CN110526772B - Antibacterial fertilizer for preventing and treating bacterial wilt of crops and preparation method thereof - Google Patents
Antibacterial fertilizer for preventing and treating bacterial wilt of crops and preparation method thereof Download PDFInfo
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- CN110526772B CN110526772B CN201910818444.9A CN201910818444A CN110526772B CN 110526772 B CN110526772 B CN 110526772B CN 201910818444 A CN201910818444 A CN 201910818444A CN 110526772 B CN110526772 B CN 110526772B
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05B—PHOSPHATIC FERTILISERS
- C05B7/00—Fertilisers based essentially on alkali or ammonium orthophosphates
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
- C05G3/00—Mixtures of one or more fertilisers with additives not having a specially fertilising activity
Abstract
The invention provides an application of rhamnolipid in preparation of a pesticide or a bacterial inhibition fertilizer for preventing and treating crop bacterial wilt. In addition, the rhamnolipid is used for preparing an antibacterial fertilizer for preventing and treating crop bacterial wilt, and is matched with humate, molasses, a nitrogen source, a phosphorus source, a potassium source, trace elements and a chelating agent in a proper proportion, so that the antibacterial effect is further improved, the rhamnolipid can be used for direct root application and leaf surface spraying and seed soaking, has a good prevention and treatment effect on the crop bacterial wilt, contains organic matters, can effectively improve the crop resistance, improves the soil condition, and can greatly improve the quality and the yield of crops.
Description
Technical Field
The invention belongs to the technical field of agricultural production, and particularly relates to an antibacterial fertilizer for preventing and treating crop bacterial wilt and a preparation method thereof.
Background
Bacterial wilt is caused by Ralstonia solanacearum (also called Ralstonia solanacearum) widely distributed in tropical, subtropical and temperate regions, and has been proved to infect hundreds of plants in 44 families up to now, including important economic crops such as peanuts, tomatoes, tobaccos, bananas and the like. Research shows that the pathogenic factors of tomato bacterial wilt disease are mainly exopolysaccharide, polygalacturonase and endoglucanase secreted by pathogenic bacteria, and especially exopolysaccharide can block smaller-bore catheters at petiole nodes and leaflets, influence and hinder water transportation in vivo, thereby causing plant wilting. The optimal temperature for the development of ralstonia solanacearum is 25-35 ℃. Ralstonia solanacearum can survive for a long time under natural conditions, germs can usually invade from the roots or the stems of plants to cause diseases, the stems and leaves of infected plants at the initial stage wither in the daytime and recover at night, the whole plants die after several days but are still greenish, when the weather is humid, the diseased plants have patches which are changed into brown from water stain shapes, the vascular bundles of diseased stems are browned, and milky bacteria liquid overflows after being squeezed by hands. The tomato ralstonia solanacearum can live for years after overwintering in soil mainly along with diseased plant residues, is mainly spread in fields through rainwater and irrigation water, and can also be spread by people, livestock, farm implements, bacteria-carrying soil, insects, nematodes and the like to cause repeated infection, so that the disease is popular and spread.
The traditional control method comprises the modes of selecting resistant varieties, controlling by biological pesticides, adopting crop rotation and the like. The resistance of the disease-resistant variety is easy to lose due to quick change of pathogenic bacteria; the chemical prevention and treatment effect is poor, and the chemical prevention and treatment agent has great negative effects on food safety and environment; the application of organic fertilizers is limited due to the existence of heavy metals, antibiotics and hormone substances. Therefore, adopting other effective measures to control the harm of bacterial wilt becomes an important problem to be solved urgently in crop production.
Disclosure of Invention
Based on the above, the invention aims to provide the bacteriostatic fertilizer for preventing and treating the bacterial wilt of crops and the preparation method thereof.
The specific technical scheme is as follows:
the rhamnolipid is applied to the preparation of pesticides or bacterial inhibiting fertilizers for preventing and treating crop bacterial wilt.
