CN102108335A - Rhizosphere growth-promoting bacterial fertilizer based on ditch millet azotobacter and preparation method thereof - Google Patents

Rhizosphere growth-promoting bacterial fertilizer based on ditch millet azotobacter and preparation method thereof Download PDF

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CN102108335A
CN102108335A CN2009102560990A CN200910256099A CN102108335A CN 102108335 A CN102108335 A CN 102108335A CN 2009102560990 A CN2009102560990 A CN 2009102560990A CN 200910256099 A CN200910256099 A CN 200910256099A CN 102108335 A CN102108335 A CN 102108335A
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substratum
rhizosphere
bacterial manure
sorbent material
bacterial
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陈宏坤
范玲超
葛雨明
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Shandong Kingenta Ecological Engineering Co Ltd
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Shandong Kingenta Ecological Engineering Co Ltd
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Abstract

The invention discloses a rhizosphere growth-promoting bacterial fertilizer which is prepared by adding 130-170 ml of bacterial suspension into every 500-700g of adsorbent, and the viable count of the ditch millet azotobacter is 0.5*10<8>-2*10<8> cfu/g. The product disclosed in the invention does not contain components of inorganic fertilizers, and can improve the soil instead of polluting the soil when being used for a long time.

Description

Based on the short bacterial manure and preparation method thereof of giving birth to of the rhizosphere of ditch millet vinelandii
Technical field
The invention belongs to biological fertilizer field, relate to a kind of bacterial manure, also relate to the production method of this bacterial manure simultaneously, relate in particular to a kind of efficient rhizosphere bacterial manure that wheat class gramineous crop is had fixed nitrogen, molten phosphorus, growth-promoting functions and preparation method thereof.
Background technology
Plant rhizosphere is urged living bacterium (Plant Growth Promoting Rhizobacteria, be called for short PGPR) and is meant that free living can promote plant-growth at soil or the class that is colonizated in roots of plants table, Gen Nei or cauline leaf, prevents and treats disease, increase the beneficial microorganism of crop yield.Be often referred to have fixed nitrogen, molten phosphorus, the bacterium that produces plant hormone, secretion microbiotic and produce abilities such as promoting the plant-growth physiologically active substance or have one of them ability at least.It can increase the phytomass of plant, improves the plant nutrition quality, and the inducing plant system produces many effects such as disease resistance.PGPR is a kind of bacterial manure that grows up the eighties, it is wider that it uses area, be mainly used in Gramineae, greengrocery, it can produce growth hormone, suppress pathogenic bacteria, fixed nitrogen and decomposition phosphide, the effect of PGPR is not single, inoculate a large amount of advantage beneficial bacterias in crop rhizosphere, growth-promoting functions is comprehensive result, and it is not a certain effect of common little fertilizer but a kind of combined effect.The mechanism of action: secretion plant growth-promoting material, someone is from 50 isolates that can promote the insoluble phosphorus conversion, find that 43 produce plant growth hormones, 29 produce Plant hormones regulators,gibberellins, 45 produce the Kinetins material, in these PGPR, also secrete multivitamin, carefully support one's family B1, B6, H, B12 etc., what also have can secrete amino acid; Promotion is sprouted, the regulation and control soil-borne disease.Alleviate soil-borne disease, reduce the number of times that takes place.PGPR can produce siderophore, it can have superpower complexing power to Fe3+, the rhizosphere bacteria that can produce a large amount of siderophores will be preponderated when the harmful microorganism very little with not producing siderophore or output competed the nutrition of iron element, thereby suppress the harmful microbe growth and breeding, show the biological control effect.
