CN104130050A - Multi-element compound microorganism fertilizer and production method thereof - Google Patents
Multi-element compound microorganism fertilizer and production method thereof Download PDFInfo
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- CN104130050A CN104130050A CN201410324985.3A CN201410324985A CN104130050A CN 104130050 A CN104130050 A CN 104130050A CN 201410324985 A CN201410324985 A CN 201410324985A CN 104130050 A CN104130050 A CN 104130050A
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Abstract
The invention discloses a multi-element compound microorganism fertilizer. The multi-element compound microorganism fertilizer is a potassium-rich multi-element compound microorganism fertilizer which is made by using potassium-oxide-rich potassic shale and humic acid as main raw materials, using bacillus mucilaginosus SCUP-76-8 as a potassium-solubilizing strain, using bacillus megaterium as a phosphate-solubilizing strain, using Frankia fgc6 as a nitrogen-fixing strain and performing fermental cultivation on the potassic shale and the humic acid. A production method of the multi-element compound microorganism fertilizer comprises the following steps: preparing potassic shale powder, humic acid powder, castor cake powder, a potassium-solubilizing bacteria fermentation liquor, a phosphate-solubilizing bacteria fermentation liquor and a nitrogen-fixing bacteria fermentation liquor respectively, uniformly mixing the potassic shale powder, the potassium-solubilizing fermentation liquor and the phosphate-solubilizing bacteria fermentation liquor in the mass volume ratio of 1:1:0.8, aerobically fermenting the mixture under a certain condition for 7-9 days, and naturally drying the fermented mixture; uniformly mixing the humic acid powder, the castor cake powder and the nitrogen-fixing bacteria fermentation liquor in the mass volume ratio of (5-8):1:1, aerobically fermenting the mixture under a certain condition for 10-12 days, and naturally drying the fermented mixture; and uniformly drying the activated potassic shale powder and the nitrogen-fixing bacteria humic acid as well as the castor cake powder in the mass ratio of (58-63):(37-42), and drying the mixture to obtain a finished product.
Description
Technical field
The present invention relates to a kind of composite fertilizer, particularly a kind of take potassium felspar sand as main raw material, be rich in to the multiple element composite microbe fertilizer of the efficient potassium decomposing of potassium felspar sand, molten phosphorus and nitrogen-fixing microorganism with and production method.
Background technology
Growth and development of plants needs the different element of multiple quantity, especially needs a large amount of nitrogen, phosphorus, potassium element.For meeting people's demand growing to each kind of plant, traditional fertilizer obviously cannot meet the demand of plant, and therefore, a large amount of nitrogen, phosphorus, the potassium chemical fertilizer of suitability for industrialized production is considered to realize this purpose main path always.Yet, continuous increase along with fertilizer application amount, its negative impact is day by day obvious, and outstanding behaviours is that the hardening soil, Soil degradation, soil and underground water severe contamination, Soil structure and micro-flora and diversity are suffered to destroy, crops quality declines, jeopardize food safety etc. when serious.So many restrictions have been made to the use of chemical fertilizer by the World Food Programme, some developed countries and area even start to ban use of chemical fertilizer.
Along with scientific and technological development and country more and more add attention to the control of environment, China has also done a large amount of work in the research and development of novel plant fertilizer.The various new-type fertilizers of having reported and having used are a lot, have and take humic acids as main fertilizer, have the composite microbe fertilizer of one or more the composition of the microorganisms, have biological and mineral compound fertilizer etc.The invention belongs to one of biological and mineral compound fertilizer.CN100343202C discloses a kind of biological multi element phosphorus potassium ferbilizer, after will potassium shale pulverizing, add the secondary fermentation of P and K decomposing bacterium to make, and the defect of this fertilizer is that to contain element less, and effectively potassium content is also not enough.Applicant discloses a kind of biological mineral compound fertilizer and production method (CN101475417A) thereof, by the potassium shale powder of 70%-80% and the humic acids of 20%-30%, after potassium decomposing, phosphorus decomposing and enzymatic microorganism fermentation, made, owing to there being the problems such as the ability of potassium solubilizing bacteria is weak and the accounting of potassium shale in composite fertilizer is larger, cause effective potassium of discharging from potassium shale lower, the over-all properties of improving soil of composite fertilizer is less, and range of application is subject to certain restrictions.For further improving the utilization ratio of potassium shale, improve fertilizer efficiency, improve the over-all properties of composite fertilizer, broadened application scope, applicant is from mining area screening wild-type indigenous strain, and carried out selection by mutation and fermentation technique research, found the bacterial strain that ability of dissolving potassium is powerful, and this application and development has been gone out to multielement composite microbe fertilizer.