The invention also provides an antibacterial fertilizer for preventing and treating the bacterial wilt of crops, and the specific technical scheme is as follows:
the raw materials comprise rhamnolipid, humate, molasses, a nitrogen source, a phosphorus source, a potassium source, trace elements and a chelating agent.
In some embodiments, the raw materials comprise, in parts by weight:
the invention also provides a preparation method of the bacterial inhibiting fertilizer for preventing and treating the bacterial wilt of crops, and the specific technical scheme is as follows:
the preparation method of the bacterial inhibiting fertilizer for controlling the bacterial wilt of crops comprises the following steps:
sequentially adding trace elements and a chelating agent, which are helpful for improving the stability of the fertilizer, into water, adding humate while stirring, and adding molasses after the humate is completely dissolved; adding a nitrogen source, a phosphorus source and a potassium source while stirring; adding rhamnolipid, and mixing.
Based on the technical scheme, the invention has the following beneficial effects:
the inventor of the invention discovers that the rhamnolipid has a good inhibition effect on ralstonia solanacearum, can be used as a novel inhibitor for inhibiting ralstonia solanacearum, is applied to preparation of an antibacterial fertilizer for preventing and treating crop bacterial wilt, can effectively reduce crop bacterial wilt diseases and promote crop growth.
In addition, the rhamnolipid is used for preparing an antibacterial fertilizer for preventing and treating crop bacterial wilt, and is matched with humate, molasses, a nitrogen source, a phosphorus source, a potassium source, trace elements and a chelating agent in a proper proportion, so that the antibacterial effect is further improved, the rhamnolipid can be used for direct root application and leaf surface spraying and seed soaking, has a good prevention and treatment effect on the crop bacterial wilt, contains organic matters, can effectively improve the crop resistance, improves the soil condition, and can greatly improve the quality and the yield of crops. Compared with the traditional bacterial wilt prevention and control method, the method adopts environment-friendly biogenic antibacterial components, is nontoxic and harmless to human bodies, is more environment-friendly than traditional chemical agents, and is beneficial to improving the economic value of crops and promoting the development of organic agriculture.
Drawings
FIG. 1 shows the result of the rhamnolipid bacteriostatic effect test;
FIG. 2 shows the disease rate of tomato plants using the bacteriostatic fertilizer for controlling crop bacterial wilt according to the invention;
fig. 3 shows the control effect of the bacteriostatic fertilizer for controlling crop bacterial wilt on tomato plant bacterial wilt in embodiment 5 of the present invention, and the bacteriostatic fertilizers used from left to right are respectively negative control, embodiment 1, embodiment 2, embodiment 3 and normal plant control.
Detailed Description
In order that the invention may be more readily understood, reference will now be made to the following more particular description of the invention, examples of which are set forth below. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. These embodiments are provided so that this disclosure will be thorough and complete. It is to be understood that the experimental procedures in the following examples, where specific conditions are not noted, are generally in accordance with conventional conditions, or with conditions recommended by the manufacturer. The various reagents used in the examples are commercially available.
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. The terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. As used herein, the term "and/or" includes any and all combinations of one or more of the associated listed items.
The invention relates to an antibacterial fertilizer for preventing and treating crop bacterial wilt, which comprises rhamnolipid, humate, molasses, a nitrogen source, a phosphorus source, a potassium source, trace elements and a chelating agent.
Preferably, the raw materials comprise, by weight:
optionally, the nitrogen source comprises one or more of uric acid, ammonium nitrate, potassium nitrate, ammonium polyphosphate and ammonium dihydrogen phosphate. Preferably, the nitrogen source is selected from one or two of ammonium polyphosphate and ammonium nitrate.
Optionally, the source of phosphorus comprises one or more of phosphoric acid, ammonium dihydrogen phosphate, ammonium hydrogen phosphate, ammonium polyphosphate, potassium hydrogen phosphate, and dipotassium hydrogen phosphate. Preferably, the phosphorus source is selected from one or two of ammonium polyphosphate and dipotassium hydrogen phosphate.
Optionally, the potassium source comprises one or more of potassium chloride, potassium hydrogen phosphate, dipotassium hydrogen phosphate and potassium nitrate. Preferably, the potassium source comprises dipotassium hydrogen phosphate.
