CN110226606B - Prebiotic formula product for regulating soil microorganisms, increasing plant system resistance and reducing morbidity - Google Patents
Prebiotic formula product for regulating soil microorganisms, increasing plant system resistance and reducing morbidity Download PDFInfo
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- CN110226606B CN110226606B CN201910600748.8A CN201910600748A CN110226606B CN 110226606 B CN110226606 B CN 110226606B CN 201910600748 A CN201910600748 A CN 201910600748A CN 110226606 B CN110226606 B CN 110226606B
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01B—SOIL WORKING IN AGRICULTURE OR FORESTRY; PARTS, DETAILS, OR ACCESSORIES OF AGRICULTURAL MACHINES OR IMPLEMENTS, IN GENERAL
- A01B79/00—Methods for working soil
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N37/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids
- A01N37/02—Saturated carboxylic acids or thio analogues thereof; Derivatives thereof
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N37/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids
- A01N37/44—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids containing at least one carboxylic group or a thio analogue, or a derivative thereof, and a nitrogen atom attached to the same carbon skeleton by a single or double bond, this nitrogen atom not being a member of a derivative or of a thio analogue of a carboxylic group, e.g. amino-carboxylic acids
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
- A01N43/34—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one nitrogen atom as the only ring hetero atom
- A01N43/36—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one nitrogen atom as the only ring hetero atom five-membered rings
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
- A01N43/34—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one nitrogen atom as the only ring hetero atom
- A01N43/36—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one nitrogen atom as the only ring hetero atom five-membered rings
- A01N43/38—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one nitrogen atom as the only ring hetero atom five-membered rings condensed with carbocyclic rings
Abstract
The invention discloses a prebiotics formula product for regulating soil microorganisms, increasing plant system resistance and reducing morbidity. The soil microorganism prebiotic composition consists of a long-chain fatty acid substance and an amino acid substance; the fatty acid substances are selected from one or more of capric acid, lauric acid, tridecylic acid, pentadecylic acid, palmitoleic acid, palmitic acid, stearic acid and arachidic acid; the amino acid substances are selected from one or more of isoleucine, leucine, methionine, proline, tryptophan, ornithine, alanine and aspartic acid. After the application, the quantity of pseudomonas in soil can be increased by more than two orders of magnitude, the plant system resistance is obviously improved, and the morbidity is reduced. The formula product is suitable for being applied to herbaceous crops such as fruits and vegetables.
Description
Technical Field
The invention belongs to the technical field of agricultural chemicals, and relates to a soil microbial community conditioner and a using method thereof.
Background
The planting area of economic crops in China is increased year by year, and heavy load is brought to soil while considerable economic value is brought to continuous cropping and intensive planting. The most remarkable of the problems are that soil continuous cropping obstacles frequently occur, soil-borne diseases are serious, and exogenous beneficial bacteria are difficult to add and survive; the root of the method is the unbalance of soil microbial communities and the reduction of the abundance of beneficial bacteria. Some farmers continue continuous cropping to obtain higher economic value, and only need to invest a large amount of agricultural chemicals, so that the damage of the soil microbial community is aggravated while food safety accidents are frequent, and the continuous cropping obstacle enters a vicious circle. In recent years, the development of microbial agents, particularly bioorganic fertilizers, provides certain help for the sustainable development of facility agriculture. However, the microbial product still has the phenomena of unstable effect and the like caused by difficult colonization of exogenous strains. Compared with the addition of exogenous microorganisms to help crops resist diseases, the beneficial microorganisms in the soil are mobilized, so that the disease resistance efficiency is greatly improved, and the cost is reduced.
