CN114836325B - Algae composite repair liquid for improving quick-acting phosphorus content of saline-alkali soil and application thereof - Google Patents
Algae composite repair liquid for improving quick-acting phosphorus content of saline-alkali soil and application thereof Download PDFInfo
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- 239000002689 soil Substances 0.000 title claims abstract description 138
- 241000195493 Cryptophyta Species 0.000 title claims abstract description 63
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 title claims abstract description 56
- 229910052698 phosphorus Inorganic materials 0.000 title claims abstract description 55
- 239000011574 phosphorus Substances 0.000 title claims abstract description 55
- 239000007788 liquid Substances 0.000 title claims abstract description 47
- 239000003513 alkali Substances 0.000 title claims abstract description 43
- 239000002131 composite material Substances 0.000 title claims abstract description 43
- 230000008439 repair process Effects 0.000 title claims description 36
- 241000894006 Bacteria Species 0.000 claims abstract description 30
- 241000195649 Chlorella <Chlorellales> Species 0.000 claims abstract description 25
- 238000000034 method Methods 0.000 claims description 20
- 241000190967 Rhodospirillum Species 0.000 claims description 17
- 241000192656 Nostoc Species 0.000 claims description 16
- 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
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 9
- 239000001963 growth medium Substances 0.000 claims description 8
- 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
- 239000000203 mixture Substances 0.000 claims description 7
- 238000005286 illumination Methods 0.000 claims description 6
- 239000003621 irrigation water Substances 0.000 claims description 6
- 238000002360 preparation method Methods 0.000 claims description 5
- 230000007613 environmental effect Effects 0.000 claims description 4
- 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
- 238000012258 culturing Methods 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
- 235000015097 nutrients Nutrition 0.000 claims description 3
- 235000019319 peptone Nutrition 0.000 claims description 3
- 239000000126 substance Substances 0.000 claims description 3
- 230000001580 bacterial effect Effects 0.000 claims description 2
- BHEPBYXIRTUNPN-UHFFFAOYSA-N hydridophosphorus(.) (triplet) Chemical compound [PH] BHEPBYXIRTUNPN-UHFFFAOYSA-N 0.000 claims 3
- 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 10
- 229910052757 nitrogen Inorganic materials 0.000 abstract description 10
- 229920001282 polysaccharide Polymers 0.000 abstract description 10
- 239000005017 polysaccharide Substances 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
- 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
- 235000021317 phosphate Nutrition 0.000 description 14
- 239000010452 phosphate Substances 0.000 description 12
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 11
- 239000002253 acid Substances 0.000 description 9
- 102000004160 Phosphoric Monoester Hydrolases Human genes 0.000 description 8
- 108090000608 Phosphoric Monoester Hydrolases Proteins 0.000 description 8
- 150000003839 salts Chemical class 0.000 description 7
- 150000001768 cations Chemical class 0.000 description 5
- 244000005700 microbiome Species 0.000 description 5
- 239000002686 phosphate fertilizer Substances 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
- 230000002053 acidogenic effect Effects 0.000 description 3
- 230000006872 improvement Effects 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 229920002444 Exopolysaccharide Polymers 0.000 description 2
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 2
- 150000001875 compounds Chemical class 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
- 150000003013 phosphoric acid derivatives Chemical class 0.000 description 2
- 230000029553 photosynthesis Effects 0.000 description 2
- 238000010672 photosynthesis Methods 0.000 description 2
- 238000004321 preservation Methods 0.000 description 2
- 238000005067 remediation Methods 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
- 241000222120 Candida <Saccharomycetales> Species 0.000 description 1
- 241000893563 Dysosma Species 0.000 description 1
- 238000008157 ELISA kit Methods 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 241000168525 Haematococcus Species 0.000 description 1
- 241000605861 Prevotella Species 0.000 description 1
- 241000736131 Sphingomonas Species 0.000 description 1
- 238000012271 agricultural production Methods 0.000 description 1
- 230000002353 algacidal effect Effects 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
- 230000009286 beneficial 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
- 238000013329 compounding Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 210000003608 fece Anatomy 0.000 description 1
- 230000004720 fertilization Effects 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 229910052742 iron Inorganic materials 0.000 description 1
- 239000010871 livestock manure Substances 0.000 description 1
- 238000004519 manufacturing process Methods 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
- 239000005416 organic matter Substances 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
- 230000001737 promoting effect Effects 0.000 description 1