Wherein, when the humate is potassium humate, the humate can also be used as a potassium source. The ammonium polyphosphate contains nitrogen elements and phosphorus elements and can play the roles of both nitrogen sources and phosphorus sources. The dipotassium hydrogen phosphate contains phosphorus element and potassium element, and has the functions of phosphorus source and potassium source. The bacteriostatic fertilizer for preventing and treating the crop bacterial wilt contains higher-content phosphorus elements and a part of polymeric phosphorus (ammonium polyphosphate), and the ammonium polyphosphate can chelate trace elements in the fertilizer on one hand and increase the storage time of the bacteriostatic fertilizer; on the other hand, the mobility of the ammonium polyphosphate in the soil is better than that of a common phosphorus source, and the ammonium polyphosphate is beneficial to the absorption of the tomato on phosphorus. In the seedling stage of the tomatoes, the higher phosphorus element is also beneficial to the growth of the tomatoes and improves the disease resistance of the tomatoes. The bacteriostatic fertilizer for the crop bacterial wilt contains nitrogen and potassium elements with proper contents, and can meet the requirements in the production process of tomatoes.
Optionally, the trace elements include one or more of copper sulfate, manganese sulfate, zinc sulfate, boric acid, EDTA-Zn, sodium octaborate tetrahydrate, EDTA-Mn, EDTA-Fe, EDTA-Cu. Preferably, the trace elements comprise one or more of EDTA-Zn, sodium octaborate tetrahydrate, EDTA-Mn, EDTA-Fe, EDTA-Cu. More preferably, the trace elements include EDTA-Zn, sodium octaborate tetrahydrate, EDTA-Mn, EDTA-Fe, and EDTA-Cu. In the bacteriostatic fertilizer for preventing and treating the bacterial wilt of crops, the addition of trace elements in proper proportion is beneficial to the growth of tomatoes, and the application of the trace elements Mn, Zn and B is beneficial to the early flowering of the tomatoes, so that the content of vitamin C in the tomatoes is increased, the quality of the tomatoes is improved, and the effect of improving yield and quality is achieved.
Optionally, the chelating agent comprises one or more of EDTA-2Na, citric acid, amino acids, malic acid. Preferably, the chelating agent is EDTA-2 Na.
Preferably, the bacteriostatic fertilizer for preventing and treating the crop bacterial wilt comprises the following raw materials in parts by weight:
preferably, the bacteriostatic fertilizer for preventing and treating the crop bacterial wilt comprises the following raw materials in parts by weight:
in the antibacterial fertilizer for preventing and treating the bacterial wilt of crops, ammonium polyphosphate is selected to be reasonably matched with rhamnolipid and each component in the antibacterial fertilizer, and the ammonium polyphosphate exists as a nitrogen source and a phosphorus source in the antibacterial fertilizer to help plant growth on one hand, and on the other hand, the ammonium polyphosphate is selected to improve the antibacterial effect of the antibacterial fertilizer in a proper proportion range, and the ammonium polyphosphate is selected to improve the stability of the fertilizer.
The preparation method of the bacterial inhibiting fertilizer for controlling the bacterial wilt of crops, disclosed by the invention, comprises the following steps of:
sequentially adding trace elements and a chelating agent, which are helpful for improving the stability of the fertilizer, into water, adding humate while stirring, and adding molasses after the humate is completely dissolved; adding a nitrogen source, a phosphorus source and a potassium source while stirring; adding rhamnolipid, and mixing.
The present invention will be described in detail with reference to specific examples.