How to mobilize beneficial microorganisms in soil is always a difficult problem for the soil scientist and the plant protection scientist. The key to solve the problem is to screen and find materials or preparations capable of improving the abundance of beneficial bacteria in the soil. In fact, when plants in the nature invade pathogenic bacteria, the chemical environment of the rhizosphere is changed, so that rhizosphere beneficial bacteria are recruited, and the beneficial bacteria assist the plants to resist the invasion of the pathogenic bacteria in the modes of forming a biological film on the root surface, secreting antibacterial substances, competing nutrition with the pathogenic bacteria and the like. Root exudates are a general term for all organic and inorganic ions released by plants to the rhizosphere through the root system, and are an important means for the plants to change the chemical environment of the rhizosphere. Therefore, the plant root exudates can be used as a resource library for excavating the prebiotic preparation, namely, the soil microbial community is directionally regulated and controlled by searching substances capable of promoting the growth of beneficial microbial groups, so that the plant root exudates have the function of helping the plant to resist the invasion of pathogenic bacteria.
Pseudomonas is a common microorganism in soil, is also the most common microorganism group of plant rhizosphere, and has the characteristics of wide distribution, large quantity, simple nutrition, quick propagation and strong competitive colonization. Many pseudomonas strains reported at present can produce active substances including 2, 4-diacetyl benzene trisphenol (2,4-DAPG) to resist various plant diseases, can induce plant system resistance and improve the capability of crops to resist pathogenic bacteria.
Disclosure of Invention
The invention aims to develop a rhizosphere prebiotic formula for increasing the abundance of pseudomonas in soil to improve plant disease resistance aiming at the problems of unbalanced soil microbial community, reduced abundance of beneficial bacteria and difficult survival of exogenous beneficial bacteria. After the fertilizer is applied, the number of the pseudomonads in the soil is increased by two orders of magnitude, and the disease incidence of plants is obviously reduced.
A soil microorganism prebiotic composition for increasing systemic resistance in plants, said soil microorganism prebiotic composition comprised of long chain fatty acids and amino acids; the fatty acid substances are selected from one or more of capric acid, lauric acid, tridecylic acid, pentadecylic acid, palmitoleic acid, palmitic acid, stearic acid and arachidic acid; the amino acid substances are selected from one or more of isoleucine, leucine, methionine, proline, tryptophan, ornithine, alanine and aspartic acid.
The fatty acid substances are preferably one or more of tridecanoic acid, pentadecanoic acid, palmitoleic acid, hexadecanoic acid, octadecanoic acid and arachidic acid; the amino acid is preferably one or more of isoleucine, leucine, methionine, proline, tryptophan and ornithine.
The soil microorganism prebiotic composition is preferably composed of isoleucine, leucine, methionine, proline, tryptophan, ornithine, tridecanoic acid, pentadecanoic acid, palmitoleic acid, hexadecanoic acid, octadecanoic acid and arachidic acid.
The soil microorganism prebiotic composition is preferably composed of leucine, methionine, proline, tryptophan, ornithine, pentadecanoic acid, palmitoleic acid, hexadecanoic acid, octadecanoic acid and arachidic acid.
The soil microorganism prebiotic composition is preferably composed of leucine, methionine, proline, tryptophan, ornithine, aspartic acid, dodecanoic acid, tridecanoic acid, pentadecanoic acid, hexadecanoic acid, octadecanoic acid and arachidic acid.
The soil microorganism prebiotic composition preferably consists of capric acid, lauric acid, tridecanoic acid, pentadecanoic acid, palmitoleic acid, palmitic acid, stearic acid, arachidic acid, isoleucine, leucine, methionine, proline, tryptophan, ornithine, alanine, and aspartic acid.
The concentration of each fatty acid or amino acid in the soil microorganism prebiotic composition is preferably 5-20 mmol/L, and more preferably 10 mmol/L.
The preparation method of the prebiotic composition disclosed by the invention is characterized in that the components in the formula are prepared into an aqueous solution, and the concentration of each fatty acid or amino acid in the aqueous solution is 5-20 mmol/L, preferably 10 mmol/L.
Application of prebiotic composition in improving abundance of pseudomonas in soil, enhancing plant system resistance and reducing plant morbidity
The application is preferably that the prebiotic composition is applied before seedling transplantation or sowing for one month, 12-18 kg of prebiotic composition is applied to each mu, and the prebiotic composition is applied for 1-2 times per week for four weeks continuously.
