent, heavy metal Ion adsorbent, flocculant, slow releasing agent and pharmaceutical carrier etc., lead in environmental conservation, food, medicine, agricultural, control of desert etc. There is application in domain.
Chinese patent literature CN101851123A (application number 201010183268.5) disclose a kind of high concentration be combined micro- Bio-bacterial manure and its production method, prepared high-concentration compound microorganism bacterial fertilizer living bacteria count be maintained at 5,000,000,000/mL with On.
Chinese patent literature CN101863685A (application number 201010192330.7) discloses a kind of making microorganism fertilizer The method of material high-concentration bacterial powder, is finally reached the side spore of production, potassium spore mycopowder viable bacteria content up to 20,000,000,000/gram.
At present, low containing bacteria concentration for microbial manure, single varieties, soil field planting rate are low, in-convenience in use, transport is not Convenient the shortcomings of, the living bacteria count that above-mentioned patent methods described is obtained, are compared current actual production and are improved to some extent, But the also rising space, how to obtain stable higher concentration microbial bacterial agent be present invention mainly solves problem.
Content of the invention
The present invention is directed to the deficiencies in the prior art, there is provided a kind of high concentration microorganism microbial inoculum containing polyglutamic acid and its system Preparation Method and application.
Summary of the invention
The preparation method and application of the high concentration microorganism microbial inoculum containing polyglutamic acid of the present invention, wherein microbial bacteria Agent is by bacillus subtilises (Bacillus subtilis), Brevibacillus laterosporus (Bacillus laterosporus), lichens Bacillus cereuss (Bacillus licheniformis), colloid bacillus cereus (Paenibacillus mucilaginosus), huge Bacterium anthracoides (Bacillus megaterium) are obtained through liquid, solid combined ferment.Living bacteria count can reach >=100,000,000,000 Individual/gram, every kind of bacterium is not less than 5,000,000,000/gram, and adds water-retaining agent polyglutamic acid, it is possible to increase the effective stability of viable bacteria and fixed Plant rate, with preferably loosening the soil, water conservation, anti-compaction, preventing disease and pest, potassium decomposing, phosphorus decomposing, the effect for increasing the improved soils such as fertilizer efficiency.
Detailed description of the invention
Technical solution of the present invention is as follows:
A kind of high concentration microorganism microbial inoculum containing polyglutamic acid, component are as follows, are weight portion:
15~25 parts of mixed microorganism yeast powder, 55~70 parts of precipitated calcium carbonate, 5~10 parts of polyglutamic acid, magnesium sulfate 3~ 5 parts, 3 parts of boric acid, 2 parts of zinc sulfate;
The polyglutamic acid is poly-gamma-glutamic acid, bonded by amide by D types glutamic acid and L-type glutamic acid, each Glutamic acid all leaves a free carboxyl.
According to currently preferred, in the mixed microorganism yeast powder, contain bacillus subtilises (Bacillus Subtilis), Brevibacillus laterosporus (Bacillus laterosporus), Bacillus licheniformis (Bacillus Licheniformis), colloid bacillus cereus (Paenibacillus mucilaginosus), bacillus megaterium The ratio of the number of viable of (Bacillus megaterium) is 1:(2.5~3.5):(0.5~1.5):(0.5~1.5):(1.5 ~2.5), effective total viable count reaches >=100,000,000,000/gram, and wherein every kind of bacterium living bacteria count is not less than 5,000,000,000/gram.
According to currently preferred, bacillus subtilises (the Bacillus subtilis) is micro- from Chinese agriculture Biological inoculum preservation administrative center, bacterium numbering ACCC 10242;Brevibacillus laterosporus (Bacillus laterosporus) come Come from Chinese agriculture Microbiological Culture Collection administrative center, bacterium numbering ACCC 11079;Bacillus licheniformis (Bacillus Licheniformis) Chinese agriculture Microbiological Culture Collection administrative center, bacterium numbering ACCC 02002 are derived from;Colloid bud Spore bacillus (Paenibacillus mucilaginosus) derives from Chinese agriculture Microbiological Culture Collection administrative center ACCC10012;Bacillus megaterium (Bacillus megaterium) is in the management of Chinese industrial Microbiological Culture Collection The heart, bacterium numbering CICC 21694.
According to currently preferred, molecular weight >=1,000,000 of the polyglutamic acid.
The preparation method of the above-mentioned high concentration microorganism microbial inoculum containing polyglutamic acid, step is:Mixed microorganism is fermented Powder, precipitated calcium carbonate, polyglutamic acid, magnesium sulfate, boric acid, zinc sulfate are mixed in proportion, and obtain final product.
