A bio-ingredient suspension seed coating agent for preventing and treating peanut root rot and application thereof

Abstract

This invention relates to the field of plant disease control technology, and discloses a biological suspension seed coating agent for controlling peanut root rot and its application. The composition of the agent is: Bacillus amyloliquefaciens: 2 × 10⁻⁶ 7 ~1×10 8 CFU, chitosan: 100-150 g / L, glycerol adjuvant: 30-50 g / L, CMC-Na stabilizer: 20-30 g / L. This invention utilizes the combined use of Bacillus amyloliquefaciens, chitosan, glycerol adjuvant, and stabilizer to eradicate peanut root rot using a suspension seed coating agent. This results in a more effective cure for peanut root rot, increasing the number of seedlings and reducing the impact of rot on seedling emergence, thus improving the seedling rate. Each treatment was repeated in 30 pots, cultured in an artificial climate chamber at 28°C and 95% relative humidity. The disease incidence was assessed 25 days after treatment, and the degree of vascular bundle browning was observed by dissecting the plant stems to calculate the relative efficacy. The composition was then applied to the roots of artificially cultivated peanut seedlings in a net-house for root rot prevention, followed by inoculation with peanut root rot pathogens.

Description

Technical Field

[0001] This invention relates to the field of plant disease control technology, specifically to a biological suspension seed coating agent for controlling peanut root rot and its application. Background Technology

[0002] Peanuts (Arachis hypogaea L.) are the world's fourth largest oilseed and thirteenth largest food source crop, and an important source of high-quality plant protein and oil. They are also a significant economic resource for some developing countries in Asia, South America, and Africa. According to statistics from the Food and Agriculture Organization of the United Nations (FAO), global peanut production in 2017 totaled over 40 million tons, while China's total peanut production that year was over 17 million tons, accounting for 40% of the world's total, ranking first globally. This is six times the total peanut production of the United States and five times that of India. Peanuts are an important economic and oilseed crop in my country. Peanuts are a popular food, rich in fat, protein, dietary fiber, and various micronutrients. The most important nutrient in peanut kernels is fat, which accounts for 39% to 62%. Peanuts contain a variety of fatty acids, including oleic acid, linoleic acid, arachidic acid, palmitic acid, stearic acid, arachidic acid, and behenic acid. Among them, unsaturated fatty acids mainly include oleic acid and linoleic acid, which together account for about 80% of the total fatty acid content. Oleic acid can effectively reduce the level of low-density lipoprotein in the blood, which is harmful to the human body, while maintaining the level of high-density lipoprotein, which is beneficial to the human body, thus protecting cardiovascular health.

[0003] Peanut root rot, also known as "rat's tail" or "root rot," is a fungal soil-borne disease. It occurs throughout my country, severely impacting peanut yield and quality. Peanut root rot is primarily caused by fungi of the genus *Fusarium*, such as *Fusarium solani* and *F. oxysporum*. Root rot can occur throughout the peanut's growth cycle, mainly affecting the roots and vascular bundles. The roots turn brown and rot, subsequently causing the vascular bundles to turn brown and rot, leading to poor plant growth and eventually the death of the entire plant. Later infection can damage the pods, causing pod rot, resulting in the death of most or all of the plant, leading to missing seedlings and gaps in the rows. The incidence rate is generally between 5% and 10%, but can reach over 30% in severe cases, causing significant economic losses and severely hindering the development of my country's peanut industry. ] .

[0004] For a long time, some scholars at home and abroad have conducted multi-angle and multi-field research on peanut root rot and its pathogens, and have made some progress. However, there has been no major breakthrough in disease-resistant breeding. More and more people have begun to shift their attention to the research on the occurrence and integrated control technology of peanut root rot, and have achieved gratifying results. This article focuses on reviewing the latest research progress on the occurrence and integrated control of peanut root rot from three aspects: disease symptoms, occurrence patterns and control technology. The aim is to provide a theoretical basis for in-depth research and effective control of peanut root rot. Therefore, in the process of peanut planting, it is necessary to use a biological suspension seed coating agent for the control of peanut root rot and its application.