Has the advantages that:
the invention prepares the screened substances for improving the abundance of the pseudomonas in the soil into the prebiotic preparation. The prebiotic preparation can rapidly improve the abundance of pseudomonas in soil so as to induce plants to generate systemic resistance and reduce the incidence of plant invasion caused by pathogenic bacteria.
The rhizosphere prebiotics product produced by the invention has the advantages of convenient use, low cost and good disease prevention effect, and is suitable for large-area use in planting of economic crops such as fruits, vegetables, flowers and the like. The application of the invention has important significance for ensuring the sustainable development of facility agriculture, reducing the input of chemicals such as pesticides and the like, protecting the ecological environment and improving the quality of agricultural products.
Compared with the current products on the market, the invention has the following advantages:
1) the invention reduces the cost of the production raw materials of the product and improves the market competitiveness of the product.
2) The invention uses common chemical products, and the raw materials are easy to obtain.
3) The product of the invention is convenient to use and can be used together with water and fertilizer.
4) The product of the invention is used before seedling transplanting, the quantity of pseudomonas in soil is increased by more than two orders of magnitude, and the crop morbidity can be obviously reduced.
Description of the drawings:
FIG. 1 Effect of different combinations of substances on the amount of Pseudomonas in soil
A is amino acid, B is fatty acid, C is organic acid, and D is saccharide
FIG. 2 Effect of prebiotic administration on the amount of Pseudomonas in soil
FIG. 3 application of prebiotics to help Arabidopsis thaliana in combating overground diseases
FIG. 4 shows that Pseudomonas cultured with prebiotics helps Arabidopsis thaliana resist overground diseases
FIG. 5 administration of prebiotics to help tomato resist Ralstonia solanacearum invasion
The specific implementation mode is as follows:
example 1 obtainment of prebiotic formulation
Inoculating pathogenic bacteria to arabidopsis thaliana leaves, and carrying out infection culture. The root exudate metabolome of healthy plants and plants infected by pathogenic bacteria was determined one week later. As a result, it was found that 28 species of substance components of root exudates, including capric acid, lauric acid, tridecanoic acid, pentadecanoic acid, palmitoleic acid, palmitic acid, stearic acid, arachidic acid, isoleucine, leucine, methionine, proline, tryptophan, ornithine, aspartic acid, alanine, oxaloacetic acid, citric acid, aconitic acid, ketoglutaric acid, succinic acid, malic acid, maltose, ribose, glucose, sucrose, fructose, and xylose, were different after inoculation with pathogenic bacteria. After subdivision, these 28 substances were found to belong to class 4, namely long chain fatty acids (decanoic acid, dodecanoic acid, tridecanoic acid, pentadecanoic acid, palmitoleic acid, hexadecanoic acid, octadecanoic acid and arachidic acid), amino acids (isoleucine, leucine, methionine, proline, tryptophan, ornithine, alanine and aspartic acid), organic acids (oxaloacetic acid, citric acid, aconitic acid, ketoglutaric acid, succinic acid and malic acid), and sugars (maltose, ribose, glucose, sucrose, fructose and xylose). Further research shows that the contents of the four types of substances, namely fatty acid and amino acid in a pathogen inoculation sample are obviously higher than those of a healthy sample, and the contents of saccharide and organic acid in the healthy sample are obviously higher than those of a disease sample. Therefore, the four types of substances are used for the next screening individually or in combination of two types or three types or four types.