- 230000000630 rising effect Effects 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
- 230000002195 synergetic 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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- 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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- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K17/00—Soil-conditioning materials or soil-stabilising materials
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- 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
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- C09K2109/00—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE pH regulation
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Abstract
The invention belongs to the field of soil treatment, and particularly relates to an algae composite restoration liquid for improving quick-acting phosphorus content of saline-alkali soil and application thereof. Restoring acidified nitrate polluted cultivated soil by using the chlorella, wherein the chlorella can grow and reproduce by using nitrate as a nitrogen source, the nitrate is converted into an organic nitrogen compound, a slow-release organic fertilizer is provided, and meanwhile, extracellular polysaccharide secreted by the chlorella can be used as a carbon source of denitrifying bacteria, so that the denitrification performance of the denitrifying bacteria is improved, the nitrate is converted into nitrogen through denitrification, and the nitrate pollution is reduced; the chlorella excretes alkaline metabolites to raise the pH value of the soil to be neutral, and is especially suitable for the 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 repair liquid in improving quick-acting phosphorus content of salinized soil.
Background
The disclosure of this background section is only intended to increase the 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 already known to those of ordinary skill in the art.
The total phosphorus content in the soil is usually high, the total phosphorus content in each kilogram of soil can reach thousands of milligrams, but more than 95% of phosphorus in the soil exists in the form of organic phosphorus or indissolvable phosphate and cannot be directly absorbed and utilized by plants. The content of soluble quick-acting phosphorus in soil which is easy to be absorbed and utilized by crops is usually only 10-20 mg/kg.
There are large areas of salinized soil worldwide, including both primary salinized soil and secondary salinized soil due to excessive fertilization and irrational utilization. The content of cations such as calcium, aluminum, iron and the like in the salinized soil is 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 quick-acting phosphorus in the salinized soil can be reduced to below 5mg/kg.
In the agricultural production process, although the quick-acting phosphorus content of the soil can be improved in a short period by applying the phosphate fertilizer, only 10-25% of the phosphate fertilizer can be effectively utilized, and if the salinization phenomenon exists in the soil, the utilization rate of the phosphate fertilizer is further reduced. Phosphate which is not utilized in time in the phosphate fertilizer is fixed by cations in soil to form insoluble phosphate, so that the salinization degree of the soil is further increased, and the phosphate is reciprocated to form vicious circle. Therefore, the capability of releasing endogenous phosphorus elements of the soil is improved, the application of the phosphate fertilizer 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 the blue algae or bacteria and other microorganisms, but the improvement effect is reduced or even eliminated along with the rising of the salinity and the pH value of the soil.
Disclosure of Invention
In order to solve the defects in the prior art, the invention provides application of an algae-bacteria composite repair liquid in improving the quick-acting phosphorus content of salinized soil. The algae bacteria composite repairing liquid consists of chlorella, nostoc, rhodospirillum and water. The chlorella and the nostoc are nontoxic blue algae with better tolerance to the salt and alkali degree, do not generate blue algae toxin, 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; the nostoc has the function of fixing nitrogen, and can provide a nitrogen source for the growth of the nostoc and the haematococcus; the rhodospirillum can realize the heterotrophic life by taking soil organic matters and blue algae exopolysaccharide as carbon sources, the pH value of the soil is reduced by fermenting and producing acid, a good environment is provided for the survival of the blue algae, and the rhodospirillum can perform self-cultivation with the blue algaePhotosynthesis of nourishing CO 2 Is converted into soil organic matters, provides carbon sources for growth of phosphorus-dissolving bacteria, and improves activity of the phosphorus-dissolving bacteria in the soil, thereby promoting release of phosphorus in the soil. The synergistic effect of the chlorella, the nostoc and the rhodospirillum 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 an algae composite repair liquid for improving the quick-acting phosphorus content of saline-alkali soil, which comprises the following components: nostoc, chroococcus, rhodochrous and water.
