CN1789214A - Biological formulation for soybean and its production method - Google Patents
Biological formulation for soybean and its production method Download PDFInfo
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Abstract
The invention relates to the applied technology of agricultural microorganism, which in detail is a biological agent for soybean and its preparing method, comprising organic carrier, soybean nodule bacteria, autogen azotobacter and actinomyces, mixing the A and B sort fermentation accordiong to the proportion of 1:1; the weight/volume ratio of the organic carrier with microorganism seed solution in sort A is: organic carrier: soybean nodule bacteria: autogen azotobacter=100:15-25:5-15, the viable bacteria number in the soybean nodule bacteria seed solution and autogen azotobacter seed solution is more than 0.8 billion per ml; the weight/volume ratio of the organic carrier with microorganism seed solution in sort B is: organic carrier: actinomyces=100:25-35, the viable bacterial plate count in the actinomyces seed solution is more than 20 million per ml.
Description
Technical field
The present invention relates to agriculture microorganism application technology, a kind of specifically biological formulation for soybean and production method thereof.
Background technology
China's microbial fertilizer research can be traced back to the thirties in last century, and the famous soil microbiology expert Zhang Xianwu professor of China delivers first piece of rihizobium japonicum research and use article nineteen thirty-seven, existing so far nearly 80 years history.The early 1950s scientific research personnel of Inst. of Forestry and Soil, Chinese Academy of Sciences leads down professor Zhang Xianwu, and big area is promoted rihizobium japonicum Inoculant technology on 1,500,000 hectares of soils, area northeastward, on average increases production soybean more than 10%.Former Soviet Union microorganism scholar has also obtained breakthrough in the research work of vinelandii, phosphorus bacteria, silicate bacteria, and large scale application in the improvement of soil fertility soil fertility, has obtained good result.The China's agricultural development outline was classified bacterial fertilizer as an important agrotechnical measure in 1958, to the research and the application of microbial fertilizer, played active promoting function.
Current, the research Showed Very Brisk of the short living bacterium PGPR of plant rhizosphere.People such as Broalbent separation in 1977 obtains a bacillus subtilis (Bacillus.subtilis A13) and successfully uses.U.S. Weller in 1988 has reported that subtilis A13 can suppress plant pathogenic fungi, and promotes the various plants growth, can improve Radix Dauci Sativae output 48%, amount of oat yield 33%, peanut 37%, uses area and has reached 4,000,000 hectares.China field is used the PGPR biotechnological formulation and is started from 1979, uses area to yield increasing fungus in 1994 and has reached 3,930 ten thousand hectares.With the PGPR biotechnological formulation that marine actinomycete MB-97 makes, use and start from 1997, using on soybean has had 20,000 hectares, on average increases production 10.0～20.0%.With the PGPR biotechnological formulation that marine bacteria 9912 is made, use and start from 2000 (field test demonstration phase), in economic class crops such as soybean, vegetables, use, on average increase production 10.0～18.9%.
Biological formulation for soybean is a kind of efficient, nontoxic, pollution-free, the non-harmful PGPR type bio-feritlizer that grows up on the microbial fertilizer basis.In developed countries such as the U.S., Brazil, Russia; the product that this respect has been arranged; they consider from environment protection, resource regeneration, culture fertility with to the aspects such as specificity of crop; in conjunction with microorganism liquid-solid two-phase fermentation technique; the ight soil of domestic waste, large-scale plant and active sludge etc. are carried out harmless treatment, manufacture high benefit, free of contamination various tailored version microbial preparations.
Microbial fertilizer research, since nineteen thirty-seven rihizobium japonicum research and application, to vinelandii, phosphorus bacteria, silicate bacteria, actinomycetic application, urge research and the application of living bacterium PGPR to the plant rhizosphere of today, people are long usually from the angle of applied research, microorganism and plant are considered not only many bacterial strains, multi-functional but also develop and produce some obligatory type fertilizer at some characteristic of crop as an organic whole.People such as king's book brocade in 1979, Xue Delin area northeastward separate and obtain many strains and give birth to type soon and give birth to type soybean nodulation bacteria strain slowly, and further investigate at aspects such as the nodulation and nitrogen fixation of soybean, increasing yield, and, carried out large-area applications in area, the Three River Plain, Heilungkiang and obtained good effect of increasing production the high nitrogenase activity rihizobium japonicum QB1130 that separation obtains.People such as nineteen eighty-three king book brocade, Xue Delin adopt bioactivation soil fertility on the culture fertility of low humidity arable land, the Three River Plain, Heilungkiang, large-area applications vinelandii, phosphorus bacteria, silicate bacteria have been obtained good effect of increasing production on soybean, paddy rice, wheat, corn.King's book brocade, people such as Hu Jiangchun, Xue Delin used the double effects that marine actinomycete has been obtained the not underproduction of continuous cropping, met batch diseases prevention attenuation that can increase production and increase soybean yields in control meeting next batch of soybean disease in 1997.
