CN103805204A - Farmland soil microorganism water-holding product and preparation method thereof - Google Patents

Farmland soil microorganism water-holding product and preparation method thereof Download PDF

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CN103805204A
CN103805204A CN201310307877.0A CN201310307877A CN103805204A CN 103805204 A CN103805204 A CN 103805204A CN 201310307877 A CN201310307877 A CN 201310307877A CN 103805204 A CN103805204 A CN 103805204A
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陈立华
邵孝侯
程晋
金斌斌
崔智伟
蒋韵妮
毛欣宇
邓伯均
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Hohai University HHU
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Abstract

The invention relates to a farmland soil microorganism water-holding product as well as method for preparing the product through a function microbial strain for synthesizing and a function microbial strain for synthesizing gamma-poly glutamic acid, and belongs to the technical field of microorganism. The microbial strains include IAE ((i) Brevibacterium (/i) (i) flavum (/i) IAE) and IAE ((i) Bacillus amyloliquefaciens (/i) IAE), and are separated from saline-alkali soil of coastal mud flat of Jiangsu. The microorganism water-holding product is produced from the two strains by taking pig manure compost, rice straw and (NH4)2SO4 as solid fermentation substrate, and the product contains effective water-holding substance higher than 19.6kg-1 dry material weight. Experiments of the product's water holding capacity on land soil show that the microorganism organic fertilizer, applied to soil, can reduce soil bulk density, increase water hydraulic conductivity and soil saturated water content, and can increase plant height, fresh weight, dry weight and yield.

Description

A kind of agricultural land soil microorganism water conservation goods and preparation method thereof
one, technical field
The microorganism water conservation goods that the present invention relates to prepare the microorganism strains of agricultural soil water-keeping material and produce by solid fermentation, belong to microbial technology field.
two, background technology
Agricultural is China water rich and influential family, its water consumption accounts for 70% left and right of national water total amount, approximately 30,000,000,000 cubes of agricultural year lacks of water, develop the technical development of efficiently retaining and conserveing soil moisture and account for approximately 56% dry farming of territory land area, make full use of precipitation and underground water (comprising the soil water) Developing Water-saving Agriculture, be the fundamental way of agricultural modernization development, application water-keeping material is retained and conserved soil moisture and made full use of water resources is to realize one of water-saving agriculture important channel.
The microbial organic fertilizer of the synthetic water suction water storage high molecular polymer of microbial solid fermentation, having advantages of degradable, environmental friendliness and can significantly reduce widespread pollution from the overuse of fertilizers and pesticides in rural area by high added valueization processing solid organic castoff, is a study hotspot in water-saving agriculture field.Microorganism fermentation synthetic product γ-polyglutamic acid (poly-γ-glutamic acid) is a kind of macromole long-chain polypeptide, molecular weight is at 30 000D ~ 50 000D, water absorbent rate can reach 3500 times, and repetition good water absorption, is widely used in the fields such as food, makeup, life water-absorbing material.
Most of γ-PGA high yield microorganism is L-glutamic acid (Glu) dependent form bacterial strain, and solid fermentation matrix need to be added and is rich in glutamic acid-type material, and as bean powder, casein food grade, L-glutamic acid or residue of monosodium glutamate etc., production cost is higher.γ-polyglutamic acid is a kind of natural macromolecular substance, and easily degraded in soil, causes water conservation effect to reduce.
Microorganism can utilize inorganic nitrogen-sourced (urea, ammonium sulfate etc.) synthesizes glutamic acid-type material, combined ferment (co-fermentation) solid organic castoff by synthetic L-glutamic acid microorganism and synthetic γ-polyglutamic acid microorganism is made biological water-keeping goods, raw materials cost is low, and the L-glutamic acid synthesized micro-organism that enters soil by carrying continues synthetic L-glutamic acid and can effectively maintain the stability of soil γ-polyglutamic acid in soil; γ-PGA the biological organic fertilizer that simultaneously adopts microbial solid fermentation agricultural organic waste and inorganic nitrogenous source to produce, can continue supply microorganism energy by organic matter and reduce the extinction of functional microorganism in soil, the water conservation effect that extends goods.By combined ferment explained hereafter microbe soil water conservation goods, can high added value utilize agricultural wastes, can effectively increase soil moisture conservation, be conducive to improve the utilising efficiency of agricultural water resources, made microorganism water conservation goods will have good application prospect.
