CN114836325A - Phycomycetes composite remediation liquid for increasing quick-acting phosphorus content of saline-alkali soil and application - Google Patents
Phycomycetes composite remediation liquid for increasing quick-acting phosphorus content of saline-alkali soil and application Download PDFInfo
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- 239000002689 soil Substances 0.000 title claims abstract description 135
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 title claims abstract description 55
- 229910052698 phosphorus Inorganic materials 0.000 title claims abstract description 55
- 239000011574 phosphorus Substances 0.000 title claims abstract description 55
- 239000003513 alkali Substances 0.000 title claims abstract description 44
- 239000002131 composite material Substances 0.000 title claims abstract description 41
- 239000007788 liquid Substances 0.000 title claims abstract description 40
- 238000005067 remediation Methods 0.000 title claims abstract description 21
- 230000001965 increasing effect Effects 0.000 title claims description 23
- 241000894006 Bacteria Species 0.000 claims abstract description 35
- 241000195493 Cryptophyta Species 0.000 claims abstract description 29
- 238000000034 method Methods 0.000 claims abstract description 21
- 241000192656 Nostoc Species 0.000 claims description 16
- 241000190967 Rhodospirillum Species 0.000 claims description 16
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 10
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 claims description 9
- 238000002360 preparation method Methods 0.000 claims description 8
- 239000001963 growth medium Substances 0.000 claims description 7
- 238000003973 irrigation Methods 0.000 claims description 7
- 230000002262 irrigation Effects 0.000 claims description 7
- 238000009630 liquid culture Methods 0.000 claims description 7
- 238000005286 illumination Methods 0.000 claims description 6
- 239000003621 irrigation water Substances 0.000 claims description 6
- 239000000203 mixture Substances 0.000 claims description 6
- 230000008569 process Effects 0.000 claims description 6
- 238000001914 filtration Methods 0.000 claims description 4
- 239000002344 surface layer Substances 0.000 claims description 4
- 241001219477 Chroococcus Species 0.000 claims description 3
- 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 claims description 3
- 239000001888 Peptone Substances 0.000 claims description 3
- 108010080698 Peptones Proteins 0.000 claims description 3
- FRHBOQMZUOWXQL-UHFFFAOYSA-L ammonium ferric citrate Chemical compound [NH4+].[Fe+3].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O FRHBOQMZUOWXQL-UHFFFAOYSA-L 0.000 claims description 3
- 230000003321 amplification Effects 0.000 claims description 3
- 150000001875 compounds Chemical class 0.000 claims description 3
- 238000012258 culturing Methods 0.000 claims description 3
- 229960004642 ferric ammonium citrate Drugs 0.000 claims description 3
- 239000008103 glucose Substances 0.000 claims description 3
- 239000004313 iron ammonium citrate Substances 0.000 claims description 3
- 235000000011 iron ammonium citrate Nutrition 0.000 claims description 3
- 238000003199 nucleic acid amplification method Methods 0.000 claims description 3
- 235000015097 nutrients Nutrition 0.000 claims description 3
- 235000019319 peptone Nutrition 0.000 claims description 3
- 239000000126 substance Substances 0.000 claims description 3
- 239000012530 fluid Substances 0.000 claims 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 abstract description 20
- 150000004676 glycans Chemical class 0.000 abstract description 11
- 229920001282 polysaccharide Polymers 0.000 abstract description 11
- 239000005017 polysaccharide Substances 0.000 abstract description 11
- 229910052757 nitrogen Inorganic materials 0.000 abstract description 10
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 abstract description 8
- 229910052799 carbon Inorganic materials 0.000 abstract description 8
- 230000008439 repair process Effects 0.000 abstract description 5
- 239000002207 metabolite Substances 0.000 abstract description 2
- 229910002651 NO3 Inorganic materials 0.000 abstract 6
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 abstract 6
- 230000007935 neutral effect Effects 0.000 abstract 1
- 239000000618 nitrogen fertilizer Substances 0.000 abstract 1
- 239000003895 organic fertilizer Substances 0.000 abstract 1
- 150000002897 organic nitrogen compounds Chemical class 0.000 abstract 1
- 210000004027 cell Anatomy 0.000 description 22
- 230000000694 effects Effects 0.000 description 15
- 229910019142 PO4 Inorganic materials 0.000 description 14
- 239000010452 phosphate Substances 0.000 description 14
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 13
- 239000002253 acid Substances 0.000 description 9
- 102000004160 Phosphoric Monoester Hydrolases Human genes 0.000 description 7
- 108090000608 Phosphoric Monoester Hydrolases Proteins 0.000 description 7
- 241000192700 Cyanobacteria Species 0.000 description 5
