Application of 2-amino-5-benzoylbenzimidazole in prevention and treatment of wheat powdery mildew and promotion of wheat growth

Abstract

The invention belongs to the technical field of biological agriculture, and particularly relates to application of 2-amino-5-benzoylbenzimidazole in prevention and treatment of wheat powdery mildew and promotion of wheat growth. The 2-amino-5-benzoyl benzimidazole with powdery mildew resistance activity is obtained by deeply researching the Trichosporidium tilletii 17wy1 and screening and analyzing metabolites of the Trichosporidium tilletii 17wy1, and in-vitro leaf segment tests and pot tests prove that the 2-amino-5-benzoyl benzimidazole has a good prevention and treatment effect on wheat powdery mildew and has a good application prospect in prevention and treatment of powdery mildew of wheat. Particularly, when the concentration is 4.0 mg / mL, the prevention effect on wheat powdery mildew reaches 86.34%, and the treatment effect reaches 78.56%; meanwhile, the 2-amino-5-benzoyl benzimidazole with low concentration is found to promote the growth of the wheat. The invention provides more technical means for prevention and control of plant powdery mildew and plant growth regulators, and has a good application prospect.

Description

Technical Field

[0001] This invention belongs to the field of bio-agricultural technology, specifically relating to the application of 2-amino-5-benzoylbenzimidazole in the prevention and control of wheat powdery mildew and the promotion of wheat growth. Background Technology

[0002] Powdery mildew is a plant disease caused by obligate parasitic fungi, with a very high incidence and wide range of damage in agricultural production. Many important crops, such as wheat, barley, grapes, strawberries, and cucumbers, are susceptible to it. This disease can infect the green tissues of plants, such as leaves, stems, and fruits. In the early stages, chlorotic yellow spots appear on the leaf surface, followed by the development of a white powdery mold layer. As the disease progresses, the mold layer gradually thickens and changes color from white to gray, eventually leading to yellowing, curling, and shedding of leaves, stunted stem growth, and poor fruit development, deformities, or even rot.

[0003] Taking wheat powdery mildew as an example, as one of the major diseases in wheat-producing areas worldwide, it is highly susceptible to outbreaks and epidemics under conditions such as warm and humid climates, high planting density, and excessive application of nitrogen fertilizer. When the disease is severe, it can reduce wheat yield by 30% to 50%, seriously affecting global food security and agricultural economic stability.

[0004] Currently, chemical pesticides remain the primary means of controlling powdery mildew in plants. However, with the widespread and long-term use of chemical pesticides, their drawbacks, such as pesticide residues and the development of pesticide resistance in diseases, are becoming increasingly prominent. Faced with the numerous problems brought about by chemical control, biological control, as a green and sustainable disease control strategy, is gradually becoming a research hotspot in the agricultural field. Biological control utilizes beneficial organisms or their metabolites to inhibit the growth, reproduction, and infection of pathogens, and regulates the interaction between plants and pathogens. It has significant advantages such as not polluting the environment, not inducing pesticide resistance, and being safe for humans and animals. It can effectively compensate for the shortcomings of chemical control and provide a more environmentally friendly and healthy way of disease control for agricultural production.

[0005] *Saccharomyces cerevisiae* 17wy1 is a yeast strain successfully isolated from abnormally growing conidia of powdery mildew fungi on wheat seedling leaves during our research on green control technologies for plant powdery mildew. Previous studies have shown that *Saccharomyces cerevisiae* 17wy1 has efficacy in controlling powdery mildew. However, its core components are unknown. Screening for and obtaining its core components for powdery mildew resistance and applying it to the control of plant powdery mildew is of great significance. Summary of the Invention

[0006] To address the aforementioned technical problems, this invention conducts in-depth research on the fermentation broth and filtrate of *Saccharomyces cerevisiae* 17wy1 at different fermentation stages. Metabolic components of the fermentation filtrate from *Saccharomyces cerevisiae* 17wy1 at different fermentation stages are studied. Through screening, a compound, 2-amino-5-benzoylbenzimidazole, with control effects against wheat powdery mildew is obtained. Further research on this compound reveals its excellent control effect against wheat powdery mildew. Simultaneously, 2-amino-5-benzoylbenzimidazole is found to promote wheat growth.

[0007] On the one hand, the present invention provides the application of 1,2-amino-5-benzoylbenzimidazole in the prevention and control of wheat powdery mildew.

[0008] Furthermore, in the aforementioned applications, the concentration of the 2-amino-5-benzoylbenzimidazole is 0.01 mg / mL to 4.0 mg / mL.

[0009] Furthermore, in the aforementioned application, the concentration of the 2-amino-5-benzoylbenzimidazole is 4.0 mg / mL.