Rihizobium japonicum is to utilize microbial life activity and meta-bolites thereof, improves dross and the nitrogen fixing capacity of soybean, promotes soybeans they grow for soybean provides nitrogen nutrition; Azotobacter is by nitrogen in the fixed air, and the supply crop nitrogen nutrition promotes plant growth; Actinomycetes are by thalline and meta-bolites excretory microbiotic thereof, diseases prevention, attenuation, stimulation plant growth.The PGPR soybean tailored version biotechnological formulation that three kinds of bacterium are combined and make than single formulation obvious effect of increasing production, and has disease-resistant, attenuation, improves the crop quality effect.Along with the deep and range of application of microbial fertilizer research work is widened, people do as a whole research to microorganism, fertilizer and environment, and single culture transforms to composite bacteria; Simple bacteria agent transforms to compound PGPR type bio-fertilizer; Single formulation transforms to polynary formulation; In fact microbial fertilizer by rudimentary to senior, poor efficiency to efficiently, develop to comprehensive, obligatory type, intensive industrialization direction.
Summary of the invention
The object of the present invention is to provide a kind of required bacterial classification number few, raw material sources are extensive, and technological operation is simple, the biological formulation for soybean of low production cost and production method thereof.
For achieving the above object, the technical solution used in the present invention is:
Biological formulation for soybean, by organic carrier, rihizobium japonicum, azotobacter, actinomycetes are formed, and divide A, B two groups of fermentations, mix by 1: 1, make biological formulation for soybean.The by weight/volume of A group organic carrier and microorganism species seed liquor is organic carrier: rihizobium japonicum: azotobacter=100: 15-25: 5-15, every milliliter viable count 〉=800,000,000 in rihizobium japonicum seed liquor and the azotobacter seed liquor; The by weight/volume of B group organic carrier and microorganism species seed liquor is organic carrier: actinomycetes=100: 25-35, every milliliter of viable bacteria plate count 〉=2,000 ten thousand in the actinomycetes seed liquor.The water content of raw material should be 20-30%, C: N=20～25: 1.
Organic carrier is turfy soil and/or humic acid;
The preparation of biological formulation for soybean and production method thereof, concrete steps are: with organic carriers such as turfy soil, humic acids is raw material, adding known batch production two-stage fermentation production rihizobium japonicum, azotobacter and actinomycetes seed liquor is that microorganism species carries out the heavy dose inoculation to sterilized organic carrier, at ambient temperature, cultivate through solid fermentation, that manufactures high density has high nitrogenase activity and the biological formulation for soybean of prophylaxis effect is arranged.
In the solid fermentation process, the by weight/volume of A group raw material and microorganism species seed liquor is organic carrier: rihizobium japonicum: azotobacter=100: 15-25: 5-15.The by weight/volume of B group raw material and microorganism species seed liquor is organic carrier: actinomycetes=100: 25-35; The water content of raw material should be 20-40% behind the sterilization, C: N=20～25: 1, and according to the difference of raw material, add different carbon source and nitrogenous source, control potential of hydrogen, control oxygen level (ventilation overturning);
The water content of described raw material should be 20-30%; Every milliliter viable count is respectively 800,000,000 at least in viable count rihizobium japonicum seed liquor and the vinelandii seed liquor.Every milliliter of viable bacteria plate count 〉=2,000 ten thousand in the actinomycetes seed liquor.(sampling and measuring at any time during fermentor cultivation adopts counting method of blood cell direct calculating bacterium number under opticmicroscope);
The rihizobium japonicum seed liquor is carried out fermentation culture, and the weight of substratum consists of: glucose 1-3%, sal epsom 0.01-0.05%, sodium-chlor 0.01-0.05%, lime carbonate 0.1-0.3%, surplus are water, PH7.0-7.2; Air flow is 1: 0.3-0.6, and culture temperature is 28 ℃, the time is 48 hours; Preferable weight consists of: glucose 1%, dipotassium hydrogen phosphate 0.05%, sal epsom 0.02%, yeast extract paste 0.1%, sodium-chlor 0.01%, lime carbonate 0.2%, 0.1% boric acid 0.2ml, 0.1% Sodium orthomolybdate 0.2ml, surplus are water, PH7.0; Air flow is 1: 0.5.