- 150000001768 cations Chemical class 0.000 description 5
- 241000186660 Lactobacillus Species 0.000 description 4
- 239000013522 chelant Substances 0.000 description 4
- 229940039696 lactobacillus Drugs 0.000 description 4
- 244000005700 microbiome Species 0.000 description 4
- 239000002686 phosphate fertilizer Substances 0.000 description 4
- 230000002053 acidogenic effect Effects 0.000 description 3
- 230000007613 environmental effect Effects 0.000 description 3
- 230000029553 photosynthesis Effects 0.000 description 3
- 238000010672 photosynthesis Methods 0.000 description 3
- 150000003839 salts Chemical class 0.000 description 3
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 238000013329 compounding Methods 0.000 description 2
- 238000000855 fermentation Methods 0.000 description 2
- 230000004151 fermentation Effects 0.000 description 2
- 239000003337 fertilizer Substances 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 239000005416 organic matter Substances 0.000 description 2
- 230000000413 phospholytic effect Effects 0.000 description 2
- 230000001737 promoting effect Effects 0.000 description 2
- 230000002195 synergetic effect Effects 0.000 description 2
- 241000304886 Bacilli Species 0.000 description 1
- 239000002028 Biomass Substances 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- BHPQYMZQTOCNFJ-UHFFFAOYSA-N Calcium cation Chemical compound [Ca+2] BHPQYMZQTOCNFJ-UHFFFAOYSA-N 0.000 description 1
- 238000008157 ELISA kit Methods 0.000 description 1
- 229920002444 Exopolysaccharide Polymers 0.000 description 1
- 241001478892 Nostoc sp. PCC 7120 Species 0.000 description 1
- 102000045595 Phosphoprotein Phosphatases Human genes 0.000 description 1
- 108700019535 Phosphoprotein Phosphatases Proteins 0.000 description 1
- 241000605861 Prevotella Species 0.000 description 1
- 241000190984 Rhodospirillum rubrum Species 0.000 description 1
- 241000736131 Sphingomonas Species 0.000 description 1
- 238000012271 agricultural production Methods 0.000 description 1
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 description 1
- 229910052782 aluminium Inorganic materials 0.000 description 1
- 230000001651 autotrophic effect Effects 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 229910001424 calcium ion Inorganic materials 0.000 description 1
- 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
- 210000005056 cell body Anatomy 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000004720 fertilization Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 229910052742 iron Inorganic materials 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- -1 organic matters Substances 0.000 description 1
- OQUKIQWCVTZJAF-UHFFFAOYSA-N phenol;sulfuric acid Chemical compound OS(O)(=O)=O.OC1=CC=CC=C1 OQUKIQWCVTZJAF-UHFFFAOYSA-N 0.000 description 1
- 230000000243 photosynthetic effect Effects 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 239000004016 soil organic matter Substances 0.000 description 1
- 239000007921 spray Substances 0.000 description 1
- 238000010561 standard procedure Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 239000003053 toxin Substances 0.000 description 1
- 231100000765 toxin Toxicity 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
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/12—Unicellular algae; Culture media therefor
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- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K17/00—Soil-conditioning materials or soil-stabilising materials
- C09K17/14—Soil-conditioning materials or soil-stabilising materials containing organic compounds only
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- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- C09K2109/00—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE pH regulation
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- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
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Abstract
The invention belongs to the field of soil treatment, and particularly relates to an algae bacterium composite remediation liquid for improving the content of available phosphorus in saline-alkali soil and application thereof. The method is characterized in that the crophaera is adopted to repair acidified nitrate polluted cultivated soil, the crophaera can grow and reproduce by taking nitrate as a nitrogen source, the nitrate is converted into an organic nitrogen compound to provide a slow-release organic fertilizer, meanwhile, extracellular polysaccharide secreted by the crophaera can be used as a carbon source of denitrifying bacteria to improve the denitrifying performance of the denitrifying bacteria, and the nitrate is converted into nitrogen to be removed through the denitrifying action, so that the nitrate pollution is reduced; the chromococcus secretes alkaline metabolites to increase the pH value of the soil to be neutral, and is particularly suitable for an acidified soil environment with accumulated nitrate caused by applying a large amount of nitrogen fertilizer.