At present, the production of research of domestic PGPR bacterium and relevant ecological bacterial manure report is few.Abroad, particularly extensive studies has been done by countries such as India, Pakistan, Brazil, the U.S., Australia, has the ecological bacterial fertilizer products of PGPR of oneself, constantly occurs with stylish research field and new technology.Because of root system of plant, the cane in the different habitats specific short living bacteria strain of surviving, therefore we can only use for reference external successful experience, from the specific weather of China, isolate the PGPR bacterial strain in the habitat, to produce the bio-bacterial manure that is fit to China's weather, habitat, to aspect theoretical, do deep especially research simultaneously.At present, domestic leguminous plants bio-feritlizer is more, but Gramineae and other non-leguminous plant bio-feritlizer vacancies.The gramineous crop cultivated area is bigger in the world wide, has important economic value and ecological functions.Their great majority are main nutritive element with nitrogen, phosphorus all.The experimental results shows that PGPR obtains separating at grass rhizospheres such as wheat, corn, paddy rice, sugarcanes, and the research range of PGPR and content are also constantly being expanded and goed deep into.PGPR particularly has fixed nitrogen, molten phosphorus concurrently, produces plant hormone, secretion microbiotic bacterial classification, is valuable Biological resources, and their development and use will be played very important effect in the research and development of the development of bio-bacterial manure and grass.So consider that from aspects such as the market requirement and the enhancings of people's environmental consciousness the research of ecological bacterial manure and exploitation thereof have important practical significance and vast potential for future development.
Summary of the invention
At the deficiencies in the prior art, the present invention seeks to, a kind of have fixed nitrogen, molten phosphorus, the short bacterial manure of giving birth to the plant hormone effect of secretion are provided, this fertilizer adaptability is strong, stable performance, has multiple functions such as fixed nitrogen, molten phosphorus, promotes growth.Simultaneously, the present invention also aims to provide the preparation method of a grow wheat gramineous crop with efficient rhizosphere bacterial manure.The present invention realizes by following technical measures:
The screening of bacterial strain ditch millet vinelandii:
1. soil sample collection
Sample picks up from Linyi Prefecture's wheat rhizosphere settling, pack into behind the sample collecting sterilization seal in the polyethylene bag cryopreservation.
2. substratum
(1) LB substratum: peptone 10g, yeast extract paste 5g, NaCl 10g, Agar 20g, cumulative volume 1000ml (distilled water is supplied), pH 7.0-7.5;
(2) NFM substratum: oxysuccinic acid 5g, K 2PHO 40.5g, MgSO 47H 2O 0.2g, CaCl 22H 2O 0.02g, NaCl 0.1g, NaMoO 42H 2O 0.002g, KOH 4.5g, Bromothymol Blue (BTB 0.5%) 5ml, Biotine10ug, Agar 2% (solid medium), cumulative volume 1000ml (distilled water is supplied), PH7.0-7.5;
(3) CCM substratum: KH 2PO 40.2g, NaCl 0.1g, K 2HPO 40.8g, Na 2FeEDTA28mg, Sodium orthomolybdate 25mg, yeast extract 100mg, N.F,USP MANNITOL 5g, sucrose 5g, Sodium.alpha.-hydroxypropionate 0.5ml, distilled water 900ml.
3. bacterial strain screening method
Sample thief 1g is made into 10 with 0.85% physiological saline under aseptic condition -4, 10 -5, 10 -6Three extent of dilution, each dilution suspension are inoculated in the serum test tube bottle that fills semi-solid NFM, CCM substratum, behind the 28-30 ℃ of cultivation 48h, inject 1mlC 2H 2, place 28-30 ℃ to cultivate 24h-48h again after, to C is arranged 2H 4The sample that produces is incubated at it respectively on LB, NFM, the CCM plate culture medium with plate streak again, places 28-30 ℃ incubator to cultivate 24-48h.The different typical single bacterium colony of picking respectively then, and be inoculated among the CCM, NFM serum bottle of new preparation, its nitrogenase activity measured.Pick out growth is fast, circle is big bacterium colony again after the separation and purification, change the 4 ℃ of preservations in inclined-plane.
4, bacterial strain antagonism reaction test
Respectively with the bacterial strain streak inoculation of culture purified in the same culture dish that fills the LB solid medium, place 28-30 ℃ of incubator, cultivate 4-8d, and the bacterial growth situation in routine observation bacterium point of crossing, judge that it has or not restraining effect.If it is poor that this point of crossing bacterium does not grow or grows, antagonistic action when being described, is arranged in these two kinds of bacterium coexistences; Otherwise,, illustrate that these two kinds of bacteriums can coexist, and do not have restraining effect therebetween if this point of crossing bacterial growth is good.Each bacterial strain suspension that the antagonism reaction does not take place is each other mixed, otherwise use separately.