And preparing four kinds of substances into aqueous solutions with the concentration of 10mmol/L respectively by using two kinds of substances or three kinds or four kinds of substances. Each solution was added to soil to a final concentration of 1. mu. mol/kg. Added twice a week for four consecutive weeks. Control treatment equal amounts of sterile water were added. After the treatment is finished, extracting soil DNA, and detecting the number of the pseudomonad samples in the added prebiotics and control soil samples by using a quantitative PCR technology. As shown in fig. 1, the number of pseudomonads in the soil after the combined treatment of fatty acid and amino acid was significantly higher than that of the other treatments, and was increased by one order of magnitude compared to the control. While the treatment of organic acids, sugars and combinations of organic acids is significantly lower than other treatments. It was also found that fatty acids alone or amino acids alone did not increase the number of pseudomonas in the soil. Therefore, it was preliminarily determined that prebiotic products consist of both these fatty acids and amino acids.
Example 2 further optimization of prebiotic formulations
Any one or more of 8 kinds of amino acids and any one or more of 8 kinds of fatty acids are combined to prepare a solution, and then the solution is added into soil according to the requirements. The results show that any of the above amino acids in combination with any of the above fatty acids can increase the abundance of Pseudomonas solanacearum. When the types of amino acids and fatty acids contained in the prebiotics reach more than 5 types (including 5 types) respectively, the effect is not obviously different from the effect of all 16 substances, and as shown in figure 2, three preferable formulas (three formulas are shown in table 1) containing more than 10 types of substances and the improvement effect of all 16 types of substances on the pseudo monospore abundance in the soil are realized.
Table 1 three preferred prebiotic formulations
Example 3 soil after prebiotics application helps plants to fight pathogen invasion
3.1 the soil after administration of prebiotics helps Arabidopsis to defend against the invasion of pathogens from the overground part
And respectively planting the arabidopsis thaliana bred in advance in the soil to which the prebiotics formula is applied, and inoculating pathogenic bacteria to the leaf parts of the arabidopsis thaliana after two weeks. Morbidity was calculated 7 days after inoculation. The results are shown in figure 3, where 4 prebiotic soils were applied, the incidence of the plants was around 20%, whereas the incidence of the control treatments without the product was above 60%. The product is shown to help plants resist the invasion of pathogenic bacteria and obviously reduce the morbidity.
3.2 isolation of Pseudomonas in soil after prebiotic administration
12 strains of pseudomonas were isolated and purified by dilution and coating. These 12 strains were inoculated back to the root of Arabidopsis thaliana for culture. The disease resistance of the plants is induced by the test according to the morbidity rate so as to resist the invasion of pathogenic bacteria. As shown in FIG. 4, the 12 isolated strains all helped reduce the incidence of disease by less than 40%, with the incidence of disease after treatment of two strains being as low as less than 10%, and the incidence of disease of the control being about 60%. The pseudomonas soil cultured by the prebiotic product can effectively help plants to resist the invasion of pathogenic bacteria.
Example 4 soil after prebiotic administration helps tomatoes to fight the invasion of Ralstonia solanacearum
The previously cultivated arabidopsis thaliana was planted in the soil to which the 4 prebiotics formulations were applied, respectively, and the tomato roots were inoculated with a bacterial wilt pathogen (lakellla solanacearum) one week later. The number of pathogens was counted 14 days after inoculation. The results are shown in fig. 5, where the number of pathogenic bacteria was reduced by one order of magnitude compared to the control after tomato planting in soil with both prebiotics. The two prebiotic products are shown to help tomatoes resist the invasion of the Laurella solanacearum and obviously reduce the number of rhizosphere pathogenic bacteria.
Soil treatment experiments show that the prebiotics formula can obviously promote the number of pseudomonas in soil, and the promoted pseudomonas can improve the plant system resistance and reduce the morbidity. Meanwhile, the incidence of tomato bacterial wilt can be obviously reduced by applying the prebiotic product. The study on the diseases of the overground part of the model crop Arabidopsis thaliana and the underground part of the tomato by verifying the prebiotic product shows that the prebiotic product can achieve the purpose of reducing the plant morbidity by increasing the number of pseudomonas in soil.
The fatty acids are cis-fatty acids, and the amino acids are L-amino acids.
