CN113774050B - Microbial agent for promoting nutrient absorption - Google Patents
Microbial agent for promoting nutrient absorption Download PDFInfo
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- CN113774050B CN113774050B CN202111064947.5A CN202111064947A CN113774050B CN 113774050 B CN113774050 B CN 113774050B CN 202111064947 A CN202111064947 A CN 202111064947A CN 113774050 B CN113774050 B CN 113774050B
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- 230000000813 microbial effect Effects 0.000 title claims abstract description 55
- 230000001737 promoting effect Effects 0.000 title claims abstract description 16
- 235000015816 nutrient absorption Nutrition 0.000 title claims abstract description 14
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 52
- QJZYHAIUNVAGQP-UHFFFAOYSA-N 3-nitrobicyclo[2.2.1]hept-5-ene-2,3-dicarboxylic acid Chemical compound C1C2C=CC1C(C(=O)O)C2(C(O)=O)[N+]([O-])=O QJZYHAIUNVAGQP-UHFFFAOYSA-N 0.000 claims abstract description 22
- 244000005700 microbiome Species 0.000 claims abstract description 18
- 239000004021 humic acid Substances 0.000 claims abstract description 17
- 150000001875 compounds Chemical class 0.000 claims abstract description 15
- 239000002068 microbial inoculum Substances 0.000 claims abstract description 7
- 239000002994 raw material Substances 0.000 claims abstract description 5
- 241000194107 Bacillus megaterium Species 0.000 claims description 24
- 241000228245 Aspergillus niger Species 0.000 claims description 21
- 241000589516 Pseudomonas Species 0.000 claims description 21
- 238000004321 preservation Methods 0.000 claims description 21
- 238000002156 mixing Methods 0.000 claims description 20
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 claims description 18
- 238000003756 stirring Methods 0.000 claims description 16
- 239000001963 growth medium Substances 0.000 claims description 13
- 229940010698 activated attapulgite Drugs 0.000 claims description 12
- 238000001035 drying Methods 0.000 claims description 12
- 238000002360 preparation method Methods 0.000 claims description 12
- 229960000892 attapulgite Drugs 0.000 claims description 10
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- 229910052625 palygorskite Inorganic materials 0.000 claims description 10
- 241000894006 Bacteria Species 0.000 claims description 9
- GRYLNZFGIOXLOG-UHFFFAOYSA-N Nitric acid Chemical compound O[N+]([O-])=O GRYLNZFGIOXLOG-UHFFFAOYSA-N 0.000 claims description 9
- 229910017604 nitric acid Inorganic materials 0.000 claims description 9
- 238000001354 calcination Methods 0.000 claims description 8
- 238000001816 cooling Methods 0.000 claims description 8
- 238000010438 heat treatment Methods 0.000 claims description 8
- 238000000034 method Methods 0.000 claims description 8
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- 238000005406 washing Methods 0.000 claims description 8
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- 108010020346 Polyglutamic Acid Proteins 0.000 claims description 3
- 239000002250 absorbent Substances 0.000 claims description 3
- 229920002643 polyglutamic acid Polymers 0.000 claims description 3
- 239000011347 resin Substances 0.000 claims description 3
- 229920005989 resin Polymers 0.000 claims description 3
- 229920002472 Starch Polymers 0.000 claims description 2
- 239000008107 starch Substances 0.000 claims description 2
- 235000019698 starch Nutrition 0.000 claims description 2
- 239000002689 soil Substances 0.000 abstract description 22
- 239000003337 fertilizer Substances 0.000 abstract description 14
- 230000012010 growth Effects 0.000 abstract description 8
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 abstract description 8
- 229910019142 PO4 Inorganic materials 0.000 abstract description 7
- 239000010452 phosphate Substances 0.000 abstract description 7
- 230000008901 benefit Effects 0.000 abstract description 2
- 230000035558 fertility Effects 0.000 abstract description 2
- 241000196324 Embryophyta Species 0.000 description 15
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 12
- 238000009629 microbiological culture Methods 0.000 description 12
- 229910052698 phosphorus Inorganic materials 0.000 description 12
- 239000011574 phosphorus Substances 0.000 description 12
- 238000012360 testing method Methods 0.000 description 10
- 230000000694 effects Effects 0.000 description 9
- 241000227653 Lycopersicon Species 0.000 description 7
- 235000007688 Lycopersicon esculentum Nutrition 0.000 description 7
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 6
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 6
- 230000000052 comparative effect Effects 0.000 description 6
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- 241000228212 Aspergillus Species 0.000 description 3
- 241000282414 Homo sapiens Species 0.000 description 3
- 239000001888 Peptone Substances 0.000 description 3
- 108010080698 Peptones Proteins 0.000 description 3
- 241001259677 Pseudomonas guguanensis Species 0.000 description 3
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 3
- 238000010521 absorption reaction Methods 0.000 description 3
- 230000036782 biological activation Effects 0.000 description 3
- 238000011161 development Methods 0.000 description 3