According to currently preferred, mixed microorganism yeast powder is prepared as follows:
(1) by bacillus subtilises (Bacillus subtilis) through slant culture after, be obtained inclined-plane kind A;By inclined-plane kind A transfers in liquid seed culture medium, through shake-flask culture, liquid seeds A is obtained;
Obtained liquid seeds A is inoculated in liquid fermentation medium, control ph 7~8, ventilation ratio is 1: (0.5~ 1), 100~200 revs/min of rotating speed, cultivate 36~48h at 30~37 DEG C, and fermentation liquid A is obtained;Then by fermentation liquid A by 6%~ 10% mass ratio is forwarded in solid fermentation culture medium, in 35~38 DEG C of temperature, tank pressure 0.02~0.06Mpa condition bottom fermentations Cultivate to microscopy spore rate be 90% when, through filter pressing, flash distillation, dries pulverizing, mycopowder A is obtained;
(2) by Brevibacillus laterosporus (Bacillus laterosporus) through slant culture after, be obtained inclined-plane kind B;Will be tiltedly Face kind B is transferred in liquid seed culture medium, through shake-flask culture, liquid seeds B is obtained;
Obtained liquid seeds B is inoculated in liquid fermentation medium, control ph 6~7, ventilation ratio is 1: (0.5~ 1), 100~200 revs/min of rotating speed, cultivate 36~48h at 30~37 DEG C, and fermentation liquid B is obtained;Then by fermentation liquid B by 6%~ 10% mass ratio is forwarded in solid fermentation culture medium, in 30~35 DEG C of temperature, tank pressure 0.02~0.06Mpa condition bottom fermentations Cultivate to spore rate be 90% when, through filter pressing, flash distillation, dries pulverizing, mycopowder B is obtained;
(3) by Bacillus licheniformis (Bacillus licheniformis) through slant culture after, be obtained inclined-plane kind C;Will Inclined-plane kind C is transferred in liquid seed culture medium, through shake-flask culture, liquid seeds C is obtained;
Obtained liquid seeds C is inoculated in liquid fermentation medium, control ph 7~8, ventilation ratio is 1: (0.3~ 0.8), 80~150 revs/min of rotating speed, cultivate 36~48h at 30~37 DEG C, be obtained fermentation liquid C then by fermentation liquid C by 6%~ 10% mass ratio is forwarded in solid fermentation culture medium, in 32~40 DEG C of temperature, tank pressure 0.02~0.06Mpa condition bottom fermentations Cultivate to spore rate be 90% when, through filter pressing, flash distillation, dries pulverizing, mycopowder C is obtained;
(4) by colloid bacillus cereus (Paenibacillus mucilaginosus) through slant culture after, be obtained inclined-plane kind D;Inclined-plane kind D is transferred in liquid seed culture medium, through shake-flask culture, liquid seeds D is obtained;
Obtained liquid seeds D is inoculated in liquid fermentation medium, control ph 7~7.5, ventilation ratio is 1: (0.3 ~0.8), 190~250 revs/min of rotating speed, 36~48h are cultivated at 30~37 DEG C, fermentation liquid D is obtained;Then fermentation liquid D is pressed 6% ~10% mass ratio is forwarded in solid fermentation culture medium, and in 28~36 DEG C of temperature, tank pressure 0.02~0.06Mpa conditions are issued When ferment culture to spore rate is 90%, through filter pressing, flash distillation, dries pulverizing, mycopowder D is obtained;
(5) by bacillus megaterium (Paenibacillus mucilaginosus) through slant culture after, be obtained inclined-plane kind E;Inclined-plane kind E is transferred in liquid seed culture medium, through shake-flask culture, liquid seeds E is obtained;
Obtained liquid seeds E is inoculated in liquid fermentation medium, control ph 8~9, ventilation ratio is 1: (0.5~ 1), 100~280 revs/min of rotating speed, cultivate 36~48h at 30~37 DEG C, and fermentation liquid E is obtained;Then by fermentation liquid E by 6%~ 10% mass ratio is forwarded in solid fermentation culture medium, in 25~35 DEG C of temperature, tank pressure 0.02~0.06Mpa condition bottom fermentations Cultivate to spore rate be 90% when, through filter pressing, flash distillation, dries pulverizing, mycopowder E is obtained;
(6) the mycopowder A for preparing step (1) to (5), mycopowder B, mycopowder C, mycopowder D, mycopowder E are according to mass ratio 1:(2.5 ~3.5):(0.5~1.5):(0.5~1.5):(1.5~2.5) mix, mixed microorganism yeast powder is obtained.
According to currently preferred, in step (1) to (5), slant culture condition is:In 30~37 DEG C of temperature Under, 24~48h is cultivated in slant medium;Every liter of component of the slant medium is as follows:
6~9g of peptone, 3~5g of yeast powder, 2~4g of Carnis Bovis seu Bubali cream, 3~5g of sodium chloride, 10~25g of agar, biphosphate 0.5~1g of potassium, 0.5~1g of dipotassium hydrogen phosphate.
