Use of chlorindole hydrazide in seed treatment

Abstract

The present application belongs to the technical field of seed treatment, and provides the use of chlorindole hydrazide in seed treatment, and specifically provides a seed treatment agent with chlorindole hydrazide as an effective component, and the dosage form of the treatment agent is a flowable concentrate. The seed treatment agent can overcome the disadvantage that long-term single use of a pesticide is prone to resistance, prolong the service life of the pesticide, and has important significance for the comprehensive management of resistance of diseases such as wheat scab, sheath blight, leaf blight, tobacco virus disease, tomato virus disease, and pepper virus disease. Meanwhile, the seed treatment with chlorindole hydrazide can also improve the germination rate and promote the growth of seedlings. The present application provides a new application direction for chlorindole hydrazide and fills the corresponding blank.

Description

Technical Field

[0001] This invention belongs to the field of seed treatment technology and provides the use of chloroindolehydrazine in seed treatment. Background Technology

[0002] During crop growth and development, crops are susceptible to various diseases, especially fungal diseases, which are numerous and cause serious damage. For a long time, chemical control has been the main and effective means of controlling these diseases.

[0003] Common fungal diseases include wheat scab, wheat glume blight, and wheat take-all. These diseases severely impact wheat growth. Currently, the main fungicides used to control these diseases include difenoconazole, azoxystrobin, and carbendazim. These pesticides have been used continuously for many years, and pathogens have developed resistance to them, resulting in adverse effects such as reduced seed germination rates (paper: Effects of different seed dressing agents on wheat germination and seedling growth; Hou Wenbang et al., 2011). Moreover, based on existing technologies, Huang Tingting and Zhou Mingguo from the College of Plant Protection at Nanjing Agricultural University shared at the 2011 Annual Meeting of the Chinese Society for Plant Pathology that the commonly used fungicide carbendazim can induce resistance and upregulate the expression of DON toxin synthesis genes in plants. Increased expression of DON toxin (vomitoxin, which has certain harmful effects on humans and is classified as a Group 3 carcinogen by the European Union) greatly increases the risk of wheat grain toxin contamination. Therefore, existing commonly used control agents have shown many drawbacks, and even with increased dosage, the control effect is still not ideal.

[0004] Viral diseases, commonly known as "dragon head disease," are diseases affecting a wide range of crops, including tomatoes, cucumbers, peppers, eggplants, watermelons, and cabbages. They are widespread, cause serious damage, and are difficult to control. Once infected, they can lead to reduced yields or even total crop failure, earning them the nickname "plant cancer." There are many types of viral diseases, primarily affecting the leaves, growing points, fruits, and stems of crops. Infected leaves curl and fail to flatten, or exhibit mosaic patterns, severely reducing photosynthesis and even causing death. Infected fruits develop uneven surfaces, inconsistent coloring, and spoiled flesh, rendering them inedible and commercially worthless. Infected stems develop long, brown lesions, leading to stem necrosis in severe cases. Infected growing points cause slow growth, distortion, and even death.

[0005] During seed production, if the plant becomes infected with a viral disease, the virus can enter and survive inside the seed. When seeds infected with the virus are sown, the plant becomes a carrier. Since seeds themselves carry many pathogens and viruses, pre-sowing disinfection can significantly reduce the probability of disease development after sowing. Common disinfection methods include warm water soaking and chemical disinfection. However, their effectiveness is limited, and seed-borne viruses are often a key cause of viral diseases in the field. Therefore, a method that addresses the root cause is needed. Currently, there is a severe shortage of agents for seed treatment to control viral diseases, posing a significant challenge to seed propagation and subsequent cultivation. Summary of the Invention

[0006] This invention addresses the aforementioned shortcomings of existing technologies by providing a use of chloroindolehydrazine in seed treatment. Specifically, it provides a seed treatment agent with chloroindolehydrazine as the active ingredient, in the form of a suspension seed coating agent. Using this seed coating agent to treat plant seeds overcomes the drawback of resistance development from long-term use of single agents, extends the agent's lifespan, and is of great significance for the integrated management of resistance to diseases such as wheat scab, sheath blight, glume blight, tobacco virus diseases, tomato virus diseases, and pepper virus diseases. Furthermore, chloroindolehydrazine seed treatment can improve germination rate and promote seedling growth. This invention provides a completely new application direction for chloroindolehydrazine, filling a relevant gap in the market.

[0007] N-(4-chlorophenylmethylene)-1-methyl-2,3,4,9-tetrahydropyrido[3,4-b]indole-3-carboxylhydrazine, abbreviated as chloroindolehydrazine, has the following structural formula:

[0008]

[0009] This compound was jointly developed by Nankai University and the applicant. After years of research, foliar spraying of chloroindolehydrazine suspension onto plants has shown that it effectively controls various fungal and viral diseases. It exhibits excellent biological activity, is easily absorbed by plants, readily degrades in the environment, and has good environmental compatibility. The applicant was surprised to discover during experiments that seed treatment with chloroindolehydrazine can promote seed germination and seedling growth, and effectively control various fungal and viral diseases. This is of great significance for crop growth and the prevention of fungal and viral diseases.

[0010] The specific technical solution of the present invention is as follows:

[0011] Uses of chloroindolehydrazine in seed treatment.

[0012] Chloroindolehydrazine, as an active ingredient in seed treatment agents, can promote seed germination, seedling growth, and / or prevent plant viral or bacterial / fungal diseases.

[0013] The plant virus mentioned is tobacco mosaic virus, pepper virus, or tomato virus, and the bacterial / fungal disease is wheat scab, wheat take-all, wheat glume blight, or rice sheath blight.