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- 229910052757 nitrogen Inorganic materials 0.000 description 3
- 235000015097 nutrients Nutrition 0.000 description 3
- 235000019319 peptone Nutrition 0.000 description 3
- 235000010958 polyglycerol polyricinoleate Nutrition 0.000 description 3
- 239000000843 powder Substances 0.000 description 3
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- 230000001954 sterilising effect Effects 0.000 description 3
- 229920001817 Agar Polymers 0.000 description 2
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 2
- 239000000589 Siderophore Substances 0.000 description 2
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- 230000008635 plant growth Effects 0.000 description 2
- 239000003375 plant hormone Substances 0.000 description 2
- ZZZCUOFIHGPKAK-UHFFFAOYSA-N D-erythro-ascorbic acid Natural products OCC1OC(=O)C(O)=C1O ZZZCUOFIHGPKAK-UHFFFAOYSA-N 0.000 description 1
- VGGSQFUCUMXWEO-UHFFFAOYSA-N Ethene Chemical compound C=C VGGSQFUCUMXWEO-UHFFFAOYSA-N 0.000 description 1
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- 229930191978 Gibberellin Natural products 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 102000018997 Growth Hormone Human genes 0.000 description 1
- 108010051696 Growth Hormone Proteins 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- 229930003268 Vitamin C Natural products 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 238000012271 agricultural production Methods 0.000 description 1
- 238000005904 alkaline hydrolysis reaction Methods 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
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- 239000001506 calcium phosphate Substances 0.000 description 1
- 229910000389 calcium phosphate Inorganic materials 0.000 description 1
- 235000011010 calcium phosphates Nutrition 0.000 description 1
- 238000010668 complexation reaction Methods 0.000 description 1
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- 230000002349 favourable effect Effects 0.000 description 1
- IXORZMNAPKEEDV-UHFFFAOYSA-N gibberellic acid GA3 Natural products OC(=O)C1C2(C3)CC(=C)C3(O)CCC2C2(C=CC3O)C1C3(C)C(=O)O2 IXORZMNAPKEEDV-UHFFFAOYSA-N 0.000 description 1
- 239000003448 gibberellin Substances 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 239000003673 groundwater Substances 0.000 description 1
- 239000000122 growth hormone Substances 0.000 description 1
- 229910001385 heavy metal Inorganic materials 0.000 description 1
- 238000003837 high-temperature calcination Methods 0.000 description 1
- 238000003384 imaging method Methods 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 229910010272 inorganic material Inorganic materials 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 238000009630 liquid culture Methods 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 239000002686 phosphate fertilizer Substances 0.000 description 1
- 230000008979 phosphorus utilization Effects 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 239000006041 probiotic Substances 0.000 description 1
- 235000018291 probiotics Nutrition 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 239000004576 sand Substances 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 230000003248 secreting effect Effects 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
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- 239000000725 suspension Substances 0.000 description 1
- 230000002195 synergetic effect Effects 0.000 description 1
- 230000009044 synergistic interaction Effects 0.000 description 1
- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 description 1
- 235000019154 vitamin C Nutrition 0.000 description 1
- 239000011718 vitamin C Substances 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
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- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/14—Enzymes or microbial cells immobilised on or in an inorganic carrier
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/20—Bacteria; Substances produced thereby or obtained therefrom
- A01N63/22—Bacillus
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
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- A01N63/20—Bacteria; Substances produced thereby or obtained therefrom
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- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/30—Microbial fungi; Substances produced thereby or obtained therefrom
- A01N63/34—Aspergillus
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Abstract
The invention discloses a microbial agent for promoting nutrient absorption, and belongs to the technical field of microorganisms. The microbial agent disclosed by the invention is prepared from the following raw materials in parts by weight: 20-30 parts of microorganism carrier, 15-20 parts of compound microorganism microbial inoculum, 10-15 parts of humic acid and 3-5 parts of water-retaining agent. The microbial agent can continuously and efficiently play a role in phosphate dissolving and growth promotion after being applied into soil, improve the micro-ecological environment of the soil, continuously maintain the soil fertility, remarkably improve the utilization rate of fertilizer, promote the yield and quality of crops and have remarkable economic benefit.