Claims (11)
1. A soil microorganism prebiotic composition for increasing systemic resistance in plants, characterized in that the soil microorganism prebiotic composition comprises long chain fatty acids and amino acids; the fatty acid substances are selected from one or more of capric acid, lauric acid, tridecylic acid, pentadecylic acid, palmitoleic acid, palmitic acid, stearic acid and arachidic acid; the amino acid substances are selected from one or more of isoleucine, leucine, methionine, proline, tryptophan, ornithine, alanine and aspartic acid.
2. The prebiotic composition according to claim 1, wherein the fatty acids are selected from one or more of tridecanoic acid, pentadecanoic acid, palmitoleic acid, hexadecanoic acid, octadecanoic acid, and arachidic acid, and the amino acids are selected from one or more of isoleucine, leucine, methionine, proline, tryptophan, and ornithine.
3. The prebiotic composition of claim 2, wherein said soil microorganism prebiotic composition is comprised of isoleucine, leucine, methionine, proline, tryptophan, ornithine, tridecanoic acid, pentadecanoic acid, palmitoleic acid, hexadecanoic acid, octadecanoic acid, arachidic acid.
4. The prebiotic composition of claim 2, wherein said soil microorganism prebiotic composition comprises leucine, methionine, proline, tryptophan, ornithine, pentadecanoic acid, palmitoleic acid, hexadecanoic acid, octadecanoic acid, arachidic acid.
5. The prebiotic composition of claim 1, wherein said soil microorganism prebiotic composition is comprised of leucine, methionine, proline, tryptophan, ornithine, aspartic acid, dodecanoic acid, tridecanoic acid, pentadecanoic acid, hexadecanoic acid, octadecanoic acid, arachidic acid.
6. The prebiotic composition according to any of claims 1 to 5, characterized in that the concentration of each fatty acid or amino acid in the soil microorganism prebiotic composition is 5 to 20 mmol/L.
7. The prebiotic composition according to claim 6, characterized in that the soil microorganism prebiotic composition has a concentration of 10mmol/L of each fatty acid or amino acid.
8. The method of claim 1, wherein the components of the formulation are formulated as an aqueous solution, wherein the concentration of each fatty acid or amino acid in the aqueous solution is 5 to 20 mmol/L.
9. The method according to claim 8, wherein the concentration of each of the fatty acid or amino acid in the aqueous solution is 10 mmol/L.
10. Use of the prebiotic composition of claim 1 to increase pseudomonas abundance in soil, enhance plant systemic resistance, reduce plant morbidity.
11. The use of claim 10, wherein the prebiotic composition is applied one month before seedling transplantation or sowing, at 12-18 kg per acre, 1-2 times per week for four weeks.
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| CN201910600748.8A CN110226606B (en) | 2019-07-04 | 2019-07-04 | Prebiotic formula product for regulating soil microorganisms, increasing plant system resistance and reducing morbidity |
| CN202011111666.6A CN112369416B (en) | 2019-07-04 | 2019-07-04 | Soil microorganism prebiotic composition for increasing plant system resistance and application thereof |
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| CN1925748A (en) * | 2004-03-03 | 2007-03-07 | 伊萨格罗股份公司 | Mixtures and methods for the induction of resistance in plants |
| CN101426369A (en) * | 2006-02-27 | 2009-05-06 | 西北大学 | Composition in the form of a microemulsion containing free fatty acids and/or free fatty acid derivatives |
| CN107124994A (en) * | 2017-05-26 | 2017-09-05 | 庆阳敦博科技发展有限公司 | A kind of prevention and controls of duck wheat brown spot |
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| CN1925748A (en) * | 2004-03-03 | 2007-03-07 | 伊萨格罗股份公司 | Mixtures and methods for the induction of resistance in plants |
| CN101426369A (en) * | 2006-02-27 | 2009-05-06 | 西北大学 | Composition in the form of a microemulsion containing free fatty acids and/or free fatty acid derivatives |
| CN107124994A (en) * | 2017-05-26 | 2017-09-05 | 庆阳敦博科技发展有限公司 | A kind of prevention and controls of duck wheat brown spot |
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