[0014] The seeds mentioned are any one of wheat seeds, corn seeds, chili pepper seeds, tomato seeds, tobacco seeds, and rice seeds.

[0015] The present invention provides a seed treatment agent, wherein the active ingredient of the seed treatment agent is chloroindolehydrazine, and the content of chloroindolehydrazine is 1-30 wt%, more preferably 10-20 wt%.

[0016] The preferred formulation of the above-mentioned seed treatment agent is a suspension seed coating agent. In addition, the treatment agent also contains one or more of the following: film-forming agent, wetting and dispersing agent, defoamer, thickener, antifreeze agent, preservative, and warning color. All of the above adjuvants are known substances and are various adjuvants commonly used in pesticide formulations. They may vary depending on different situations and are not particularly limited.

[0017] In a specific implementation method

[0018] The film-forming agent is one or a mixture of two of the following: non-water-soluble chitin derivatives, polyvinyl alcohol, and cellulose acetate ester;

[0019] The wetting and dispersing agent is one or a mixture of several of the following: alkylbenzene sulfonate, fatty alcohol polyoxyethylene ether, phenethylphenol polyoxypropylene polyoxyethylene ether, castor oil polyoxyethylene ether, phenethyl polyoxyethyl polyoxypropylene ether, phenethylphenol polyoxyethylene ether, alkylphenol polyoxyethylene ether phosphate, lignin and its derivative sulfonate, polycarboxylate, fatty acid ethane adduct phosphate, alkylphenol polyvinyl ether sulfonate, naphthalene or alkylnaphthalene formaldehyde condensate and its salts;

[0020] The thickener is xanthan gum or magnesium aluminum silicate;

[0021] The antifreeze agent is one or a mixture of several of glycerin, ethylene glycol, propylene glycol, and sorbitol;

[0022] The defoamer is one or a mixture of several of the following: silicone oil, ethanol, epoxidized soybean oil, amides, and phosphate esters;

[0023] The preservative is one or a mixture of two of methylparaben or sodium benzoate;

[0024] The warning color is one or a mixture of two of the dyes: bright red or disperse red.

[0025] After obtaining the above seed treatment agent, conventional methods can be used for seed dressing.

[0026] Preferably, after treating the seeds with the above-mentioned seed treatment agent, the wheat seeds can also be uniformly treated with 70wt% imidacloprid wet seed dressing agent to control wheat aphids and other underground pests. This invention is not limited to imidacloprid as a single insecticide seed treatment agent. The amount of water used for seed dressing is 3% of the seed weight (refer to DB34 / T 2908-2017 Field Efficacy Test Guidelines for Seed Treatment Agents). However, imidacloprid is preferred because it is a neonicotinoid insecticide with strong systemic activity and high activity, possessing both stomach poison and contact action, primarily controlling wheat aphids. Suitable crops include wheat, barley, rice, sugar beets, rapeseed, legumes, vegetables, grapes, and apples. At the recommended dosage, it is safe for crops and causes no phytotoxicity.

[0027] In summary, the seed treatment agent provided in this application exhibits significant control effects within a certain ratio range, and is significantly improved compared to other agents. As a seed treatment agent, it broadens its fungicidal spectrum, overcomes the disadvantage of resistance easily generated by long-term single use of agents, and extends the service life of the agent. It is of great significance for the integrated management of resistance to diseases such as wheat scab, take-all, glume blight, tobacco mosaic virus, pepper virus, and tomato virus, and ensures crop safety. At the same time, it can significantly improve seed germination rate and promote seedling growth. Detailed Implementation

[0028] The following detailed description of specific embodiments further illustrates the above-described content of the present invention, but it should not be construed as limiting the scope of the subject matter of the present invention to the following examples. All technologies implemented based on the above-described content of the present invention fall within the scope of the present invention. All percentages below are by weight, and all additives used are conventional additives in the art.

[0029] Formulation Example 1: 10% Chloroindolehydrazine suspension seed coating agent

[0030] Raw material composition: 10% chloroindolehydrazine technical grade, 1% polycarboxylate GY-D07 dispersant (Beijing Guangyuan Yinong), 4% sodium naphthalenesulfonic acid formaldehyde condensate dispersant, 4% ethylene glycol antifreeze, 1.5% magnesium aluminum silicate thickener, 0.2% xanthan gum thickener, 0.5% sodium benzoate preservative, 3% polyvinyl alcohol film-forming agent, 0.2% organosilicon defoamer, 5% bright red dye, 2% non-water-soluble chitin derivative, and deionized water to 100%.

[0031] The above-mentioned suspension seed coating agent preparation method is as follows: after mixing and shearing the above materials evenly, they are added to a sand mill for grinding so that the particle size D90 is controlled below 5 μm, and then filtered to obtain 10% chloroindolehydrazine suspension seed coating agent.

[0032] Formulation Example 2: 20% Chloroindolehydrazine Suspension Seed Coating

[0033] Chloroindolehydrazine technical grade 20%, dispersant GY-D07 (Beijing Guangyuan Yinong) 2%, dispersant sodium naphthalenesulfonic acid formaldehyde condensate 4%, antifreeze ethylene glycol 4%, thickener magnesium aluminum silicate 1%, thickener xanthan gum 0.15%, preservative sodium benzoate 0.5%, film-forming agent polyvinyl alcohol 5%, defoamer organosilicon 0.2%, dye bright red 5%, non-water-soluble chitin derivative 2%, deionized water to 100%.

[0034] The method for preparing the suspension seed coating agent is as follows: after mixing and shearing the above materials evenly, they are added to a sand mill for grinding so that the particle size D90 is controlled below 5 μm, and then filtered to obtain 20% chloroindolehydrazine suspension seed coating agent.