Summary of the invention
Technical problem to be solved by this invention is to provide a kind of multielement composite microbe fertilizer and production method thereof, and this multielement composite microbe fertilizer has improved the utilization ratio of potassium shale, has improved the over-all properties of composite fertilizer, has improved fertilizer efficiency, has expanded range of application.
For solving the problems of the technologies described above, the potassium shale that is rich in potassium oxide and humic acids that the applicant location abounds with are raw material, the colloid bacillus cereus that the powerful deposit number of the ability of dissolving potassium of take is CGMCCNo.8481 (
bacillus mucilaginosus) the HSCUP-76-8 bacillus megaterium (Bacillus megaterium) that is CGMCC No.4296 as potassium decomposing bacterial classification, deposit number that the phosphorus decomposing ability of usining is strong frankia (Frankia.sp) the fgc6 bacterial strain that is CCTCC M207197 as phosphorus decomposing bacterial classification, the deposit number of usining is as fixed nitrogen bacterial classification, respectively potassium shale and humic acids are carried out after fermentation culture, make to be rich in and take potassium as main multielement composite microbe fertilizer.
Multielement composite microbe fertilizer of the present invention is to be prepared from according to the following steps:
(1) prepare potassium shale powder and choose potassium oxide content at more than 8% potassium shale, be ground into 150-250 order, standby;
(2) prepare humic acid powder and get organic content at more than 55% humic acids, grind to form 100-150 order, standby;
(3) prepare Semen Ricini cake powder and get Semen Ricini cake and be ground into 80-100 order, standby;
(4) prepare potassium solubilizing bacteria fermented liquid go bail for hide be numbered CGMCCNo.8481 colloid bacillus cereus (
bacillus mucilaginosus) HSCUP-76-8 carries out high density fermentation preparation as potassium solubilizing bacteria, concrete technology step is:
A is inoculated in slant medium by colloid bacillus cereus HSCUP-76-8 and activates, and temperature is controlled at 29-33 ℃, cultivates 24-48 hour, obtain original inclined-plane seed, again original inclined-plane seed is inoculated in to slant medium, the same terms is cultivated, and obtains production inclined-plane seed;
B is inoculated in production inclined-plane seed in liquid nutrient medium, packs the liquid nutrient medium of 20%-45% in the container of 200-500mL into, 29-33 ℃ of shaking table culture temperature, rotating speed 160-220r/min,, incubation time 8-12 hour, obtain seed liquor;
C is inoculated in seed tank culture base by seed liquor, coefficient 0.7-0.8,121 ℃ of steam sterilizings 20 minutes, inoculum size 5%-10%, culture temperature 29-33 ℃, air flow is 1.0:0.8-1.0(v/vmin), with mixing speed, control Nutrient solution DO value 10%-30%, pH is controlled at 7.2 ± 0.2, cultivates 8～12 hours, obtains seeding tank seed liquor;
D is inoculated in seeding tank seed liquor in fermentation basic medium, coefficient 0.7-0.8,121 ℃ of steam sterilizings 20 minutes, inoculum size 5%-10%, control parameter: temperature 29-33 ℃, pH value 7.0-7.2, air flow 1:0.8-1.2(v/vmin), with mixing speed, control Nutrient solution DO value 20%-30%, take vegetables oil as defoamer control foam; Get sample one time at interval of 4 hours, make immediately microscopy and observe bacterium number and thalli morphology, chemical examination sugar, ammonia-state nitrogen, according to the in good time feed supplement of assay, to control sugared dosage be 2g/L-3g/L, with NH