  
three, summary of the invention
goal of the invention
The object of the present invention is to provide the microorganism strains of combined ferment agricultural wastes and inorganic nitrogen-sourced production soil water-retaining goods and the soil water-retaining goods that fermentation is made thereof, for agriculture production utilization, development high-efficiency water-saving modern agriculture.
technical scheme
A kind of agricultural land soil microorganism of the present invention water conservation goods and preparation method thereof are realized by following scheme:
A kind of agricultural land soil microorganism water conservation goods, is characterized in that described a kind of agricultural land soil microorganism water conservation goods are pig manure, rice straw powder and the (NH of the synthetic corynebacterium glutamicum strain of a strain and the synthetic strain-combined fermentation maturity of γ-polyglutamic acid of a strain 4) 2sO 4the water conservation goods that obtain;
Wherein said corynebacterium glutamicum strain is brevibacterium flavum IAE( brevibacterium flavumiAE), on March 15th, 2013 is preserved in Chinese Typical Representative culture collection center, and preservation address is Luo Jia Shan, wuchang, wuhan, and culture presevation number is CCTCC NO:M 2013087; Referred to as brevibacterium flavum CCTCC NO:M 2013087;
Described γ-many corynebacterium glutamicum strains are bacillus amyloliquefaciens IAE( bacillus amyloliquefaciensiAE), on March 15th, 2013 is preserved in Chinese Typical Representative culture collection center, and preservation address is Luo Jia Shan, wuchang, wuhan, and culture presevation number is CCTCC NO:M 2013086, referred to as bacillus amyloliquefaciens CCTCC NO:M 2013086.
In these goods, γ-polyglutamic acid content is higher than 19.6g kg -1dry products, contains 0.5 × 10 8cfu g -1above brevibacterium flavum CCTCC NO:M 2013087 colony-forming units and 0.5 × 10 8cfu g -1above bacillus amyloliquefaciens CCTCC NO:M 2013086 colony-forming units, quality of organic matter is 30～35% than content, water ratio is lower than 30%.
The culture condition of described brevibacterium flavum CCTCC NO:M 2013087: be inoculated in the 250ml triangular flask that 100ml nutrient solution is housed with transfering loop picking growth bacterium of 24 hours from culture dish, 37 ℃, 170 rmp cultivate 48 hours, the centrifugal 20min of bacterium liquid supernatant 10000rpm, measuring supernatant liquor content of glutamic acid is 26.6g L -1.
Brevibacterium flavum CCTCC NO:M 2013087 nutrient solution compound method used is, to prepare 1L substratum as example: glucose 50g, corn steep liquor 10ml, urea 5g, KH 2pO 43g, MgSO 40.8g, distilled water 1000ml, pH value nature, 115 ℃ of sterilizing 30min.
The culture condition of described bacillus amyloliquefaciens CCTCC NO:M 2013086: be inoculated in the 250ml triangular flask that the synthetic L-glutamic acid nutrient solution of 100ml is housed with transfering loop picking growth bacterium of 24 hours from culture dish, 37 ℃, 170rmp are cultivated 48 hours, the centrifugal 20min of bacterium liquid supernatant 10000rpm, measuring supernatant liquor γ-polyglutamic acid content is 21.8g L -1.
Described bacillus amyloliquefaciens CCTCC NO:M 2013086 liquid nutrient medium compound method used is, to prepare 1L substratum as example: glucose 80g, Sodium Glutamate 50g, (NH 4) 2sO 48g, NaCl 5g, K 2hPO 43H 2o 2g, MnSO 47H 2o 0.25g, MnSO 4h 2o 0.03 g, distilled water 1000 ml, pH value nature, 115 ℃ of sterilizing 30min.