[0035] Application Example 1: Chloroindolehydrazine suspension seed coating agent promotes seed germination and seedling growth in potted wheat.

[0036] Variety: Jimai 22 (disease-free seeds);

[0037] Experimental location: Jingbo Agricultural Chemical Experimental Base, Boxing County, Shandong Province.

[0038] Test reagents:

[0039] 1. Formulation Example 1: Chloroindolehydrazine suspension seed coating agent;

[0040] 2. 0.1% S-inducer soluble solution;

[0041] 3. 0.01% 24-Brassinolide soluble concentrate;

[0042] Seed dressing method:

[0043] Wheat seeds were treated with various pesticides, then dried in a cool place for 3-4 hours before being uniformly treated with 70% imidacloprid wet seed treatment agent. This method is used to control wheat aphids and other underground pests. This invention is not limited to imidacloprid as a single insecticide seed treatment agent. The amount of water used for seed treatment is 3% of the seed weight (refer to DB34 / T 2908-2017 Field Efficacy Test Guidelines for Seed Treatment Agents).

[0044] Table 1. Experimental Design of Seed Treatment for Jimai 22

[0045]

[0046] During the protected cultivation (greenhouse) experiment, the temperature was (30℃-20℃)±7℃; pot sowing: each pot contained loam soil with consistent soil quality and water and fertilizer. On September 16, 2021, 30 seeds were sown in each pot.

[0047] Experimental replication: Each treatment was repeated 6 times.

[0048] Survey time and number of times:

[0049] Two investigations were conducted in total;

[0050] First survey: September 22, 2021, visual inspection of seed conditions;

[0051] Second survey: October 2, 2021, to investigate seed germination rate, plant height, root length and fresh weight.

[0052] Germination rate calculation method:

[0053] All data from the experiment were analyzed using Excel. Growth indicators such as germination were investigated and statistically analyzed for different treatments.

[0054] The germination rate and root-to-stem ratio are calculated using formulas (1) and (2), respectively.

[0055] Germination rate = M1 / M × 100% (1)

[0056] Root-to-stem ratio = R / S × 100% (2)

[0057] Where M1 is the total number of germinated grains; M is the number of seeds tested; R is the wheat root length; and S is the wheat plant height.

[0058] Growth Indicator Survey Methods

[0059] The number of plants, plant height (cm), root length (cm), and fresh weight (g) of each treatment potted plant were recorded. SPSS 22.0 (Duncan) was used to analyze the significant differences in the efficacy of each treatment after the application of the drug.

[0060] Results of the first survey: Six days after sowing in pots, wheat in all treatments began to germinate and grow normally.

[0061] The results of the second survey:

[0062] The results of the wheat seed dressing experiment 16 days after sowing are shown in Table 2. The results of the wheat seed dressing experiment 16 days after sowing are shown in Table 3. The results of the wheat seed dressing experiment 16 days after sowing are shown in Table 4.

[0063] Table 2. Results of wheat seed dressing experiment 16 days after sowing emergence rate

[0064]

[0065] As shown in Table 2:

[0066] 1. Compared with the water control group, the germination rate of the chloroindolehydrazine treatment group was significantly improved, indicating that chloroindolehydrazine seed treatment can significantly improve the seed germination rate.

[0067] 2. The effect of chloroindolehydrazine (15g / 10kg) on ​​promoting seed germination rate is basically the same as that of S-inducer (10g / 10kg) on ​​promoting seed germination rate, indicating that chloroindolehydrazine has broad application prospects in seed treatment.

[0068] Table 3. Results of wheat seed dressing experiment 16 days after sowing.

[0069]

[0070]

[0071] As shown in Table 3:

[0072] 1. Compared with the water control group, the chloroindolehydrazine treatment group showed a significant increase in plant height and root length, and a significant increase in root-to-shoot ratio, indicating that chloroindolehydrazine seed treatment can promote wheat seedling growth.

[0073] 2. The promoting effect of chloroindolehydrazine (15g / 10kg) on ​​plant height and root length is basically the same as that of S-inducer (10g / 10kg) on ​​plant height and root length, indicating that chloroindolehydrazine has broad application prospects in seed treatment.

[0074] Table 4. Results of fresh weight survey of wheat seed dressing experiment 16 days after sowing

[0075]

[0076] As shown in Table 4:

[0077] 1. Compared with the water control group, the fresh weight of the chloroindolehydrazine treatment group was significantly increased, indicating that chloroindolehydrazine seed treatment can promote the growth of wheat seedlings.

[0078] 2. The effect of chloroindolehydrazine (15g / 10kg) on ​​increasing fresh weight is basically equivalent to that of S-inducer (10g / 10kg), indicating that chloroindolehydrazine has broad application prospects in seed treatment.

[0079] Application Example 2: Chloroindolehydrazine suspension seed coating agent promotes wheat seed germination and seedling growth in the field and reduces the incidence of fungal diseases.

[0080] Seed treatment was conducted in the field to verify the uniformity of potted plant results, and the incidence of wheat stem rot was investigated.

[0081] Field sowing at Jingbo Agricultural Chemicals Experimental Base: The soil quality and water and fertilizer conditions were consistent across the experimental plots. On October 26, 2021, each treatment covered 30 square meters.

[0082] Experimental replication: Each treatment was repeated 4 times.

[0083] Methods for investigating wheat stem base rot

[0084] For each treatment, a five-point sampling method was used, with 10 plants randomly sampled at each point. The number of diseased plants in each treatment was recorded, and the disease incidence rate was calculated. SPSS 22.0 (Duncan) was used to perform a significance analysis of the differences in disease incidence rates among the treatments.