Description
Technical Field
The invention belongs to the technical field of microorganisms, and particularly relates to a microbial agent for promoting nutrient absorption.
Background
The fertilizer is an indispensable nutrient supply body for human indirectly depending on survival, and is derived from human beings and also acts on human beings, and plants can directly absorb and grow the fertilizer. Chemical fertilizer is the biggest material input in agricultural production, and the plant is limited to fertilizer's absorptivity, and its absorption and loss coexist, and the effect that plays to increasing production accounts for 40-60%, and on the one hand the fertilizer that loses causes farmland, groundwater's pollution and the waste of resource, on the other hand crop leads to output and quality decline because of unable high-efficient absorption fertilizer.
Therefore, the control of the fertilizer dosage is a realistic requirement for promoting cost saving, efficiency improvement, energy saving and emission reduction, and has very important significance for guaranteeing national grain safety, agricultural product quality safety and agricultural ecological safety. The research and development of the functional microbial fertilizer and the screening and compounding of the agricultural probiotics are one of the important means for realizing fertilizer reduction.
Plant growth promoting bacteria (PGP) are a group of self-growing bacteria in soil around plant rhizosphere, inhibit or reduce adverse effects on plants caused by plant diseases, heavy metals, salt stress and the like by means of producing antibiotics, secreting siderophores and the like, and directly stimulate and regulate plant growth by means of phosphate dissolving, nitrogen fixing, plant hormone production, ethylene level reduction by enzymolysis and the like. The development, development and application of the novel PGPR composition and production process composite microbial inoculum are the directions of one hot spot of the current microbial fertilizer. The aim of the complexation is to achieve complementation of the pro-active effect and amplification of the efficacy between strains including PGPR, and the combination of PGPR with other beneficial flora.
How to utilize rhizosphere microorganisms fully so as to promote the maximum utilization of fertilizer nutrients, reduce the hardening caused by excessive soil nutrition and the like, and promote the yield increase of crops is the most popular agricultural research direction at present.
Disclosure of Invention
The invention provides a compound microbial flora, which has synergistic effect, and can realize full decomposition of insoluble phosphorus in soil for crop absorption and utilization, and can secrete plant hormone with high efficiency to promote crop growth.
In order to achieve the technical purpose, the invention adopts the following technical scheme:
the microbial agent for promoting nutrient absorption is prepared from the following raw materials in parts by weight: 20-30 parts of microorganism carrier, 15-20 parts of compound microorganism microbial inoculum, 10-15 parts of humic acid and 3-5 parts of water-retaining agent.
Further, the microbial carrier is activated attapulgite, the attapulgite is added into a mixed solution of nitric acid with the weight percentage of 20% and hydrogen peroxide with the weight percentage of 5%, and the mixture is prepared into slurry with the weight percentage of 20%, and the activated attapulgite is obtained through heating, stirring, cooling, washing, drying and calcining.
Furthermore, the compound microbial agent is obtained by mixing bacillus megatherium, pseudomonas glutinosa and aspergillus niger cultures according to the volume ratio of 1:1:1.