The azotobacter seed liquor is carried out fermentation culture, and the weight of substratum consists of: N.F,USP MANNITOL 1-2%, dipotassium hydrogen phosphate 0.01-0.05%, sal epsom 0.01-0.05%, sodium-chlor 0.01-0.05%, lime carbonate 0.2-0.8%, calcium sulfate 0.01-0.03%, surplus are water, PH7.0-7.2; Air flow is 1: 0.3-0.5, and culture temperature is 28 ℃, the time is 48 hours; Preferable weight consists of: N.F,USP MANNITOL 1%, dipotassium hydrogen phosphate 0.02%, sal epsom 0.02%, sodium-chlor 0.02%, lime carbonate 0.5%, calcium sulfate 0.02%, surplus are water, PH7.0; Air flow is 1: 0.5.
The actinomycetes seed liquor is carried out fermentation culture, and the weight of substratum consists of: Zulkovsky starch 1～3%, sal epsom 0.02～0.08%, potassium primary phosphate 0.03～0.08%, sodium-chlor 0.03～0.08%, saltpetre 0.1～0.5%%, surplus are water; PH7.0～7.2; Air flow is 1: 0.3～0.5, and the fermentation culture temperature is 28 ℃, and the time is 48 hours; Preferable weight consists of: Zulkovsky starch 2%, sal epsom 0.05%, potassium primary phosphate 0.05%, sodium-chlor 0.05%, saltpetre 0.1%, surplus are that water, PH7.0, air flow are 1::3.5.
The present invention has following advantage:
1. simple to operate, cost is low, is easy to promote.The present invention draws the advantage of related microorganism fertiliser production method, adopting bacterial classifications such as known rihizobium japonicum, azotobacter and actinomycetes is microorganism species, with organic carriers such as turfy soil, humic acids is raw material, through the liquid-solid two-phase fermentation, the turfy soil of step-down value, humic acid are the high-quality soybean special biological agent that has high nitrogenase activity and have prophylaxis effect.
2. with short production cycle, the quality height.The present invention adopts the microbial liquid fermentation technique that combines with solid, to the heavy dose of microorganism species that inserts of raw material, can better solve in the solid fermentation process, and fermentation period is long, pollution easily, and various fermentation conditions are difficult to problems such as control.And can make full use of carbon source, nitrogenous source in the matrix, adapting under the microorganism growth condition, guaranteed quality product.
3. product application is effective.Microorganism species and bacterial metabolism product thereof that the present invention adopts, crop dross rate is improved, and nitrogen fixing capacity improves and can secrete the various plants hormone, can promote plant growth, strengthen crop anti-adversity, improve crop yield, improve crop quality, and have prophylaxis effect; Simultaneously, again can culture fertility, preserve the ecological environment, be a kind of green micro organism organic fertilizer that human health requires that meets; Show through a large amount of field test results, can make soybean yield-increasing more than 10%.
In a word, the raw material sources of the present invention's employing is extensive, cheap; Adopt preferred high nitrogenase activity rihizobium japonicum, azotobacter and plant rhizosphere to urge living bacterium PGPR actinomadura roseorufa micro organisms flora, effect of increasing production is obvious; Liquid that adopts and the fermentation of solid two-phase have guaranteed quality product; Making the biological formulation for soybean field test results of producing shows: existing production-increasing function has prophylaxis effect again; The soil fertility of both having fostered and apply fertilizer has been improved ecotope again; Belong to green organic manure, be easy to apply.
Embodiment
1. rihizobium japonicum (can symbiosis with soybean, produce root nodule, nitrogen in the fixed air, the supply crop nitrogen nutrition produces the hormonal stimulation plant growth) flora liquid seeds fermentation step is as follows:
The weight of seed liquor fermention medium consists of: glucose 1-3%, sal epsom 0.01-0.05%, sodium-chlor 0.01-0.05%, lime carbonate 0.2-0.6%, surplus are water, PH7.0-7.2; Air flow is 1: 0.3-0.6, and culture temperature is 28 ℃, the time is 48 hours.
1) plants female activation: rihizobium japonicum (Rhizobium japonicum) is planted mother activate; Planting female activation consists of with slant medium: glucose 10g, dipotassium hydrogen phosphate 0.5g, sal epsom 0.2g, sodium-chlor 0.2g, lime carbonate 2.0g, agar 18g, distilled water 1000mL, PH7.0; The sterilization in 18 minutes of a normal atmosphere;
2) fermentor tank inoculation and fermentation culture: the rihizobium japonicum fermentation enters logarithmic growth and jar indicates for putting during vigorous period, the microscopy thalline is neat, healthy and strong, pollution-free, containing viable bacteria can reach more than 800,000,000/ml, (adopting counting method of blood cell direct viewing under opticmicroscope to calculate the bacterium number) nutrient solution free from extraneous odour that is creamy white can be used for enlarging fermentation culture;
3) examine under a microscope the rihizobium japonicum cellular form, can see the shaft-like thalline that two terminal circle is blunt, its size is between the μ m of (4.3 * 0.7)～(1.7 * 0.6), aerobic, Gram-negative, slowly give birth to 1 flagellum of type rihizobium japonicum adnation, give birth to type rihizobium japonicum 2-5 root or many flagellums soon, on Congo red substratum, can finely grow; Growth was observed in 3-4 days in the plate substratum, and single bacterium colony is rounded, protuberance, white, moistening and glossy, cultivate with the passing of time, colony colour becomes oyster white gradually, the sticking mucous membrane in bacterium colony surface, Congo red do not dye or dye extremely shallow.