[0085] Germination rate calculation method:

[0086] All data from the experiment were analyzed using Excel. Germination, plant height, and other growth indicators were investigated and statistically analyzed for different treatments. Germination was recorded by counting, and plant height was measured using a ruler. The growth rate was calculated.

[0087] The germination rate and growth rate are calculated as shown in formulas (3) and (4), respectively.

[0088] Germination rate = M1 / M × 100% (3)

[0089] Where M1 represents the total number of germinated seeds; M represents the number of seeds tested.

[0090] Growth rate (%) = (Plant height in the treated area - Plant height in the blank control area) / Plant height in the blank control area × 100 (4)

[0091] Survey time and number of times:

[0092] A total of four investigations were conducted;

[0093] First survey: November 2, 2021, visual inspection of seed germination;

[0094] Second survey: November 10, 2021, to investigate seed germination rate.

[0095] Third survey: March 20, 2022, surveying wheat plant height.

[0096] Fourth survey: April 15, 2022, to investigate the incidence of wheat stem rot during the peak period.

[0097] The results of the wheat seed treatment experiment on germination potential and germination rate are shown in Table 5; the results of the plant height survey are shown in Table 6; and the results of the disease incidence rate during the high incidence period of wheat basal rot are shown in Table 7.

[0098] Table 5. Germination potential and germination rate of wheat seed dressing experiment

[0099]

[0100] As shown in Table 5, the wheat emergence rate after seed treatment with the chloroindolehydrazine suspension seed coating agent of Formulation Example 1 was significantly better than that of the water control. Moreover, the wheat emergence rate when the seed treatment dosage of the chloroindolehydrazine suspension seed coating agent of Formulation Example 1 was 10 g / Kg and 20 g / Kg was even higher than that when seed treatment was performed with 0.01% 24-brassinolide soluble concentrate and 0.1% S-inducer soluble concentrate. Seed treatment with chloroindolehydrazine suspension seed coating agent can promote seed germination.

[0101] Table 6. Plant height and survey results of wheat treated with seed dressing 130 days after sowing

[0102]

[0103]

[0104] As shown in Table 6, after seed dressing with the chloroindolehydrazine suspension seed dressing agent of Formulation Example 1, the average plant height of wheat was significantly better than that of the water control. Moreover, when the seed dressing dosage of the chloroindolehydrazine suspension seed dressing agent of Formulation Example 1 was 20 g / Kg and 30 g / Kg, the average plant height of wheat was even higher than that of wheat treated with 0.01% 2,4-brassinolide soluble concentrate and 0.1% S-inducer soluble concentrate. Seed dressing with chloroindolehydrazine suspension seed dressing agent can promote the growth of wheat seedlings.

[0105] Table 7. Percentage of wheat stem rot

[0106]

[0107] The results of the field trials in Table 7 show that:

[0108] In Example 1, the incidence rates of wheat stem rot were 24.50%, 18.50%, 16.50%, and 14.50% respectively for different dosages of the chloroindolehydrazine suspension seed dressing agent. The incidence rates for 0.01% 24-brassinolide soluble concentrate and 0.1% S-inducer soluble concentrate were 28.00% and 27.50% respectively. Treatment 2 showed no significant difference from treatments 3 and 4, but was highly significant compared to other treatments. Treatments 1, 5, and 6 showed highly significant differences from treatment 7, with significantly higher incidence rates than other treatments. Example 1, chloroindolehydrazine suspension seed dressing agent, used for seed treatment, effectively prevents wheat stem rot, achieving prevention rather than control, which is of great significance to agricultural production.

[0109] Taking into account seedling emergence rate, plant height, and wheat stem base rot rate, the seed dressing dosage of chloroindolehydrazine suspension seed dressing agent in Formulation Example 1 can be selected as 15g / 10Kg-20g / 10Kg.

[0110] During the experiment, it was observed that the test agent, within the specified dosage range, did not have any adverse effects on wheat growth, nor did it have any significant adverse effects on other organisms.

[0111] Application Example 3: Seed dressing treatment of wheat seeds infected with pathogens such as Fusarium head blight and / or take-all and / or glume blight using chloroindolehydrazine.

[0112] Select diseased seeds;

[0113] Source of diseased seeds: The variety of the crop was Jimai 22, which was planted on farmers' own land. Due to the lack of effective pesticide control in the early stage, diseases such as Fusarium head blight, take-all, and glume blight occurred on a large scale during the crop growth period. Seeds carrying the diseases were selected for subsequent experiments.

[0114] 3.1 Pot Experiment

[0115] Experimental location: Potted seedlings in a greenhouse in Pangjia Village, Boxing County, Binzhou City, Shandong Province. The experimental soil was the substrate, with uniform fertility; all experimental environmental cultivation conditions (soil type, water and fertilizer management, planting density) were uniform and consistent, and conformed to local Good Agricultural Practices (GAP).

[0116] Experimental design: Formulation Example 1 was a 10% chlorindolehydrazine suspension seed coating agent; Formulation Example 2 was a 20% chlorindolehydrazine suspension seed coating agent; Control agent ① was a 10% difenoconazole suspension seed coating agent; Control agent ② was a 10% azoxystrobin suspension seed coating agent; 0.01% brassinolide soluble concentrate, 0.1% styraxin soluble concentrate, and blank control (water control).

[0117] Seed treatment was performed by adding 3% water by seed weight on December 15, 2020. After treatment, the seeds were dried indoors. Before sowing, each pot was watered with 300ml of water and 30 diseased wheat seeds were sown in each pot. Each treatment was repeated 6 times.