Further, the preservation number of the bacillus megaterium is CGMCC1.16094, the preservation number of the pseudomonas glutinosa is CGMCC 1.15627, and the preservation number of the aspergillus niger is CCTCC HF 2008599.
The bacillus megatherium (Bacillus megaterium) is purchased from China general microbiological culture collection center (China General Microbiological Culture Collection Center, CGMCC), and has the following addresses: beijing city, chaoyang district, north Chen Xili No. 1, 3, date of preservation: 2017, 3, 11 days, deposit number: CGMCC1.16094.
The Pseudomonas glutinosa (Pseudomonas guguanensis) is purchased from China general microbiological culture collection center (China General Microbiological Culture Collection Center, CGMCC), and has the following addresses: beijing city, chaoyang district, north Chen Xili No. 1, 3, date of preservation: 25 days of 2 months 2015, deposit number: CGMCC 1.15627.
Aspergillus niger (Aspergillus) of the invention is purchased from China center for type culture Collection (university of Wuhan), eight-way 299 No. of Wuchang district of Wuhan, hubei province, and the preservation number is CCTCC HF 2008599.
Further, the preparation method of the compound microbial agent comprises the following steps: inoculating Bacillus megaterium, pseudomonas glutinosa and Aspergillus niger into LB culture medium, shake culturing at 28-30deg.C until the bacterial concentration is OD600 ≡2.5, and mixing at volume ratio of 1:1:1.
Further, the composition of the LB culture medium is as follows: 10g of peptone, 5g of yeast powder, 10g of NaCl, 1000mL of distilled water, pH value of 7.2-7.4 and sterilizing at 121 ℃ for 19min.
Further, the humic acid is the humic acid which is subjected to biological activation, and the activation method comprises the following steps: regulating humic acid humidity to 20-30%, uniformly spraying cellulose enzyme solution with mass concentration of 20%, uniformly stirring, and standing for 6-10 h.
Further, the water-retaining agent is starch, polyglutamic acid or water-absorbent resin.
A preparation method of a microbial agent for promoting nutrient absorption comprises the following steps:
(1) Preparation of a microbial carrier: adding attapulgite into a mixed solution of nitric acid with the weight percentage content of 20% and hydrogen peroxide with the weight percentage content of 5%, preparing slurry with the weight percentage concentration of 20%, heating, stirring, cooling, washing, drying and calcining to obtain activated attapulgite;
(2) Inoculating bacillus megatherium, pseudomonas glutinosa and aspergillus niger into an LB culture medium respectively, carrying out shake culture at 28-30 ℃ until the concentration of the bacteria is OD600 apprxeq 2.5, and then mixing according to the volume ratio of 1:1:1 to obtain a compound microbial agent;
(3) And fully mixing the microbial carrier and the composite microbial agent, then adding the water-retaining agent and the activated humic acid, fully mixing and uniformly stirring, granulating and drying.
The microbial agent of the invention is used in an amount of 30-50 kg/mu.
Unlike nitrogen sources, there is no source of phosphorus in the atmosphere and it is not bioavailable to convert to available phosphorus. Therefore, the lack of available phosphorus in the soil severely restricts the growth of crops and affects the yield of crops.
Under different soil and climatic conditions, the application of the phosphate solubilizing microorganism is considered as a measure which is economical and favorable for sustainable development of agriculture, because the application of the phosphate solubilizing microorganism can reduce the use of phosphate fertilizer and improve the utilization efficiency of phosphorus in soil. Therefore, phosphate-solubilizing microorganisms become a key to improving the efficiency of phosphorus utilization in soil.