4cl controls nitrogenous source 0.5g/L-0.1g/L, cultivates 12-14 hour, stops mending nitrogen, then adjust and control parameter: temperature 33-37 ℃, pH value 7.2-8.9, air flow is 1:1.0-0.8(v/vmin), with mixing speed, control Nutrient solution DO value 5%～10%, promote sporulation, get sample one time at interval of 4 hours, make immediately microscopy and observe thalli morphology, when the thalli morphology in the microscopy visual field more than 98% is gemma, stop fermentation culture, gemma number reaches 1.8 * 10
9cfu/mL-2.0 * 10
9cfu/mL, gemma transformation efficiency is more than 75%, standby;
The moiety of described slant medium is: sucrose 5-6g, NaH
2pO
41-1.5g, MgSO
47H
2o 0.3-0.5g, FeCl
30.004-0.005g, CaCO
30-0.1g, agar 20-30g, distilled water 1000mL, pH 7.0-7.4;
The moiety of described liquid nutrient medium is: sucrose 5-10g, NH
4cl 0.5-1g, NaH
2pO
41-1.5g, MgSO
47H
2o 0.5-1g, FeCl
30.005g, CaCO
30.1-0.3g, distilled water 1000mL, pH 7.0-7.4;
The moiety of described seed tank culture base is: sucrose 2-5g, starch 3-10g, (NH
4)
2sO
41-2g/ NH
4cl 0.5-1g, yeast soak powder 1-2g, NaH
2pO
41-1.5g, MgSO
47H
2o 0.5-1g, FeCl
30.005g, distilled water 1000mL, pH 7.0-7.4;
The moiety of described fermentation basic medium is: sucrose 5g, (NH
4)
2sO
41-2g/ NH
4cl 0.5-1g, yeast soak powder 1-2g, NaH
2pO
41-2g, MgSO
47H
2o 0.5-1g, FeCl
30.005g, distilled water 1000mL, pH 7.0-7.4; Wherein said sucrose can be used any one or above equivalent replacement of Semen Maydis powder, starch;
(5) preparing phosphate solubilizing bacteria fermented liquid goes bail for and hides the bacillus megaterium (Bacillus megaterium) be numbered CGMCC No.4296 and carry out high density fermentation preparation as phosphate solubilizing bacteria, the moiety of phosphate solubilizing bacteria substratum is: glucose 8-10g, (NH
4)
2sO
40.5-0.8g, NaCl 0.3-0.5g, MgSO
47H
2o 0.3-0.5g, KCl 0.3-0.5g, FeSO
47H
2o 0.03-0.05g, MnSO
44H
2o 0.03-0.05g, Ca
3(PO
4)
25-6g, distilled water 1000mL, pH7.0-7.2; Fermentation time 32-48 hour reaches 4.5 * 10
9more than cfu/mL, stop fermentation culture, standby;
(6) prepare vinelandii fermented liquid go bail for hide be numbered CCTCC M207197 frankia (Frankia.sp) fgc6 bacterial strain as vinelandii, the moiety of the substratum of fermentation preparation high-density bacterium liquid is by total mass mark 100%, glucose 0.03%-0.08%, tween 80 0.003%-0.008%, Sodium.alpha.-ketopropionate 0.03%-0.08%, (NH4) 2SO40.05%-0.1%, KH2PO4 0.095%, K2HPO4 0.0435%, MgSO47H2O 0.0096%, CaCl2 0.001%, FeSO4 0.00056%, NaEDTA 0.00075%, trace element water-soluble liquid 0.1%, the VITAMIN aqueous solution 0.1%, surplus is water, trace element water-soluble liquid wherein forms by total mass mark 100%, is H3BO3 0.286%, and MnCl2 0.181%, and ZnSO4 0.022%, and CuSO4 0.008%, and Na2MoO4 0.0025%, and CoCl2 0.0025% and surplus are water, the VITAMIN aqueous solution forms by total mass mark 100%, is vitamin V B1 0.001%, nicotinic acid 0.005%, and vitamin H 0.0015%, folic acid 0.001%, calcium pantothenate 0.001%, riboflavin 0.001% and surplus are water, first cultivate after 10-15 days at 28-30 ℃, then at 28-30 ℃, cultivate and within 72-96 hour, obtain dry cell weight and be fermented liquids more than 300 μ g/mL, standby,
