CN102153381A - Composite microbial fertilizer and low temperature embedding preparation method thereof - Google Patents
Composite microbial fertilizer and low temperature embedding preparation method thereof Download PDFInfo
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- CN102153381A CN102153381A CN2010105902780A CN201010590278A CN102153381A CN 102153381 A CN102153381 A CN 102153381A CN 2010105902780 A CN2010105902780 A CN 2010105902780A CN 201010590278 A CN201010590278 A CN 201010590278A CN 102153381 A CN102153381 A CN 102153381A
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Abstract
The invention relates to the field of bio-fertilizer, in particular to a composite microbial fertilizer and a low temperature embedding preparation method thereof. The composite microbial fertilizer contains a leavening of a composite microbe, an embedding medium and bonding powder; and the leavening, the embedding medium and the bonding powder are embedded in a low temperature plasmid and dried to obtain the composite microbial fertilizer; the composite microbial fertilizer contains the following components in percentage by weight: 50-70% of the leavening of the composite microbe, 25-45% of the embedding medium and 3-8% of the bonding powder, wherein the raw materials are pelletized at a low temperature of 8-20 DEG C and dried at a low temperature of 10-20 DEG C; and the leavening of the composite microbe is obtained by compounding a coculturing material of Candida utilis, Lactococcus lactis, Lactobacillus acidophilus and bacillus subtilis with a single culture solution of Bradyrhizobium japonicum, Azospirillum lipoferum, Rhodopseudomonas palustris, Bacillus megaterium, Bacillus mucilaginosus and Azotobocter chroococcum. The fertilizer provided by the invention is prepared in a dry state so that the activity of the microbe can be kept at more than 90% and the quality guarantee period reaches more than 18 months.
Description
Technical field
What the present invention relates to is biological fertilizer field, a kind of specifically composite microbiological fertilizer and low temperature embedding preparation method thereof.
Background technology
By granulation technique microniological proudcts are made particulate product, have multiple advantages such as the product of raising keeping quality, storage property and transportation property, using and circulating in being convenient to produce.But traditional granulating process causes than macrolesion microbial activity in granulation and drying process, can significantly reduce the result of use of microbial product.Therefore, the efficient granulation technique of microbial activity that keeps of research and development is significant to the production of microbial product.
Studies show that the microbe species difference is to the tolerance of pelleting temperature difference to some extent, for example, lactobacillus is relatively more fragile, the ability of bearing the adverse circumstance environment, and the bacillus category resistance is stronger relatively, the tolerance of microorganisms such as yeast is usually mediate.But the high temperature that tradition is granulated all can affect greatly it, particularly when complex microorganism is granulated, because different microorganisms is to the tolerance difference of environment, cause vigor decline degree difference to some extent, product after the granulation often is difficult to keep original proportion compatibility and having complementary functions property, significantly reduce result of use, can't reach set granulation target.
Summary of the invention
The purpose of this invention is to provide a kind of composite microbiological fertilizer and low temperature embedding preparation method thereof.
For achieving the above object, the technical solution used in the present invention is:
A kind of composite microbiological fertilizer, form by composite bacteria, embedding medium and bonding powder, it is characterized in that: composite microbiological fertilizer is made up of fermented product, embedding medium and the bonding powder of complex microorganism, granulates and drying by low temperature after the embedding medium embedding, promptly gets composite microbiological fertilizer; Composite microbiological fertilizer by weight percentage, the fermented product of complex microorganism is 50%-70%, embedding medium is 25%-45%; Bonding powder accounts for the 3-8% that is of fertilizer total amount; Wherein granulate, 10-20 ℃ cryodrying at 8-20 ℃ low temperature; The fermented product of described complex microorganism is to be composited for the co-cultivation thing of Candida utilis, streptococcus acidi lactici, Lactobacterium acidophilum, subtilis and slow raw soybean root nodule bacterium, the single nutrient solution of giving birth to fat azospirillum, swamp Rhodopseudomonas, bacillus megaterium, colloid bacillus cereus and azotobacter chroococcum.