The invention provides the short bacterial manure of giving birth to of a kind of rhizosphere, is to make ditch millet vinelandii viable count 0.5x10 by adding the 130-170ml bacteria suspension in every 500-700g sorbent material 8-2x10 8Cfu/g.
The short bacterial manure of giving birth to of aforesaid rhizosphere, preferred scheme is: described sorbent material is peat, activated carbon, vermiculite or mixture wherein.That described sorbent material is preferably is nontoxic to the PGPR bacterial strain, the water-absorbing-retaining performance is good, the pH value between 6.5-7.0, particle is tiny and easily (drying, be crushed to 100 orders with seed adhesion, prevented from caking, cheap peat.) for inoculating carrier.
Invention also provides rhizosphere the short preparation method who gives birth to bacterial manure, may further comprise the steps:
[1] preparation of bacteria suspension:
(1) ditch millet vinelandii inclined-plane: 28-30 ℃, cultivates 2-3d;
(2) shake-flask seed is cultivated: culturing bottle is contained substratum 300ml, 121 ℃ of sterilization 20-30min, and 1ml inclined-plane bacterial strain is inserted in the cooling back, places 28 ℃ of shaking culture, shaking speed 125rpm/min, incubation time 2-3d;
(3) enlarge fermentation: the 300L fermentor tank substratum 150L that packs into, 121 ℃ of sterilizations are placed on 2-3d under the room temperature, pollution-free on inspection after, insert the 15L seed culture fluid, place the track shaking table under 28 ℃, rotating speed is 125rpm/min, incubation time 3-4d, gained is bacteria suspension;
[2] processing of sorbent material:
With sorbent material drying, pulverizing, cross 100 mesh sieves, 121 ℃ of sterilization 2-3h, packing 500g/ bag is standby;
[3] bacterial manure inoculation and cultivation:
Measure the 150ml bacterial suspension inoculation in the sorbent material of 500g sterilization with sterilized graduated cylinder, under aseptic technique, pack into and seal immediately in the plastic packaging bag, and with bacteria suspension and sorbent material mixing, in the middle of reaching around the plastics bag, prick several holes with the sterilization pin again, place 28-30 ℃ to cultivate 6-7d, the ditch millet vinelandii are further bred, and the viable bacteria number average is 10 8Individual/g, packing back normal temperature is preserved down, is the short bacterial manure of giving birth to of rhizosphere.
Aforesaid preparation method, the substratum of described step (1) is: peptone 10g, yeast extract paste 5g, NaCl 10g, Agar 20g, distilled water 1000ml, PH 7.0-7.5.
Aforesaid preparation method, the substratum of described step (2) and step (3) is: syrup 10g, yeast extract paste 5g, CMC-Na5g, KH 2PO 40.5g, MgSO 40.25g, distilled water 1000ml, PH 7.0-7.5.
The present invention compared with prior art has following outstanding technique effect:
1, the present invention adopt the basic method of specific culture (LB, NFM, CCM) to isolate to have fixed nitrogen from wheat rhizosphere, secretion plant-growth class material and hold the azotobacter strain of phosphorus effect;
2, bacterial strain of the present invention obtains from China's wheat rhizosphere, and adaptability is strong, stable performance, can produce multiple wheat growth and promote material, wheat plant robust growth, output, output value height;
3, product of the present invention does not contain the composition of inorganic fertilizer, and life-time service can contaminated soil, and can improve the soil.
Embodiment
Describe technical scheme of the present invention in detail below in conjunction with embodiment, experimental example, but protection domain is not by this restriction.
Used peptone, yeast extract paste, NaCl, K among the embodiment 2PHO 4Be all homemade analytical pure etc. conventional medicine, all can buy preparation from market.
The screening of embodiment 1 bacterial strain ditch millet vinelandii.
1. soil sample collection
Sample picks up from Linyi Prefecture's wheat rhizosphere settling, pack into behind the sample collecting sterilization seal in the polyethylene bag cryopreservation.