The preparation method of described soil microorganisms water conservation goods, is characterized in that realizing as follows:
1) brevibacterium flavum CCTCC NO:M 2013087 bacterium are inoculated into liquid seed culture medium and cultivate, culture condition is: 37 ℃ of culture temperature, and shaking speed 150rpm, incubation time 24 hours, seed bacteria containing amount is greater than 1 × 10 9cfu ml -1, seed is inoculated into fermentor tank according to 5% ratio and carries out liquid fermenting production, and the condition of its fermentative production is: 28 ~ 33 ℃ of culture temperature, dissolved oxygen air flow scope is 30～100%, 150 ~ 220rpm, colony-forming unit>=1 × 10 of this bacterial strain of fermentation later stage fermentation liquid 9cfu ml -1.The formula of the seed culture medium of liquid used is to prepare according to 1L substratum: extractum carnis 3g, peptone 10g, NaCl 10g, tap water 1000ml, pH scope 7.0-7.5,121 ℃ of sterilizing 20min; Liquid fermentation medium formula used is to prepare according to 1L substratum: glucose 50g, corn steep liquor 10ml, urea 5g, KH 2pO 43g, MgSO 40.8g, distilled water 1000ml, pH value nature, 115 ℃ of sterilizing 30min;
2) bacillus amyloliquefaciens CCTCC NO:M 2013086 is inoculated into liquid seed culture medium and cultivates, culture condition is: 37 ℃ of culture temperature, and shaking speed 130 ~ 160rpm, incubation time 24 hours, seed bacteria containing amount is greater than 1 × 10 9cfu ml -1, seed is inoculated into fermentor tank according to 8% ratio and carries out liquid fermenting production, and the condition of its fermentative production is: 28 ~ 35 ℃ of culture temperature, dissolved oxygen air flow scope is 30～100%, 120 ~ 160rpm, fermentation later stage part bacterial strain forms gemma, colony-forming unit>=1 × 10 of this bacterial strain of fermented liquid 9cfu ml -1, the retrogradation of fermented liquid concentration; The formula of the seed culture medium of liquid used is to prepare according to 1L substratum: extractum carnis 3g, peptone 10g, NaCl 10g, tap water 1000ml, pH scope 7.0-7.5,121 ℃ of sterilizing 20min.Liquid fermentation medium formula used is to prepare according to 1L substratum: glucose 80g, Sodium Glutamate 50g, (NH 4) 2sO 48g, NaCl 5g, K 2hPO 43H 2o 2g, MnSO 47H 2o 0.25g, MnSO 4h 2o 0.03 g, distilled water 1000ml, pH value nature, 115 ℃ of sterilizing 30min;
3) by brevibacterium flavum CCTCC NO:M 2013087 fermented liquids, bacillus amyloliquefaciens CCTCC NO:M 2013086 fermented liquids, (NH 4) 2sO 4, rice straw powder and the pig manure that becomes thoroughly decomposed be according to 5 ~ 8:6 ~ 10:7 ~ 13:25 ~ 35:40 ~ 55(w:w:w:w:w:w) stirring and evenly mixing thoroughly, the fermentation of banking up of bar buttress formula, leavening temperature is 30 ~ 50 ℃, turning in every 3 days 1 time in fermenting process, ferment and finish after 15 ~ 20 days, make brevibacterium flavum content reach 0.5 × 10 8cfu g -1above, bacillus amyloliquefaciens content reaches 0.5 × 10 8cfu g -1above, bacterial strain stable content, γ-polyglutamic acid content is higher than 19.6g kg -1, the goods of acquisition are agricultural soil microorganism water conservation goods;
Described microbe soil water conservation goods can be used for agricultural crops planting soil, increase water retention in soil, improve water resource utilization efficiency, increase plant biomass.