The weight of the rihizobium japonicum seed liquor fermention medium optimum formula that this example adopts consists of: glucose 1%, dipotassium hydrogen phosphate 0.05%, sal epsom 0.02%, yeast extract paste 0.1%, sodium-chlor 0.01%, lime carbonate 0.2%, 0.5% Congo red 0.5ml, 0.1% boric acid 0.2ml, 0.1% Sodium orthomolybdate 0.2ml, surplus are water, PH7.0; Air flow is 1: 0.5, and culture temperature is 28 ℃, and the time is 48 hours.
4) rihizobium japonicum is that with the difference of vinelandii the height of nitrogenase activity is relevant with strain properties, also whether coordinate relevant with the symbiotic relationship of soybean, be that soybean is relevant with the avidity power of root nodule bacterium, therefore excellent work is selected in the pairing of rihizobium japonicum syntaxial system, to screening effective symbiosis nodulation and nitrogen fixation system, it is significant to improve soybean yields, and its preferred result sees Table 1, table 2:
The dross of table 1. different soybean-root nodule bacterium syntaxial system and the mensuration of fixed nitrogen performance
*
| Bacterial strain | Iron rich 8 | Shen Nong 25104 | Open and educate 8 | Jilin 20 | Long farming 8 | Breathe out 7799 | Close rich 25 |
| (giving birth to type QB113 soon) | ++/++ | +++/+++ | ++/+ | ++/++ | +++/+ | +++/++ | +++/+++ |
| (giving birth to type B16-11C slowly) | +++/++ | +++/+ | +/+ | ++/+++ | +++/+++ | +++/+++ | +++/+++ |
| (giving birth to type 61A76 slowly) | ++/+ | +/+ | +++/+++ | ++/+ | ++/+++ | ++/+ | +++/+++ |
*+++: is strong dross and strong nitrogen: the florescence is measured, and every strain dross number on average reaches more than 50, and nitrogenase activity reaches 50C
2H
4/ strain. more than the branch;
*+++: is strong dross and strong nitrogen: the florescence is measured, and every strain dross number on average reaches more than 30, and nitrogenase activity reaches 30C
2H
4/ strain. more than the branch;
*+++: is strong dross and strong nitrogen: the florescence is measured, and every strain dross number on average reaches more than 10, and nitrogenase activity reaches 10C
2H
4/ strain. more than the branch;
*-: be non-nodulating or fixed nitrogen not: the florescence is measured, no dross phenomenon or the minority root nodule is arranged but less than with the nitrogen activity.
Table 2. soybean～root nodule bacterium syntaxial system is to the influence of economical character, fixed nitrogen performance and output thereof
| Project | Plant height (cm) | Stem joint number (individual/strain) | Strain pod number (individual/strain) | Strain grain number (individual/strain) | Biomass (g dry weight/strain) | Root nodules nitrogenase activity (nmolC2H4/g. fresh weight .h) | Output (kg/ mu) | Stimulation ratio (%) |
| Do not inoculate (ck) | 79.9 | 14.3 | 21.4 | 45.2 | 16.6 | 1138.4 | 123.4 | 100.0 |
| Inoculation QB113 | 78.4 | 15.2 | 24.5 | 45.5 | 18.4 | 4036.8 | 142.8 | 116.0 |
| Inoculation B16-11C | 77.3 | 15.4 | 24.4 | 51.2 | 17.1 | 4876.2 | 144.2 | 117.0 |
For examination bacterial classification: QB113, B16-11C; Experimental cultivar: breathe out 7799
2. vinelandii (nitrogen in the fixed air, the supply crop nitrogen nutrition produces the hormonal stimulation plant growth) flora liquid seeds fermentation step is as follows:
1) plants female activation: vinelandii (azotobacter Azotobacter vinelandl) are planted female, move respectively on the Kolle flask inclined-plane that the vinelandii substratum is housed that is connected to new preparation, activation culture 24～48 hours, 28 ℃ of temperature, the thalline in the Kolle flask after the activation maturation is called seed; Planting female activation consists of with slant medium: N.F,USP MANNITOL 10g, sal epsom 0.2g, potassium primary phosphate 0.2g, sodium-chlor 0.2g, lime carbonate 5.0g, calcium sulfate 0.1g, agar 18g, distilled water 1000mL, PH7.0; The sterilization in 18 minutes of a normal atmosphere.
