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 PDF

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CN114836325B
CN114836325B CN202210610328.XA CN202210610328A CN114836325B CN 114836325 B CN114836325 B CN 114836325B CN 202210610328 A CN202210610328 A CN 202210610328A CN 114836325 B CN114836325 B CN 114836325B
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刘莹
刘帅彤
宋岳
蔡勇
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Shandong University
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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

Algae composite repair liquid for improving quick-acting phosphorus content of saline-alkali soil and application thereof
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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