Advantageous effects
(1) According to the invention, the attapulgite is subjected to activation treatment, nitric acid and hydrogen peroxide pretreatment, and under the combined action of high-temperature calcination, the porosity and specific surface area of the attapulgite are greatly improved, the composite microbial agent can be fully adsorbed, and the microorganism is protected, and meanwhile, the continuous and efficient action of the microorganism in the soil is assisted;
(2) According to the invention, three bacteria including bacillus megatherium, pseudomonas glutinosa and aspergillus niger are compounded to form the compound microbial agent, after the three bacteria are applied to the soil, the phosphorus dissolving effect is doubled, the secreted organic acid substances can convert indissolvable phosphorus in the soil into phosphate ions which can be absorbed and utilized by plants, and meanwhile, growth hormone, gibberellin, synthetic siderophores, antibiotics and disease-resistant active substances can be secreted, so that the stress resistance of crops is improved while nutrition is provided for the crops, and the three bacteria are used simultaneously to play a role of synergistic interaction;
(3) The invention also adds the humic acid activated by enzyme, so that the effective substances are fully dissolved out, and the invention can provide abundant nutrient substances for soil and activate the soil; a proper amount of water-retaining agent is added, so that soil moisture loss is reduced; the microbial agent can continuously and efficiently play a role in phosphate dissolving and growth promotion after being applied into soil, improve the micro-ecological environment of the soil, continuously maintain the soil fertility, remarkably improve the utilization rate of fertilizer, promote the yield and quality of crops and have remarkable economic benefit.
Drawings
FIG. 1 is a graph showing the effect of dissolving phosphorus in examples 1-2 of the present invention.
Detailed Description
The technical scheme of the present invention is further described below with reference to specific examples, but is not limited thereto.
Example 1
The microbial agent for promoting nutrient absorption is prepared from the following raw materials in parts by weight: 20 parts of microorganism carrier, 15 parts of compound microorganism microbial inoculum, 10 parts of humic acid and 3 parts of water-retaining agent.
The microbial carrier is activated attapulgite, the attapulgite is added into a mixed solution of nitric acid with the weight percentage of 20% and hydrogen peroxide with the weight percentage of 5%, the mixed solution is prepared into slurry with the weight percentage of 20%, and the activated attapulgite is obtained through heating, stirring, cooling, washing, drying and calcining.
The compound microbial agent is obtained by mixing bacillus megatherium, pseudomonas glutinosa and aspergillus niger cultures according to the volume ratio of 1:1:1.
The preservation number of the bacillus megaterium is CGMCC1.16094, the preservation number of the pseudomonas glutinosa is CGMCC 1.15627, and the preservation number of the aspergillus niger is CCTCC HF 2008599.
The bacillus megatherium (Bacillus megaterium) is purchased from China general microbiological culture collection center (China General Microbiological Culture Collection Center, CGMCC), and has the following addresses: beijing city, chaoyang district, north Chen Xili No. 1, 3, date of preservation: 2017, 3, 11 days, deposit number: CGMCC1.16094.
The Pseudomonas glutinosa (Pseudomonas guguanensis) is purchased from China general microbiological culture collection center (China General Microbiological Culture Collection Center, CGMCC), and has the following addresses: beijing city, chaoyang district, north Chen Xili No. 1, 3, date of preservation: 25 days of 2 months 2015, deposit number: CGMCC 1.15627.
Aspergillus niger (Aspergillus) of the invention is purchased from China center for type culture Collection (university of Wuhan), eight-way 299 No. of Wuchang district of Wuhan, hubei province, and the preservation number is CCTCC HF 2008599.
The preparation method of the composite microbial agent comprises the following steps: inoculating Bacillus megaterium, pseudomonas glutinosa and Aspergillus niger into LB culture medium, shake culturing at 28-30deg.C until the bacterial concentration is OD600 ≡2.5, and mixing at volume ratio of 1:1:1.
The composition of the LB culture medium is as follows: 10g of peptone, 5g of yeast powder, 10g of NaCl, 1000mL of distilled water, pH value of 7.2-7.4 and sterilizing at 121 ℃ for 19min.
The humic acid is the humic acid which is subjected to biological activation, and the activation method comprises the following steps: regulating humic acid humidity to 20-30%, uniformly spraying cellulose enzyme solution with mass concentration of 20%, stirring uniformly, and standing for 6 h.
The water-retaining agent is polyglutamic acid.