[0005] The inventors discovered that the existing technology suffers from at least the following unresolved problems: 1. When treating peanut root rot, ordinary root-curing suspensions are ineffective against peanut roots, resulting in fewer seedlings and thus lowering the germination rate; 2. When using suspension seed coating agents, they only target the germination rate, failing to address root rot effectively and prevent its spread, thus reducing the functionality of the suspension seed coating agents; 3. When using suspension seed coating agents, it is impossible to test their preventive effect and application in fields severely affected by peanut root rot, thus reducing the comprehensiveness of their use. Therefore, this invention designs a biological suspension seed coating agent for the prevention and control of peanut root rot and its application. Summary of the Invention

[0006] The purpose of this invention is to provide a biological component suspension seed coating agent for the prevention and control of peanut root rot and its application, addressing the shortcomings of the prior art. Ordinary root-curing suspension agents have poor efficacy against peanut roots, resulting in fewer seedlings and a lower emergence rate. Furthermore, the application of suspension seed coating agents only targets the emergence rate, failing to address root rot and reduce their overall effectiveness. Moreover, the effectiveness and application status of suspension seed coating agents in severely affected peanut root rot fields cannot be assessed, thus limiting their comprehensive application. Therefore, this invention designs a biological component suspension seed coating agent for the prevention and control of peanut root rot and its application.

[0007] To achieve the above objectives, the present invention provides the following technical solution: a biological component suspension seed coating agent for preventing and controlling peanut root rot, the composition of which is as follows:

[0008] Bacillus amyloliquefaciens: 2×10 7 ~1×10 8 CFU;

[0009] Chitosan: 100-150 g / L;

[0010] Additive glycerin: 30-50 g / L;

[0011] Stabilizer CMC-Na: 20-30 g / L.

[0012] Preferably, formulation 1 contains 2 × 10⁶ Bacillus amyloliquefaciens. 7 CFU, chitosan 100 g / L, glycerol 30 g / L (auxiliary agent), and CMC-Na 20 g / L (stabilizer).

[0013] Preferably, the component of formulation 2 is 1×10⁻⁶ Bacillus amyloliquefaciens. 8 CFU, chitosan 150 g / L, glycerol 50 g / L (auxiliary agent), and CMC-Na 30 g / L (stabilizer).

[0014] Preferably, component 3 of formulation is 2×10⁻⁶ Bacillus amyloliquefaciens. 7 CFU, chitosan 150 g / L, glycerol 40 g / L (auxiliary agent), and CMC-Na 25 g / L (stabilizer).

[0015] Preferably, Bacillus amyloliquefaciens is a Gram-positive spore-forming bacillus that is highly related to Bacillus subtilis and is effective against anthrax, gray mold, and Fusarium.

[0016] Preferably, CMC-Na is an important water-soluble polymeric stabilizer that maintains the homogeneity and physical stability of food and industrial products through thickening, emulsification, and film-forming mechanisms.

[0017] An application of a biological suspension seed coating agent for the prevention and control of peanut root rot, the preparation method of which includes the following steps:

[0018] Step 1: The experimenters prepared samples from formulations 1, 2 and 3 for testing. Then, they prepared the 350 g / L metalaxyl seed treatment emulsion, 25 g / L fludioxonil suspension seed coating agent and 10% azoxystrobin suspension seed coating agent for comparison. Then, they prepared the experimental site and screened the seeds for the experiment to ensure they were defect-free and consistent.

[0019] Step 2: The experimenters replicated the seeds in 3 pots for each treatment, and placed each pot in a constant temperature net room at 28℃. The number of seedlings was investigated 20 days after sowing, and the germination rate was calculated.

[0020] Step 3: The researchers replicated each seed treatment in 30 pots. The disease incidence was investigated 25 days after treatment. The degree of browning of the vascular bundles was observed by dissecting the stems of the plants, and the relative control efficacy was calculated.

[0021] Step 4: The preventive effect of the composition after drenching the roots of artificially cultivated peanut seedlings in net houses and then inoculating them with peanut root rot pathogens was verified. Next, the therapeutic effect of inoculating artificially cultivated peanut seedlings with peanut root rot pathogens and then drenching the roots with the medicine was verified. Finally, the control effect of the composition on fields with severe peanut root rot in previous years was verified under field conditions.

[0022] Step 5: Calculate the collected data, plot it in a table, compare the data, draw the experimental results, and modify the formula accordingly.

[0023] Preferably, in step one, each pot is placed in a constant temperature net room at 28°C.

[0024] Preferably, in step two, the plants are cultured in an artificial climate chamber at 28°C and 95% relative humidity.