[0118] Germination potential, germination rate, and growth rate are calculated using formulas (5), (6), and (7), respectively.

[0119] Germination potential (Gv) = M1 / M × 100% .......................................(5)

[0120] Germination rate (Gp) = M² / M × 100% .......................................(6)

[0121] Where M1 is the number of normally germinated seeds within the germination potential period (9 days); M2 is the total number of normally germinated seeds (15 days); and M is the number of seeds tested.

[0122] Growth rate (%) = (Plant height in the treated area - Plant height in the blank control area) / Plant height in the blank control area × 100 (7)

[0123] A total of three surveys were conducted:

[0124] First survey: December 24, 2020 (9 days after seed treatment), to investigate the number of germinations;

[0125] Second survey: December 30, 2020 (15 days after seed dressing), to investigate the number of germinated seeds;

[0126] Third survey: January 13, 2020 (29 days after seed treatment), plant height was measured;

[0127] All data from the experiment were analyzed using Excel and SPSS 22.0. Germination rates, plant height, and other growth indicators were investigated and statistically analyzed for different treatments; the results are shown in Tables 8-10.

[0128] Table 8. Effects of different seed dressing treatments on different physiological indicators of wheat.

[0129]

[0130] Table 9. Survey and significance analysis of plant height (cm) 29 days after seed treatment with different pesticides.

[0131]

[0132]

[0133] Table 10 Effects of different seed dressing treatments on wheat plant height (cm)

[0134]

[0135] As shown in Tables 8-10, for seeds infected with wheat scab and / or wheat take-all and / or wheat glume blight pathogens selected in the experiment, the seeds treated with the chloroindolehydrazine suspension seed dressing agents of Formulation Examples 1 and 2 of this invention had significantly higher germination rates than the control group and the blank group; at the same time, the plant height in the later stage was significantly higher than that of the control group and the blank group, with significant differences; and the growth rate was significantly higher than that of the control group. This indicates that the seed dressing treatment with chloroindolehydrazine suspension seed dressing agent can significantly improve the emergence rate and achieve the effect of uniform and strong seedlings.

[0136] Further research will be conducted on field seed treatment to control diseases and improve yield, and to determine the feasibility of this invention in wheat seed treatment.

[0137] 3.2 Field Trial: Control Efficacy and Yield Determination of Diseased Seeds from Formulation Examples 1 and 2

[0138] Test location: Dongyang Village, east of Boxing County, Shandong Province.

[0139] Seed treatment process: Wheat seeds were treated with the experimental agent, then dried in a cool place for 3-4 hours before being uniformly treated with 70% imidacloprid wet seed treatment agent to control wheat aphids and other underground pests. This invention is not limited to imidacloprid as a single insecticide seed treatment agent. The amount of water used for seed treatment is 3% of the seed weight (refer to DB34 / T 2908-2017 Field Efficacy Test Guidelines for Seed Treatment Agents).

[0140] Test reagents:

[0141] The 10% chloroindolehydrazine suspension seed coating agent in Formulation Example 1 was used at a dosage of 60 g / 15 kg of seeds for seed treatment.

[0142] The 20% chloroindolehydrazine suspension seed coating agent in Formulation Example 2 was applied at a dosage of 30 g / 15 kg of seeds for seed treatment.

[0143] Control agent ① 10% difenoconazole suspension seed coating agent, dosage 60 g / 15 kg of seeds;

[0144] Control agent ② 10% azoxystrobin suspension seed coating agent, dosage 60 g / 15 kg of seeds;

[0145] And a blank control (water control).

[0146] Sowing time: October 16, 2021.

[0147] Treatment area and number of repetitions: Area: 40 square meters, 4 repetitions per group;

[0148] All test treatment areas were conducted in accordance with the DB34 / T 2908-2017 guidelines for field efficacy trials of seed treatment agents.

[0149] Fertilization: In the early stage, corn straw was returned to the field, which can provide some potassium fertilizer. In addition, 40 kg of compound fertilizer per mu was used as base fertilizer. During the wheat greening period, 40 kg of potassium dihydrogen phosphate compound fertilizer per mu was used as top dressing.

[0150] Irrigation: Irrigation was carried out during the four main water-demand periods: freezing, greening, tillering, and grain-filling. Post-emergence herbicides were applied by foliar spraying with a mixture of quizalofop-P-ethyl and clopyralid to control annual grasses such as barnyard grass, foxtail, purslane, and amaranth, as well as broadleaf weeds such as cleavers, purslane, amaranth, field bindweed, chickweed, and shepherd's purse. Insecticides were applied using a mixture of pyridaben and acetamiprid to prevent aphids and spider mites throughout the wheat's growth period. To ensure the scientific rigor of the data, no disease control was performed, and no fungicides were used during the experiment.

[0151] Survey method: 100 wheat plants were selected in each plot for regional marking. Later, the number of diseased wheat plants in the marked area of ​​each treatment plot was investigated and the disease rate was calculated.

[0152] Survey period: Take-all disease of wheat (November 21, 2021, 36 days after sowing, during the peak period of take-all disease); Fusarium head blight of wheat (April 19, 2022, during the peak period of Fusarium head blight); Gluten blight of wheat (May 9, 2022, during the peak period of glume blight).

[0153] Methods for calculating drug efficacy

[0154] The efficacy is calculated according to formulas (8) and (9);

[0155] Disease incidence rate (%) = Number of diseased plants investigated / Total number of plants investigated × 100...........................(8)

[0156] Prevention and control effect (%) = (CK-PT) / CK×100.................................(9)

[0157] In the formula: CK—the disease incidence rate in the blank control area;

[0158] PT – Disease incidence rate in the treated area.