The application adopts a blue algae and bacteria compound addition method, can improve the tolerance of microorganisms to the soil salt alkalinity, and ensures the restoration effect in the salinized soil.
The second aspect of the invention provides a preparation method of an algae composite repair liquid for improving the quick-acting phosphorus content of saline-alkali soil, which comprises the following steps:
inoculating nostoc, chlorella and rhodospirillum into a liquid culture medium;
culturing the algae bacteria mixture to total cell density of 8X10 by using an expansion culture device 9 ～10×10 9 cells/mL, and filtering to collect the algae bacterial cells;
and re-suspending the filtered and collected algae cells in irrigation water to prepare the algae composite repairing liquid.
The third aspect of the invention provides a method for improving the content of quick-acting phosphorus in saline-alkali soil by using the algae composite repair liquid, which comprises the following steps:
the algae bacteria composite repairing liquid is added to the saline-alkali soil surface layer, and repairing is carried out under natural illumination.
One or more embodiments of the present invention have at least the following beneficial effects:
(1) The chlorella and the nostoc secrete extracellular polysaccharide to chelate cations in soil, so that the salinity is reduced, the generation of insoluble phosphate is inhibited, and the release of phosphorus is promoted; the rhodospirillum heterotrophically ferments blue algae extracellular polysaccharide and soil organic matter to produce acid to reduce the pH value of soil; the chlorella, the nostoc and the rhodospirillum fix carbon through photosynthesis, provide carbon sources for acid-producing bacteria in the soil, promote the growth and propagation of the acid-producing bacteria in the soil, and especially promote the propagation of lactobacillus in the soil, and the lactobacillus not only can produce acid to dissolve phosphate, but also can secrete phosphatase to hydrolyze organic phosphorus in the soil. After the algicidal composite preparation is applied, the activity of soil phosphatase is greatly increased, the pH value and the salinity are greatly reduced, and the quick-acting phosphorus content of the soil is obviously increased.
(2) The preparation method provided by the invention is simple, convenient to operate, high in practicability and easy to popularize.
Detailed Description
It should be noted that the following detailed description is exemplary and is intended to provide further explanation of the invention. 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 quick-acting phosphorus content of the soil is improved by adding a single type of microorganism, the improvement effect is reduced or even eliminated along with the increase of the salinity and the pH value of the soil. Therefore, the invention adopts a method of adding blue algae and bacteria in a compounding way, can improve the tolerance of microorganisms to the soil salt and alkalinity and ensures the restoration effect in the salinized soil.
In order to solve the technical problems, the first aspect of the invention provides an application of an algae-bacteria composite repair liquid in improving the quick-acting phosphorus content of salinized soil.
Blue algae can promote soil to release quick-acting phosphorus through direct and indirect two ways, namely, excreting extracellular polysaccharide to directly chelate cations in indissolvable phosphates, and providing carbon sources for phosphorus-dissolving bacteria through photosynthetic carbon fixation to promote the activity of the phosphorus-dissolving bacteria. However, when the salt and alkali content of the soil exceeds the tolerance range of blue algae, the blue algae cannot grow in the soil and even die, and the phosphor release function cannot be exerted.