2) seeding tank inoculation with cultivate: 1 bottle in the seed that will activate the vinelandii in the Kolle flask of getting well, wash to the aseptic triangular flask of 500mL with the 200mL sterilized water, be inoculated into flame inoculation method differential pressure in the seeding tank (or seeding tank of 100 liters) of 50 liters, liquid amount can not surpass 70%, aeration-agitation is cultivated, air flow is 1: 0.3-0.5, temperature is 28～30 ℃, incubation time is 48 hours, the motion of microscopically microscopy thalline is active, neatly, pollution-free, can use for seed fermentation jar expanding propagation.
3) fermentor tank inoculation and fermentation culture (determining substratum and fermentating condition test): with the kind liquid in the sophisticated seeding tank, adopt pressure differential method, be inoculated in the fermentor tank (or fermentor tank of 1000 liters) of 500 liters, liquid amount is no more than 70%, aeration-agitation is cultivated, air flow is 1: 0.3-0.5, temperature is 28-30 ℃, incubation time is 24-48 hour, as adopt azotobacter vinelandii, for putting a jar sign, microscopy bacterium thalline is neat when 8 fonts then occurring, healthy and strong, pollution-free, 8 fonts account for 50%, contain viable bacteria and can reach more than 800,000,000/ml, (adopt counting method of blood cell calculate bacterium number) nutrient solution free from extraneous odour that is creamy white, can for enlarge fermentation culture with or give over to the liquid seeds liquid that solid fermentation is used.
The weight of the vinelandii seed liquor fermention medium optimum formula that adopts in this example consists of: N.F,USP MANNITOL 1%, dipotassium hydrogen phosphate 0.02%, sal epsom 0.02%, sodium-chlor 0.02%, lime carbonate 0.5%, calcium sulfate 0.02%, surplus are water, PH7.0; Air flow is 1: 0.5, and culture temperature is 28 ℃, and the time is 48 hours.
5) vinelandii are used in mixing, rihizobium japonicum bacterium liquid is obvious to the soybean yield-increasing effect, than the better effects if of using vinelandii separately.(ginseng is shown in Table 3)
Table 3. vinelandii, rihizobium japonicum bacterium liquid are to the influence (850 farms, Heilongjiang Province) of soybean yields
| Handle | Soybean seeds 100-grain weight (g) | Soybean yields | |
| (g/10 strain) | (％) | ||
| CK | 19.9 | 97.75 | 100.0 |
| Each 500ml/ mu of azotobacteria bacterial liquid | 20.3 | 113.41 | 112.6 |
| Each 500ml/ mu of rihizobium japonicum liquid | 20.3 | 110.31 | 112.8 |
| Vinelandii+each 500ml/ mu of rihizobium japonicum bacterium liquid | 20.2 | 116.45 | 119.1 |
*Repeat mean number three times;
Actinomycetes (disease-resistant, attenuation, stimulation plant growth, promote crop yield, improve crop quality) flora liquid seeds fermentation step is with 1, wherein difference is:
1) plants female activation:, move to respectively on the Kolle flask inclined-plane of substratum of new preparation and carry out activation culture with the unwrapping wire bacterial classification mother of laboratory preservation.Planting female activation counts by weight percentage with potato-sucrose nutrient agar (PDA) composition: 20% potato leach liquor 1000mL; Sucrose 20g, agar 18g, PH7.0, the sterilization in 20 minutes of a normal atmosphere;
2) seeding tank inoculation and cultivation: air flow is 1: 0.5, and temperature is 28～30 ℃, and incubation time is 48 hours, and it is netted, pollution-free that mycelia is group during sediments microscope inspection, can use for the further expanding propagation of the fermentor tank of seed;
3) fermentor tank inoculation and fermentation culture: aeration-agitation, air flow is 1: 0.3～0.5, temperature is 28～30 ℃, incubation time is 48 hours, jar indicate for putting when occurring that mycelia is painted to shoal, stalwartness, pollution-free, every milliliter of viable bacteria plate count 〉=2,000 ten thousand promptly can be used as the seed liquor that solid fermentation is used.
The weight of the actinomycetes seed liquor fermention medium optimum formula that present embodiment adopts consists of: Zulkovsky starch 2%, sal epsom 0.05%, potassium primary phosphate 0.05%, sodium-chlor 0.05%, saltpetre 0.1%, surplus are water, PH7.0; Air flow is 1: 0.3～0.5, the training fermentation culture, and culture temperature is 28 ℃, the time is 48 hours;
4) bacteriostatic test and field effect of increasing production thereof
Bacteriostatic test adopts cylinder plate method.Is example with marine actinomycete MB-97 bacterial strain to the restraining effect (antagonistic effect) of phytopathogen, the experimental implementation process is: pathogenic bacteria bacteria suspension plate is smoothened, put the Oxford cup, in drum rim, splash into actinomycete fermentation filtrate (perhaps put 4 fritter plates and cultivate the bacterium piece), cultivate after 3-5 days, obvious inhibition zone appears on every side in drum rim (bacterium piece), thereby proves that this bacterial strain has bacteriostatic action.Application marine actinomycete MB-97 biotechnological formulation sees Table 4 to the field test results of secondary soybean prevention effect.