(7) by the mass volume ratio of potassium page rock powder ︰ potassium decomposing fermented liquid ︰ phosphate solubilizing bacteria fermented liquid, be 1 ︰ 1 ︰ 0.8, phosphate solubilizing bacteria liquid mixing and stirring prepared by potassium solubilizing bacteria liquid prepared by potassium shale powder prepared by (1), (4), (5), under 34-36 ℃, humidity 65%-75% condition, aerobic fermentation 7-9 days, then natural air drying, standby;
(8) by the mass volume ratio of humic acids Fen ︰ Semen Ricini cake Fen ︰ vinelandii fermented liquid, be 5-8 ︰ 1 ︰ 1, vinelandii mixing and stirring prepared by Semen Ricini cake powder prepared by humic acid powder prepared by (2), (3), (7), under 28-30 ℃, humidity 65%-75% condition, aerobic fermentation 10-12 days, then natural air drying, standby;
(9) vinelandii humic acids prepared by the activation potassium shale powder of being prepared by (7) and (8) and Semen Ricini cake powder in mass ratio 58-63 ︰ 37-42 mix, and be dried to below moisture be less than or equal to 8% at 35 ℃, are finished product.
After testing, the multielement composite microbe fertilizer of producing according to method of the present invention contains following composition: organic 10%-12%, potassium oxide 5%-7%, sulphur 0.7%-0.8%, calcium 5%-8%, magnesium 0.4%-0.5%, boron 0.1%-0.3%, iron 1.5%-3%, copper 0.2%-0.3%, zinc 0.1%-0.3%, manganese 0.2%-0.4%, molybdenum 0.3%-0.4 $, potassium solubilizing bacteria 1.4-1.7%, phosphate solubilizing bacteria 1.2%-1.4%, vinelandii 1.5%-1.9%, water 7%-8%, surplus are silicon-dioxide.
Multi-elements composite fertilizer of the present invention is selected the potassium shale of rich potassium and is rich in organic humic acids and Semen Ricini cake powder as raw material, under the effect of potassium solubilizing bacteria fermented liquid, phosphate solubilizing bacteria fermented liquid and vinelandii fermented liquid, utilize biotechnology that the potassium of potassium shale and soil nonabsorable, phosphorus etc. are discharged to unit and usually for crop, directly absorb respectively.The a large amount of various microorganisms that contain in multi-elements composite fertilizer of the present invention promote exchange of substance and the benign cycle of field biological circle soil, plant, animal and microorganism, the effect of reach improvement soil, optimizing the environment.
Multi-elements composite fertilizer of the present invention has the feature of many nutrition, plyability, contained nutrient big or middle, micro-and that microorganism fixed nitrogen, potassium decomposing, phosphate solubilizing bacteria discharge can meet crop at the nutritional needs of each growth period, reduce the generation of disease and pest, improved the quality and yield of agricultural-food.It is disposable employed that multi-elements composite fertilizer of the present invention is made base manure, reduced peasant's operating cost.Multi-elements composite fertilizer environmental protection of the present invention, energy-saving low-carbon, utilizes biogenic emission technique in process of production, there is no " three wastes " discharge, fermenting process carries out under normal temperature and thermophilic condition, there is no the objectionable constituent such as acid group in chemical fertilizer, base, can effectively improve soil and environment.Use multi-elements composite fertilizer of the present invention can reach the conditional request of Organic food.Through big area agricultural experiment statistic data, show, use multi-elements composite fertilizer field crop volume increase of the present invention more than 15%, gourd, fruit and vegetable increases production more than 30%.