beneficial effect
The invention provides and a kind ofly can adapt to production method arid lean soil environment, that can synthesize L-glutamic acid and can synthesize microorganism strains and the agricultural soil water conservation goods thereof of γ-polyglutamic acid.By long-term lack of water arid and synthetic L-glutamic acid in lean soil environment and separating of γ-polyglutamic acid microorganism are obtained to the synthetic corynebacterium glutamicum strain of a plant height effect and efficient synthetic γ-polyglutamic acid bacterial strain, utilize fermentation and the stability program of this two strains bacterial strain in agricultural solid residue, make the microbial product with water retention, its goods and market goods relatively have the following advantages:
1. this bacterial strain has the ability that adapts to arid and lean soil environment, test-results shows, use the beach saline-alkali edatope of these goods, after arid winter and 6 months spring, in soil, brevibacterium flavum bacterial strain and synthetic γ-polyglutamic acid Bacillus amyloliquefaciens strain survival volume of synthetic L-glutamic acid are greater than respectively 60% and 80%, can continue synthetic water conservation material, compared to soil application single γ-polyglutamic acid compound or single bacillus amyloliquefaciens water conservation goods, the water conservation effect time length is more of a specified duration, water retention capacity is stronger.
2. these goods contain and enrich water conservation material, can increase soil macro aggregate quantity, reduce the soil weight, reduce top layer skinning, increase soil saturation/unsaturated hydraulic conductivity, the validity that moisturizes, increase specific water capacity.
3. owing to being biological products, completely less than the series of problems bringing because of the use of chemicals, the Sustainable development that is conducive to improve agricultural water use efficiency.
four, accompanying drawing explanation
Fig. 1 is brevibacterium flavum bacterial strain electromicroscopic photograph.
Fig. 2 is Bacillus amyloliquefaciens strain electromicroscopic photograph.
Fig. 3 is γ-polyglutamic acid soaking effect figure after purifying.
Fig. 4 is the moisture-preservation growth-promoting design sketch of water conservation goods.
five, embodiment
embodiment 1
the separation of bacterial strain and evaluation
Gather the plant rhizosphere surrounding soil cryopreservation of in the exposed beach of tidal flat of Jiangsu Province, sporadicly growing, adopt basic beef-protein medium to separate L-glutamic acid synthesized micro-organism bacterial strain to Soil Microorganism, its substratum preparation according to 1L amount compound method is: extractum carnis 3g, peptone 10g, NaCl 10g, tap water 1000ml, pH scope 7.0-7.5,121 ℃ of sterilizing 20min.
Further measure the efficiently synthetic bacterial strain of concentration screening of synthetic L-glutamic acid by liquid culture.Culture condition: inoculation culture ware is cultivated the bacterial strain of 24h in the 250ml triangular flask of 100ml liquid nutrient medium is housed, inoculum size 10 4cfu ml -1, 37 ℃, 150rmp are cultivated 7 days, and the centrifugal 20min of bacterium liquid supernatant 10000rpm measures the aminoglutaric acid concentration in nutrient solution, obtains the maximum bacterial strain of L-glutamic acid resultant quantity, fermented liquid aminoglutaric acid concentration 26.6g L -1.This bacterial strain belongs to brevibacterium flavum IAE( brevibacterium flavumiAE), be preserved in Chinese Typical Representative culture collection center on March 15th, 2013, preservation address is Luo Jia Shan, wuchang, wuhan, and culture presevation number is CCTCC NO:M 2013087, and Main Biological is: bacterium colony is faint yellow, rod-short, Gram-positive, amphimicrobian does not move, atrichia, without gemma, the electromicroscopic photograph of bacterial strain as shown in Figure 1; Through the evolutionary analysis of 16s rDNA sequence, result be shown as brevibacterium flavum ( b. flavum).
Above-mentioned synthetic L-glutamic acid liquid culture based formulas is: glucose 50g, corn steep liquor 10ml, urea 5g, KH 2pO 43g, MgSO 40.8g, distilled water 1000ml, pH value nature, 115 ℃ of sterilizing 30min.