Table 4. secondary soybean is used the effect of increasing production (853 farms, Heilongjiang Province) of MB-97 biotechnological formulation
| Handle | Mu strain number (strain) | 100-grain weight (g) | Per mu yield (kg) | Stimulation ratio (%) |
| Continuous cropping regulation and control in 4 years | 9444.9 | 19.43 | 240.4 | 113.9 |
| Continuous cropping contrast in 4 years | 10000.5 | 18.98 | 211.1 | 100.0 |
| Continuous cropping regulation and control in 5 years | 9444.9 | 19.28 | 249.5 | 131.6 |
| Continuous cropping contrast in 5 years | 10000.5 | 19.07 | 189.6 | 100.0 |
Use marine actinomycete MB-97 biotechnological formulation, unusual effect is arranged, show the soybean plant strain strong stress resistance overcoming the soybean continuous cropping obstacle, soybean quality makes moderate progress, and net effects such as soil fertility raising are through a large amount of field tests, statistical study is determined, can make the secondary soybean volume increase more than 13.9%;
4. rihizobium japonicum, vinelandii and actinomycetes seed liquor product solid fermentation operation steps are as follows:
1) be raw material with organic carriers such as turfy soil, humic acids, adopt bacterial classifications such as the preferred high nitrogenase activity rihizobium japonicum of known process, vinelandii actinomycetes be microorganism species (through different strain and the pairing of different soybean varieties select excellent, determine high nitrogenase activity rihizobium japonicum), the batch production two-stage fermentation is produced root nodule bacterium, vinelandii and actinomycetes seed liquor, organic carriers such as the turfy soil of sterilizing, humic acid are carried out the heavy dose inoculation, carry out solid fermentation and cultivate;
2) with microorganism species such as the high-quality high nitrogenase activity rihizobium japonicum of above-mentioned 1,2,3 step gained, azotobacter and actinomycetes, through batch production two-stage fermentation seeding liquid, with organic carriers such as turfy soil, humic acids is raw material, and the water content behind the high-temperature sterilization in the raw material must be in the 20-30% scope.A group and B group inoculum size all are controlled between the 25-35%, water content before the inoculation secondary fermentation is in the 60-70% scope (" holding agglomerating; land and just loose " no free-water oozes out and is appropriateness) greatly, C: N=20: 1-25: 1, PH7.0-7.2, turn evenly, natural solid fermentation in cleaning ambient is measured the solid fermentation temperature every day 2 times, starting temperature is more than 20 ℃, when rising to 50-60 ℃, need stir 1-2 time, in the solid fermentation process along with the solid fermentation temperature, the A group needs fermentation 48 hours, the B group needs fermentation 96～120 hours, and PH keeps fermenting to terminal point in the 6.8-7.2 scope.
3) after solid fermentation finishes, A is organized rihizobium japonicum and azotobacter solid fermentation product and B group actinomycete fermentation product, by the air-dry or oven dry (bake out temperature is less than 50 ℃) in mixing back in 1: 1, moisture content can not surpass 15%, through pulverizing, sieve, quality inspection, packing, manufacture having of high density biological formulation for soybean product high nitrogenase activity and that have prophylaxis effect, be respectively 800,000,000 at least in the every gram product of contained rihizobium japonicum and azotobacter viable count in the biological formulation for soybean.Be at least 100,000,000 in the every gram product of actinomycetes viable count.Formulation is mainly powdery, also can carry out low temperature disk rolling granulation.
Present embodiment is cultivated material and is consisted of: 200 kilograms of raw materials, 20 kilograms of rihizobium japonicums, 10 kilograms of azotobacters, 30 kilograms in actinomycetes.And dose different carbon sources and nitrogenous source etc. according to the difference of raw material, (cultivate material and consist of raw material: rihizobium japonicum: vinelandii: actinomycetes=200: 20: 10: 30; The water content of raw material should be 20-30%).