According to currently preferred, in step (1) to (5), shake flask culture conditions are:Under the conditions of 30~37 DEG C, 36~48h of culture;Every liter of component of the liquid seed culture medium is as follows:
6~9g of peptone, 5~8g of glucose, 3~5g of yeast powder, 2~4g of Carnis Bovis seu Bubali cream, 3~5g of sodium chloride, biphosphate 0.5~1g of potassium, 0.5~1g of dipotassium hydrogen phosphate.
According to currently preferred, in step (1) to (5), every liter of component of liquid fermentation medium is as follows:
10~15g of Semen Maydis powder, 10~15g of soybean cake powder, 5~10g of glucose, 5~10g of ammonium sulfate, 3~5g of yeast powder, phosphorus Acid dihydride 0.5~0.8g of potassium, 0.1~0.25g of magnesium sulfate, 0.1~0.25g of manganese sulfate, 0.1~0.2g of ferrous chloride, adjust pH Value 7~8.
According to currently preferred, in step (1) to (5), solid fermentation nutrient media components is as follows, is weight Part:
20~30 parts of wheat bran, 10~20 parts of soybean cake powder, 10~20 parts of Semen Maydis powder, 10~15 parts of Carnis Bovis seu Bubali cream, peptone 5~10 Part, 0.5~1 part of sodium chloride, 0.5~1 part of ammonium sulfate, 0.2~0.5 part of Calcium Carbonate, 0.05~0.1 part of potassium dihydrogen phosphate, sulphuric acid 0.01~0.025 part of magnesium, 0.01~0.025 part of manganese sulfate, 0.01~0.02 part of ferrous chloride, 0.01~0.02 part of zinc sulfate, In water material mass ratio 1: (1: 1~1.2) ratio adds water, pH value 7~8 is adjusted.
Application of the above-mentioned high concentration microorganism microbial inoculum containing polyglutamic acid in improved soil.
Above-mentioned application, step are as follows:
The high concentration microorganism microbial inoculum containing polyglutamic acid is uniformly sprinkling upon in soil during site preparation, is stirred with soil by ploughing Mix uniform;Or, punching of watering is applied;Every mu of amount of application is 3~5kg.
Beneficial effect
1st, the present invention with the addition of polyglutamic acid in microbial bacterial agent, substantial amounts of superpower hydrophilic due to containing in polyglutamic acid Group-carboxyl, can fully keep soil moisture, improve porosity and the bulkiness of soil, improve the retain water and nutrients energy of soil Power;The addition of polyglutamic acid can be greatly improved the field planting rate of microorganism simultaneously;
2nd, it is obtained after the present invention is mixed with polyglutamic acid and other trace element using high concentration mixed microorganism microbial inoculum, high Concentration mixed microorganism microbial inoculum is by bacillus subtilises, Brevibacillus laterosporus, Bacillus licheniformis, colloid bacillus cereus, huge Bacillus cereuss combination is obtained, and polyglutamic acid and precipitated calcium carbonate can be good at thalline steady load;In soil microorganism area In system's composition, above species microorganism not only has and mutually promotes, the effect for mutually cooperateing with, and can strengthen phosphorus decomposing, potassium decomposing Ability, increases soil fertility, without any side effects to soil crop, while supplementing crop calcium constituent, improves plant disease-resistant insect pest Ability;
3rd, the polyglutamic acid added in microbial bacterial agent of the present invention is easy to degrade, and catabolite is glutamic acid, It is a kind of nutrient substance, while increasing crop to other Nutrient Absorption abilities, increases fertilizer efficiency;
4th, high concentration microorganism microbial inoculum of the present invention is obtained by liquid, solid combined fermentation method, and the end-product for obtaining has Effect viable count can reach >=100,000,000,000/gram, greatly better than the viable count in current actual production;And add water-retaining agent polyglutamic Acid, strengthens survival ability of the viable bacteria in soil.
Specific embodiment
Technical scheme is described further with reference to embodiment, but institute's protection domain of the present invention is not limited to This.
Microbe-derived
Bacillus subtilises (Bacillus subtilis) derive from Chinese agriculture Microbiological Culture Collection administrative center, Bacterium numbering ACCC 10242;
Brevibacillus laterosporus (Bacillus laterosporus) are in the management of Chinese agriculture Microbiological Culture Collection The heart, bacterium numbering ACCC 11079.
Bacillus licheniformis (Bacillus licheniformis) are managed from Chinese agriculture Microbiological Culture Collection Center, bacterium numbering ACCC 02002
Colloid bacillus cereus (Paenibacillus mucilaginosus) derive from Chinese agriculture Microbiological Culture Collection Administrative center ACCC 10012
Bacillus megaterium (Bacillus megaterium) is in the management of Chinese industrial Microbiological Culture Collection The heart, bacterium numbering CICC 21694
Embodiment 1
A kind of high concentration microorganism microbial inoculum containing polyglutamic acid, component are as follows, are weight portion:
20 parts of mixed microorganism yeast powder, 63 parts of precipitated calcium carbonate, 8 parts of polyglutamic acid, 4 parts of magnesium sulfate, 3 parts of boric acid, sulfur 2 parts of sour zinc;
Contain bacillus subtilises (Bacillus in the high concentration microorganism microbial inoculum containing polyglutamic acid Subtilis), Brevibacillus laterosporus (Bacillus laterosporus), Bacillus licheniformis (Bacillus Licheniformis), colloid bacillus cereus (Paenibacillus mucilaginosus), bacillus megaterium The ratio of the number of viable of (Bacillus megaterium) is 1: 2.5:0.8:1:2.5, effective total viable count can reach 130,000,000,000 Individual/gram.