A preparation method of a microbial agent for promoting nutrient absorption comprises the following steps:
(1) Preparation of a microbial carrier: adding attapulgite into a mixed solution of nitric acid with the weight percentage content of 20% and hydrogen peroxide with the weight percentage content of 5%, preparing slurry with the weight percentage concentration of 20%, heating, stirring, cooling, washing, drying and calcining to obtain activated attapulgite;
(2) Inoculating bacillus megatherium, pseudomonas glutinosa and aspergillus niger into an LB culture medium respectively, carrying out shake culture at 28-30 ℃ until the concentration of the bacteria is OD600 apprxeq 2.5, and then mixing according to the volume ratio of 1:1:1 to obtain a compound microbial agent;
(3) And fully mixing the microbial carrier and the composite microbial agent, then adding the water-retaining agent and the activated humic acid, fully mixing and uniformly stirring, granulating and drying.
Example 2
The microbial agent for promoting nutrient absorption is prepared from the following raw materials in parts by weight: 30 parts of microorganism carrier, 20 parts of composite microorganism microbial inoculum, 15 parts of humic acid and 5 parts of water-retaining agent.
The microbial carrier is activated attapulgite, the attapulgite is added into a mixed solution of nitric acid with the weight percentage of 20% and hydrogen peroxide with the weight percentage of 5%, the mixed solution is prepared into slurry with the weight percentage of 20%, and the activated attapulgite is obtained through heating, stirring, cooling, washing, drying and calcining.
The compound microbial agent is obtained by mixing bacillus megatherium, pseudomonas glutinosa and aspergillus niger cultures according to the volume ratio of 1:1:1.
The preservation number of the bacillus megaterium is CGMCC1.16094, the preservation number of the pseudomonas glutinosa is CGMCC 1.15627, and the preservation number of the aspergillus niger is CCTCC HF 2008599.
The bacillus megatherium (Bacillus megaterium) is purchased from China general microbiological culture collection center (China General Microbiological Culture Collection Center, CGMCC), and has the following addresses: beijing city, chaoyang district, north Chen Xili No. 1, 3, date of preservation: 2017, 3, 11 days, deposit number: CGMCC1.16094.
The Pseudomonas glutinosa (Pseudomonas guguanensis) is purchased from China general microbiological culture collection center (China General Microbiological Culture Collection Center, CGMCC), and has the following addresses: beijing city, chaoyang district, north Chen Xili No. 1, 3, date of preservation: 25 days of 2 months 2015, deposit number: CGMCC 1.15627.
Aspergillus niger (Aspergillus) of the invention is purchased from China center for type culture Collection (university of Wuhan), eight-way 299 No. of Wuchang district of Wuhan, hubei province, and the preservation number is CCTCC HF 2008599.
The preparation method of the composite microbial agent comprises the following steps: inoculating Bacillus megaterium, pseudomonas glutinosa and Aspergillus niger into LB culture medium, shake culturing at 28-30deg.C until the bacterial concentration is OD600 ≡2.5, and mixing at volume ratio of 1:1:1.
The composition of the LB culture medium is as follows: 10g of peptone, 5g of yeast powder, 10g of NaCl, 1000mL of distilled water, pH value of 7.2-7.4 and sterilizing at 121 ℃ for 19min.
The humic acid is the humic acid which is subjected to biological activation, and the activation method comprises the following steps: regulating humic acid humidity to 20-30%, uniformly spraying cellulose enzyme solution with mass concentration of 20%, stirring uniformly, and standing for 10 h.
The water-retaining agent is water-absorbent resin.
A preparation method of a microbial agent for promoting nutrient absorption comprises the following steps:
(1) Preparation of a microbial carrier: adding attapulgite into a mixed solution of nitric acid with the weight percentage content of 20% and hydrogen peroxide with the weight percentage content of 5%, preparing slurry with the weight percentage concentration of 20%, heating, stirring, cooling, washing, drying and calcining to obtain activated attapulgite;
(2) Inoculating bacillus megatherium, pseudomonas glutinosa and aspergillus niger into an LB culture medium respectively, carrying out shake culture at 28-30 ℃ until the concentration of the bacteria is OD600 apprxeq 2.5, and then mixing according to the volume ratio of 1:1:1 to obtain a compound microbial agent;
(3) And fully mixing the microbial carrier and the composite microbial agent, then adding the water-retaining agent and the activated humic acid, fully mixing and uniformly stirring, granulating and drying.