Gather the plant rhizosphere surrounding soil cryopreservation of in the exposed beach of tidal flat of Jiangsu Province, sporadicly growing, adopt L-glutamic acid substratum separating gamma-polyglutamic acid synthesized micro-organism bacterial strain.The standard of screening is that bacterium colony forms high protuberance on substratum, and convex protrusion has very strong viscosity and absorptive bacterial strain.The strongest bacterial strain of ability of the synthetic γ-polyglutamic acid of screening liquid fermenting, it is 21.8g L that this bacterial strain synthesizes γ-polyglutamic acid amount -1; This bacterial strain is bacillus amyloliquefaciens IAE( bacillus amyloliquefaciensiAE), on March 15th, 2013 is preserved in Chinese Typical Representative culture collection center, and preservation address is Luo Jia Shan, wuchang, wuhan, culture presevation number is CCTCC NO:M 2013086, and Main Biological is: thalline is shaft-like, even dyeing, tool mobility, amphimicrobian, gemma ovalize, Gram-positive, the bacterium colony opaque colony that is white in color on substratum, edge is irregular, has protuberance, surface folding, bacterial strain electromicroscopic photograph is as shown in Figure 2; Through the evolutionary analysis of 16s rDNA sequence, result be shown as bacillus amyloliquefaciens ( b. amyloliquefaciens).
The formula of above-mentioned substratum is to prepare according to preparation 1L amount: glucose 80g, Sodium Glutamate 50g, (NH 4) 2sO 48g, NaCl 5g, K 2hPO 43H 2o 2g, MnSO 47H 2o 0.25g, MnSO 4h 2o 0.03 g, distilled water 1000ml, pH value nature, 115 ℃ of sterilizing 30min.
microbial inoculum is produced
Brevibacterium flavum CCTCC NO:M 2013087 is inoculated into liquid seed culture medium and cultivates, culture condition is: 37 ℃ of culture temperature, and shaking speed 150rpm, incubation time 24 hours, seed bacteria containing amount is greater than 1 × 10 9cfu ml -1, seed is inoculated into fermentor tank according to 5% ratio and carries out liquid fermenting production, and the condition of its fermentative production is: 28 ~ 33 ℃ of culture temperature, dissolved oxygen air flow scope is 30～100%, 150 ~ 220rpm, colony-forming unit>=1 × 10 of this bacterial strain of fermentation later stage fermentation liquid 9cfu ml -1; The seed culture medium of liquid used and the formula of fermention medium are to prepare according to 1L substratum: extractum carnis 3g, peptone 10g, NaCl 10g, tap water 1000ml, pH scope 7.0-7.5,121 ℃ of sterilizing 20min.
Bacillus amyloliquefaciens CCTCC NO:M 2013086 is inoculated into liquid seed culture medium and cultivates, culture condition is: 37 ℃ of culture temperature, and shaking speed 130 ~ 160rpm, incubation time 24 hours, seed bacteria containing amount is greater than 1 × 10 9cfu ml -1, seed is inoculated into fermentor tank according to 8% ratio and carries out liquid fermenting production, and the condition of its fermentative production is: 28 ~ 35 ℃ of culture temperature, dissolved oxygen air flow scope is 30～100%, 120 ~ 160rpm, fermentation later stage part bacterial strain forms gemma, colony-forming unit>=1 × 10 of this bacterial strain of fermented liquid 9cfu ml -1, the retrogradation of fermented liquid concentration.The formula of liquid seed culture medium used is to prepare according to 1L substratum: extractum carnis 3g, peptone 10g, NaCl 10g, tap water 1000ml, pH scope 7.0-7.5,121 ℃ of sterilizing 20min.Liquid fermentation medium formula used is to prepare according to 1L substratum: glucose 80g, Sodium Glutamate 50g, (NH 4) 2sO 48g, NaCl 5g, K 2hPO 43H 2o 2g, MnSO 47H 2o 0.25g, MnSO 4h 2o 0.03 g, distilled water 1000ml, pH value nature, 115 ℃ of sterilizing 30min.