The preparation method of the above-mentioned high concentration microorganism microbial inoculum containing polyglutamic acid, comprises the steps:
(1) by bacillus subtilises (Bacillus subtilis) through slant culture after, be obtained inclined-plane kind A;By inclined-plane kind A transfers in liquid seed culture medium, through shake-flask culture, liquid seeds A is obtained;
Obtained liquid seeds A is inoculated in liquid fermentation medium, control ph 7.5, ventilation ratio is 1: 1, rotating speed , 42h is cultivated at 37 DEG C, fermentation liquid A is obtained by 100 revs/min;Then fermentation liquid A is forwarded to solid fermentation by 10% mass ratio In culture medium, in 37 DEG C of temperature, when under the conditions of tank pressure 0.04Mpa, fermentation culture to microscopy spore rate is 90%, through filter pressing, sudden strain of a muscle Steaming, dries pulverizing, are obtained mycopowder A;
(2) by Brevibacillus laterosporus (Bacillus laterosporus) through slant culture after, be obtained inclined-plane kind B;Will be tiltedly Face kind B is transferred in liquid seed culture medium, through shake-flask culture, liquid seeds B is obtained;
Obtained liquid seeds B is inoculated in liquid fermentation medium, control ph 6.5, ventilation ratio is 1: 0.5, turns 200 revs/min of speed, cultivates 48h at 30 DEG C, and fermentation liquid B is obtained;Then fermentation liquid B is forwarded to solid fermentation by 6% mass ratio In culture medium, in 35 DEG C of temperature, when under the conditions of tank pressure 0.06Mpa, fermentation culture to spore rate is 90%, through filter pressing, flash distillation, baking Dry grinding, is obtained mycopowder B;
(3) by Bacillus licheniformis (Bacillus licheniformis) through slant culture after, be obtained inclined-plane kind C;Will Inclined-plane kind C is transferred in liquid seed culture medium, through shake-flask culture, liquid seeds C is obtained;
Obtained liquid seeds C is inoculated in liquid fermentation medium, control ph 7.5, ventilation ratio is 1: 0.3, turns 150 revs/min of speed, cultivates 36h at 35 DEG C, fermentation liquid C is obtained and then fermentation liquid C is forwarded to solid fermentation by 10% mass ratio In culture medium, in 40 DEG C of temperature, when under the conditions of tank pressure 0.02Mpa, fermentation culture to spore rate is 90%, through filter pressing, flash distillation, baking Dry grinding, is obtained mycopowder C;
(4) by colloid bacillus cereus (Paenibacillus mucilaginosus) through slant culture after, be obtained inclined-plane kind D;Inclined-plane kind D is transferred in liquid seed culture medium, through shake-flask culture, liquid seeds D is obtained;
Obtained liquid seeds D is inoculated in liquid fermentation medium, control ph 7, ventilation ratio is 1: 0.8, rotating speed , 36h is cultivated at 37 DEG C, fermentation liquid D is obtained by 190 revs/min;Then fermentation liquid D is forwarded to solid fermentation training by 8% mass ratio In foster base, in 32 DEG C of temperature, when under the conditions of tank pressure 0.05Mpa, fermentation culture to spore rate is 90%, through filter pressing, flash distillation, drying Crush, mycopowder D is obtained;
(5) by bacillus megaterium (Paenibacillus mucilaginosus) through slant culture after, be obtained inclined-plane kind E;Inclined-plane kind E is transferred in liquid seed culture medium, through shake-flask culture, liquid seeds E is obtained;
Obtained liquid seeds E is inoculated in liquid fermentation medium, control ph 8.5, ventilation ratio is 1: 0.8, turns 180 revs/min of speed, cultivates 45h at 35 DEG C, and fermentation liquid E is obtained;Then fermentation liquid E is forwarded to solid fermentation by 8% mass ratio In culture medium, in 30 DEG C of temperature, when under the conditions of tank pressure 0.04Mpa, fermentation culture to spore rate is 90%, through filter pressing, flash distillation, baking Dry grinding, is obtained mycopowder E;
(6) the mycopowder A for preparing step (1) to (5), mycopowder B, mycopowder C, mycopowder D, mycopowder E are according to mass ratio 1:2.5: 0.8:1:2.5 mixing, are obtained mixed microorganism yeast powder.