Described embedding medium is seniority in the family ratio meter by weight, the gelatin of the sodium alginate of 2-3%, the starch of 0.5-1%, 1-2%, and surplus is a water.The feed adhesive that described bonding powder is used always for those skilled in the art.
The low temperature embedding preparation method of composite microbiological fertilizer:
1) Candida utilis, streptococcus acidi lactici, Lactobacterium acidophilum and subtilis co-fermentation are cultured to total count reach 109/more than the mL, again with the slow raw soybean root nodule bacterium that are cultured to logarithmic phase respectively, give birth to fat azospirillum, swamp Rhodopseudomonas, bacillus megaterium, colloid bacillus cereus and azotobacter chroococcum mix the fermented product of complex microorganism, blending ratio accounts for 40% for the co-fermentation thing, and every single culturing bacterium accounts for 10% respectively;
2) preparation embedding medium is with the sodium alginate of 2-3%, the starch of 0.5-1%, the gelatin of 1-2%, the mixing solutions of making soluble in water, autoclaving.
3) by weight percentage, get fermented product, the 25%-45% step 2 of the complex microorganism after the 50%-70% step 1) is cultivated) embedding medium and the bonding powder mixing embedding of 3-8%, then granulate down at 8-20 ℃, the back of granulating promptly obtains composite microbiological fertilizer in 10-20 ℃ of drying.
The particle diameter of described step 3) granulation gained embedded particles is 2-3mm.
Above-mentioned bacterial strains is all available from Chinese agriculture microbial strains preservation center, and its Candida utilis is in the ACCC20060 that is numbered at Chinese agriculture microbial strains preservation center; Streptococcus acidi lactici is in the ACCC10653 that is numbered at Chinese agriculture microbial strains preservation center; Lactobacterium acidophilum is in the CICC6005 that is numbered at Chinese industrial microbial strains preservation center; Subtilis is in the CICC20037 that is numbered at Chinese industrial microbial strains preservation center; Slow raw soybean root nodule bacterium are in the ACCC15402 that is numbered at Chinese agriculture microbial strains preservation center; Give birth to the fat azospirillum in the ACCC10481 that is numbered at Chinese agriculture microbial strains preservation center; Swamp Rhodopseudomonas is in the ACCC10649 that is numbered at Chinese agriculture microbial strains preservation center; Bacillus megaterium is in the ACCC02991 that is numbered at Chinese agriculture microbial strains preservation center; Colloid bacillus cereus is in the ACCC02983 that is numbered at Chinese agriculture microbial strains preservation center; Azotobacter chroococcum is in the ACCC10098 that is numbered at Chinese agriculture microbial strains preservation center.
The present invention has following advantage:
1. the present invention adopts low temperature granulation technique and dry technology to finish granulation, has realized low temperature granulation, cryodrying, has guaranteed that composite microbiological fertilizer has function originally.
2. fertilizer of the present invention is by making under drying regime, and then can keep microbic activity to reach more than 90%, and the quality guaranteed period reached more than 18 months.
Description of drawings
Fig. 1 is the process flow sheet of the embodiment of the invention.
Embodiment
Embodiment 1
A kind of composite microbiological fertilizer is made up of compound microorganism ferments thing, embedding medium and bonding powder, and by weight percentage, the compound microorganism ferments thing is 50%, and embedding medium is 45%; What bonding powder accounted for the fertilizer total amount is 5%.
The preparation method:
1) Candida utilis, streptococcus acidi lactici, Lactobacterium acidophilum and subtilis are carried out activation culture respectively to logarithmic phase according to ordinary method, Candida utilis, streptococcus acidi lactici, Lactobacterium acidophilum and subtilis after then will activating 1: 1: 1 by volume: 1 mixes, then will mix the back bacterial classification and be inoculated in compound cultivation in the liquid fermentation medium according to weight percent 4%, under 30 ℃, fermentation culture 70h, fermented product pH value is 4.5-7.5, to total count reach 109/more than the mL; Wherein liquid fermentation medium is a glucose 2%, peptone 1%, extractum carnis 1%, yeast extract paste 0.5%, surplus is a distilled water, according to a conventional method with slow raw soybean root nodule bacterium, give birth to fat azospirillum, swamp Rhodopseudomonas, bacillus megaterium, colloid bacillus cereus and azotobacter chroococcum respectively at shake-flask culture 48-72h in the substratum separately, respectively enlarged culturing to microbe population reach 108/more than the mL, promptly be activated to and be in logarithmic phase.