Embodiment
Multielement composite microbe fertilizer of the present invention take be rich in potassium oxide potassium shale, be rich in organic humic acids and Semen Ricini cake is raw material, the colloid bacillus cereus that the deposit number of take is CGMCCNo.8481 (
bacillus mucilaginosus) the HSCUP-76-8 bacillus megaterium (Bacillus megaterium) that is CGMCC No.4296 as potassium decomposing bacterial classification, the deposit number of usining frankia (Frankia.sp) the fgc6 bacterial strain that is CCTCC M207197 as phosphorus decomposing bacterial classification, the deposit number of usining is as fixed nitrogen bacterial classification, respectively potassium shale and humic acids are carried out after fermentation culture, make to be rich in and take potassium as main multielement composite microbe fertilizer.
Multielement composite microbe fertilizer of the present invention is to be prepared from according to the following steps:
(1) prepare potassium shale powder and choose potassium oxide content at more than 8% potassium shale, be ground into 150-250 order, standby;
(2) prepare humic acid powder and get organic content at more than 55% humic acids, grind to form 100-150 order, standby;
(3) prepare Semen Ricini cake powder and get Semen Ricini cake and be ground into 80-100 order, standby;
(4) prepare potassium solubilizing bacteria fermented liquid and go bail for and hide the colloid bacillus cereus HSCUP-76-8 be numbered CGMCCNo.8481 and carry out high density fermentation preparation as potassium solubilizing bacteria, concrete technology step is:
A is inoculated in slant medium by colloid bacillus cereus HSCUP-76-8, and (moiety of slant medium is: sucrose 5-6g, NaH
2pO
41-1.5g, MgSO
47H
2o 0.3-0.5g, FeCl
30.004-0.005g, CaCO
30-0.1g, agar 20-30g, distilled water 1000mL, pH 7.0-7.4) activate, temperature is controlled at 29-33 ℃, cultivates 24-48 hour, obtains original inclined-plane seed, again original inclined-plane seed is inoculated in to slant medium, the same terms is cultivated, and obtains production inclined-plane seed;
B is inoculated in liquid nutrient medium by production inclined-plane seed, and (moiety of liquid nutrient medium is: sucrose 5-10g, NH
4cl 0.5-1g, NaH
2pO
41-1.5g, MgSO
47H
2o 0.5-1g, FeCl
30.005g, CaCO
30.1-0.3g, distilled water 1000mL in pH 7.0-7.4, pack the liquid nutrient medium of 20%-45% in the container of 200-500mL, 29-33 ℃ of shaking table culture temperature, rotating speed 160-220r/min,, incubation time 8-12 hour, obtain seed liquor;
C is inoculated in seed tank culture base by seed liquor, and (moiety of seed tank culture base is: sucrose 2-5g, starch 3-10g, (NH
4)
2sO
41-2g/ NH
4cl 0.5-1g, yeast soak powder 1-2g, NaH
2pO
41-1.5g, MgSO
47H
2o 0.5-1g, FeCl
30.005g, distilled water 1000mL, pH 7.0-7.4, coefficient 0.7-0.8,121 ℃ of steam sterilizings 20 minutes, inoculum size 5%-10%, culture temperature 29-33 ℃, air flow is 1.0:0.8-1.0(v/vmin), with mixing speed, control Nutrient solution DO value 10%-30%, pH is controlled at 7.2 ± 0.2, cultivate 8～12 hours, obtain seeding tank seed liquor;
D is inoculated in fermentation basic medium by seeding tank seed liquor, and (moiety of fermentation basic medium is: sucrose 5g, (NH
4)
2sO
41-2g/ NH
4cl 0.5-1g, yeast soak powder 1-2g, NaH
2pO
41-2g, MgSO
47H
2o 0.5-1g, FeCl
30.005g, distilled water 1000mL, pH 7.0-7.4; Wherein said sucrose can be used any one or above equivalent replacement of Semen Maydis powder, starch, coefficient 0.7-0.8,121 ℃ of steam sterilizings 20 minutes, inoculum size 5%-10%, control parameter: temperature 29-33 ℃, pH value 7.0-7.2, air flow 1:0.8-1.2(v/vmin), with mixing speed, control Nutrient solution DO value 20%-30%, take vegetables oil as defoamer control foam; Get sample one time at interval of 4 hours, make immediately microscopy and observe bacterium number and thalli morphology, chemical examination sugar, ammonia-state nitrogen, according to the in good time feed supplement of assay, to control sugared dosage be 2g/L-3g/L, with NH