Inhibitory effect of rhamnolipids on ralstonia solanacearum
A growth inhibition rate method is adopted to prepare a PDA solid culture medium (200 g of potatoes, 20g of glucose, 20g of agar and 1000mL of water, the pH is natural), after the PDA solid culture medium is cooled to about 45 ℃, proper amounts of rhamnolipid solutions after filtration and sterilization are respectively added, after the rhamnolipid solutions are mixed with the culture medium, the final concentrations of the rhamnolipid in the culture medium are 0.1 wt%, 0.15 wt% and 0.2 wt%, a negative control is a flat plate added with an equal amount of sterile water, and the bacterial wilt is not reliable and effective at present, so that a positive control is not available. Using a puncher with the diameter of 8.6mm to punch bacterial blocks of ralstonia solanacearum, moving the bacterial blocks to the center of each processing plate, repeating the same processing for 4 times, placing the same processing plate in a constant-temperature incubator at 28 ℃, measuring and recording the diameter of bacterial colonies by using a vernier caliper by adopting a cross method when the bacterial colonies on a control plate are covered with about two thirds of a culture dish, and taking an average value. The results are shown in FIG. 1 and Table 1.
TABLE 1 bacteriostatic effect of rhamnolipid with different concentrations on ralstonia solanacearum
As can be seen from the results in the table above, the rhamnolipid has a better bacteriostatic effect on ralstonia solanacearum, and the bacteriostatic effect on ralstonia solanacearum is close to that of a positive control.
Examples 1 to 3
The bacteriostatic fertilizer for preventing and treating the crop bacterial wilt comprises the components in parts by weight shown in Table 2.
TABLE 2 bacteriostatic fertilizer for preventing and treating crop bacterial wilt
| Example 1 | Example 2 | Example 3 | Comparative example 1 | |
| |
20 | 15 | 10 | 0 |
| Ammonium polyphosphate | 15 | 15 | 15 | 15 |
| Ammonium nitrate | 10 | 10 | 10 | 10 |
| Dipotassium hydrogen phosphate | 10 | 10 | 10 | 10 |
| |
5 | 5 | 5 | 5 |
| Molasses for health protection | 10 | 10 | 10 | 10 |
| EDTA-Zn | 0.1 | 0.1 | 0.1 | 0.1 |
| Sodium octaborate tetrahydrate | 0.1 | 0.1 | 0.1 | 0.1 |
| EDTA-Mn | 0.1 | 0.1 | 0.1 | 0.1 |
| EDTA-Fe | 0.1 | 0.1 | 0.1 | 0.1 |
| EDTA-Cu | 0.1 | 0.1 | 0.1 | 0.1 |
| EDTA-2Na | 1 | 1 | 1 | 1 |
| Water (W) | 28.5 | 33.5 | 38.5 | 48.5 |
The preparation method comprises the following steps:
sequentially dissolving EDTA-Zn, sodium octaborate tetrahydrate, EDTA-Mn, EDTA-Fe, EDTA-Cu and EDTA-2Na in water; adding potassium humate while stirring, and adding molasses after the potassium humate is completely dissolved; sequentially adding ammonium polyphosphate, ammonium nitrate and dipotassium hydrogen phosphate while stirring; adding rhamnolipid, and mixing.
Example 4
The same amount of the bacterial manure for preventing and treating the crop bacterial wilt, which is disclosed in the embodiments 1-3 and the comparative example 1 of the invention, is prepared into a PDA solid culture medium (200 g of potatoes, 20g of glucose, 20g of agar, 1000mL of water and natural pH) by adopting a growth inhibition rate method, after the PDA solid culture medium is cooled to about 45 ℃, the bacterial manure, which is disclosed in the embodiments 1-3 and the comparative example 1 after filtration and sterilization, is respectively added, wherein the volume ratio of the bacterial manure to the culture medium is 1:9, and the contrast is a flat plate added with the same amount of clear water. Using a puncher with the diameter of 8.6mm to punch bacterial blocks of ralstonia solanacearum, moving the bacterial blocks to the center of each processing plate, repeating the same processing for 4 times, placing the processing plates in a constant temperature incubator at 28 ℃, and measuring and recording the diameters of bacterial colonies by using a vernier caliper by a cross method when the bacterial colonies on a control plate are about two thirds full of a culture dish, wherein the results are shown in table 3.