The chlorella can resist extreme environments such as high salinity, low temperature, drought and the like, is blue algae which can live in saline-alkali soil, has a good phosphorus release effect, and has limitations when the chlorella is applied to the saline-alkali soil. Firstly, although the chlorella can live in saline-alkali soil, alkaline metabolites can be secreted to gradually increase the pH value of the saline-alkali soil, so that the phosphorus release effect is reduced; secondly, saline-alkali soil is usually very barren, the nitrogen content in soil is very low, a nitrogen source is required to be consumed in the growth process of the chlorella, but the chlorella does not have a nitrogen fixation function, and after the nitrogen in the soil is consumed by the chlorella, the chlorella gradually dies.
The inventor finds that the candida and rhodospirillum as well as the chlorella are prepared into a compound microbial preparation, which can remarkably improve the content of soluble quick-acting phosphorus in saline-alkali soil, and is specifically expressed as follows:
the nostoc can provide a nitrogen source for the growth of the chlorella through the nitrogen fixation, secrete blue algae extracellular polysaccharide together with the chlorella to reduce the salinity of soil and chelate the cations in indissolvable phosphates, and promote the release of free phosphate radicals;
the rhodospirillum can heterotrophically ferment soil organic matters and blue algae exopolysaccharide, and the acid production reduces the pH of the soil, thereby creating a good environment for the growth of blue algae;
the three components cooperate to fix carbon through photosynthesis, increase the organic matter content of soil, promote the reproduction of acidogenic phosphorus-producing bacteria represented by lactobacillus in the soil, reduce the pH value of the soil, promote the phosphatase activity of the soil, and promote the conversion of insoluble phosphate and organic phosphorus in the soil into soluble quick-acting phosphorus.
Further, the saline-alkali soil includes primary saline-alkali soil and secondary saline-alkali soil.
Further, the salt content in the soil with the surface layer of 0-30 cm 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 10mg/kg.
The invention provides an algae bacteria composite repairing liquid, which is prepared by the following steps:
(1) Inoculating nostoc, chlorella and rhodospirillum into liquid culture medium, wherein the initial cell density of the nostoc, chlorella 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, optimally, all three initial cell densities are2×10 6 cells/mL；
(2) Culturing the algae-bacteria mixture to a total cell density of 8×10 under the culture conditions of 25-30deg.C and illumination intensity of 2000-2500 lx with an expansion culture device 9 ～10×10 9 cells/mL, and filtering and collecting the algae cells by using a 0.22 μm filter membrane;
(3) Re-suspending the filtered and collected algae cells in irrigation water to obtain algae cells with total cell density of 1-2×10 8 The algae bacteria composite repairing liquid is cell/mL.
Furthermore, the liquid culture medium is prepared by adding nutrient substances into irrigation water, and the nutrient substance concentration in the culture medium is as follows: k (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 ammonium ferric citrate 2 0.001g/L、Na 2 CO 3 0.02g/L, 5g/L glucose and 5g/L peptone.
Further, the algae strain expansion culture device comprises a conical flask, an open cell body or a photobioreactor.
The third aspect of the invention provides a method for improving the content of quick-acting phosphorus in saline-alkali soil by adopting the algae composite repair liquid, which specifically comprises the following steps:
the algae-bacteria composite repairing liquid is added on the surface layer of saline-alkali soil, and repairing is carried out under natural illumination.
The algae-bacteria composite repairing liquid is added by a liquid manure machine, drip irrigation, spray irrigation or manual irrigation.
Further, the environmental temperature in the repair process is 5-35 ℃, preferably 20-30 ℃, and more preferably 20-25 ℃.
Further, the repair time is 40-80 days.
Further, adding 0.5-1.5 mL of the algae bacteria composite repair liquid on each square centimeter of soil;
preferably, 0.8-1.2 mL of the algae bacteria composite repair liquid is added to each square centimeter of soil;
further preferably, 1.0mL of the algae composite repair liquid is added to each square centimeter of soil.
Furthermore, after the algae composite restoration liquid is added to the soil surface, the water content of the soil is kept between 20% and 30% through irrigation.