Description
Technical Field
The invention belongs to the field of soil treatment, and particularly relates to application of an algae-bacteria composite remediation liquid in improving the content of available phosphorus in saline-alkali soil.
Background
The information in this background section is only for enhancement of understanding of the general background of the invention and is not necessarily to be construed as an admission or any form of suggestion that this information forms the prior art that is already known to a person of ordinary skill in the art.
The total phosphorus content in soil is usually high, and can reach thousands of milligrams per kilogram of soil, but more than 95 percent of phosphorus in the soil exists in the form of organic phosphorus or insoluble phosphate and cannot be directly absorbed and utilized by plants. The content of soluble quick-acting phosphorus easily absorbed and utilized by crops in soil is usually only 10-20 mg/kg.
Large areas of salinized soil exist worldwide, including both primary salinized soil and secondary salinized soil due to over fertilization and abuse. The contents of calcium, aluminum, iron and other cations in the saline-alkali soil are increased, the binding capacity with phosphate radicals is enhanced, the content of insoluble phosphate is increased, the content of quick-acting phosphorus in the soil is further reduced, and the content of the quick-acting phosphorus in the saline-alkali soil can be reduced to be below 5 mg/kg.
Although the quick-acting phosphorus content of soil can be improved in a short period by applying a phosphate fertilizer in the agricultural production process, only 10-25% of the phosphate fertilizer can be effectively utilized, and if the soil has a salinization phenomenon, the utilization rate of the phosphate fertilizer is further reduced. Phosphate radicals which are not utilized in time in the phosphate fertilizer are fixed by cations in the soil to form insoluble phosphate, so that the salinization degree of the soil is further increased, and the steps are repeated to form a vicious circle. Therefore, the capability of releasing endogenous phosphorus elements in the soil is improved, the application of phosphate fertilizers is reduced, the method has important significance for preventing and treating the salinized soil, and the sustainable utilization of the cultivated soil is facilitated.
The quick-acting phosphorus content of the soil can be improved by independently adding microorganisms such as blue algae or bacteria, but the improving effect is reduced or even disappears along with the increase of the salinity and the pH value of the soil.
Disclosure of Invention
In order to solve the defects of the prior art, the invention provides aThe application of the phycomycete composite repairing liquid in improving the content of available phosphorus in saline-alkali soil. The algae-bacteria composite repairing liquid consists of chromococcus, nostoc, rhodospirillum and water. The chromococcus and nostoc are nontoxic blue algae with good tolerance to salinity and alkalinity, do not produce blue algae toxin, and can secrete extracellular polysaccharide to chelate calcium ions in soil after being applied to the soil, so that phosphate ions are released from insoluble calcium phosphate, and the salinity of the soil can be reduced; nostoc has nitrogen fixation function and can provide nitrogen source for growth of Nostoc and Chroococci; the rhodospirillum has health-preserving function, on one hand, heterotrophic life is carried out by taking soil organic matters and blue-green algae exopolysaccharide as carbon sources, acid is produced through fermentation to reduce the pH value of soil, and a good environment is provided for the survival of the blue-green algae, on the other hand, autotrophic photosynthesis can be carried out together with the blue-green algae, and CO is generated 2 The phosphorus-dissolving bacteria are converted into soil organic matters to provide a carbon source for the growth of the phosphorus-dissolving bacteria, and the activity of the phosphorus-dissolving bacteria in the soil is improved, so that the release of phosphorus in the soil is promoted. The chromococcus, nostoc and rhodospirillum have synergistic effect, and can effectively improve the content of soluble quick-acting phosphorus in the salinized soil.