2. substratum
(1) LB substratum: peptone 10g, yeast extract paste 5g, NaCl 10g, Agar 20g, cumulative volume 1000ml (distilled water is supplied), PH 7.0-7.5;
(2) NFM substratum: oxysuccinic acid 5g, K 2PHO 40.5g, MgSO 47H 2O 0.2g, CaCl 22H 2O 0.02g, NaCl 0.1g, NaMoO 42H 2O 0.002g, KOH 4.5g, Bromothymol Blue (BTB 0.5%) 5ml, Biotine10ug, Agar 2% (solid medium), cumulative volume 1000ml (distilled water is supplied), PH7.0-7.5;
(3) CCM substratum: KH 2PO 40.2g, NaCl 0.1g, K 2HPO 40.8g, Na 2FeEDTA28mg, Sodium orthomolybdate 25mg, yeast extract 100mg, N.F,USP MANNITOL 5g, sucrose 5g, Sodium.alpha.-hydroxypropionate 0.5ml, distilled water 900ml.
3. bacterial strain screening method
Sample thief 1g is made into 10 with 0.85% physiological saline under aseptic condition -4, 10 -5, 10 -6Three extent of dilution, each dilution suspension are inoculated in the serum test tube bottle that fills semi-solid NFM, CCM substratum, behind the 28-30 ℃ of cultivation 48h, inject 1mlC 2H 2, place 28-30 ℃ to cultivate 24h-48h again after, to C is arranged 2H 4The sample that produces is incubated at it respectively on LB, NFM, the CCM plate culture medium with plate streak again, places 28-30 ℃ incubator to cultivate 24-48h.The different typical single bacterium colony of picking respectively then, and be inoculated among the CCM, NFM serum bottle of new preparation, its nitrogenase activity measured.Pick out growth is fast, circle is big bacterium colony again after the separation and purification, change the 4 ℃ of preservations in inclined-plane.
4, bacterial strain antagonism reaction test
Respectively with the bacterial strain streak inoculation of culture purified in the same culture dish that fills the LB solid medium, place 28-30 ℃ of incubator, cultivate 4-8d, and the bacterial growth situation in routine observation bacterium point of crossing, judge that it has or not restraining effect.If it is poor that this point of crossing bacterium does not grow or grows, antagonistic action when being described, is arranged in these two kinds of bacterium coexistences; Otherwise,, illustrate that these two kinds of bacteriums can coexist, and do not have restraining effect therebetween if this point of crossing bacterial growth is good.Each bacterial strain suspension that the antagonism reaction does not take place is each other mixed, otherwise use separately.
The short making of giving birth to bacterial manure of embodiment 2 rhizospheres.
The short bacterial manure of giving birth to of present embodiment rhizosphere is to make ditch millet vinelandii viable count 2x108cfu/g by adding the 130ml bacteria suspension in the 500g sorbent material.Sorbent material is a peat, preferably nontoxic to the PGPR bacterial strain, the water-absorbing-retaining performance is good, the pH value between 6.5-7.0, particle is tiny and easily (drying, be crushed to 100 orders with seed adhesion, prevented from caking, cheap peat.)
The short preparation method who gives birth to bacterial manure of this rhizosphere may further comprise the steps:
[1] preparation of bacteria suspension:
(1) ditch millet vinelandii inclined-plane: 28-30 ℃, cultivates 2-3d;
(2) shake-flask seed is cultivated: culturing bottle is contained substratum 300ml, 121 ℃ of sterilization 20-30min, and 1ml inclined-plane bacterial strain is inserted in the cooling back, places 28 ℃ of shaking culture, shaking speed 125rpm/min, incubation time 2-3d;
(3) enlarge fermentation: the 300L fermentor tank substratum 150L that packs into, 121 ℃ of sterilizations are placed on 2-3d under the room temperature, pollution-free on inspection after, insert the 15L seed culture fluid, place the track shaking table under 28 ℃, rotating speed is 125rpm/min, incubation time 3-4d, gained is bacteria suspension;
[2] processing of sorbent material:
With sorbent material drying, pulverizing, cross 100 mesh sieves, 121 ℃ of sterilization 2-3h, packing 500g/ bag is standby;
[3] bacterial manure inoculation and cultivation:
Measure the 150ml bacterial suspension inoculation in the sorbent material of 500g sterilization with sterilized graduated cylinder, under aseptic technique, pack into and seal immediately in the plastic packaging bag, and with bacteria suspension and sorbent material mixing, in the middle of reaching around the plastics bag, prick several holes with the sterilization pin again, place 28-30 ℃ to cultivate 6-7d, the ditch millet vinelandii are further bred, and the viable bacteria number average is 10 8Individual/g, packing back normal temperature is preserved down, is the short bacterial manure of giving birth to of rhizosphere.