In step (1) to (5), slant culture condition is:At a temperature of 35 DEG C, cultivate in slant medium 36h;Every liter of component of the slant medium is as follows:
Peptone 8g, yeast powder 4g, Carnis Bovis seu Bubali cream 3g, sodium chloride 4g, agar 17g, potassium dihydrogen phosphate 0.8g, dipotassium hydrogen phosphate 0.8g.
According to currently preferred, in step (1) to (5), shake flask culture conditions are:Under the conditions of 35 DEG C, culture 42h;Every liter of component of the liquid seed culture medium is as follows:
Peptone 9g, glucose 5g yeast powder 5g, Carnis Bovis seu Bubali cream 2g, sodium chloride 5g, potassium dihydrogen phosphate 0.5g, dipotassium hydrogen phosphate 1g.
In step (1) to (5), every liter of component of liquid fermentation medium is as follows:
Semen Maydis powder 12g, soybean cake powder 13g, glucose 8g, ammonium sulfate 6g, yeast powder 4g, potassium dihydrogen phosphate 0.7g, magnesium sulfate 0.2g, manganese sulfate 0.15g, ferrous chloride 0.15g, adjust pH value 7.4.
In step (1) to (5), solid fermentation nutrient media components is as follows, is weight portion:
25 parts of wheat bran, 15 parts of soybean cake powder, 15 parts of Semen Maydis powder, 13 parts of Carnis Bovis seu Bubali cream, 8 parts of peptone, 0.8 part of sodium chloride, sulphuric acid 0.8 part of ammonium, 0.4 part of Calcium Carbonate, 0.08 part of potassium dihydrogen phosphate, 0.02 part of magnesium sulfate, 0.02 part of manganese sulfate, ferrous chloride 0.015 Part, 0.015 part of zinc sulfate adds water in 1: 1.1 ratio of water material mass ratio, adjusts pH value 7.4.
Embodiment 2
The preparation method of the high concentration microorganism microbial inoculum containing polyglutamic acid as described in Example 1, difference are:
Mycopowder A, mycopowder B, mycopowder C, mycopowder D, mycopowder E by weight 1:3:1:1:2 ratio mix homogeneously, is obtained mixing Fermentable powder;Then press mixed microorganism yeast powder 20kg, precipitated calcium carbonate 63kg, polyglutamic acid 8kg, magnesium sulfate 4kg, Boric acid 3kg, zinc sulfate 2kg mix homogeneously, are obtained the high concentration microorganism microbial inoculum containing polyglutamic acid.
Data are as shown in table 1 after testing:
Table 1
Test of 3 polyglutamic acid of embodiment in the field planting rate for increasing Soil Microorganism:
Method as described in embodiment 1, by mycopowder A, mycopowder B, mycopowder C, mycopowder D, mycopowder E by weight 1:3:1:1:2 Ratio mix homogeneously, mixed microorganism yeast powder is obtained;Again by 20 parts of mixed microorganism yeast powder, 63 parts of precipitated calcium carbonate, 2 parts of 4 parts of magnesium sulfate, 3 parts of boric acid and zinc sulfate are uniformly mixed and made into composite high concentration microbial bacterial agent;
The mixed microorganism yeast powder of above-mentioned preparation is added different types of water-retaining agent respectively to apply by 5 grams/m of punchings, Punching is manured into soil, detection soil microorganism, available phosphoruss and quick-acting potassium content after two weeks:
Treatment group one:Composite high concentration microbial bacterial agent;
Treatment group two:+ 8 parts of composite high concentration microbial bacterial agent (8%) polyglutamic acid;
Treatment group three:+ 8 parts of composite high concentration microbial bacterial agent (8%) polyacrylamide.
Testing result is as shown in table 2:
Impact of the 2 different disposal group of table to soil microorganism field planting rate
Impact of the 3 different disposal group of table to soil available phosphorus and quick-acting potassium content
As can be seen from Table 2, adding water-retaining agent can increase the field planting rate of Soil Microorganism microbial inoculum, add polyglutamic acid and Matched group compared by polyacrylamide can increase bacteria planting rate 25.05%, 14.05% respectively.Add polyglutamic acid simultaneously to soil In earth, original actinomycetes and funguses also have certain growth promoting function.
As can be seen from Table 3, adding water-retaining agent can increase the phosphorus decomposing of microorganism, ability of dissolving potassium, add polyglutamic acid and gather Matched group compared by acrylamide can increase available phosphorus content 18.34%, 15.22% in soil respectively, increase quick-acting potassium content 15.16%th, 7.33%.
The application of high concentration microorganism microbial inoculum of the embodiment 4 containing polyglutamic acid:
In October, 2014 in June, 2015, moisten hundred standing grain Vegetable Bases in Laiwu City, experimental cultivar is Fructus Lycopersici esculenti, MAOFEN 802, Greenhouse cultivation.