2) co-fermentation is cultured to total count and reaches the above compound culture of Candida utilis, streptococcus acidi lactici, Lactobacterium acidophilum, subtilis of 109/mL, with the slow raw soybean root nodule bacterium that are cultured to logarithmic phase respectively, give birth to fat azospirillum, swamp Rhodopseudomonas, bacillus megaterium, colloid bacillus cereus and azotobacter chroococcum mix the fermented product of complex microorganism, blending ratio accounts for 40% for the co-fermentation thing, and single culturing bacterium accounts for 10% respectively.
Above-mentioned bacterial strains all can be respectively cultivated according to ordinary method, and a kind of in the numerous culture condition of the knowledge that present embodiment provides is respectively:
The substratum of Candida utilis: potato is soaked powder 3g, glucose 20g, and agar powder 15g, distilled water adds to 1000mL, 121 ℃ of autoclavings 20 minutes.Candida utilis multiplication culture condition is cultivated 24-48h for 28 ℃.
The substratum of streptococcus acidi lactici: glucose 2%, peptone 1%, extractum carnis 1%, yeast extract paste 0.5%, dipotassium hydrogen phosphate 0.2%, sodium acetate 0.5%, ammonium citrate 0.2%, sal epsom 0.058%, manganous sulfate 0.025%, agar 1.5-2%, pH6.5.121 ℃ of autoclavings 20 minutes.Streptococcus acidi lactici multiplication culture condition is cultivated 48-72h for 30 ℃.
The substratum of Lactobacterium acidophilum: glucose 2%, peptone 1%, extractum carnis 1%, yeast extract paste 0.5%, dipotassium hydrogen phosphate 0.2%, sodium acetate 0.5%, ammonium citrate 0.2%, sal epsom 0.058%, manganous sulfate 0.025%, agar 1.5-2%, pH6.5.121 ℃ of autoclavings 20 minutes.Lactobacterium acidophilum multiplication culture condition is cultivated 48-72h for 37 ℃.
The substratum of subtilis: peptone 10g, agar 20g, sodium-chlor 5g, meat soak powder 10g, and water adds to 1000mL, pH7.2.121 ℃ of autoclavings 20 minutes.Subtilis multiplication culture condition is cultivated 48-72h for 30 ℃.
The substratum of slow raw soybean root nodule bacterium: N.F,USP MANNITOL 10g, K2HPO40.5g, yeast powder 0.4g, MgSO47H2O0.2g, NaCl0.1g, CaCl26H2O0.1g, distilled water 1000mL, pH6.8-7.0.121 ℃ of autoclavings 20 minutes.Slow raw soybean root nodule bacterium multiplication culture condition is cultivated 48-72h for 28 ℃.
Give birth to the substratum of fat azospirillum: sucrose 10g, K
2HPO
40.5g, NaCl0.1g, yeast extract paste 1g, ironic citrate (1%) 1mL, CaSO
40.2g, MgSO
47H2O0.2g, MnSO
4(1%) 1mL, NaMoO
4(1%) 1mL, boric acid (1%) 1mL, distilled water 1000mL, pH6.8-7.0; 121 ℃ of autoclavings 20 minutes.Give birth to fat azospirillum multiplication culture condition under 25-30 ℃ of condition, cultivate 48-72h.
Have a liking for the substratum of swamp Rhodopseudomonas: yeast powder 0.3%, peptone 0.3%, sal epsom 0.05%, calcium chloride 0.03%, distilled water 1000mL, pH6.8-7.0; 121 ℃ of autoclavings 20 minutes.Have a liking for swamp Rhodopseudomonas multiplication culture condition under 30 ℃ of conditions, cultivate 72-96h.