[0010] Finally, the present invention also provides the application of 2-amino-5-benzoylbenzimidazole in promoting wheat growth, wherein the final concentration of 2-amino-5-benzoylbenzimidazole is 0.01 mg / mL to 0.1 mg / mL.

[0011] Furthermore, in the application of 2-amino-5-benzoylbenzimidazole to promote wheat growth, the final concentration of 2-amino-5-benzoylbenzimidazole is 0.05 mg / mL.

[0012] Compared with the prior art, the technical solution provided by the present invention has at least the following beneficial effects or advantages: (1) By studying the powdery mildew resistance of fermentation broth from *Saccharomyces cerevisiae* 17wy1 at different fermentation stages, the fermentation time was optimized and the optimal fermentation time was determined. Fermentation broths at different fermentation stages were screened, and metabolomic differential compounds were screened. Finally, 2-amino-5-benzoylbenzimidazole was selected for in-depth research to explore its role in the control of powdery mildew in plants. Through indoor experiments on detached leaf segments and potted seedlings, it was verified that 2-amino-5-benzoylbenzimidazole has a good control effect on wheat powdery mildew. In particular, when the concentration of 2-amino-5-benzoylbenzimidazole is 4.0 mg / mL, the preventive efficacy against wheat powdery mildew reaches 86.34%, and the curative efficacy against wheat powdery mildew is 78.56%. This provides a new, biological control method for wheat powdery mildew, which helps to reduce the use of chemical pesticides, reduce environmental pollution, and improve the safety and quality of agricultural products.

[0013] (2) It was found that low concentrations of 2-amino-5-benzoylbenzimidazole promote plant growth. Through experiments with wheat seedlings in petri dishes, it was verified that low concentrations of 2-amino-5-benzoylbenzimidazole promote wheat growth, while high concentrations inhibit wheat growth. The concentration of 2-amino-5-benzoylbenzimidazole at 0.01 mg / mL to 0.1 mg / mL promotes wheat growth. Attached Figure Description

[0014] Figure 1 Correlation heatmap of metabolomics samples of Saccharomyces cerevisiae 17wy1.

[0015] Figure 2 Principal component analysis (PCA) diagram of the metabolome of *Saccharomyces cerevisiae* 17wy1.

[0016] Figure 3 Volcano plot of metabolome differences in Saccharomyces cerevisiae 17wy1.

[0017] Figure 4 Classification of KEGG compounds for differential metabolome of Saccharomyces cerevisiae 17wy1.

[0018] Figure 5 This study identified key differential pathways in the metabolome of *Saccharomyces cerevisiae* 17wy1, including the differentially expressed compounds.

[0019] Figure 6 These are important differential metabolites from the fermentation broth of *Saccharomyces cerevisiae* 17wy1 fermented for 7 days.

[0020] Figure 7 Wheat powdery mildew under different treatments ( Blumeria graminis f. sp. tritici , Bgt )Statistical results of conidial germination. A represents the sterile water treatment group; B represents the 1 mg / mL ABZMI treatment group.

[0021] Figure 8 The figures show the agronomic traits of detached leaf segments of wheat under different treatments for powdery mildew. A represents the sterile water treatment group; B represents the 0.01 mg / mL ABZMI treatment group; C represents the 0.05 mg / mL ABZMI treatment group; D represents the 0.1 mg / mL ABZMI treatment group; E represents the 0.5 mg / mL ABZMI treatment group; and F represents the 1 mg / mL ABZMI treatment group.

[0022] Figure 9 The graph shows the statistical results of the therapeutic effect of ABZMI on wheat powdery mildew. Different letters indicate significant differences in experimental treatments (p<0.05).

[0023] Figure 10The graph shows the statistical results of the preventive effect of ABZMI on wheat powdery mildew. Different letters indicate significant differences in experimental treatments (p<0.05).

[0024] Figure 11 The figures show the agronomic traits of wheat under different treatments. A represents the sterile water treatment group; B represents the 0.01 mg / mL ABZMI treatment group; C represents the 0.05 mg / mL ABZMI treatment group; D represents the 0.1 mg / mL ABZMI treatment group; E represents the 0.5 mg / mL ABZMI treatment group; and F represents the 1 mg / mL ABZMI treatment group. Detailed Implementation

[0025] The technical solution of the present invention will be described below with reference to the embodiments. However, the present invention is not limited to the following embodiments.

[0026] To enable those skilled in the art to better understand and implement the technical solutions of the present invention, the present invention will be further described below in conjunction with specific embodiments and accompanying drawings. However, the embodiments described are not intended to limit the present invention.

[0027] Unless otherwise specified, the experimental and detection methods described in the following embodiments are conventional methods; unless otherwise specified, the reagents and materials are commercially available.

[0028] The *Saccharomyces cerevisiae* 17wy1, with accession number CGMCC No:14148, has been disclosed in the applicant's patent application CN 108004154 B filed in 2017.