In order to achieve the above purpose, the present invention specifically provides the following technical solutions:
the invention provides a phycomycete composite repairing liquid for improving quick-acting phosphorus content of saline-alkali soil, which comprises the following components: nostoc, chromococcus, rhodospirillum and water.
The method for adding blue algae and bacteria in a compounding manner can improve the tolerance of microorganisms to the soil salinity and alkalinity and ensure the restoration effect in the salinized soil.
The second aspect of the invention provides a preparation method of an algae bacterium composite remediation liquid for increasing the content of available phosphorus in saline-alkali soil, which comprises the following steps:
inoculating nostoc, chromococcus and rhodospirillum to a liquid culture medium;
culturing the mixture of phycomycetes with an amplification culture device until the total cell density is 8 × 10 9 ～10×10 9 cells/mL, filtering and collecting algal cells;
and (4) resuspending the filtered and collected phycomycete cells in irrigation water to prepare the phycomycete composite repairing liquid.
The third aspect of the invention provides a method for increasing the content of available phosphorus in saline-alkali soil by using the phycomycete composite repairing liquid, which comprises the following steps:
and adding the phycomycete composite repairing liquid to the surface layer of saline-alkali soil, and repairing under natural illumination.
One or more embodiments of the present invention have at least the following advantageous effects:
(1) the chromococcus and nostoc secrete extracellular polysaccharide to chelate cations in soil, reduce salinity, inhibit the generation of insoluble phosphate and promote the release of phosphorus; the rhodospirillum heterotrophic fermentation blue algae extracellular polysaccharide and soil organic matter produce acid to reduce the pH value of the soil; the chromococcus, nostoc and rhodospirillum fix carbon through photosynthesis, provide carbon sources for acid-producing bacteria in soil, promote the growth and the propagation of the acid-producing bacteria in the soil, particularly promote the propagation of lactobacillus in the soil, and the lactobacillus can not only produce acid and dissolve phosphate, but also secrete phosphatase to hydrolyze organic phosphorus in the soil. After the algae bacterium compound preparation is applied, the activity of the soil phosphatase is greatly increased, the pH value and the salinity are greatly reduced, and the content of the quick-acting phosphorus in the soil is obviously increased.
(2) The preparation method is simple, convenient to operate, high in practicability and easy to popularize.
Detailed Description
It is to be understood that the following detailed description is exemplary and is intended to provide further explanation of the invention as claimed. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.
As described in the background art, when the content of the available phosphorus in the soil is increased by adding a single type of microorganism, the increasing effect is reduced or even disappears along with the increase of the salinity and the pH value of the soil. Therefore, the method for adding the blue algae and the bacteria in a compounding manner can improve the tolerance of the microorganisms to the soil salinity and alkalinity and ensure the remediation effect in the salinized soil.
In order to solve the technical problems, the invention provides an application of an algae-bacteria composite remediation solution in improving the content of available phosphorus in saline-alkali soil.
The blue algae can promote soil to release quick-acting phosphorus through two ways, namely direct and indirect, firstly secrete extracellular polysaccharide to directly chelate cations in insoluble phosphate, and secondly provide a carbon source for phosphorus-dissolving bacteria through photosynthetic carbon fixation, so that the activity of the phosphorus-dissolving bacteria is improved. However, when the salinity of the soil exceeds the tolerance range of the blue algae, the blue algae cannot grow or even die in the soil and cannot play a role in releasing phosphorus.
The chromococcus can tolerate extreme environments such as high salinity, low temperature, drought and the like, is blue algae capable of living in saline-alkali soil, and has a good phosphorus release effect, but the chromococcus has limitations when being applied to the saline-alkali soil. Firstly, although the chromococcus can live in the saline-alkali soil, the chromococcus can secrete alkaline metabolites to gradually increase the pH value of the saline-alkali soil and reduce the phosphorus release effect; secondly, saline-alkali soil is generally quite barren, the nitrogen content in the soil is very low, a nitrogen source needs to be consumed in the growth process of the chromococcus, but the chromococcus itself does not have the nitrogen fixation function, and the chromococcus will die gradually when the nitrogen in the soil is completely consumed by the chromococcus.