EXPERIMENTAL DESIGN:
Process 1:High concentration microorganism microbial inoculum containing polyglutamic acid prepared by fertilising+embodiment 1;
Process 2:Conventional fertilizer application+by embodiment 1 prepare the high concentration microorganism bacterium containing polyglutamic acid for lacking mycopowder A Agent;
Process 3:Conventional fertilizer application+fire extinguishing substrate;
Process 4:Conventional fertilizer application;
Four treatment groups, 8 row areas.It is repeated 3 times, 0.05 mu of plot area.Random alignment, Cell Setup protect 1 meter of row, respectively Cell list fills single, applies 3000 kilograms of farmyard manure by per mu during site preparation, tomato growth stage, with the net chemical control elder brother of 1500 times of flea beetles Worm, other cultivation management measures are consistent with land for growing field crops.
Fertilizing method:
Process 1:On the basis of conventional fertilizer application, when transplanting, mu applies 2kg, applies with soil pit by 1: 100;During tomato growth Microbial inoculum is watered pouring by 1: 1000 times, was imposed once every 15 days, impose 3 times altogether;
Process 2:By high concentration microorganism microbial inoculum (the administration side containing polyglutamic acid for lacking mycopowder A prepared by embodiment 1 1) method is same to be processed;
Process 3:Microbial inoculum is processed as substrate of putting out a fire (application process is with process 1);
Process 4:Conventional fertilizer application, mu carbamide 20kg, calcium superphosphate 100kg, potassium sulfate 50kg during plant, is starting to harvest When, topdressed carbamide 20kg/ mus every 20 days, potassium sulfate 30kg/ mus.
Impact of each fertilizer treatment to tomato yield is counted, the full phosphorus of measurement each group plant, full potassium content after off-test, Gather each group soil sample simultaneously and determine soil physical and chemical property.Method adopts soil weight application core cutter method, organic matter application dichromic acid Potassium capacity method determines (referring to NY/T 1121.4-2006 standards).
Testing result:
Impact of the 4 each fertilizer treatment of table to tomato yield
Impact of the 5 each fertilizer treatment of table to plant content of tatal phosphorus
Impact of the 6 each fertilizer treatment of table to the full potassium content of plant
Impact of the 7 each fertilizer treatment of table to the soil weight
Impact of the 8 each fertilizer treatment of table to the soil organism
Can be shown as can be seen that applying the high concentration microorganism microbial inoculum containing polyglutamic acid of the present invention by upper table 4 The yield for improving Fructus Lycopersici esculenti is write, conventional fertilizer application group mu rate of growth 17.99%, obvious effect of increasing production is compared.
Found out by upper table 5 and table 6, applying the high concentration microorganism microbial inoculum containing polyglutamic acid of the present invention can show Write and improve the full phosphorus of plant, full potassium content, comparing conventional fertilizer application group increases by 44.64%, 20.04% respectively, it can be seen that this The described high concentration microorganism microbial inoculum containing polyglutamic acid of invention can dramatically increase the absorption of plant pair phosphorus, potassium.
Can be shown as can be seen that applying the high concentration microorganism microbial inoculum containing polyglutamic acid of the present invention by upper table 7 Write and improve the soil weight, comparing the conventional fertilizer application group soil weight increases by 2.4%.By upper table 8 as can be seen that applying of the present invention The high concentration microorganism microbial inoculum containing polyglutamic acid can significantly improve soil organic matter content, compare conventional fertilizer application group soil Organic matter increases by 3.0%.It can be seen that the high concentration microorganism microbial inoculum containing polyglutamic acid of the present invention is to soil Improvement has positive effect.

Claims (10)

1. a kind of high concentration microorganism microbial inoculum containing polyglutamic acid, it is characterised in that component is as follows, is weight portion:
15~25 parts of mixed microorganism yeast powder, 55~70 parts of precipitated calcium carbonate, 5~10 parts of polyglutamic acid, 3~5 parts of magnesium sulfate, 3 parts of boric acid, 2 parts of zinc sulfate;
The polyglutamic acid is poly-gamma-glutamic acid, by D types glutamic acid and L-type glutamic acid by amide bonded, each paddy ammonia Acid all leaves a free carboxyl.
2. high concentration microorganism microbial inoculum as claimed in claim 1, it is characterised in that contain in the mixed microorganism yeast powder Bacillus subtilises (Bacillus subtilis), Brevibacillus laterosporus (Bacillus laterosporus), lichens spore Bacillus (Bacillus licheniformis), colloid bacillus cereus (Paenibacillus mucilaginosus), huge bud The ratio of the number of viable of spore bacillus (Bacillus megaterium) is 1:(2.5~3.5):(0.5~1.5):(0.5~ 1.5):(1.5~2.5), effective total viable count reach >=100,000,000,000/gram, and wherein every kind of bacterium living bacteria count is not less than 5,000,000,000 Individual/gram.