[0029] 2-Amino-5-benzoylbenzimidazole, CAS: A593760, purchased from Toronto Research Chemicals.

[0030] Example 1 This example describes the preparation of fermentation filtrate of Saccharomyces cerevisiae 17wy1.

[0031] PDA medium: 200 g / L potato, 20 g / L glucose, 20 g / L agar, natural pH, autoclaved at 121°C for 30 min.

[0032] YPB medium: yeast extract 10g / L, peptone 20g / L, glucose 20g / L, natural pH, autoclaved at 121℃ for 30min.

[0033] Take out the preserved *Saccharomyces cerevisiae* strain 17wy1, transfer it to PDA solid medium, and incubate it upside down at 25°C for 3 days. Pick the well-grown *Saccharomyces cerevisiae* strain 17wy1 from the PDA solid medium, inoculate it into the prepared YPB liquid medium, and incubate it in a shaker at 180 rpm and 25°C for 7 days. Centrifuge and collect the filtrate after filtering out the cells to obtain the fermentation filtrate of *Saccharomyces cerevisiae* strain 17wy1, which is ready for use.

[0034] Example 2 This embodiment describes the preparation of a spore suspension of *Saccharomyces cerevisiae* 17wy1. The fermentation broth of *Saccharomyces cerevisiae* 17wy1 cultured for 7 days was diluted with sterile water to achieve a viable cell concentration of 5 × 10⁻⁶. 7 CFU / mL available for use.

[0035] Example 3 This embodiment is a test experiment on the inhibition of wheat powdery mildew conidia germination by fermentation broth of Saccharomyces cerevisiae 17wy1.

[0036] Take out the preserved *Saccharomyces cerevisiae* strain 17wy1, transfer it to PDA solid medium, and incubate it upside down at 25°C for 3 days. Pick the well-grown *Saccharomyces cerevisiae* strain 17wy1 on the PDA solid medium and inoculate it into the prepared YPB liquid medium. Incubate it in a shaker at 180 rpm and 25°C. Take the fermentation broth after 3 days (3d), 7 days (7d), and 11 days (11d) for later use.

[0037] Wheat powdery mildew conidia were cultured by spreading culture medium on glass slides, and the effect of the fermentation broth of an antagonistic yeast (Saccharomyces cerevisiae 17wy1) on the germination of wheat powdery mildew conidia was determined.

[0038] The specific steps are as follows: Prepare 0.5% water agar medium. Using a pipette, take 450 μL of uncooled 0.5% water agar and an equal volume of antagonistic yeast fermentation broth, mix them in a 2 mL centrifuge tube, and spread evenly onto a glass slide. Four treatments were set up: fermentation broth of *Saccharomyces cerevisiae* 17wy1 cultured for 3, 7, and 11 days, and CK (water agar medium). Each treatment was repeated three times. Fresh wheat powdery mildew conidia were evenly shaken onto the cooled medium. The slide was then placed in a plastic box with absorbent paper to maintain moisture, and incubated in the dark at 18°C ​​for 24 hours. The germination of wheat powdery mildew conidia was observed and recorded using a regular optical microscope. The spore germination rate and the inhibition rate of *Saccharomyces cerevisiae* 17wy1 fermentation broth on spore germination were calculated using the following formula: Spore germination rate (%) = (Number of germinating spores / Total number of spores) × 100 Spore germination inhibition rate (%) = (control germination rate - treatment germination rate) / control germination rate × 100.

[0039] Table 1. Results of the test on the inhibition of powdery mildew conidia germination by fermentation broth of *Saccharomyces cerevisiae* 17wy1.

[0040] The results are shown in Table 1. The fermentation broth of *Saccharomyces cerevisiae* 17wy1 after 3 days and 11 days of fermentation inhibited the germination of powdery mildew conidia by 0.32% and -1.48%, respectively, indicating no inhibitory effect. The fermentation broth of *Saccharomyces cerevisiae* 17wy1 after 7 days of fermentation inhibited the germination of powdery mildew conidia by 81.80%, indicating that the 7-day fermentation broth had a strong inhibitory effect on the germination of wheat powdery mildew conidia.

[0041] Example 4 This example describes the analysis and screening of metabolites from *Saccharomyces cerevisiae* 17wy1.