[0159] The yield increase rate is calculated according to formula (10):

[0160] Yield increase rate (%) = (Yield of the treated area - Yield of the blank control area) / Yield of the blank control area × 100 (10)

[0161] Direct impact on crops

[0162] Observe whether the pesticide causes phytotoxicity to the crop, and record the type and extent of the phytotoxicity. In addition, record any other beneficial effects on the crop (such as promoting ripening, stimulating growth, etc.).

[0163] Record pesticide damage using the following methods:

[0164] a) If pesticide damage can be measured or calculated, it should be expressed in absolute values, such as plant height.

[0165] b) In other cases, the severity and frequency of phytotoxicity can be estimated using the following two methods:

[0166] 1) Record the pesticide damage situation of each plot according to the pesticide damage classification method, and use -, +, ++, +++, ++++ to represent it.

[0167] Methods for classifying phytotoxicity:

[0168] - No pesticide damage;

[0169] +: Mild pesticide damage, does not affect normal crop growth;

[0170] ++: Significant pesticide damage, recoverable, and will not cause crop spillage;

[0171] +++: Severe pesticide damage, affecting normal crop growth and causing a certain degree of loss in crop yield and quality, generally requiring compensation for some economic losses;

[0172] ++++: Severe pesticide damage, resulting in stunted crop growth and significant losses in crop yield and quality; economic losses should be compensated.

[0173] The experimental results are shown in Table 11.

[0174] Table 11 Average control efficacy and yield increase rate of wheat take-all disease, Fusarium head blight, and glume blight.

[0175]

[0176] Table 11 shows that when wheat seeds were treated with the chloroindolehydrazine suspension seed dressing agents of Examples 1 and 2 of this invention, the incidence and control efficacy of wheat diseases were investigated during the growing season. Specifically, for wheat take-all disease, the disease incidence rates of Examples 1 and 2 were 11.25% and 9.50%, respectively, with control efficiencies of 72.05% and 76.40%, respectively; while the disease incidence rates of control agents ① and ② were 15.50% and 14.00%, respectively, with control efficiencies of 61.49% and 65.22%, respectively. For wheat scab, the disease incidence rates of Examples 1 and 2 were 9.75% and 5.25%, respectively, with control efficiencies of 81.43% and 90.00%, respectively; while the disease incidence rates of control agents ① and ② were 25.00% and 16.25%, respectively, with control efficiencies of 52.38% and 69.05%, respectively. For wheat glume blight, the disease incidence rates of Examples 1 and 2 were... The disease incidence rates of the two formulations were 3.25% and 1.50%, respectively, with control efficacy of 88.60% and 94.74%. The disease incidence rates of control agents ① and ② were 8.50% and 6.75%, respectively, with control efficacy of 70.18% and 76.32%. Yields were measured at harvest. The yields of formulations 1 and 2 were 625.65 kg / mu and 695.98 kg / mu, respectively, with yield increases of 24.79% and 38.82%. The control agents ① and ② showed similar efficacy. ② The yields per mu were 546.89 kg and 562.55 kg, respectively, with yield increases of 9.08% and 12.21%, respectively; significantly higher than the control group and the blank group. The yield increase rate was significantly higher than that of the control group, indicating that the treatment of seeds with chloroindolehydrazine can effectively inhibit the occurrence of wheat diseases such as Fusarium head blight, glume blight, and take-all. It can not only significantly reduce the incidence of diseased plants, but also effectively prevent the incidence of Fusarium head blight, glume blight, and take-all, which fully proves that the invention has good prevention and yield increase effects.

[0177] The above examples demonstrate that, with reduced or equivalent dosages, the seed treatment agents of this invention, specifically Formulation Examples 1 and 2, generally achieved at 30-60 grams per 15 kg of wheat seeds, yielded optimal results. Their control efficacy was significantly better than the control agents 10% difenoconazole suspension seed dressing and 10% azoxystrobin suspension seed dressing. Furthermore, this wheat seed treatment trial proved that Formulation Examples 1 and 2 significantly promoted germination and growth of diseased wheat seeds compared to control agents ① and ②. The use of this seed treatment agent improves actual disease control efficacy and significantly promotes wheat yield. It also overcomes the disadvantage of long-term single-use of pesticides leading to resistance, extending the pesticide's lifespan. This technology offers advantages such as convenient application and environmental friendliness, showing promising application prospects. It helps improve wheat quality and reduces the environmental pressure caused by repeated use of chemical pesticides. It is also of great significance for the comprehensive management of resistance to wheat diseases such as Fusarium head blight, take-all, and glume blight.

[0178] Application Example 4: A bioassay experiment on the treatment of pepper seeds with formulations 1 and 2 to control viral diseases.

[0179] Experiment location: Pangjia Greenhouse, Boxing County, Binzhou City, Shandong Province.

[0180] The experimental soil was neutral loam (pH 6.9) with uniform fertility; all experimental environments had uniform cultivation conditions (soil type, water and fertilizer management, planting density) and conformed to local scientific agricultural practices (GAP).

[0181] Source of virus-infected seeds: self-saved seeds of pepper mosaic virus from the Pangjia experimental site in the early stage;

[0182] Experimental Design:

[0183] Treatment 1: Pepper mosaic virus self-saved seeds were treated with formulation Example 1 (10% chloroindolehydrazine suspension seed dressing agent);

[0184] Treatment 2: Treatment of self-saved seeds of pepper mosaic virus with formulation Example 2 (20% chloroindolehydrazine suspension seed dressing);

[0185] Treatment 3: Seed treatment with 36% oligosaccharide bifenthiamethoxam suspension for pepper mosaic virus self-saved seeds;

[0186] Treatment 4: Water control

[0187] The three experimental agents were used to treat seeds at a ratio of 1:50, as well as a blank control (water control).