4cl controls nitrogenous source 0.5g/L-0.1g/L, cultivates 12-14 hour, stops mending nitrogen, then adjust and control parameter: temperature 33-37 ℃, pH value 7.2-8.9, air flow is 1:1.0-0.8(v/vmin), with mixing speed, control Nutrient solution DO value 5%～10%, promote sporulation, get sample one time at interval of 4 hours, make immediately microscopy and observe thalli morphology, when the thalli morphology in the microscopy visual field more than 98% is gemma, stop fermentation culture, gemma number reaches 1.8 * 10
9cfu/mL-2.0 * 10
9cfu/mL, gemma transformation efficiency is more than 75%, standby;
(5) prepare phosphate solubilizing bacteria fermented liquid and go bail for and hide the bacillus megaterium be numbered CGMCC No.4296 and carry out high density fermentation preparation as phosphate solubilizing bacteria, the moiety of phosphate solubilizing bacteria substratum is: glucose 8-10g, (NH
4)
2sO
40.5-0.8g, NaCl 0.3-0.5g, MgSO
47H2O 0.3-0.5g, KCl 0.3-0.5g, FeSO
47H2O 0.03-0.05g, MnSO
44H
2o 0.03-0.05g, Ca
3(PO
4)
25-6g, distilled water 1000mL, pH7.0-7.2; Fermentation time 32-48 hour reaches 4.5 * 10
9more than cfu/mL, stop fermentation culture, standby;
(6) prepare vinelandii fermented liquid go bail for hide be numbered CCTCC M207197 frankia fgc6 bacterial strain as vinelandii, the moiety of the substratum of fermentation preparation high-density bacterium liquid is by total mass mark 100%, glucose 0.03%-0.08%, tween 80 0.003%-0.008%, Sodium.alpha.-ketopropionate 0.03%-0.08%, (NH4) 2SO40.05%-0.1%, KH2PO4 0.095%, K2HPO4 0.0435%, MgSO47H2O 0.0096%, CaCl2 0.001%, FeSO4 0.00056%, NaEDTA 0.00075%, trace element water-soluble liquid 0.1%, the VITAMIN aqueous solution 0.1%, surplus is water, trace element water-soluble liquid wherein forms by total mass mark 100%, is H3BO3 0.286%, and MnCl2 0.181%, and ZnSO4 0.022%, and CuSO4 0.008%, and Na2MoO4 0.0025%, and CoCl2 0.0025% and surplus are water, the VITAMIN aqueous solution forms by total mass mark 100%, is vitamin V B1 0.001%, nicotinic acid 0.005%, and vitamin H 0.0015%, folic acid 0.001%, calcium pantothenate 0.001%, riboflavin 0.001% and surplus are water, first cultivate after 10-15 days at 28-30 ℃, then at 28-30 ℃, cultivate and within 72-96 hour, obtain dry cell weight and be fermented liquids more than 300 μ g/mL, standby,
(7) by the mass volume ratio of potassium page rock powder ︰ potassium decomposing fermented liquid ︰ phosphate solubilizing bacteria fermented liquid, be 1 ︰ 1 ︰ 0.8, phosphate solubilizing bacteria liquid mixing and stirring prepared by potassium solubilizing bacteria liquid prepared by potassium shale powder prepared by (1), (4), (5), under 34-36 ℃, humidity 65%-75% condition, aerobic fermentation 7-9 days, then natural air drying, standby;
(8) by the mass volume ratio of humic acids Fen ︰ Semen Ricini cake Fen ︰ vinelandii fermented liquid, be 5-8 ︰ 1 ︰ 1, vinelandii mixing and stirring prepared by Semen Ricini cake powder prepared by humic acid powder prepared by (2), (3), (7), under 28-30 ℃, humidity 65%-75% condition, aerobic fermentation 10-12 days, then natural air drying, standby;
(9) vinelandii humic acids prepared by the activation potassium shale powder of being prepared by (7) and (8) and Semen Ricini cake powder in mass ratio 58-63 ︰ 37-42 mix, and be dried to below moisture be less than or equal to 8% at 35 ℃, are finished product.