The substratum of described step (1) is: peptone 10g, yeast extract paste 5g, NaCl 10g, Agar 20g, distilled water 1000ml, PH 7.0-7.5.
The substratum of described step (2) and step (3) is: syrup 10g, yeast extract paste 5g, CMC-Na5g, KH 2PO 40.5g, MgSO 40.25g, distilled water 1000ml, PH 7.0-7.5.
The short making of giving birth to bacterial manure of embodiment 3 rhizospheres.The short bacterial manure of giving birth to of present embodiment rhizosphere is to make ditch millet vinelandii viable count 0.5x108cfu/g by adding the 170ml bacteria suspension in the 700g sorbent material.Sorbent material is an activated carbon.The short preparation method of bacterial manure that gives birth to of this rhizosphere is with embodiment 2.
Experimental example 1 experiment place: Linshu County, Shandong; Experimental technique: 1, experiment material: for trying wheat breed: agricultural university 664; For trying soil: topsoil (0-20cm) soil nutrient content before testing, organic matter 1.57%, full nitrogen 0.099%, phosphorus 0.072%, potassium 1.08% entirely entirely; Alkali-hydrolyzable nitrogen 96.37mg/kg, rapid available phosphorus 21.40mg/kg, available potassium 107.33mg/kg.2, experiment is handled: each handles 4 mu of experiment sub-district areas, adopts district's group arrangement at random, and each processing is provided with 3 repetitions.The condition of all experimental plots (as factors such as field management measure such as soil, irrigation, fertilizer, breeding time and distance between rows and hills and illumination) is consistent, and meets the agricultural planting practice of local science.
Test design is 4 processing, and it is handled the title code name and thes contents are as follows:
1. the short bacterial manure of giving birth to of rhizosphere is joined for 60 jin/mu and is executed 10 jin/mu of phosphorus two ammoniums;
2. the short bacterial manure of giving birth to of rhizosphere is joined for 40 jin/mu and is executed 20 jin/mu of phosphorus two ammoniums;
3. the short bacterial manure of giving birth to of rhizosphere is joined for 30 jin/mu and is executed 30 jin/mu of phosphorus two ammoniums;
4. phosphorus two ammoniums are 50 jin/mu.
Each handles the unanimity of topdressing, unified 40 jin/mu in the urea that imposes.
1, test result analysis
1.1 the short influence of giving birth to bacterial manure of embodiment 2 rhizospheres to the wheat proterties
Table 1 different treatment is to the influence of the main economic characters of wheat
Project is handled Mu spike number (ten thousand) Plant height (cm) Spike length (cm) Spikelet number (individual) Grain number per spike (grain) Thousand seed weight (gram)
① 41.14 81.6 9 14.2 32.8 39.0
② 41.17 82 8.7 14.5 31.9 38.8
③ 41.25 83.6 9 14.2 33.8 39.2
④ 40.58 83.1 8.5 13.6 31.3 38.4
Table 1 projects are measured and are shown, it is different to the main economic characters influence of wheat that wheat is used bacterial manure different treatment of the present invention, handles significant difference.Handle 3. promptly the short bacterial manure of giving birth to of mu rhizosphere and join for 30 jin and execute 30 jin of two ammoniums and reach conspicuous level, increase by 0.873 fringe to fertilize than single respectively than handling output three elements mu spike number, grain number per spike, the thousand seed weight difference of 4. promptly singly executing 50 jin of/mu wheats of two ammoniums, 2.5,0.8 gram; Handling 1. promptly mu executes the short bacterial manure of giving birth to of rhizosphere and joins for 60 jin and execute 10 jin of two ammoniums than the mu spike numbers of handling 4. promptly singly to execute 50 jin of/mu wheats of two ammoniums, grain number per spike, thousand seed weight difference reaches conspicuous level, and wherein the mu spike number increases by 0.56 ten thousand fringe, and grain number per spike increases by 1.5, thousand seed weight increase by 0.6 gram.