[0025] Compared with the prior art, the beneficial effects of the present invention are:

[0026] 1. This invention utilizes the combined use of Bacillus amyloliquefaciens, chitosan, glycerol, and stabilizer when treating peanut root rot with a suspension seed coating agent. This combination effectively eradicates peanut root rot, increases the number of peanut seedlings, reduces the impact of rot on peanut seedling emergence, and thus improves the peanut seedling emergence rate.

[0027] 2. This invention involves repeating each treatment in 30 pots, culturing them in an artificial climate chamber at 28°C and 95% relative humidity, investigating the disease incidence 25 days after treatment, observing the degree of vascular bundle browning by dissecting the plant stems, calculating the relative preventive effect, and then applying the composition to the roots of artificially cultivated peanut seedlings in a net-house for root rot prevention and treatment. This allows for the testing of the effect of suspension seed coating agents on the prevention and treatment of rot, thus improving the multifunctionality of suspension seed coating agents for peanuts.

[0028] 3. This invention utilizes 2×10 Bacillus amyloliquefaciens 7 After treating peanuts with CFU, chitosan 100 g / L, adjuvant glycerol 30 g / L, and stabilizer CMC-Na 20 g / L, the control effect of the combination on peanut root rot in fields with severe occurrences in previous years was obtained under field conditions. This allows us to obtain the control effect of suspension seed coating agents on actual fields and improves the authenticity of seed coating agent application results. Attached Figure Description

[0029] Figure 1 This is a schematic diagram of the process for the suspension seed coating agent of the present invention;

[0030] Figure 2 This is an experimental diagram showing the effect of the composition of the present invention on peanut emergence rate. Detailed Implementation

[0031] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0032] See Figure 1-2 The present invention provides a technical solution: a biological component suspension seed coating agent for preventing and controlling peanut root rot.

[0033] Application Example 1:

[0034] This application example aims to verify the effect of the described composition on peanut germination rate. The test formulations used were samples prepared in Examples 1, 2, and 3.

[0035] Each treatment was replicated in triplicate, with each pot placed in a 28℃ constant-temperature greenhouse. The number of seedlings was assessed 20 days after sowing, and the germination rate was calculated.

[0036] Table 1. Data on the impact of peanut emergence rate

[0037] Example 1 300 256 85.33 Example 2 300 267 89.00 Example 3 300 243 81.00 Comparative Example 1 300 187 62.33 Comparative Example 2 300 223 74.33 Comparative Example 3 300 198 66.00 CK (Shimizu) 300 137 45.67

[0038] Example 1: Bacillus amyloliquefaciens 2×10 7 CFU; chitosan 100 g / L; glycerol 30 g / L; stabilizer CMC-Na 20 g / L.

[0039] Example 2: Bacillus amyloliquefaciens 1×10 8 CFU; chitosan 150 g / L; glycerol 50 g / L; stabilizer CMC-Na 30 g / L.

[0040] Example 3: Bacillus amyloliquefaciens 2×10 7 CFU; chitosan 150 g / L; glycerol 40 g / L; stabilizer CMC-Na 25 g / L.

[0041] Comparative Example 1: 350 g / L metalaxyl seed treatment emulsion.

[0042] Comparative Example 2: 25 g / L fludioxonil suspension seed coating agent.

[0043] Comparative Example 3: 10% azoxystrobin suspension seed coating agent.

[0044] Application Example 2:

[0045] This application example aims to verify the efficacy of the composition in controlling peanut root rot. Each treatment was replicated in 30 pots and cultured in an artificial climate chamber at 28°C and 95% relative humidity. Disease incidence was assessed 25 days after treatment, and the degree of browning of the vascular bundles was observed by dissecting the plant stems to calculate the relative control efficacy.

[0046] Table 2. Data on the effects of peanut stem base vascular bundles and their control efficacy.

[0047] Example 1 15.64 83.08 Example 2 17.72 80.83 Example 3 16.33 82.34 Comparative Example 1 38.71 58.13 Comparative Example 2 41.95 54.63 Comparative Example 3 34.74 62.43 CK (Shimizu) 92.46 —

[0048] Example 1: Bacillus amyloliquefaciens 2×10 7 CFU; chitosan 100 g / L; glycerol 30 g / L; stabilizer CMC-Na 20 g / L.

[0049] Example 2: Bacillus amyloliquefaciens 1×10 8 CFU; chitosan 150 g / L; glycerol 50 g / L; stabilizer CMC-Na 30 g / L.