agricultural soil microorganism water conservation production of articles
By brevibacterium flavum CCTCC NO:M 2013087 fermented liquids, bacillus amyloliquefaciens CCTCC NO:M 2013086 fermented liquids, (NH 4) 2sO 4, rice straw powder and the pig manure that becomes thoroughly decomposed be according to 5 ~ 8:6 ~ 10:7 ~ 13:25 ~ 35:40 ~ 55(w:w:w:w:w:w) stirring and evenly mixing thoroughly, the fermentation of banking up of bar buttress formula, leavening temperature is 30 ~ 50 ℃, turning in every 3 days 1 time in fermenting process, ferment and finish after 15 ~ 20 days, make brevibacterium flavum CCTCC NO:M 2013087 content reach 0.5 × 10 8cfu g -1above, bacillus amyloliquefaciens CCTCC NO:M 2013086 content reach 0.5 × 10 8cfu g -1above, bacterial strain stable content, γ-polyglutamic acid content is higher than 19.6g kg -1, the goods of acquisition are agricultural soil microorganism water conservation goods.
agricultural soil water conservation goods water conservation compliance test result
Being aleuritic texture soil for examination soil, is rape for studying thing.
Test set handling is as follows: process 1. contrasts; Process 2. application of organic fertilizers; Process 3. and use microorganism water conservation goods.Process 1 soil for contrast, process 2 soil and add fertilizer according to mass ratio 10%, process 3 soil and add microorganism water conservation goods according to mass ratio 10%, fertilizer, water conservation goods all and soil stir.The amount that three processing are watered occurs water saturated time stopping watering with reference to control treatment soil, finds that processing plant for three all shows when lack of water is wilted and start to water.Measure plant height, fresh weight and the dry weight of capsicum interior plant breeding time, the soil weight, hydraulic conductivity, saturated aqueous rate after capsicum results.
Experimental result shows, rapeseed cultivation is after 60 days, and average plant height, fresh weight and the dry weight of control treatment plant is 17.6cm, 98.9g, 6.7g; Average plant height, fresh weight and dry weight that application of organic fertilizers is processed plant are 18.2cm, 100.5g, 7.2g; Average plant height, fresh weight and dry weight that microorganism water conservation goods are processed plant are 22.8cm, 125.7g, 9.8g; Compared to control treatment 1, average plant height, fresh weight and the dry weight of using microorganism water conservation goods processing plant increase respectively 27.8%, 27.1% and 36.1%; Compared to processing 2, average plant height, fresh weight and the dry weight of using microorganism water conservation goods processing plant increase respectively 25.3%, 25.1% and 46.2%.Contrast, fertilizer and microorganism water conservation goods are processed the soil weight and are respectively 1.32g/cm 3, 1.27g/cm 3, 1.22g/cm 3, soil hydraulic conductivity is respectively 5.2mm/h, 5.8mm/h, 6.7mm/h, and saturation moisture content is respectively 36%, 39%, 46%, compared to control treatment, microorganism water conservation goods are processed the soil weight and have been declined 7.5%, and soil hydraulic conductivity has increased by 28.8%, and saturation moisture content increases by 27.8%.
  

Claims (7)

1. agricultural land soil microorganism water conservation goods, is characterized in that described a kind of agricultural land soil microorganism water conservation goods are pig manure, rice straw powder and the (NH of the synthetic corynebacterium glutamicum strain of a strain and the synthetic strain-combined fermentation maturity of γ-polyglutamic acid of a strain 4) 2sO 4the water conservation goods that obtain;
Wherein said synthetic corynebacterium glutamicum strain is brevibacterium flavum IAE( brevilbacterium flavumiAE), on March 15th, 2013 is preserved in Chinese Typical Representative culture collection center, and preservation address is Luo Jia Shan, wuchang, wuhan, and culture presevation number is CCTCC NO:M 2013087;
Described synthetic γ-many corynebacterium glutamicum strains are bacillus amyloliquefaciens IAE( bacllus amyloliquefaciensiAE), on March 15th, 2013 is preserved in Chinese Typical Representative culture collection center, and preservation address is Luo Jia Shan, wuchang, wuhan, and culture presevation number is CCTCC NO:M 2013086.