The substratum of bacillus megaterium: peptone 0.5%, extractum carnis 0.3%, sodium-chlor 0.5%, distilled water 1000mL, pH7.0; 121 ℃ of autoclavings 20 minutes.Bacillus megaterium multiplication culture condition is cultivated 24-48h under 30 ℃ of conditions.
Colloid bacillus cereus substratum: sucrose 10g, yeast extract paste 0.2g, (NH
4) SO
40.5g, MgSO
40.1g, KCl0.1g, Na
2HPO
40.1g, CaCO
31.0g, distilled water 1000mL, pH7.0.121 ℃ of autoclavings 20 minutes; The multiplication culture condition is cultivated 20-40h under 32-36 ℃ of condition.
Azotobacter chroococcum substratum: glucose 10g, KH
2PO
40.2g, MgSO
47H
2O0.2g, NaCl0.2g, CaSO
47H
2O0.2g, CaCO
35.0g, distilled water 1000mL, pH7.0-7.2 sterilized 30 minutes for 113 ℃.The multiplication culture condition is cultivated 24-48h under 28-36 ℃ of condition.
Above-mentioned bacterial strains is all available from Chinese agriculture microbial strains preservation center, and cultivates separately according to prior art well known to those of ordinary skill in the art, and its Candida utilis is in the ACCC20060 that is numbered at Chinese agriculture microbial strains preservation center; Streptococcus acidi lactici is in the ACCC10653 that is numbered at Chinese agriculture microbial strains preservation center; Lactobacterium acidophilum is in the CICC6005 that is numbered at Chinese industrial microbial strains preservation center; Subtilis is in the CICC20037 that is numbered at Chinese industrial microbial strains preservation center; Slow raw soybean root nodule bacterium are in the ACCC15402 that is numbered at Chinese agriculture microbial strains preservation center; Give birth to the fat azospirillum in the ACCC10481 that is numbered at Chinese agriculture microbial strains preservation center; Swamp Rhodopseudomonas is in the ACCC10649 that is numbered at Chinese agriculture microbial strains preservation center; Bacillus megaterium is in the ACCC02991 that is numbered at Chinese agriculture microbial strains preservation center; Colloid bacillus cereus is in the ACCC02983 that is numbered at Chinese agriculture microbial strains preservation center; Azotobacter chroococcum is in the ACCC10098 that is numbered at Chinese agriculture microbial strains preservation center.
3) preparation embedding medium, the sodium alginate with 3%, 0.5% starch, 1% gelatin, the mixing solutions of making soluble in water, autoclaving.Bonding powder is a wheat gluten flour.
4) by weight percentage, get 50% step 2) fermented product, the embedding medium of 45% step 3) and 5% the bonding powder mixing embedding of complex microorganism after cultivating.With step 2) compound microorganism ferments liquid joins in the embedding medium of step 3), abundant mixing, and under stirring condition, join in the calcium chloride solution that concentration is 0.2mol/L, obtain the microbial fertilizer particle, mix the back again granulates down at 8 ℃ with tackiness agent, granulate the back in 10 ℃ of dryings, promptly obtain the composite microbiological fertilizer that particle diameter is 2-3mm.
According to colony counting method, detect and make each bacterial activity assay among the composite microbiological fertilizer storage 1-520d, referring to table 1.
Table 1 storage time is to the influence of composite microbiological fertilizer total viable count
Embodiment 2
A kind of composite microbiological fertilizer is made up of compound microorganism ferments thing, embedding medium and bonding powder, and by weight percentage, the compound microorganism ferments thing is 70%, and embedding medium is 25%; What bonding powder accounted for the fertilizer total amount is 5%.