The inventor finds that the compound microbial preparation prepared from nostoc, rhodospirillum and chromococcus can obviously improve the content of soluble and quick-acting phosphorus in saline-alkali soil, and the specific expression is as follows:
the nostoc provides a nitrogen source for the growth of the chromococcus through nitrogen fixation, secretes blue algae extracellular polysaccharide together with the chromococcus to reduce the salinity of soil and chelate cations in insoluble phosphate, and promotes the release of free phosphate;
the rhodospirillum can heterotrophically ferment soil organic matters and blue-green algae extracellular polysaccharide, produce acid and reduce the pH value of the soil, thereby creating a good environment for the growth of the blue-green algae;
the three components have synergistic effect, and can be used for fixing carbon through photosynthesis, increasing the content of organic matters in soil, promoting the reproduction of acid-producing and phosphorus-dissolving bacteria represented by lactobacillus in the soil, reducing the pH value of the soil, improving the activity of soil phosphatase, and promoting the conversion of insoluble phosphate and organic phosphorus in the soil to soluble and quick-acting phosphorus.
Further, the saline-alkali soil includes primary saline-alkali soil and secondary saline-alkali soil.
Furthermore, the salt content of the soil with the surface layer of 0-30 cm of the saline-alkali soil is 0.1-0.6%, and the pH value is 8.0-10.0.
Further, the initial quick-acting phosphorus content in the saline-alkali soil is less than 10 mg/kg.
The invention provides a algae-bacteria composite repairing liquid in a second aspect, and the specific preparation method comprises the following steps:
(1) inoculating Nostoc, Chroococcus and Rhodospirillum in liquid culture medium, wherein the initial cell density of the Nostoc, Chroococcus and Rhodospirillum is 1 × 10 6 ～3×10 6 cells/mL, preferably all three, have an initial cell density of 1X 10 6 ～2×10 6 cells/mL, most preferably, all three have an initial cell density of 2X 10 6 cells/mL；
(2) Culturing the phycomycete mixture by using an amplification culture device under the culture conditions of the temperature of 25-30 ℃ and the illumination intensity of 2000-2500 lx until the total cell density is 8 multiplied by 10 9 ～10×10 9 cells/mL, filtering and collecting phycomycete cells by using a 0.22 mu m filter membrane;
(3) resuspending the filtered and collected phycomycete cells in irrigation water to obtain a product with a total cell density of 1-2 × 10 8 cells/mL of algae bacterium composite repairing liquid.
Further, the liquid culture medium is prepared by adding nutrient substances into irrigation water, and the concentration of the nutrient substances in the culture medium is as follows: k is 2 HPO 4 0.04g/L、NaNO 3 1.5g/L、MgSO 4 0.06g/L、CaCl 2 0.03g/L, 0.006g/L citric acid, 0.006g/L, EDTANa ferric ammonium citrate 2 0.001g/L、Na 2 CO 3 0.02g/L, glucose 5g/L and peptone 5 g/L.
Furthermore, the phycomycete expanding culture device comprises a conical flask, an open cell body or a photobioreactor.
The third aspect of the invention provides a method for increasing the content of available phosphorus in saline-alkali soil by using the phycomycete composite repairing liquid, which comprises the following specific steps:
the algae bacterium composite repairing liquid is added on the surface layer of the saline-alkali soil, and the repairing is carried out under the natural illumination.
The adding mode of the phycomycete composite repairing liquid is adding by a water fertilizer machine, drip irrigation, spray irrigation or manual irrigation.
Further, in the repairing process, the environmental temperature is 5-35 ℃, preferably 20-30 ℃, and further preferably 20-25 ℃.
Further, the repairing time is 40-80 days.
Further, 0.5-1.5 mL of algae bacterium composite repairing liquid is added to each square centimeter of soil;
preferably, 0.8-1.2 mL of algae bacterium composite repairing liquid is added to each square centimeter of soil;
further preferably, 1.0mL of the algae bacterium composite repairing liquid is added to each square centimeter of soil.