4) the field effect of biological formulation for soybean
The effect test of rihizobium japonicum field, the result asks for an interview table 5, table 6, table 7:
The fast field plot trial result who gives birth to type rihizobium japonicum QB113 of watch 5 inoculations
| Project | Plant height (cm) | Strain pod (individual/strain) | 100-grain weight (g) | Strain grain number (individual/strain) | Biomass (g dry weight/strain) | Nitrogenase activity (nmolC 2H 4/ g. fresh weight .h) | Seed production (kg/ mu) | Stimulation ratio (%) |
| Do not inoculate (ck) | 47.2 | 23.8 | 10.3 | 51.0 | 26.5 | 0.63 | 9.2 | 100.0 |
| Inoculation QB113) | 53.0 | 67.2 | 18.3 | 107.8 | 61.3 | 0.902 | 20.4 | 221.7 |
| Inoculation (B16-11C) | 56.0 | 74.6 | 13.7 | 113.8 | 53.0 | 1.102 | 18.7 | 203.3 |
*Contrast enriches 2.5 kilograms of rihizobium japonicum fertilizers for conventional fertilizer application, test
The fast field effect of increasing production of giving birth to type rihizobium japonicum QB113 of watch 6. Baijiang soil areas (8 50 farm) inoculation
| Project | Plant height (cm) | Stem joint number (individual/strain) | Strain pod number (individual/strain) | Strain grain heavy (individual/strain) | 100-grain weight (g) | Invalid pod number (individual/strain) | Output (kg/ mu) | Stimulation ratio (%) |
| Do not inoculate (ck) | 80.6 | 11.7 | 15.3 | 4.5 | 16.6 | 0.8 | 123.4 | 100.0 |
| Inoculation QB13) | 84.4 | 13.6 | 17.9 | 4.9 | 16.3 | 1.0 | 358.1 | 108.6 |
| Inoculation (B16-11C) | 86.3 | 13.8 | 18.8 | 6.2 | 17.7 | 0.9 | 394.7 | 119.7 |
*Contrast enriches 2.5 kilograms of rihizobium japonicum fertilizers for conventional fertilizer application, test
Table 7. biological formulation for soybean is to soybean economical character and yield effect * (850 farms, Heilongjiang Province)
| Handle | Plant height (cm) | Joint number (individual strain) | Pod number (individual/strain) | Grain number (individual/strain) | 100-grain weight (g) | Output (kg/ mu) | Stimulation ratio (%) |
| Test | 85.7 | 17.0 | 22.0 | 60.1 | 23.15 | 192.9 | 113.3 |
| Contrast | 76.7 | 14.4 | 17.5 | 46.1 | 23.00 | 170.3 | 100.0 |
* contrast is conventional fertilizer application, and test enriches 2.5 kilograms of biological formulation for soybean; Test area is 20 mu, and the contrast area is 20 mu.
From above-mentioned test-results as can be seen, biological formulation for soybean all has influence to economical character and the output of soybean, and the result shows: can make soybean yield-increasing 13.3%-19.1%, and by variance analysis, significant difference.And using method is easy, and the effect of improving the soil is obvious, and can improve crop quality.
For improving the quality of products, shorten the production cycle, the applicant has also tried out Shenyang Inst. of Applied Ecology, Chinese Academy of Sciences's separation, purifying and the high-activity microorganism bacterial strain identified (as: give birth to type rihizobium japonicum Rhiobium japonicumB16-11c slowly, give birth to type rihizobium japonicum Rhizobium japonicumQB1130, azotobacter vinelandii Azotobacter vinelandii 230, marine actinomycete Streptomycessoseoflvus MB-97 soon), its better effects if, in Heilungkiang, ground tests such as Jilin and Liaoning use.
Claims (9)
1. biological formulation for soybean is characterized in that: by organic carrier, rihizobium japonicum, azotobacter, actinomycetes are formed, divide two groups of fermentations of A, B after, make by 1: 1 uniform mixing; The by weight/volume of A group organic carrier and microorganism species seed liquor is organic carrier: rihizobium japonicum: azotobacter=100: 15-25: 5-15, every milliliter viable count 〉=800,000,000 in rihizobium japonicum seed liquor and the azotobacter seed liquor; The by weight/volume of B group organic carrier and microorganism species seed liquor is organic carrier: actinomycetes=100: 25-35, every milliliter of viable bacteria plate count 〉=2,000 ten thousand in the actinomycetes seed liquor.
2. according to the described biological formulation for soybean of claim 1, it is characterized in that: described organic carrier is turfy soil and/or humic acid.
3. the production method of the described biological formulation for soybean of claim 1, it is characterized in that: be raw material with the organic carrier, adopting rihizobium japonicum, azotobacter, actinomycetes seed liquor is that microorganism species carries out the heavy dose inoculation to sterilized organic carrier, cultivate through solid fermentation, get the biotechnological formulation of high density; In the solid fermentation process, the by weight/volume of A group raw material and microorganism species seed liquor is, organic carrier: rihizobium japonicum: azotobacter=100: 15-25: 5-15, the by weight/volume of B group raw material and microorganism species seed liquor is organic carrier: actinomycetes=100: 25-35; The water content of raw material should be 20-30% behind the sterilization, C: N=20～25: 1.