Comparative example
The composition of the microbial agent is changed, and comparative examples are respectively set to verify the experimental effect, wherein the volume ratio of the microbial agent to the comparative examples is shown in table 1:
table 1 comparative example settings
| Comparative example | Bacillus megaterium | Pseudomonas glutinosa | Aspergillus niger |
| 1 | 1 | 2 | 1 |
| 2 | 1 | 1 | 2 |
| 3 | 2 | 1 | 1 |
| 4 | 1 | 1 | 0 |
| 5 | 1 | 0 | 1 |
| 6 | 0 | 1 | 1 |
| 7 | 1 | 0 | 0 |
| 8 | 0 | 1 | 0 |
| 9 | 0 | 0 | 1 |
Test example 1
And (3) testing the phosphate dissolving effect of the bacterial liquid:
100 mu L of the bacterial suspension obtained in the step 2 in each preparation method is sucked by a pipette, and is injected into an inorganic phosphorus culture medium, and after 7d of dark culture at 28 ℃, the diameter of the phosphate solubilizing transparent ring is measured in a crisscross manner.
Inorganic phosphorus medium: (NH) 4 ) 2 SO 4 0.5g，MgSO 4 ·7H 2 0.3g of O, KCl0.3g, naCl0.3g, 5g of calcium phosphate, 10g of glucose and MnSO 4 ·H 2 O0.3g，FeSO 4 ·7H 2 0.3g of O, 20g of agar, 1000mL of distilled water and pH7.2, and the liquid culture medium is obtained without adding agar.
TABLE 2 dephosphorization effect
| Test group | Diameter cm of transparent ring |
| 1 | 2.41±0.1bc |
| 2 | 2.52±0.1a |
| 1 | 1.91±0.22e |
| 2 | 1.95±0.3ab |
| 3 | 1.90±0.1a |
| 4 | 1.51±0.3bc |
| 5 | 1.58±0.1ab |
| 6 | 1.50±0.2d |
| 7 | 1.41±0.1d |
| 8 | 1.43±0.3ab |
| 9 | 1.44±0.1ac |
Note that: data in the table are mean ± standard deviation. The different letters in the same column indicate that the difference is significant at P < 0.05 levels as tested by LSD.
As is apparent from FIG. 1, the phosphorus solubilizing ring is evident from examples 1 (A) and 2 (B) of the present invention.
Planting test
The test is carried out in the agricultural test garden in the temporary county of Yiyi city of Shandong province in 1-6 months in 2020, the soil of the test base is mainly sand and wind soil, and the average volume weight of the soil layer of 0-100 cm is 1.58g/cm 3 The pH value is 6.5, the organic matter content is 8.63g/kg, the alkaline hydrolysis nitrogen content is 25mg/kg, the available phosphorus content is 56.6mg/kg, the quick-acting potassium content is 115mg/kg, and the conductivity value is 456 mu S/cm.
Test material
The tomato variety to be tested is nong song 520, the tomato is planted in 1 month and 1 day, the top is cut off in 20 months, the tomato is harvested in 28 months, and the seedling is pulled out in 10 months and 6 months. The planting mode is ridging and film covering, and the plant row spacing is 20cm multiplied by 40cm.
Test design
The test consisted of 10 treatment groups, each:
s1, example 2 microbial agent+conventional fertilization;
S2-S10 comparative examples 1-9 microbial inoculant+conventional fertilization;
the application amount is 30 kg/mu, and the organic-inorganic compound fertilizer (the organic matter is more than or equal to 20% and N-P) is applied by conventional fertilization 2 O 5 -K 2 15-5-10) 2000kg/hm 2 The microbial inoculum is applied along with conventional fertilization.
Content and method of measurement
The measurement of the growth index was carried out during the third ear enlargement period (25 days of 4 months) of the tomato.
(1) Plant height and stem thickness: 6 plants with consistent growth vigor are selected for each treatment, and the plant height (from the stem base to the growth point) is measured by using a ruler; the stem thickness (the stem at the first petiole from the ground) was measured with a digital vernier caliper.