2. a kind of agricultural land soil microorganism water conservation goods according to claim 1, is characterized in that in these goods that γ-polyglutamic acid content is higher than 19.6g kg -1dry products, contains 0.5 × 10 8cfu g -1above brevibacterium flavum CCTCC NO:M 2013087 colony-forming units and 0.5 × 10 8cfu g -1above bacillus amyloliquefaciens CCTCC NO:M 2013086 colony-forming units, quality of organic matter is 30～35% than content, water ratio is lower than 30%.
3. according to a kind of agricultural land soil microorganism water conservation goods described in claims 1 or 2, it is characterized in that the culture condition of brevibacterium flavum CCTCC NO:M 2013087: be inoculated in the 250ml triangular flask that 100ml nutrient solution is housed with transfering loop picking growth bacterium of 24 hours from culture dish, 37 ℃, 170 rmp cultivate 48 hours, the centrifugal 20min of bacterium liquid supernatant 10000rpm, measuring supernatant liquor content of glutamic acid is 26.6g L -1.
4. described a kind of agricultural land soil microorganism water conservation goods according to claim 3, it is characterized in that brevibacterium flavum CCTCC NO:M 2013087 nutrient solution compound method used is, to prepare 1L substratum as example: glucose 50g, corn steep liquor 10ml, urea 5g, KH 2pO 43g, MgSO 40.8g, distilled water 1000ml, pH value nature, 115 ℃ of sterilizing 30min.
5. according to a kind of agricultural land soil microorganism water conservation goods described in claim 1 or 2, it is characterized in that the culture condition of bacillus amyloliquefaciens CCTCC NO:M 2013086: be inoculated in the 250ml triangular flask that the synthetic γ-polyglutamic acid nutrient solution of 100ml is housed with transfering loop picking growth bacterium of 24 hours from culture dish, 37 ℃, 170rmp are cultivated 48 hours, the centrifugal 20min of bacterium liquid supernatant 10000rpm, measuring supernatant liquor γ-polyglutamic acid content is 21.8g L -1.
6. a kind of agricultural land soil microorganism water conservation goods according to claim 5, it is characterized in that bacillus amyloliquefaciens CCTCC NO:M 2013086 liquid nutrient medium compound method used is, take preparation 1L substratum as example: glucose 80g, Sodium Glutamate 50g, (NH 4) 2sO 48g, NaCl 5g, K 2hPO 43H 2o 2g, MnSO 47H 2o 0.25g, MnSO 4h 2o 0.03 g, distilled water 1000 ml, pH value nature, 115 ℃ of sterilizing 30min.
7. according to the preparation method of soil microorganisms water conservation goods described in claim 1 or 2, it is characterized in that realizing as follows:
1) brevibacterium flavum CCTCC NO:M 2013087 bacterium are inoculated into liquid seed culture medium and cultivate, culture condition is: 37 ℃ of culture temperature, and shaking speed 150rpm, incubation time 24 hours, seed bacteria containing amount is greater than 1 × 10 9cfu ml -1, seed is inoculated into fermentor tank according to 5% ratio and carries out liquid fermenting production, and the condition of its fermentative production is: 28 ~ 33 ℃ of culture temperature, dissolved oxygen air flow scope is 30～100%, 150 ~ 220rpm, colony-forming unit>=1 × 10 of this bacterial strain of fermentation later stage fermentation liquid 9cfu ml -1;
The formula of the seed culture medium of liquid used is to prepare according to 1L substratum: extractum carnis 3g, peptone 10g, NaCl 10g, tap water 1000ml, pH scope 7.0-7.5,121 ℃ of sterilizing 20min; Liquid fermentation medium formula used is to prepare according to 1L substratum: glucose 50g, corn steep liquor 10ml, urea 5g, KH 2pO 43g, MgSO 40.8g, distilled water 1000ml, pH value nature, 115 ℃ of sterilizing 30min;