4. according to the production method of the described biological formulation for soybean of claim 3, it is characterized in that: described rihizobium japonicum seed liquor, the weight of its fermention medium consists of: glucose 1-3%, sal epsom 0.01-0.05%, sodium-chlor 0.01-0.05%, lime carbonate 0.1-0.3%, surplus are water, PH7.0-7.2; Air flow is 1: 0.3-0.6, and culture temperature is 28 ℃, the time is 48 hours.
5. according to the production method of the described biological formulation for soybean of claim 3, it is characterized in that: the weight of described rihizobium japonicum seed liquor fermention medium consists of: glucose 1%, dipotassium hydrogen phosphate 0.05%, sal epsom 0.02%, yeast extract paste 0.1%, sodium-chlor 0.01%, lime carbonate 0.2%, 0.1% boric acid 0.2ml, 0.1% Sodium orthomolybdate 0.2ml, surplus are water, PH7.0; Air flow is 1: 0.5.
6. according to the production method of the described biological formulation for soybean of claim 3, it is characterized in that: described vinelandii seed liquor, the weight of its fermention medium consists of: N.F,USP MANNITOL 1-2%, dipotassium hydrogen phosphate 0.01-0.05%, sal epsom 0.01-0.05%, sodium-chlor 0.01-0.05%, lime carbonate 0.2-0.8%, calcium sulfate 0.01-0.03%, surplus are water, PH7.0-7.2; Air flow is 1: 0.3-0.5, and culture temperature is 28 ℃, the time is 48 hours.
7. according to the production method of the described biological formulation for soybean of claim 6, it is characterized in that: the weight of described vinelandii seed liquor fermention medium consists of: N.F,USP MANNITOL 1%, dipotassium hydrogen phosphate 0.02%, sal epsom 0.02%, sodium-chlor 0.02%, lime carbonate 0.5%, calcium sulfate 0.02%, surplus are water, PH7.0; Air flow is 1: 0.5.
8. according to the production method of the described biological formulation for soybean of claim 3, it is characterized in that: described actinomycetes seed liquor, the weight of its fermention medium consists of: Zulkovsky starch 1～3%, sal epsom 0.02～0.08%, potassium primary phosphate 0.03～0.08%, sodium-chlor 0.03～0.08%, saltpetre 0.1～0.5%, surplus are water; PH7.0～7.2; Air flow is 1: 0.3～0.5, and the fermentation culture temperature is 28 ℃, and the time is 48 hours.
9. according to the production method of the described biological formulation for soybean of claim 8, it is characterized in that: the weight of its described actinomycetes seed liquor fermention medium consists of: Zulkovsky starch 2%, sal epsom 0.05%, potassium primary phosphate 0.05%, sodium-chlor 0.05%, saltpetre 0.1%, surplus are water, PH7.0; Air flow is 1: 0.3.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103553732A (en) * | 2013-11-11 | 2014-02-05 | 山东省农业科学院农业资源与环境研究所 | Biological organic fertilizer special for soybean and preparation method thereof |
| CN103588522A (en) * | 2013-11-12 | 2014-02-19 | 深圳市新华南方生物科技股份有限公司 | Method for preparing organic active liquid quick-acting compound fertilizer |
| CN105859357A (en) * | 2016-05-04 | 2016-08-17 | 北京科技大学 | Method for preparing microbial fertilizer through by-products generated in bean vermicelli production process |
| CN111960883A (en) * | 2020-07-28 | 2020-11-20 | 沧州市农林科学院 | Special bacterial fertilizer for soybean varieties and preparation method thereof |
-
2004
- 2004-12-17 CN CN 200410100403 patent/CN1789214A/en active Pending
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103553732A (en) * | 2013-11-11 | 2014-02-05 | 山东省农业科学院农业资源与环境研究所 | Biological organic fertilizer special for soybean and preparation method thereof |
| CN103588522A (en) * | 2013-11-12 | 2014-02-19 | 深圳市新华南方生物科技股份有限公司 | Method for preparing organic active liquid quick-acting compound fertilizer |
| CN103588522B (en) * | 2013-11-12 | 2016-02-10 | 深圳市新华南方生物科技股份有限公司 | A kind of preparation method of quick-effective mixed fertilizer of organic active liquid |
| CN105859357A (en) * | 2016-05-04 | 2016-08-17 | 北京科技大学 | Method for preparing microbial fertilizer through by-products generated in bean vermicelli production process |
| CN111960883A (en) * | 2020-07-28 | 2020-11-20 | 沧州市农林科学院 | Special bacterial fertilizer for soybean varieties and preparation method thereof |
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