Claims (1)
1. a multielement composite microbe fertilizer, is characterized in that by being prepared from according to the following steps:
(1) choose potassium oxide content at more than 8% potassium shale, be ground into 150-250 order, get by mass percentage the potassium shale of 58%-63%, standby;
(2) get organic content at more than 55% humic acids, grind to form 100-150 order, get by mass percentage the humic acid powder of 33%-36%, standby;
(3) get Semen Ricini cake and be ground into 80-100 order, get by mass percentage the Semen Ricini cake powder of 4%-7%, standby;
(4) go bail for and hide the colloid bacillus cereus HSCUP-76-8 be numbered CGMCCNo.8481 and carry out high density fermentation preparation as potassium solubilizing bacteria, concrete technology step is: A. is inoculated in slant medium by colloid bacillus cereus HSCUP-76-8 and activates, temperature is controlled at 29-33 ℃, cultivate 24-48 hour, obtain original inclined-plane seed, again original inclined-plane seed is inoculated in to slant medium, the same terms is cultivated, and obtains production inclined-plane seed; The moiety of described slant medium is: sucrose 5-6g, NaH
2pO
41-1.5g, MgSO
47H
2o 0.3-0.5g, FeCl
30.004-0.005g, CaCO
30-0.1g, agar 20-30g, distilled water 1000mL, pH 7.0-7.4; B. production inclined-plane seed is inoculated in liquid nutrient medium, in the container of 200-500mL, packs the liquid nutrient medium of 20%-45% into, 29-33 ℃ of shaking table culture temperature, rotating speed 160-220r/min,, incubation time 8-12 hour, obtain seed liquor; Described liquid nutrient medium: sucrose 5-10g, NH
4cl 0.5-1g, NaH
2pO
41-1.5g, MgSO
47H
2o 0.5-1g, FeCl
30.005g, CaCO
30.1-0.3g, distilled water 1000mL, pH 7.0-7.4; C. seed liquor is inoculated in to seed tank culture base, coefficient 0.7-0.8,121 ℃ of steam sterilizings 20 minutes, inoculum size 5%-10%, culture temperature 29-33 ℃, air flow is 1.0:0.8-1.0(v/vmin), with mixing speed, control Nutrient solution DO value 10%-30%, pH is controlled at 7.2 ± 0.2, cultivates 8～12 hours, obtains seeding tank seed liquor; The moiety of described seed tank culture base is: sucrose 2-5g, starch 3-10g, (NH
4)
2sO
41-2g/ NH
4cl 0.5-1g, yeast soak powder 1-2g, NaH
2pO
41-1.5g, MgSO
47H
2o 0.5-1g, FeCl
30.005g, distilled water 1000mL, pH 7.0-7.4; D. seeding tank seed liquor is inoculated in fermentation basic medium, coefficient 0.7-0.8,121 ℃ of steam sterilizings 20 minutes, inoculum size 5%-10%, control parameter: temperature 29-33 ℃, pH value 7.0-7.2, air flow 1:0.8-1.2(v/vmin), with mixing speed, control Nutrient solution DO value 20%-30%, take vegetables oil as defoamer control foam; Get sample one time at interval of 4 hours, make immediately microscopy and observe bacterium number and thalli morphology, chemical examination sugar, ammonia-state nitrogen; According to the in good time feed supplement of assay, to control sugared dosage be 2g/L-3g/L, with NH
4cl controls nitrogenous source 0.5g/L-0.1g/L, cultivates 12-14 hour, stops mending nitrogen, then adjust and control parameter: temperature 33-37 ℃, pH value 7.2-8.9, air flow is 1:1.0-0.8(v/vmin), with mixing speed, control Nutrient solution DO value 5%～10%, promote sporulation, get sample one time at interval of 4 hours, make immediately microscopy and observe thalli morphology, when the thalli morphology in the microscopy visual field more than 98% is gemma, stop fermentation culture, gemma number reaches 1.8 * 10