TABLE 3 bacteriostatic effect of bacteriostatic fertilizer for preventing and treating crop bacterial wilt
| Average colony diameter (mm) | Bacteriostatic ratio (%) | |
| Example 1 | 0.19±1.20 | 99.78 |
| Example 2 | 19.10±0.34 | 78.18 |
| Example 3 | 47.24±1.23 | 46.04 |
| Comparative example 1 | 79.50±0.67 | 9.21 |
As can be seen, the bacteriostatic rate of the examples 1-3 is higher than that of rhamnolipid alone at the same concentration, wherein the example 1 is the best example and has the best bacteriostatic effect.
And comparative example 1, in which rhamnolipid is not added and only other components in the bacteriostatic fertilizer are contained, has the worst effect. The ammonium polyphosphate can chelate trace elements under the condition of ensuring the supply of a high-concentration nitrogen source and a phosphorus source, maintains the stability of the antibacterial fertilizer, can also ensure the antibacterial activity of the antibacterial fertilizer, and has no obvious influence on the antibacterial effect.
As can be seen by comparison, the ralstonia solanacearum growth after the crop ralstonia solanacearum biological control agents (including the surfactant component) described in examples 1-3 is remarkably inhibited, while the ralstonia solanacearum biological control agent (excluding rhamnolipid) described in comparative example 1 has a poor effect on the inhibition of the ralstonia solanacearum growth. Therefore, the crop ralstonia solanacearum biological control agent has a remarkable inhibition effect on the growth of ralstonia solanacearum.
Example 5
Selecting plump and glossy tomato seeds, sterilizing, placing in a constant temperature incubator, controlling the temperature at 28-30 ℃, and sowing when most seeds are exposed to white. The seedling raising soil is peat nutrient soil, and the seedlings are raised in an incubator. The illumination condition of the incubator is 14 h.d-1Light intensity of 200. mu. mol. m-2·s-1Day and night temperature is 28 deg.C/25 deg.C, and relative humidity is 80%. Transplanting seedlings when the seedlings grow to have two leaves and one heart, selecting tomato seedlings with consistent growth vigor, transplanting the tomato seedlings into culture cups filled with 400mL Hoagland nutrient solution, transplanting one tomato seedling in each pot, placing the seedlings in an incubator for growth, and controlling the temperature at 28 ℃/25 ℃ day and night. 2 days before inoculation, 40mL of each of inventive examples 1-3 and comparative examples was used for root irrigation, and the control was treated without inoculation. When the tomato grows to 4-5 leaf stage, the root is wounded by scissors, and the concentration of 15mL is 1 multiplied by 107CFU·mL-1The bacterial liquid is poured into the nutrient solution for inoculation. After the control was rooted, the same amount of nutrient solution was added. And 3d after inoculation, when bacterial wilt begins to develop, investigating disease indexes every day until tomatoes die, and normally managing during investigation.
The disease index evaluation method comprises the following steps:
the onset of bacterial wilt was investigated according to the method of Fangzhong (1998).
1 level, half wilting of one leaf;
7 grade, all leaves wither;
grade 9, leaf and plant withering.
Disease index ∑ (disease-grade number of plants × representative value)/(total number of plants × representative value of the most serious disease-grade) × 100%
The incidence of bacterial wilt in tomato plants 3-7 days after inoculation is shown in Table 4, and the statistical chart is shown in FIG. 3. A comparison of the status of the plants of examples 1-3, comparative example 1 and control at day 7 after inoculation is shown in FIG. 3. On day 4 post inoculation, example 1 reduced the incidence of disease by 57.06% over the control. As shown in fig. 2, examples 1, 2 and 3 significantly reduced the disease index of tomato compared to the comparative example under the ralstonia solanacearum inoculation condition. On day 7 of inoculation, the treatments of examples 1, 2, and 3 reduced the incidence of tomato bacterial wilt disease by 50.57%, 64.37%, and 68.39%, respectively, as compared to the comparative example. The silicon application can enhance the resistance of the tomato to bacterial wilt and relieve the damage of diseases to the growth of the tomato.
It can be seen that the control effect on bacterial wilt is the best in example 3 of the invention.