2) bacillus amyloliquefaciens CCTCC NO:M 2013086 is inoculated into liquid seed culture medium and cultivates, culture condition is: 37 ℃ of culture temperature, and shaking speed 130 ~ 160rpm, incubation time 24 hours, seed bacteria containing amount is greater than 1 × 10 9cfu ml -1, seed is inoculated into fermentor tank according to 8% ratio and carries out liquid fermenting production, and the condition of its fermentative production is: 28 ~ 35 ℃ of culture temperature, dissolved oxygen air flow scope is 30～100%, 120 ~ 160rpm, fermentation later stage part bacterial strain forms gemma, colony-forming unit>=1 × 10 of this bacterial strain of fermented liquid 9cfu ml -1, the retrogradation of fermented liquid concentration; The formula of the seed culture medium of liquid used is to prepare according to 1L substratum: extractum carnis 3g, peptone 10g, NaCl 10g, tap water 1000ml, pH scope 7.0-7.5,121 ℃ of sterilizing 20min;
Liquid fermentation medium formula used is to prepare according to 1L substratum: glucose 80g, Sodium Glutamate 50g, (NH 4) 2sO 48g, NaCl 5g, K 2hPO 43H 2o 2g, MnSO 47H 2o 0.25g, MnSO 4h 2o 0.03 g, distilled water 1000ml, pH value nature, 115 ℃ of sterilizing 30min;
3) by brevibacterium flavum CCTCC NO:M 2013087 fermented liquids, bacillus amyloliquefaciens CCTCC NO:M 2013086 fermented liquids, (NH 4) 2sO 4, rice straw powder and the pig manure that becomes thoroughly decomposed be according to 5 ~ 8:6 ~ 10:7 ~ 13:25 ~ 35:40 ~ 55(w:w:w:w:w:w) stirring and evenly mixing thoroughly, the fermentation of banking up of bar buttress formula, leavening temperature is 30 ~ 50 ℃, turning in every 3 days 1 time in fermenting process, ferment and finish after 15 ~ 20 days, make brevibacterium flavum content reach 0.5 × 10 8cfu g -1above, bacillus amyloliquefaciens content reaches 0.5 × 10 8cfu g -1above, bacterial strain stable content, γ-polyglutamic acid content is higher than 19.6g kg -1, the goods of acquisition are agricultural soil microorganism water conservation goods.
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Cited By (9)

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CN104744182A (en) * 2015-01-21 2015-07-01 河海大学 Tobacco water retention biological organic fertilizer and preparation method thereof
CN104762087A (en) * 2015-01-21 2015-07-08 河海大学 Microbe additive for improving beach saline alkali soil structure and its preparation method and use
CN104744182B (en) * 2015-01-21 2017-07-18 河海大学 One grows tobacco water conservation biological organic fertilizer and preparation method thereof
CN104629768A (en) * 2015-01-22 2015-05-20 河海大学 Microorganism product for reducing soil alkalinity of low beaches and saline-alkali land
CN104671432A (en) * 2015-01-27 2015-06-03 河海大学 Aquaculture water body cleaning agent and preparation method thereof
CN107058151A (en) * 2016-11-29 2017-08-18 广东森度生态农业科技有限公司 A kind of microbial bacteria with fermentation water retaining function and its application
CN107058151B (en) * 2016-11-29 2021-08-20 广东森度生态农业科技有限公司 Microbial bacterium with fermentation water retention function and application thereof
CN107236685A (en) * 2017-05-22 2017-10-10 中国科学院成都生物研究所 A kind of method that utilization sludge produces γ polyglutamic acid organic fertilizers
CN107236685B (en) * 2017-05-22 2020-06-02 中国科学院成都生物研究所 Method for producing gamma-polyglutamic acid organic fertilizer by using sludge
CN107318431A (en) * 2017-07-26 2017-11-07 安徽咱家田生态农业有限公司 A kind of method that utilization root division method cultivates Snakegourd Fruit seedling
CN110606782A (en) * 2019-09-30 2019-12-24 广东森度生态农业科技有限公司 Farmland soil water-retaining organic fertilizer and preparation method thereof
CN111296230A (en) * 2020-03-13 2020-06-19 河海大学 Ecological base material for mangrove planting engineering and preparation method and application thereof

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