[0050] Example 3: Bacillus amyloliquefaciens 2×10 7 CFU; chitosan 150 g / L; glycerol 40 g / L; stabilizer CMC-Na 25 g / L.

[0051] Comparative Example 1: 350 g / L metalaxyl seed treatment emulsion.

[0052] Comparative Example 2: 25 g / L fludioxonil suspension seed coating agent.

[0053] Comparative Example 3: 10% azoxystrobin suspension seed coating agent.

[0054] Application Example 3:

[0055] This application example aims to verify the preventive effect of the composition on peanut seedlings artificially cultivated in net houses after root irrigation and inoculation with peanut root rot pathogens.

[0056] Table 3. Data on the control effect of peanut root rot.

[0057] Example 1 4.43 79.51 Example 2 4.05 81.27 Example 3 3.71 82.84 Comparative Example 1 9.74 54.95 Comparative Example 2 8.97 58.51 Comparative Example 3 7.38 65.86 CK (Shimizu) 21.62 —

[0058] Example 1: Bacillus amyloliquefaciens 2×10 7 CFU; chitosan 100 g / L; glycerol 30 g / L; stabilizer CMC-Na 20 g / L.

[0059] Example 2: Bacillus amyloliquefaciens 1×10 8 CFU; chitosan 150 g / L; glycerol 50 g / L; stabilizer CMC-Na 30 g / L.

[0060] Example 3: Bacillus amyloliquefaciens 2×10 7 CFU; chitosan 150 g / L; glycerol 40 g / L; stabilizer CMC-Na 25 g / L.

[0061] Comparative Example 1: 350 g / L metalaxyl seed treatment emulsion.

[0062] Comparative Example 2: 25 g / L fludioxonil suspension seed coating agent.

[0063] Comparative Example 3: 10% azoxystrobin suspension seed coating agent.

[0064] Application Example 4:

[0065] This application example aims to verify the therapeutic effect of inoculating artificially cultivated peanut seedlings with peanut root rot disease and then applying pesticides through root irrigation.

[0066] Table 4. Data on the control effect of peanut root rot.

[0067] Example 1 5.13 78.08 Example 2 6.32 73.00 Example 3 4.76 79.67 Comparative Example 1 13.44 42.59 Comparative Example 2 12.65 45.96 Comparative Example 3 10.88 53.52 CK (Shimizu) 23.41 —

[0068] Example 1: Bacillus amyloliquefaciens 2×10 7 CFU; chitosan 100 g / L; glycerol 30 g / L; stabilizer CMC-Na 20 g / L.

[0069] Example 2: Bacillus amyloliquefaciens 1×10 8 CFU; chitosan 150 g / L; glycerol 50 g / L; stabilizer CMC-Na 30 g / L.

[0070] Example 3: Bacillus amyloliquefaciens 2×10 7 CFU; chitosan 150 g / L; glycerol 40 g / L; stabilizer CMC-Na 25 g / L.

[0071] Comparative Example 1: 350 g / L metalaxyl seed treatment emulsion.

[0072] Comparative Example 2: 25 g / L fludioxonil suspension seed coating agent.

[0073] Comparative Example 3: 10% Azoxystrobin Suspension Seed Coating

[0074] Application Example 5:

[0075] This application example aims to verify the control effect of the composition on peanut root rot in fields that have been severely affected in previous years under field conditions.

[0076] Table 5. Field control efficacy data for peanut root rot.

[0077] Example 1 3.47 82.43 Example 2 5.11 74.13 Example 3 4.53 77.06 Comparative Example 1 9.67 51.04 Comparative Example 2 8.93 54.78 Comparative Example 3 10.15 48.61 CK (Shimizu) 19.75 —

[0078] Example 1: Bacillus amyloliquefaciens 2×10 7 CFU; chitosan 100 g / L; glycerol 30 g / L; stabilizer CMC-Na 20 g / L.

[0079] Example 2: Bacillus amyloliquefaciens 1×10 8 CFU; chitosan 150 g / L; glycerol 50 g / L; stabilizer CMC-Na 30 g / L.

[0080] Example 3: Bacillus amyloliquefaciens 2×107 CFU; chitosan 150 g / L; glycerol 40 g / L; stabilizer CMC-Na 25 g / L.

[0081] Comparative Example 1: 350 g / L metalaxyl seed treatment emulsion.

[0082] Comparative Example 2: 25 g / L fludioxonil suspension seed coating agent.

[0083] Comparative Example 3: 10% azoxystrobin suspension seed coating agent.