The technical features of the above embodiments can be arbitrarily combined, and for the sake of brevity, all possible combinations of the technical features in the above embodiments are not described, however, as long as there is no contradiction between the combinations of the technical features, the scope of the present description should be considered as being described in the present specification.
The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the present invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.
Claims (7)
1. The application of the rhamnolipid in preparing the bacterial manure for preventing and treating the bacterial wilt of crops, the rhamnolipid is used as an inhibitor for inhibiting bacterial wilt,
the raw materials of the bacteriostatic fertilizer for preventing and treating the bacterial wilt of the crops comprise rhamnolipid, humate, molasses, a nitrogen source, a phosphorus source, a potassium source, trace elements and a chelating agent;
the bacteriostatic fertilizer for preventing and treating the crop bacterial wilt comprises the following raw materials in parts by weight:
5-35 parts of rhamnolipid;
1-20 parts of humate;
1-20 parts of molasses;
1-35 parts of a nitrogen source;
1-35 parts of a phosphorus source;
1-30 parts of a potassium source;
0.1-2 parts of trace elements;
0.1-2 parts of a chelating agent.
2. The use of the rhamnolipid according to claim 1 for the preparation of a bacterial manure inhibitor for controlling bacterial wilt of crops, characterized in that the nitrogen source comprises one or more of ammonium nitrate, potassium nitrate, ammonium polyphosphate and ammonium dihydrogen phosphate.
3. The use of the rhamnolipid according to claim 1 for the preparation of a bacterial manure inhibitor for controlling bacterial wilt of crops, characterized in that the phosphorus source comprises one or more of phosphoric acid, ammonium dihydrogen phosphate, ammonium hydrogen phosphate, ammonium polyphosphate, potassium hydrogen phosphate and dipotassium hydrogen phosphate; and/or
The potassium source comprises one or more of potassium chloride, potassium hydrogen phosphate, dipotassium hydrogen phosphate and potassium nitrate.
4. The use of the rhamnolipid according to claim 1 in the preparation of an antibacterial fertilizer for controlling crop bacterial wilt, wherein the trace elements include one or more of copper sulfate, manganese sulfate, zinc sulfate, boric acid, EDTA-Zn, sodium octaborate tetrahydrate, EDTA-Mn, EDTA-Fe, EDTA-Cu.
5. The use of the rhamnolipid according to claim 1 in the preparation of an antibacterial fertilizer for controlling bacterial wilt of crops, characterized in that the chelating agent comprises one or more of EDTA-2Na, citric acid, amino acid, malic acid.
6. The application of the rhamnolipid according to any one of claims 2 to 5 in preparing a bacterial manure for preventing and treating crop bacterial wilt, which is characterized in that the bacterial manure for preventing and treating crop bacterial wilt comprises the following raw materials in parts by weight:
15-25 parts of rhamnolipid;
10-20 parts of ammonium polyphosphate;
5-15 parts of ammonium nitrate;
8-12 parts of dipotassium phosphate;
3-8 parts of potassium humate;
8-12 parts of molasses;
0.05-0.15 part of EDTA-Zn;
0.05-0.15 parts of tetrahydrate sodium octaborate;
0.05-0.15 part of EDTA-Mn;
0.05-0.15 part of EDTA-Fe;
0.05-0.15 part of EDTA-Cu;
0.8-1.2 parts of EDTA-2 Na.
7. A preparation method of the bacteriostatic fertilizer for controlling the bacterial wilt of crops as the application of any one of claims 2 to 6, which is characterized by comprising the following steps:
sequentially dissolving the trace elements and the chelating agent in water, adding humate while stirring, and adding the molasses after the humate is completely dissolved; adding a nitrogen source, a phosphorus source and a potassium source while stirring; adding rhamnolipid, and mixing.
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Effective date of registration: 20220718 Address after: No.10, shiliugang Road, Haizhu District, Guangzhou City, Guangdong Province 510000 Patentee after: Nanfan Seed Industry Research Institute Guangdong Academy of Sciences Address before: No. 10, shiliugang Road, Guangzhou, Guangdong 510000 Patentee before: Institute of bioengineering, Guangdong Academy of Sciences |
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