9cfu/mL-2.0 * 10
9cfu/mL, gemma transformation efficiency is more than 75%, standby; The moiety of described fermentation basic medium is: sucrose 5g, (NH
4)
2sO
41-2g/ NH
4cl 0.5-1g, yeast soak powder 1-2g, NaH
2pO
41-2g, MgSO
47H
2o 0.5-1g, FeCl
30.005g, distilled water 1000mL, pH 7.0-7.4; Wherein said sucrose can be used any one or above equivalent replacement of Semen Maydis powder, starch;
(5) go bail for and hide the bacillus megaterium be numbered CGMCC No.4296 and carry out high density fermentation preparation as phosphate solubilizing bacteria, the moiety of phosphate solubilizing bacteria substratum is: glucose 8-10g, (NH
4)
2sO
40.5-0.8g, NaCl 0.3-0.5g, MgSO
47H
2o 0.3-0.5g, KCl 0.3-0.5g, FeSO
47H2O 0.03-0.05g, MnSO
44H
2o 0.03-0.05g, Ca
3(PO
4)
25-6g, distilled water 1000mL, pH7.0-7.2; Fermentation time 32-48 hour reaches 4.5 * 10
9more than cfu/mL, stop fermentation culture, standby;
(6) by potassium shale powder: potassium solubilizing bacteria liquid: the mass volume ratio of phosphate solubilizing bacteria liquid is 1:1:0.8, phosphate solubilizing bacteria liquid mixing and stirring prepared by potassium solubilizing bacteria liquid prepared by potassium shale powder prepared by (1), (4), (5), under 34-36 ℃, humidity 65%-75% condition, aerobic fermentation 7-9 days, then natural air drying, standby;
(7) go bail for hide be numbered CCTCC M207197 frankia fgc6 bacterial strain as vinelandii, the moiety of the substratum of fermentation preparation high-density bacterium liquid is by total mass mark 100%, glucose 0.03%-0.08%, tween 80 0.003%-0.008%, Sodium.alpha.-ketopropionate 0.03%-0.08%, (NH4) 2SO40.05%-0.1%, KH2PO4 0.095%, K2HPO4 0.0435%, MgSO47H2O 0.0096%, CaCl2 0.001%, FeSO4 0.00056%, NaEDTA 0.00075%, trace element water-soluble liquid 0.1%, the VITAMIN aqueous solution 0.1%, surplus is water, trace element water-soluble liquid wherein forms by total mass mark 100%, is H3BO3 0.286%, and MnCl2 0.181%, and ZnSO4 0.022%, and CuSO4 0.008%, and Na2MoO4 0.0025%, and CoCl2 0.0025% and surplus are water, the VITAMIN aqueous solution forms by total mass mark 100%, is vitamin V B1 0.001%, nicotinic acid 0.005%, and vitamin H 0.0015%, folic acid 0.001%, calcium pantothenate 0.001%, riboflavin 0.001% and surplus are water, first cultivate after 10-15 days at 28-30 ℃, then at 28-30 ℃, cultivate and within 72-96 hour, obtain dry cell weight and be fermented liquids more than 300 μ g/mL, standby,
(8) by humic acid powder, add Semen Ricini cake powder: the mass volume ratio of vinelandii is 1:1, vinelandii mixing and stirring prepared by Semen Ricini cake powder prepared by humic acid powder prepared by (2), (3), (7), under 28-30 ℃, humidity 65%-75% condition, aerobic fermentation 10-12 days, then natural air drying, standby;
(9) vinelandii humic acids and Semen Ricini cake powder prepared by the activation potassium shale powder of being prepared by (7) and (8) mix, and be dried to below moisture be less than or equal to 8% at 35 ℃, are finished product.
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