In order to enable those skilled in the art to more clearly understand the technical scheme of the present invention, the technical scheme of the present invention will be described in detail with reference to specific embodiments.
Example 1
An algae bacteria composite repairing liquid:
selecting a common chlorella (Chroococcus sp.FACHB-193) and a common Nostoc (Noston sp.PCC 7120), wherein the preservation unit is the institute of aquatic organisms of the national academy of sciences; a common rhodospirillum (Rhodospirillum) is selected, and the preservation unit is China academy of sciences of microorganisms.
The respective inoculation density of two blue algae strains and one rhodospirillum strain in the liquid culture medium is 2 multiplied by 10 6 cells/mL。
The liquid medium had the following composition: k (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 ammonium ferric citrate 2 0.001g/L、Na 2 CO 3 0.02g/L, 5g/L glucose and 5g/L peptone.
Expanding culture of the algae bacteria mixture to total cell density of 10X10 by using conical flask under culture condition of 25deg.C and illumination intensity of 2000lx 9 Filtering and collecting algae cells with 0.22 μm filter membrane, adding irrigation water, suspending blue algae and bacteria again to obtain cell density of 2×10 8 The algae bacteria composite repairing liquid is cell/mL.
Example 2
The method for improving the quick-acting phosphorus content of the saline-alkali soil by adopting the algae-bacteria composite repair liquid in the embodiment 1 comprises the following steps:
and (3) adding the algae composite repair liquid into the open-air primary saline-alkali soil for repair. The adding amount of the algae-bacteria composite repairing liquid on the soil surface is 1.0mL of repairing liquid per square centimeter of soil. The ambient temperature is 10-30 ℃, and the water content of the soil is kept between 20% and 30% by 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 80 days of restoration is 7.8, the salt content is 0.3%, and the quick-acting phosphorus content is 35mg/kg.
Example 3
The method for improving the quick-acting phosphorus content of the saline-alkali soil by adopting the algae-bacteria composite repair liquid in the embodiment 1 comprises the following steps:
and (3) adding the algae composite repairing liquid into the open-air secondary saline-alkali soil for repairing. The adding amount of the algae-bacteria composite repairing liquid on the soil surface is 1.0mL of repairing liquid per square centimeter of soil. The ambient temperature is 10-30 ℃, and the water content of the soil is kept between 20% and 30% by irrigation. The repair time was 40 days. In the repairing process, soil samples are collected every 10 days, and the activities of total phosphorus, quick-acting phosphorus, organic matters, polysaccharide content, pH, salinity and soil phosphatase in the soil are measured. Absolute abundance of bacteria in soil before and after repair was determined at the genus level using metagenomic sequencing techniques.
Soil total phosphorus, quick-acting phosphorus, organic matters and pH are measured by adopting a corresponding national standard method. The activity of the soil phosphatase was determined using an ELISA kit. The soil salinity measuring method comprises the following steps: 10g of saline-alkali soil is air-dried and sieved, 10mL of distilled water is added, the mixture is uniformly mixed, and then the mixture is kept stand, and the salinity of the supernatant is measured by a salinity meter. The polysaccharide content of the soil is measured by a phenol sulfuric acid method.
Various indexes in the repair process of the saline-alkali soil in the example 3 are changed with time as shown in table 1.
TABLE 1 repair effect of algae repair liquid on saline-alkali soil with time
The absolute abundance changes of typical acidogenic and phosphate-solubilizing bacteria before and after saline-alkali soil remediation in example 3 are summarized in table 2.