Further, after the algae bacterium composite repairing liquid is added to the soil surface, the water content of the soil is kept between 20% and 30% through irrigation.
In order to make the technical solutions of the present invention more clearly understood by those skilled in the art, the technical solutions of the present invention will be described in detail below with reference to specific embodiments.
Example 1
An algae-bacteria composite repairing liquid:
selecting a common chromococcus sp.FACHB-193 and a common Nostoc sp.PCC 7120, wherein the collection unit is the institute of aquatic organisms of Chinese academy of sciences; a common rhodospirillum (Rhodospirillumrubrum) is selected, and the preservation unit is the institute of microbiology of Chinese academy of sciences.
The respective inoculation densities of two blue algae and one rhodospirillum in a liquid culture medium are both 2 multiplied by 10 6 cells/mL。
The liquid medium had the following composition: k 2 HPO 4 0.04g/L、NaNO 3 1.5g/L、MgSO 4 0.06g/L、CaCl 2 0.03g/L, 0.006g/L citric acid, 0.006g/L, EDTANa ferric ammonium citrate 2 0.001g/L、Na 2 CO 3 0.02g/L, glucose 5g/L and peptone 5 g/L.
Culture strip with temperature of 25 ℃ and illumination intensity of 2000lxUnder the condition, the mixture of the phycomycetes is enlarged and cultured by using a conical flask until the total cell density is 10 multiplied by 10 9 cells/mL, filtering with 0.22 μm filter membrane to collect phycomycete cells, adding irrigation water to resuspend blue algae and bacteria, and making into cell with density of 2 × 10 8 cells/mL of algae bacterium composite repairing liquid.
Example 2
The method for improving the content of the available phosphorus in the saline-alkali soil by adopting the phycomycete composite remediation solution in the embodiment 1 comprises the following steps:
and adding the algae bacterium composite repairing liquid into the open-air native saline-alkali soil for repairing. The adding amount of the algae bacterium composite repairing liquid on the soil surface is that 1.0mL of repairing liquid is added on each square centimeter of soil. The environmental temperature is 10-30 ℃, and the water content of the soil is maintained between 20% and 30% through irrigation. The repair time was 80 days. The pH of the soil before restoration is 9.5, the salt content is 0.4%, the quick-acting phosphorus content is 5mg/kg, the pH of the soil after restoration for 80 days is 7.8, the salt content is 0.3%, and the quick-acting phosphorus content is 35 mg/kg.
Example 3
The method for improving the content of the available phosphorus in the saline-alkali soil by adopting the phycomycete composite remediation solution in the embodiment 1 comprises the following steps:
and adding the algae bacterium composite repairing liquid into the open secondary saline-alkali soil for repairing. The adding amount of the algae bacterium composite repairing liquid on the soil surface is that 1.0mL of repairing liquid is added on each square centimeter of soil. The environmental temperature is 10-30 ℃, and the water content of the soil is maintained between 20% and 30% through irrigation. The repair time was 40 days. In the repairing process, soil samples are collected every 10 days, and the contents of total phosphorus, quick-acting phosphorus, organic matters, polysaccharide, pH, salinity and soil phosphatase activity in the soil are measured. And (3) determining the absolute abundance of bacteria in the soil before and after remediation on a genus level by using a metagenome sequencing technology.
The soil total phosphorus, the available phosphorus, the organic matter and the pH are measured by adopting a corresponding national standard method. The activity of soil phosphatase was measured using an ELISA kit. The soil salinity measuring method comprises the following steps: after 10g of saline-alkali soil is air-dried and sieved, 10mL of distilled water is added, the mixture is uniformly mixed and then is kept stand, and the salinity in the supernatant is measured by a salinity meter. The soil polysaccharide content is measured by a phenol-sulfuric acid method.
The change of each index with time in the process of repairing the saline-alkali earth in example 3 is shown in table 1.
TABLE 1 repair effect of phycomycete repair liquid on saline-alkali soil changes with time
The changes in absolute abundance of typical acidogenic and phospholytic bacteria before and after saline-alkaline earth remediation in example 3 are summarized in table 2.