The preparation method:
1) co-fermentation is cultured to total count and reaches the above compound culture of Candida utilis, streptococcus acidi lactici, Lactobacterium acidophilum, subtilis of 109/mL, with the slow raw soybean root nodule bacterium that are cultured to logarithmic phase respectively, give birth to fat azospirillum, swamp Rhodopseudomonas, bacillus megaterium, colloid bacillus cereus and azotobacter chroococcum mix the fermented product of complex microorganism, blending ratio accounts for 40% for the co-fermentation thing, and single culturing bacterium accounts for 10% respectively.
2) preparation embedding medium, the sodium alginate with 2%, 1% starch, 2% gelatin, the mixing solutions of making soluble in water, autoclaving.Bonding powder is a wheat gluten flour.
3) get by weight percentage, fermented product, 25% step 2 of the complex microorganism after 70% step 1) is cultivated) embedding medium and 5% bonding powder mixing embedding.Step 1) compound microorganism ferments liquid is joined step 2) embedding medium in, abundant mixing, and under stirring condition, join in the calcium chloride solution that concentration is 0.2mol/L, obtain the microbial fertilizer particle, mix the back again granulates down at 8 ℃ with tackiness agent, granulate the back in 10 ℃ of dryings, promptly obtain the composite microbiological fertilizer that particle diameter is 2-3mm.
According to colony counting method, detect and make each bacterial activity assay among the composite microbiological fertilizer storage 1-520d, referring to table 2.
Table 2 storage time is to the influence of composite microbiological fertilizer total viable count
Aforesaid operations, nutrient solution, the reagent that adopts and bacterial classification be this area routine operation, nutrient solution, commercial reagent and bacterial classification if no special instructions.It is main raw material that composite microbiological fertilizer of the present invention in addition adopts functional microorganism fermented liquid, sodium alginate, gelatin etc., can be used for all soil types and farm crop, effect the best on vegetables, fruit, it can effectively improve Soil structure, improve the soil water-reataining fertilizer-keeping performance, has better volume increase and improves the effect of crop quality.
Claims (5)
1. composite microbiological fertilizer, form by composite bacteria, embedding medium and bonding powder, it is characterized in that: composite microbiological fertilizer is made up of fermented product, embedding medium and the bonding powder of complex microorganism, granulates and drying by low temperature after the embedding medium embedding, promptly gets composite microbiological fertilizer; Composite microbiological fertilizer by weight percentage, the fermented product of complex microorganism is 50%-70%, embedding medium is 25%-45%; Bonding powder accounts for the 3-8% that is of fertilizer total amount; Wherein granulate, 10-20 ℃ cryodrying at 8-20 ℃ low temperature; The fermented product of described complex microorganism is to be composited for the co-cultivation thing of Candida utilis, streptococcus acidi lactici, Lactobacterium acidophilum, subtilis and slow raw soybean root nodule bacterium, the single nutrient solution of giving birth to fat azospirillum, swamp Rhodopseudomonas, bacillus megaterium, colloid bacillus cereus and azotobacter chroococcum.
2. by the described composite microbiological fertilizer of claim 1, it is characterized in that: described embedding medium by weight percentage, the gelatin of the sodium alginate of 2-3%, the starch of 0.5-1%, 1-2%, surplus is a water.
3. by the described composite microbiological fertilizer of claim 1, it is characterized in that: described composite microbiological fertilizer: with Candida utilis, streptococcus acidi lactici, Lactobacterium acidophilum and subtilis co-fermentation be cultured to total count reach 109/more than the mL, again with the slow raw soybean root nodule bacterium that are cultured to logarithmic phase respectively, give birth to fat azospirillum, swamp Rhodopseudomonas, bacillus megaterium, colloid bacillus cereus and azotobacter chroococcum mix the fermented product of complex microorganism, blending ratio accounts for 40% for the co-fermentation thing, and every single culturing bacterium accounts for 10% respectively.