[0084] In summary, this invention proposes a biological suspension seed coating agent for the prevention and control of peanut root rot and its application. When the suspension seed coating agent is used to eradicate peanut root rot, the combined use of Bacillus amyloliquefaciens, chitosan, glycerol, and stabilizer can effectively eradicate peanut root rot, thereby increasing the number of peanut seedlings and reducing the impact of rot on peanut seedling emergence, thus improving the peanut seedling emergence rate.

[0085] It should be noted that, in this document, relational terms such as "first" and "second" are used only to distinguish one entity or operation from another, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Furthermore, the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, technology, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such process, technology, article, or apparatus.

[0086] Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims and their equivalents.

Claims

1. A biological suspension seed coating agent for controlling peanut root rot, characterized in that: The composition of this preparation is as follows: Bacillus amyloliquefaciens: 2×10 7 ~1×10 8 CFU; Chitosan: 100-150 g / L; Additive glycerin: 30-50 g / L; Stabilizer CMC-Na: 20-30 g / L.

2. The biological component suspension seed coating agent for preventing peanut root rot according to claim 1, characterized in that: Formulation 1 consists of 2 × 10⁶ Bacillus amyloliquefaciens. 7 CFU, chitosan 100 g / L, glycerol 30 g / L (auxiliary agent), and CMC-Na 20 g / L (stabilizer).

3. The biological component suspension seed coating agent for preventing peanut root rot according to claim 1, characterized in that: Formulation 2 consists of 1×10 Bacillus amyloliquefaciens. 8 CFU, chitosan 150 g / L, glycerol 50 g / L (auxiliary agent), and CMC-Na 30 g / L (stabilizer).

4. The biological component suspension seed coating agent for controlling peanut root rot according to claim 1, characterized in that: Formulation 3 contains 2×10 Bacillus amyloliquefaciens. 7 CFU, chitosan 150 g / L, glycerol 40 g / L (auxiliary agent), and CMC-Na 25 g / L (stabilizer).

5. The biological component suspension seed coating agent for controlling peanut root rot according to claim 1, characterized in that: Bacillus amyloliquefaciens is a Gram-positive spore-forming bacillus that is highly related to Bacillus subtilis and is effective against anthrax, gray mold, and Fusarium.

6. The biological suspension seed coating agent for controlling peanut root rot according to claim 1, characterized in that: CMC-Na is an important water-soluble polymeric stabilizer that maintains the homogeneity and physical stability of food and industrial products through thickening, emulsification, and film-forming mechanisms.

7. The application of a biological component suspension seed coating agent for controlling peanut root rot, based on the biological component suspension seed coating agent for controlling peanut root rot according to any one of claims 1-6, characterized in that, Includes the following steps: Step 1: The experimenters prepared samples from formulations 1, 2 and 3 for testing. Then, they prepared the 350 g / L metalaxyl seed treatment emulsion, 25 g / L fludioxonil suspension seed coating agent and 10% azoxystrobin suspension seed coating agent for comparison. Then, they prepared the experimental site and screened the seeds for the experiment to ensure they were defect-free and consistent. Step 2: The experimenters replicated the seeds in 3 pots for each treatment, and placed each pot in a constant temperature net room at 28℃. The number of seedlings was investigated 20 days after sowing, and the germination rate was calculated. Step 3: The researchers replicated each seed treatment in 30 pots. The disease incidence was investigated 25 days after treatment. The degree of browning of the vascular bundles was observed by dissecting the stems of the plants, and the relative control efficacy was calculated. Step 4: The preventive effect of the composition after drenching the roots of artificially cultivated peanut seedlings in net houses and then inoculating them with peanut root rot pathogens was verified. Next, the therapeutic effect of inoculating artificially cultivated peanut seedlings with peanut root rot pathogens and then drenching the roots with the medicine was verified. Finally, the control effect of the composition on fields with severe peanut root rot in previous years was verified under field conditions. Step 5: Calculate the collected data, plot it in a table, compare the data, draw the experimental results, and modify the formula accordingly.

8. The application of the biological component suspension seed coating agent for controlling peanut root rot according to claim 7, characterized in that: In step one, each pot is placed in a constant temperature net room at 28℃.

9. The application of the biological component suspension seed coating agent for controlling peanut root rot according to claim 7, characterized in that: In step two, the cells are placed in an artificial climate chamber at 28°C and 95% relative humidity for incubation.

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