TABLE 2 Absolute abundance Change of typical acidogenic and phospholytic bacteria before and after saline-alkali soil remediation
The content of the soil quick-acting phosphorus before the restoration is 10mg/kg, and the content of the soil quick-acting phosphorus after the restoration is 40 days is 45 mg/kg. The pH value and the salinity of the soil are obviously reduced, and the organic matter, the polysaccharide content and the phosphatase activity of the soil are obviously improved. Wherein the activity of the soil phosphatase is increased to 3.4 times of that before the remediation after 40 days of remediation. The biomass of sphingomonas, the wearer bacterium, the prevotella and two acidogenic bacilli in the soil is increased by hundreds of times, and the bacteria can ferment extracellular polysaccharide to produce acid, reduce the pH value of the soil and dissolve phosphate; the lactobacillus can produce acid and dissolve phosphate, and can secrete phosphatase to release organic phosphorus in the soil, so that the content of quick-acting phosphorus in the soil is greatly increased.
The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (10)
1. The utility model provides a promote compound restoration liquid of phycomycete of saline-alkali soil quick-acting phosphorus content which characterized in that includes: nostoc, chromococcus, rhodospirillum and water.
2. The phycomycete composite remediation fluid for increasing the available phosphorus content of saline-alkali soil as claimed in claim 1 with a total cell density of 1 x 10 8 ～2×10 8 cells/mL。
3. The phycomycete composite remediation fluid for increasing the available phosphorus content of saline-alkali soil of claim 1 wherein the water is irrigation water.
4. A preparation method of a phycomycete composite restoration liquid for improving the quick-acting phosphorus content of saline-alkali soil is characterized by comprising the following steps:
inoculating nostoc, chromococcus and rhodospirillum to a liquid culture medium;
culturing the mixture of phycomycetes with an amplification culture device until the total cell density is 8 × 10 9 ～10×10 9 cells/mL, filtering and collecting algal cells;
and (4) resuspending the filtered and collected phycomycete cells in irrigation water to prepare the phycomycete composite repairing liquid.
5. The preparation method of the phycomycete composite remediation liquid for increasing rapid-acting phosphorus content of saline-alkali soil as claimed in claim 4, wherein the concentration of nutrient substances in the liquid culture medium is as follows: k 2 HPO 4 0.04g/L、NaNO 3 1.5g/L、MgSO 4 0.06g/L、CaCl 2 0.03g/L, 0.006g/L citric acid, 0.006g/L, EDTANa ferric ammonium citrate 2 0.001g/L、Na 2 CO 3 0.02g/L, glucose 5g/L and peptone 5 g/L.
6. The method for preparing the phycomycete composite remediation liquid for increasing the available phosphorus content of saline-alkali soil as claimed in claim 4, wherein Nostoc, Chroococcus and Rhodospirillum are inoculated in the liquid culture mediumThe initial cell density is 1 × 10 6 ～3×10 6 cells/mL。
7. The preparation method of the phycomycete composite remediation liquid for increasing the available phosphorus content of saline-alkali soil as claimed in claim 4, wherein the specific conditions of the expanded culture are as follows: the temperature is 25-30 ℃, and the illumination intensity is 2000-2500 lx.
8. The method for preparing the phycomycete composite remediation liquid for improving the rapid-acting phosphorus content of saline-alkali soil as claimed in claim 4, wherein the expanding culture device comprises a conical flask, an open tank body and a photobioreactor.
9. A method for increasing the content of available phosphorus in saline-alkali soil by using the phycomycete composite remediation solution as claimed in any one of claims 1 to 3, the method comprising the following steps:
the phycomycete composite repairing liquid as set forth in any one of claims 1 to 3 is added to the surface layer of saline-alkali soil for repairing under natural illumination.
10. The method for increasing the content of available phosphorus in saline-alkali soil according to claim 9, wherein: the environment temperature in the repairing process is 5-35 ℃, preferably 20-30 ℃, and further preferably 20-25 ℃; the repairing time is 40-80 days;
or 0.5-1.5 mL of algae bacterium composite repairing liquid is added to soil per square centimeter;
or after the algae bacterium composite repairing liquid is added to the soil surface, the water content of the soil is kept between 20% and 30% through irrigation.
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