[0188] Seed dressing was performed on August 10, 2021, with 3% water added to the seeds. After dressing, the seeds were dried indoors. Before sowing, 50 kg of Shimeikang (17-17-17) was applied as a uniform base fertilizer per acre. 100 pepper seeds carrying the mosaic virus were selected for each treatment. After dressing, the seeds were first raised in 50-cell seedling trays. In the early stage, to facilitate the statistics of germination rate, 100 seeds were used to measure the germination rate. If the germination rate was less than 100%, the excess seedlings from that treatment were supplemented to make up 100 seedlings to facilitate the later statistics of plant height and disease index. After 35 days of seedling raising, the seedlings were transplanted to the experimental plot in the greenhouse for subsequent investigation of viral disease incidence.

[0189] The calculation of germination potential, germination rate and growth rate are shown in formulas (11), (12) and (13), respectively.

[0190] Germination potential (Gv) = M1 / M × 100% .......................................(11)

[0191] Germination rate (Gp) = M² / M × 100% .......................................(12)

[0192] Where M1 is the number of normally germinated seeds within the germination potential period (7 days); M2 is the total number of normally germinated seeds (15 days); and M is the number of seeds tested.

[0193] Growth rate (%) = (Plant height in the treated area - Plant height in the blank control area) / Plant height in the blank control area × 100 (13)

[0194] Viral disease investigation methods

[0195] The total number of plants surveyed and the number of diseased plants at each level were recorded on a per-plant basis.

[0196] Grading method:

[0197] Level 0: Asymptomatic;

[0198] Grade 1: Clear veins, light flowers and leaves;

[0199] Grade 3: Mottled leaves in the heart and middle leaves;

[0200] Grade 5: Mottled leaves in the heart and middle leaves, a few leaves are deformed, wrinkled or the plant is slightly stunted;

[0201] Level 7: Heavy foliage, most leaves are deformed, wrinkled or the plant is stunted;

[0202] Level 9: Heavy foliage, with obviously deformed and linear leaves, severely stunted plants, and even death.

[0203] Methods for calculating drug efficacy

[0204] The efficacy is calculated according to formulas (14) and (15);

[0205]

[0206] Prevention and control effect (%) = (1 - (CK0 × PT1) / (CK1 × PT0)) × 100..........(15)

[0207] In the formula: CK0 - disease index of the blank control area before drug administration;

[0208] CK1 - Disease index after drug administration in the blank control area;

[0209] PTO - Disease index in the treatment area before application of pesticides;

[0210] PT1 - Disease index after drug treatment in the treatment area

[0211] If the baseline disease incidence was not investigated before application of the medication, the control effect is calculated according to formula (16):

[0212] Control efficacy (%) = (CK1 - PT1) / CK1 × 100.................................(16)

[0213] A total of four surveys were conducted:

[0214] First survey: August 17, 2021 (7 days after sowing), to investigate the number of germinations;

[0215] Second survey: August 25, 2021 (15 days after sowing), to investigate the number of germinated seeds;

[0216] Third survey: September 10, 2021 (31 days after sowing), plant height was measured;

[0217] Fourth survey: October 20, 2021 (71 days after sowing), to investigate disease incidence;

[0218] All data from the experiment were analyzed using Excel and SPSS 22.0. Growth indicators such as germination, plant height, and disease incidence were investigated and statistically analyzed for different treatments.

[0219] Table 12 Effects of different seed dressing treatments on different physiological indicators of chili peppers

[0220]

[0221] Based on the germination rate statistics 15 days after sowing, the germination rate was less than 100%. Therefore, the excess seedlings treated (with the same time, method, etc.) were replenished to 100 plants to facilitate the later statistics on plant height and disease index.

[0222] Table 13 Effects of different seed dressing treatments on pepper plant height

[0223]

[0224] Table 14. Average control efficacy of different seed treatments for pepper mosaic virus in field trials.

[0225]

[0226] As shown in Table 12, Formulation Examples 1 and 2 can promote the germination of chili seeds and increase the germination rate. The germination rate effect is significantly better than that of the control agent, 36% oligosaccharide-bifenthiamethoxam seed treatment suspension. While investigating the germination rate, the plant height of chili plants was also measured (Table 13). The results were consistent with Table 11, showing a significant promoting effect on chili growth. Subsequent investigations into the incidence of chili mosaic virus (CMV) were conducted. The results (Table 14) showed that the control efficacy of Formulation Examples 1 and 2 for CMV seed treatment was 73.38% and 78.08%, respectively, while the control agent, 36% oligosaccharide-bifenthiamethoxam seed treatment suspension, had a control efficacy of only 59.51%, indicating poorer efficacy and a significant difference compared to Formulation Examples 1 and 2. These examples fully demonstrate that Formulation Examples 1 and 2 can not only improve the germination rate and accelerate plant growth of chili plants, but also enhance their disease resistance and significantly inhibit viral diseases.

[0227] Application Example 5: Biochemical test of tomato seed treatment for viral disease control using formulations 1 and 2.

[0228] Experimental location: Pangjia Greenhouse, Boxing County, Binzhou City, Shandong Province. The experimental soil was used as the substrate and had uniform fertility. All experimental environmental cultivation conditions (soil type, water and fertilizer management, planting density) were uniform and consistent, and conformed to local Good Agricultural Practices (GAP).