1.2 the short influence of giving birth to bacterial manure of embodiment 2 rhizospheres to wheat yield
The wheat yield The result of multiple comparisons is handled in table 2 test
Table 2 measurement result shows handles differences significantly or extremely remarkable.1. The result of multiple comparisons explanation is handled, 3. 4. fertilizer efficiency is higher than processing, promptly rhizosphere short give birth to bacterial manure join to fertilize fertilizer efficiency be higher than single to fertilize; 1. handle is that the short bacterial manure of giving birth to of rhizosphere joins that 2. execute 10 jin/mu of two ammoniums and handle be that the short living bacterial manure of rhizosphere joins that to execute 20 jin of/mu fertilizer efficiency of two ammoniums similar for 40 jin/mu for 60 jin/mu, and volume variance is not remarkable; 3. handle is that the short bacterial manure of giving birth to of rhizosphere joins that to execute 30 jin of/mu fertilizer efficiency of two ammoniums the highest for 30 jin/mu, 4. reaches utmost point conspicuous level than handling, and 2. reaches conspicuous level than handling, and than handling 42.5 jin of 4. promptly single mu to fertilize volume increase, stimulation ratio 4.7% is than handling 32.6 jin of 2. mu volume increase.
From above table, find out, use bacterial manure of the present invention and can promote the wheat normal growth to grow, strengthen resistance such as wheat is resistant to lodging.Bacterial manure of the present invention is joined to fertilize the mu spike number to wheat, grain number per spike, and thousand seed weight, output all have tangible increase, and output amplification is at 20-40 jin/mu.Bacterial manure consumption of the present invention with mu execute bacterial manure join for 30 jin execute 30 jin of two ammoniums, urea is optimum amount for 40 jin, bacterial manure cooperates ratio with chemical fertilizer be 30: 70.Wheat is used bacterial manure using method of the present invention to do base manure or seed manure, does seed manure with the disposable deep placement of phosphorus two ammoniums or with layering seed sower and two ammoniums while deep placement before broadcasting.

Claims (7)

1. the ditch millet vinelandii is characterized in that, obtain by following step:
A. soil sample collection: sample picks up from Linyi Prefecture's wheat rhizosphere settling, pack into behind the sample collecting sterilization seal in the polyethylene bag cryopreservation;
B. substratum:
(1) LB substratum: peptone 10g, yeast extract paste 5g, NaCl 10g, Agar 20g, cumulative volume 1000ml, pH 7.0-7.5;
(2) NFM substratum: oxysuccinic acid 5g, K 2PHO 40.5g, MgSO 47H 2O 0.2g, CaCl 22H 2O 0.02g, NaCl 0.1g, NaMoO 42H 2O 0.002g, KOH 4.5g, Bromothymol Blue 5ml, Biotine 10ug, Agar2%, cumulative volume 1000ml, pH7.0-7.5;
(3) CCM substratum: KH 2PO 40.2g, NaCl 0.1g, K 2HPO 40.8g, Na2FeEDTA28mg, Sodium orthomolybdate 25mg, yeast extract 100mg, N.F,USP MANNITOL 5g, sucrose 5g, Sodium.alpha.-hydroxypropionate 0.5ml, distilled water 900ml;
C. bacterial strain screening method: sample thief 1g is made into 10 with 0.85% physiological saline under aseptic condition -4, 10 -5, 10 -6Three extent of dilution, each dilution suspension are inoculated in the serum test tube bottle that fills semi-solid NFM, CCM substratum, behind the 28-30 ℃ of cultivation 48h, inject 1mlC 2H 2, place 28-30 ℃ to cultivate 24h-48h again after, to C is arranged 2H 4The sample that produces is incubated at it respectively on LB, NFM, the CCM plate culture medium with plate streak again, place 28-30 ℃ incubator to cultivate 24-48h, distinguish the different typical single bacterium colony of picking then, and be inoculated among the CCM, NFM serum bottle of new preparation, measure its nitrogenase activity, pick out growth is fast, circle is big bacterium colony again after the separation and purification, change the 4 ℃ of preservations in inclined-plane.
2. the short bacterial manure of giving birth to of rhizosphere is characterized in that: make ditch millet vinelandii viable count 0.5x10 by adding the 130-170ml bacteria suspension in every 500-700g sorbent material 8-2x10 8Cfu/g.