(2) Leaf Area Index (LAI): each treatment destructive 3 plants were taken, leaf area of the whole plant was measured by a high-speed imaging instrument, and Leaf Area Index (LAI) was obtained by converting the leaf area of the single plant and the occupied land area of the single plant.
(3) Blade SPAD value: 6 plants with consistent growth vigor are selected for each treatment, the petioles between the first inflorescence and the second inflorescence are selected, the SPAD value of 6 fully-unfolded leaves on the petioles is measured by a SPAD-502 analyzer, and the average value is obtained.
Method for measuring yield and quality
(1) In the mature period of the fruits, the mature fruits in each district are timely collected, the tomato yield in the whole harvest period is counted and converted into kg/hm 2 。
(2) Three normal fruits were selected for quality measurement per treatment during the full harvest period of tomato fruits. The soluble solids were measured using an ATAGO PAL-3 digital handheld glycometer, the vitamin C content was measured using a molybdenum-blue ratio method, and the experimental results are shown in table 3:
TABLE 3 tomato planting experiment results
It should be noted that the above-mentioned embodiments are merely some, but not all embodiments of the preferred mode of carrying out the invention. It is evident that all other embodiments obtained by a person skilled in the art without making any inventive effort, based on the above-described embodiments of the invention, shall fall within the scope of protection of the invention.
Claims (4)
1. The microbial agent for promoting nutrient absorption is characterized by comprising the following raw materials in parts by weight: 20-30 parts of microorganism carrier, 15-20 parts of compound microorganism microbial inoculum, 10-15 parts of humic acid and 3-5 parts of water-retaining agent;
the microbial carrier is activated attapulgite, the attapulgite is added into a mixed solution of nitric acid with the weight percentage of 20 percent and hydrogen peroxide with the weight percentage of 5 percent to prepare slurry with the weight percentage of 20 percent, and the activated attapulgite is obtained through heating, stirring, cooling, washing, drying and calcining;
the compound microbial agent is obtained by mixing bacillus megatherium, pseudomonas glutinosa and aspergillus niger cultures according to the volume ratio of 1:1:1;
the preservation number of the bacillus megatherium is CGMCC1.16094, the preservation number of the pseudomonas glutinosa is CGMCC 1.15627, and the preservation number of the aspergillus niger is CCTCC HF 2008599;
the preparation method of the composite microbial agent comprises the following steps: inoculating Bacillus megaterium, pseudomonas glutinosa and Aspergillus niger into LB culture medium, shake culturing at 28-30deg.C until the bacterial concentration is OD600 ≡2.5, and mixing at volume ratio of 1:1:1.
2. The microbial agent for promoting nutrient absorption according to claim 1, wherein the humic acid is biologically activated humic acid, and the activation method comprises the following steps: regulating humic acid humidity to 20-30%, uniformly spraying cellulose enzyme solution with mass concentration of 20%, uniformly stirring, and standing for 6-10 h.
3. The microbial agent for promoting nutrient absorption according to claim 1, wherein the water-retaining agent is one of starch, polyglutamic acid or a water-absorbent resin.
4. A method for preparing a microbial agent for promoting nutrient absorption as claimed in any one of claims 1 to 3, comprising the steps of:
(1) Preparation of a microbial carrier: adding attapulgite into a mixed solution of nitric acid with the weight percentage content of 20% and hydrogen peroxide with the weight percentage content of 5%, preparing slurry with the weight percentage concentration of 20%, heating, stirring, cooling, washing, drying and calcining to obtain activated attapulgite;
(2) Inoculating bacillus megatherium, pseudomonas glutinosa and aspergillus niger into an LB culture medium respectively, carrying out shake culture at 28-30 ℃ until the concentration of the bacteria is OD600 apprxeq 2.5, and then mixing according to the volume ratio of 1:1:1 to obtain a compound microbial agent;
(3) And fully mixing the microbial carrier and the composite microbial agent, then adding the water-retaining agent and the activated humic acid, fully mixing and uniformly stirring, granulating and drying.
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