TABLE 2 absolute abundance changes of typical acidogenic and phosphate-solubilizing bacteria before and after saline-alkali soil remediation
The quick-acting phosphorus content of the soil before restoration is 10mg/kg, and the quick-acting phosphorus content of the soil after 40 days of restoration is 45mg/kg. The pH value and the salinity of the soil are obviously reduced, and the organic matters, the polysaccharide content and the phosphatase activity of the soil are obviously increased. Wherein the soil phosphatase activity was increased to 3.4 times before restoration after 40 days of restoration. The biomass of Sphingomonas, dysosma, prevotella and two acid-producing bacilli in soil is increased hundreds times, and the bacteria can ferment extracellular polysaccharide to produce acid, reduce pH value of soil and dissolve phosphate; the lactobacillus not only can produce acid to dissolve phosphate, but also can secrete phosphatase to release organic phosphorus in the soil, so that the quick-acting phosphorus content in the soil is greatly increased.
The above description is only of the preferred embodiments of the present invention and is not intended to limit the present invention, but various modifications and variations can be made to the present invention by those skilled in the art. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (12)
1. An algae composite repairing liquid for improving the quick-acting phosphorus content of saline-alkali soil is characterized by comprising nostoc, chroococcus, rhodospirillum and water.
2. The composite algae repair liquid for increasing the quick-acting phosphorus content of saline-alkali soil according to claim 1, wherein the total cell density is 1 x 10 8 ～2×10 8 cells/mL。
3. The algae composite repair liquid for increasing the quick-acting phosphorus content of saline-alkali soil according to claim 1, wherein the water is irrigation water.
4. A preparation method of an algae composite repair liquid for improving the quick-acting phosphorus content of saline-alkali soil is characterized by comprising the following steps:
inoculating nostoc, chlorella and rhodospirillum into a liquid culture medium;
culturing the algae bacteria mixture to total cell density of 8X10 by using an expansion culture device 9 ～10×10 9 cells/mL, and filtering to collect the algae bacterial cells;
and re-suspending the filtered and collected algae cells in irrigation water to prepare the algae composite repairing liquid.
5. The method for preparing the algae composite repair liquid for improving the quick-acting phosphorus content of the saline-alkali soil according to claim 4, wherein the nutrient substance concentration in the liquid culture medium is as follows: k (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 ammonium ferric citrate 2 0.001g/L、Na 2 CO 3 0.02g/L, 5g/L glucose and 5g/L peptone.
6. The method for preparing an algae composite repairing liquid for increasing the quick-acting phosphorus content of saline-alkali soil as claimed in claim 4, wherein nostoc, chlorella and rhodospirillum are inoculated in the liquid culture medium, and the initial cell density of the three is 1×10 6 ～3×10 6 cells/mL。
7. The method for preparing the algae composite repair liquid for improving the quick-acting phosphorus content of the saline-alkali soil according to claim 4, wherein the specific conditions of the expansion culture are as follows: the temperature is 25-30 ℃, and the illumination intensity is 2000-2500 lx.
8. The method for preparing the composite algae repair liquid for improving the quick-acting phosphorus content of the saline-alkali soil according to claim 4, wherein the expansion culture device comprises a conical flask, an open pond body and a photo-bioreactor.
9. A method for increasing the content of quick-acting phosphorus in saline-alkali soil by using the algae composite repair liquid according to any one of claims 1 to 3, which is characterized by comprising the following steps:
the algae bacteria composite repairing liquid according to any one of claims 1-3 is added on the surface layer of saline-alkali soil, and repairing is carried out under natural illumination.
10. The method for increasing the rapid phosphorous content in saline-alkali soil according to claim 9, wherein: the environmental temperature of the repair process is 5-35 ℃; the repair time is 40-80 days;
or, adding 0.5-1.5 mL of the algae bacteria composite repair liquid on each square centimeter of soil;
or, after the algae composite restoration liquid is added to the soil surface, the water content of the soil is kept between 20% and 30% through irrigation.
11. The method for increasing the rapid phosphorous content in saline-alkali soil as recited in claim 10, wherein: the environmental temperature of the repair process is 20-30 ℃.
12. The method for increasing the rapid phosphorous content in saline-alkali soil as recited in claim 11, wherein: the environmental temperature of the repair process is 20-25 ℃.
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