3. the described rhizosphere of claim 2 is urged to give birth to bacterial manure, and it is characterized in that: described sorbent material is peat, activated carbon, vermiculite or their mixture.
4. the described rhizosphere of claim 3 is urged to give birth to bacterial manure, and it is characterized in that: described sorbent material is a peat.
5. a rhizosphere urgees to give birth to the preparation method of bacterial manure, it is characterized in that: may further comprise the steps:
A. the preparation of bacteria suspension:
(1) ditch millet vinelandii inclined-plane: 28-30 ℃, cultivates 2-3d;
(2) shake-flask seed is cultivated: culturing bottle is contained substratum 300ml, 121 ℃ of sterilization 20-30min, and 1ml inclined-plane bacterial strain is inserted in the cooling back, places 28 ℃ of shaking culture, shaking speed 125rpm/min, incubation time 2-3d;
(3) enlarge fermentation: the 300L fermentor tank substratum 150L that packs into, 121 ℃ of sterilizations are placed on 2-3d under the room temperature, pollution-free on inspection after, insert the 15L seed culture fluid, place the track shaking table under 28 ℃, rotating speed is 125rpm/min, incubation time 3-4d, gained is bacteria suspension;
B. the processing of sorbent material: with sorbent material drying, pulverizing, cross 100 mesh sieves, 121 ℃ of sterilization 2-3h, packing 500g/ bag is standby;
C. bacterial manure inoculation and cultivation: measure the 150ml bacterial suspension inoculation in the sorbent material of 500g sterilization with sterilized graduated cylinder, under aseptic technique, pack into and seal immediately in the plastic packaging bag, and with bacteria suspension and sorbent material mixing, in the middle of reaching around the plastics bag, prick several holes with the sterilization pin again, place 28-30 ℃ to cultivate 6-7d, the ditch millet vinelandii are further bred, and the viable bacteria number average is 10 8Individual/g, packing back normal temperature is preserved down, is the short bacterial manure of giving birth to of rhizosphere.
6. the described preparation method of claim 5, it is characterized in that: the substratum of described step (1) is: peptone 10g, yeast extract paste 5g, NaCl 10g, Agar 20g, distilled water 1000ml, pH 7.0-7.5.
7. the described preparation method of claim 5, it is characterized in that: the substratum of described step (2) and step (3) is: syrup 10g, yeast extract paste 5g, CMC-Na5g, KH 2PO 40.5g, MgSO 40.25g, distilled water 1000ml, pH 7.0-7.5.
CN2009102560990A 2009-12-29 2009-12-29 Rhizosphere growth-promoting bacterial fertilizer based on ditch millet azotobacter and preparation method thereof Pending CN102108335A (en)

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Publication number Priority date Publication date Assignee Title
CN102925389A (en) * 2012-10-31 2013-02-13 黑龙江省农业科学院草业研究所 Free-living nitrogen fixing bacteria with efficient auxin secretion ability and nitrogen fixing ability and application thereof
CN102925389B (en) * 2012-10-31 2014-03-19 黑龙江省农业科学院草业研究所 Free-living nitrogen fixing bacteria with efficient auxin secretion ability and nitrogen fixing ability and application thereof
CN104944569A (en) * 2015-06-03 2015-09-30 中国水产科学研究院渔业机械仪器研究所 Method for promoting azotification of aquatic microorganisms
CN106318877A (en) * 2016-08-29 2017-01-11 刘翔 Method for separating and purifying eurotium cristatum from fu-brick tea and liquid culture method of eurotium cristatum
CN109354217A (en) * 2018-12-21 2019-02-19 江西省科学院 A kind of river water body and bed mud comprehensive treatment agent and preparation method
CN109734502A (en) * 2019-03-20 2019-05-10 何建清 A kind of preparation method and application of black highland barley Special compound microbial fertilizer
CN109879703A (en) * 2019-04-24 2019-06-14 刘海波 A kind of environmental protection composite biological fertilizer
CN110438036B (en) * 2019-07-10 2022-05-17 西北农林科技大学 Nitrogen-fixing bacterium N24 with nitrogen-fixing effect and application thereof
CN113073060A (en) * 2021-03-30 2021-07-06 青岛万慧源环保科技有限公司 Composition for soil remediation and remediation method

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Application publication date: 20110629