3. high concentration microorganism microbial inoculum as claimed in claim 1, it is characterised in that the bacillus subtilises (Bacillus Subtilis) Chinese agriculture Microbiological Culture Collection administrative center, bacterium numbering ACCC 10242 are derived from;Brevibacillus laterosporus (Bacillus laterosporus) derives from Chinese agriculture Microbiological Culture Collection administrative center, bacterium numbering ACCC11079;Bacillus licheniformis (Bacillus licheniformis) derive from Chinese agriculture Microbiological Culture Collection pipe Reason center, bacterium numbering ACCC 02002;Colloid bacillus cereus (Paenibacillus mucilaginosus) are from China Agriculture Microbiological Culture Collection administrative center ACCC 10012;Bacillus megaterium (Bacillus megaterium) derives from Chinese industrial Microbiological Culture Collection administrative center, bacterium numbering CICC 21694.
4. high concentration microorganism microbial inoculum as claimed in claim 1, it is characterised in that molecular weight >=100 of the polyglutamic acid Ten thousand.
5. the preparation method of the high concentration microorganism microbial inoculum containing polyglutamic acid described in claim 1, it is characterised in that step For:Mixed microorganism yeast powder, precipitated calcium carbonate, polyglutamic acid, magnesium sulfate, boric acid, zinc sulfate are mixed in proportion, are obtained final product.
6. preparation method as claimed in claim 5, it is characterised in that mixed microorganism yeast powder is prepared as follows:
(1) by bacillus subtilises (Bacillus subtilis) through slant culture after, be obtained inclined-plane kind A;Inclined-plane kind A is turned It is connected in liquid seed culture medium, through shake-flask culture, liquid seeds A is obtained;
Obtained liquid seeds A is inoculated in liquid fermentation medium, control ph 7~8, ventilation ratio is 1: (0.5~1), 100~200 revs/min of rotating speed, cultivates 36~48h at 30~37 DEG C, and fermentation liquid A is obtained;Then fermentation liquid A is pressed 6%~10% Mass ratio be forwarded in solid fermentation culture medium, in 35~38 DEG C of temperature, fermentation culture under the conditions of 0.02~0.06Mpa of tank pressure When being 90% to microscopy spore rate, through filter pressing, flash distillation, dries pulverizing, mycopowder A is obtained;
(2) by Brevibacillus laterosporus (Bacillus laterosporus) through slant culture after, be obtained inclined-plane kind B;By inclined-plane kind B transfers in liquid seed culture medium, through shake-flask culture, liquid seeds B is obtained;
Obtained liquid seeds B is inoculated in liquid fermentation medium, control ph 6~7, ventilation ratio is 1: (0.5~1), 100~200 revs/min of rotating speed, cultivates 36~48h at 30~37 DEG C, and fermentation liquid B is obtained;Then fermentation liquid B is pressed 6%~10% Mass ratio be forwarded in solid fermentation culture medium, in 30~35 DEG C of temperature, fermentation culture under the conditions of 0.02~0.06Mpa of tank pressure When being 90% to spore rate, through filter pressing, flash distillation, dries pulverizing, mycopowder B is obtained;
(3) by Bacillus licheniformis (Bacillus licheniformis) through slant culture after, be obtained inclined-plane kind C;By inclined-plane Plant C to transfer in liquid seed culture medium, through shake-flask culture, liquid seeds C is obtained;
Obtained liquid seeds C is inoculated in liquid fermentation medium, control ph 7~8, ventilation ratio is 1: (0.3~ 0.8), 80~150 revs/min of rotating speed, cultivate 36~48h at 30~37 DEG C, be obtained fermentation liquid C then by fermentation liquid C by 6%~ 10% mass ratio is forwarded in solid fermentation culture medium, in 32~40 DEG C of temperature, tank pressure 0.02~0.06Mpa condition bottom fermentations Cultivate to spore rate be 90% when, through filter pressing, flash distillation, dries pulverizing, mycopowder C is obtained;
(4) by colloid bacillus cereus (Paenibacillus mucilaginosus) through slant culture after, be obtained inclined-plane kind D;Will Inclined-plane kind D is transferred in liquid seed culture medium, through shake-flask culture, liquid seeds D is obtained;
Obtained liquid seeds D is inoculated in liquid fermentation medium, control ph 7~7.5, ventilation ratio is 1: (0.3~ 0.8), 190~250 revs/min of rotating speed, cultivate 36~48h at 30~37 DEG C, and fermentation liquid D is obtained;Then by fermentation liquid D by 6%~ 10% mass ratio is forwarded in solid fermentation culture medium, in 28~36 DEG C of temperature, tank pressure 0.02~0.06Mpa condition bottom fermentations Cultivate to spore rate be 90% when, through filter pressing, flash distillation, dries pulverizing, mycopowder D is obtained;
(5) by bacillus megaterium (Paenibacillus mucilaginosus) through slant culture after, be obtained inclined-plane kind E;Will Inclined-plane kind E is transferred in liquid seed culture medium, through shake-flask culture, liquid seeds E is obtained;
Obtained liquid seeds E is inoculated in liquid fermentation medium, control ph 8~9, ventilation ratio is 1: (0.5~1), 100~280 revs/min of rotating speed, cultivates 36~48h at 30~37 DEG C, and fermentation liquid E is obtained;Then fermentation liquid E is pressed 6%~10% Mass ratio be forwarded in solid fermentation culture medium, in 25~35 DEG C of temperature, fermentation culture under the conditions of 0.02~0.06Mpa of tank pressure When being 90% to spore rate, through filter pressing, flash distillation, dries pulverizing, mycopowder E is obtained;
(6) the mycopowder A for preparing step (1) to (5), mycopowder B, mycopowder C, mycopowder D, mycopowder E are according to mass ratio 1:(2.5~ 3.5):(0.5~1.5):(0.5~1.5):(1.5~2.5) mix, mixed microorganism yeast powder is obtained.