[0229] Experimental design: Formulation Example 1 was a 10% chloroindolehydrazine suspension seed coating agent; Formulation Example 2 was a 20% chloroindolehydrazine suspension seed coating agent; the control agent was a 36% oligosaccharide bifenthiamethoxam seed treatment suspension agent. The three experimental agents were used to treat seeds at a ratio of 1:50, and a blank control (water control) was also included.

[0230] Source of virus-infected seeds: self-saved seeds from the Pangjia experimental site for tomato yellow leaf curl virus in the early stage;

[0231] Seed dressing was performed on January 15, 2022, with 3% water added to the seeds. After dressing, the seeds were dried indoors. Before sowing, 50 kg of Shimeikang (17-17-17) was applied as a uniform base fertilizer per acre. 100 tomato seeds carrying yellow leaf curl virus were selected for each treatment. After dressing, the seeds were first raised in 50-cell seedling trays. In the early stage, to facilitate the statistics of germination rate, 100 seeds were used to measure the germination rate. If the germination rate was less than 100%, the excess seedlings from that treatment were used to supplement the remaining seedlings to make up 100 seedlings for later statistics on plant height and disease index. After 38 days of seedling raising, the seedlings were transplanted to the experimental plot in the greenhouse for subsequent investigation of viral disease incidence.

[0232] The germination potential and germination rate are calculated using formulas (17) and (18), respectively.

[0233] Germination potential (Gv) = M1 / M × 100% .......................................(17)

[0234] Germination rate (Gp) = M² / M × 100% .......................................(18)

[0235] Where M1 is the number of normally germinated seeds within the germination potential period (10 days); M2 is the total number of normally germinated seeds (15 days); and M is the number of seeds tested.

[0236] Growth rate (%) = (Plant height in the treated area - Plant height in the blank control area) / Plant height in the blank control area × 100

[0237] Viral disease investigation methods

[0238] The total number of plants surveyed and the number of diseased plants at each level were recorded on a per-plant basis.

[0239] Grading method:

[0240] Level 0: Asymptomatic;

[0241] Grade 1: Clear veins, light flowers and leaves;

[0242] Grade 3: Mottled leaves in the heart and middle leaves;

[0243] Grade 5: Mottled leaves in the heart and middle leaves, a few leaves are deformed, wrinkled or the plant is slightly stunted;

[0244] Level 7: Heavy foliage, most leaves are deformed, wrinkled or the plant is stunted;

[0245] Level 9: Heavy foliage, with obviously deformed and linear leaves, severely stunted plants, and even death.

[0246] Methods for calculating drug efficacy

[0247] The efficacy is calculated according to formulas (10) and (20);

[0248]

[0249] Prevention and control effect (%) = (1 - (CK0 × PT1) / (CK1 × PT0)) × 100..........(20)

[0250] In the formula: CK0 - disease index of the blank control area before drug administration;

[0251] CK1 - Disease index after drug administration in the blank control area;

[0252] PTO - Disease index in the treatment area before application of pesticides;

[0253] PT1 - Disease index after drug treatment in the treatment area

[0254] If the baseline disease incidence was not investigated before application of the medication, the control effect is calculated according to formula (21):

[0255] Control efficacy (%) = (CK1 - PT1) / CK1 × 100.................................(21)

[0256] A total of four surveys were conducted:

[0257] First survey: January 25, 2022 (10 days after seed dressing), to investigate the number of germinations;

[0258] Second survey: January 30, 2022 (15 days after seed dressing), to investigate the number of germinated seeds;

[0259] Third survey: February 10, 2022 (26 days after seed treatment), plant height was measured;

[0260] Fourth survey: March 10, 2022 (54 days after seed treatment), to investigate disease incidence;

[0261] All data from the experiment were analyzed using Excel and SPSS 22.0. Growth indicators such as germination, plant height, and disease incidence were investigated and statistically analyzed for different treatments.

[0262] Table 15 Effects of different seed dressing treatments on different physiological indicators of tomatoes

[0263]

[0264] Based on the germination rate statistics 15 days after sowing, the germination rate was less than 100%. Therefore, the excess seedlings treated (from the same batch, with consistent time and method) were replenished to 100 plants to facilitate the later statistics on plant height and disease index.

[0265] Table 16 Effects of different seed dressing treatments on tomato plant height

[0266]

[0267]

[0268] Table 17 Average control efficacy of different seed treatments in field trials for preventing tomato yellow leaf curl virus disease.

[0269]

[0270] The above examples demonstrate that, with reduced or equivalent dosage, the seed treatment agent of this invention exhibits significantly better control efficacy against vegetable seed viral diseases than the control agent, 36% oligosaccharide bifenthiamethoxam seed treatment suspension. Furthermore, this vegetable seed treatment trial also proved that formulations 1 and 2 significantly promoted both germination and growth in vegetable crops. The use of this seed treatment agent improves actual control efficacy and is of great significance for the comprehensive prevention and control of vegetable viral diseases. This agent not only induces good resistance to viruses in crops but also has a certain inhibitory effect on plant pathogens and promotes seed germination and growth. It can be widely used to control viral diseases in various crops such as rice, tomatoes, peppers, cucumbers, tobacco, potatoes, and flowers. With prevention as the primary focus and treatment as a secondary measure, it enhances plant disease resistance and reduces the occurrence of diseases and viruses, aligning with the development direction of the green, efficient, and low-pollution pesticide industry.

Claims

1. The use of chloroindolehydrazine in seed treatment, characterized by: Chloroindolehydrazine, as an active ingredient in seed treatment agents, promotes seed germination and seedling growth.

2. The use according to claim 1, characterized in that: The seeds used for the seed treatment are any one of the following: wheat seeds, corn seeds, chili pepper seeds, tomato seeds, tobacco seeds, and rice seeds.

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