4. the low temperature embedding preparation method of the described composite microbiological fertilizer of claim 1 is characterized in that:
1) Candida utilis, streptococcus acidi lactici, Lactobacterium acidophilum and subtilis co-fermentation are cultured to total count reach 109/more than the mL, again with the slow raw soybean root nodule bacterium that are cultured to logarithmic phase respectively, give birth to fat azospirillum, swamp Rhodopseudomonas, bacillus megaterium, colloid bacillus cereus and azotobacter chroococcum mix the fermented product of complex microorganism, blending ratio accounts for 40% for the co-fermentation thing, and every single culturing bacterium accounts for 10% respectively;
2) preparation embedding medium is with the sodium alginate of 2-3%, the starch of 0.5-1%, the gelatin of 1-2%, the mixing solutions of making soluble in water, autoclaving.
3) by weight percentage, get fermented product, the 25%-45% step 2 of the complex microorganism after the 50%-70% step 1) is cultivated) embedding medium and the bonding powder mixing embedding of 3-8%, then granulate down at 8-20 ℃, the back of granulating promptly obtains composite microbiological fertilizer in 10-20 ℃ of drying.
5. by the low temperature embedding preparation method of the described composite microbiological fertilizer of claim 1, it is characterized in that: the particle diameter of described step 3) granulation gained embedded particles is 2-3mm.
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| CN104642539A (en) * | 2013-11-25 | 2015-05-27 | 内蒙古蒙牛乳业（集团）股份有限公司 | Probiotic granules and preparation method thereof, and long-shelf-life fermented milk containing probiotic granules and preparation method thereof |
| CN109321466A (en) * | 2018-11-17 | 2019-02-12 | 华中农业大学 | A kind of auxiliary material and application improving rhizobium liquid bacterial agent viable count and nodulation and nitrogen fixation effect |
| CN110128202A (en) * | 2019-05-31 | 2019-08-16 | 襄阳农博源农业有限公司 | A kind of biological organic fertilizer and preparation method thereof |
| CN110257298A (en) * | 2019-07-04 | 2019-09-20 | 临沂大学 | A kind of microorganism mixed bacterial promoting photosynthesis of plant |
| CN110577429A (en) * | 2018-06-08 | 2019-12-17 | 广东唐博生物科技有限公司 | double-layer embedded composite microbial fertilizer and preparation method thereof |
| CN112514918A (en) * | 2020-12-23 | 2021-03-19 | 湖南恒圣创新建筑科技有限公司 | Embedding body composite insecticidal probiotic agent and preparation method and application thereof |
| CN114736077A (en) * | 2022-03-30 | 2022-07-12 | 中诚国联（河南）生物科技有限公司 | Compound microbial agent for acid soil remediation and preparation method and application thereof |
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104642539A (en) * | 2013-11-25 | 2015-05-27 | 内蒙古蒙牛乳业（集团）股份有限公司 | Probiotic granules and preparation method thereof, and long-shelf-life fermented milk containing probiotic granules and preparation method thereof |
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| CN109321466A (en) * | 2018-11-17 | 2019-02-12 | 华中农业大学 | A kind of auxiliary material and application improving rhizobium liquid bacterial agent viable count and nodulation and nitrogen fixation effect |
| CN109321466B (en) * | 2018-11-17 | 2021-10-08 | 华中农业大学 | Auxiliary material for improving viable count, nodulation and nitrogen fixation effects of rhizobium liquid microbial inoculum and application thereof |
| CN110128202A (en) * | 2019-05-31 | 2019-08-16 | 襄阳农博源农业有限公司 | A kind of biological organic fertilizer and preparation method thereof |
| CN110257298A (en) * | 2019-07-04 | 2019-09-20 | 临沂大学 | A kind of microorganism mixed bacterial promoting photosynthesis of plant |
| CN112514918A (en) * | 2020-12-23 | 2021-03-19 | 湖南恒圣创新建筑科技有限公司 | Embedding body composite insecticidal probiotic agent and preparation method and application thereof |
| CN114736077A (en) * | 2022-03-30 | 2022-07-12 | 中诚国联（河南）生物科技有限公司 | Compound microbial agent for acid soil remediation and preparation method and application thereof |
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| CN102153381B (en) | 2013-06-19 |
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