7. preparation method as claimed in claim 6, it is characterised in that in step (1) to (5), slant culture condition is: At a temperature of 30~37 DEG C, 24~48h is cultivated in slant medium;Every liter of component of the slant medium is as follows:
6~9g of peptone, 3~5g of yeast powder, 2~4g of Carnis Bovis seu Bubali cream, 3~5g of sodium chloride, 10~25g of agar, potassium dihydrogen phosphate 0.5 ~1g, 0.5~1g of dipotassium hydrogen phosphate;
Preferably, in step (1) to (5), shake flask culture conditions are:Under the conditions of 30~37 DEG C, 36~48h is cultivated;Institute State every liter of component of liquid seed culture medium as follows:
6~9g of peptone, 5~8g of glucose, 3~5g of yeast powder, 2~4g of Carnis Bovis seu Bubali cream, 3~5g of sodium chloride, potassium dihydrogen phosphate 0.5 ~1g, 0.5~1g of dipotassium hydrogen phosphate.
8. preparation method as claimed in claim 6, it is characterised in that in step (1) to (5), liquid fermentation medium Every liter of component is as follows:
10~15g of Semen Maydis powder, 10~15g of soybean cake powder, 5~10g of glucose, 5~10g of ammonium sulfate, 3~5g of yeast powder, di(2-ethylhexyl)phosphate Hydrogen 0.5~0.8g of potassium, 0.1~0.25g of magnesium sulfate, 0.1~0.25g of manganese sulfate, 0.1~0.2g of ferrous chloride, adjustment pH value 7~ 8;
Preferably, in step (1) to (5), solid fermentation nutrient media components is as follows, is weight portion:
20~30 parts of wheat bran, 10~20 parts of soybean cake powder, 10~20 parts of Semen Maydis powder, 10~15 parts of Carnis Bovis seu Bubali cream, 5~10 parts of peptone, 0.5~1 part of sodium chloride, 0.5~1 part of ammonium sulfate, 0.2~0.5 part of Calcium Carbonate, 0.05~0.1 part of potassium dihydrogen phosphate, magnesium sulfate 0.01~0.025 part, 0.01~0.025 part of manganese sulfate, 0.01~0.02 part of ferrous chloride, 0.01~0.02 part of zinc sulfate are pressed Water material mass ratio 1: (1: 1~1.2) ratio adds water, adjusts pH value 7~8.
9. application of the high concentration microorganism microbial inoculum containing polyglutamic acid in improved soil described in claim 1.
10. application as claimed in claim 9, it is characterised in that step is as follows:
The high concentration microorganism microbial inoculum containing polyglutamic acid is uniformly sprinkling upon in soil during site preparation, by ploughing with soil stirring Even;Or, punching of watering is applied;Every mu of amount of application is 3~5kg.
CN201610954700.3A 2016-10-27 2016-10-27 A kind of high concentration microorganism microbial inoculum containing polyglutamic acid and preparation method and application Pending CN106495951A (en)

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CN109400378A (en) * 2018-12-07 2019-03-01 山东土木启生物科技有限公司 Improve the complex micro organism fungicide and preparation method of crop soil available phosphorus, potassium
CN110724008A (en) * 2019-11-28 2020-01-24 吉林省嘉博生物科技有限公司 Long-acting microbial agent for planting vegetation for slope treatment
CN111018612A (en) * 2019-12-20 2020-04-17 德州市元和农业科技开发有限责任公司 Full-water-soluble drip-irrigation microbial inoculant particles and preparation method thereof
CN110878271A (en) * 2019-12-27 2020-03-13 天津开发区坤禾生物技术有限公司 Bacillus licheniformis MES816 and product and application thereof
CN110878271B (en) * 2019-12-27 2021-06-18 天津开发区坤禾生物技术有限公司 Bacillus licheniformis MES816 and product and application thereof
CN113234446A (en) * 2021-04-02 2021-08-10 宁夏农林科学院农业生物技术研究中心(宁夏农业生物技术重点实验室) Biomass saline-alkali soil conditioner

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