[0042] Fermentation broths from *Saccharomyces cerevisiae* 17wy1 fermented for 3 and 7 days, as described in Example 3, were filtered using a 2µM filter and then mailed on dry ice to Shanghai Meiji Biotechnology Co., Ltd. for non-target metabolomics analysis to compare the metabolic differences of fermentation products at the two time points. Six replicates were set for each group. Quality control, data normalization, QC verification, and data transformation were performed to obtain qualified experimental data. Based on the differences in metabolite content in different samples, correlation heatmap analysis and principal component analysis (PCA) were conducted on the *Saccharomyces cerevisiae* 17wy1 metabolomics samples to evaluate the similarity of samples within groups and the differences between samples between groups. The correlation heatmap of the *Saccharomyces cerevisiae* 17wy1 metabolomics samples is shown below. Figure 1 As shown, Figure 1 The right and bottom sides of the graph show the sample names. Each cell in the graph represents the correlation between two samples. Different colors represent the relative magnitude of the correlation coefficient between samples. The length of the clustering branches indicates the relative distance between samples; samples on the same branch have high similarity. The PCA principal component analysis diagram of the metabolome of *Saccharomyces cerevisiae* 17wy1 is shown below. Figure 2 As shown, Figure 2 After dimensionality reduction analysis, the samples had relative coordinate points on principal components P1 and P2. The distance between these coordinate points represented the degree of clustering and dispersion among the samples; closer distances indicated higher sample similarity, while greater distances indicated greater sample differences. PCA analysis allowed observation of the inter-group separation trend in the experimental model and the presence of outliers. It also reflected the variability between and within groups from the raw data. The confidence ellipse represented the distribution of "true" samples within a 95% confidence level; samples outside this area were considered potentially outliers. The results showed good reproducibility of metabolic samples within different groups of *Saccharomyces cerevisiae* 17wy1, making them suitable for subsequent analysis.

[0043] Subsequently, differential analysis was performed on the metabolites of *Saccharomyces cerevisiae* 17wy1, and a metabolomic volcano plot of *Saccharomyces cerevisiae* 17wy1 was constructed. The results are as follows: Figure 3 As shown, Figure 3 The horizontal axis represents the fold change in metabolite expression between the two groups, i.e., log₂FC; the vertical axis represents the statistical test value of the difference in metabolite expression levels, i.e., -log₂FC. 10 The p-value represents the expression level; a higher p-value indicates a more significant difference in expression. Each dot in the graph represents a specific metabolite, and the size of the dot represents the Vip value. Dots on the left represent metabolites with downregulated expression, and dots on the right represent metabolites with upregulated expression. The further to the left, right, and top of the dots, the more significant the expression difference.

[0044] Further KEGG classification of the differential compounds yielded the following results: Figure 4 As shown. Figure 4 The vertical axis represents the KEGG compound secondary classification categories: Glycolipids, Lipids, Phospholipids, Bases, Nucleic Acids, Nucleosides, Nucleotides, Peptides, and Amino Acids. The horizontal axis represents the number of metabolites annotated to each category. Based on the biological functions of the metabolites, the results show that these differentially metabolites are mainly divided into four categories: Vitamins and Cofactors, Peptides, Nucleic Acids, and Lipids.

[0045] Further KEGG enrichment and classification of differential compounds yielded the following results: Figure 5 As shown, Figure 5The horizontal axis represents the pathway name, and the vertical axis represents the enrichment rate, which is the ratio of the number of metabolites enriched in the pathway to the number of metabolites annotated in the pathway. A higher ratio indicates a greater degree of enrichment. The bar color gradient indicates the significance of enrichment; by default, darker colors represent more significant enrichment of the KEGG term. P-values ​​or FDR < 0.001 are marked with ***, P-values ​​or FDR < 0.01 with **, and P-values ​​or FDR < 0.05 with *. The results show that the differentially expressed compounds are mainly involved in metabolic and membrane transport biological processes. Differential compounds play a leading role in biological processes such as proline metabolism, ABC transporter, lysine degradation, arginine biosynthesis, pantothenate and CoA biosynthesis, glycerol phospholipid metabolism, and β-alanine metabolism.

[0046] Further clustering and important metabolite analysis were performed on the fermentation broth of *Saccharomyces cerevisiae* 17wy1 after 7 days of fermentation to identify key differentially expressed metabolites in the 7-day fermentation broth. The results are as follows: Figure 6 As shown, Figure 6 The Y-axis represents the metabolite name, and the X-axis represents the average relative abundance of the metabolite in different groups. Different colored bars represent different groups; the rightmost bar represents... P Value, *0.01< P ≤0.05, ** 0.001< P ≤0.01, *** PThe results, ≤0.001, showed that the fermentation broth of *Saccharomyces cerevisiae* 17wy1 after 7 days contained eight important metabolites, including glucosaminic acid; mebendazole metabolite (2-Amino-5-benzoylbenzimidazole); adenosine monophosphate; lysyl-proline; hydroxy-1,4-benzoquinone; 2-hydroxy-1,4-benzoquinone; 2-hydroxycinnamic acid; cresol sulfate; and glycerophosphocholine. These metabolites are presumably closely related to the antagonistic effect of *Saccharomyces cerevisiae* 17wy1 on powdery mildew.

[0047] Based on extensive literature review and our laboratory's previous experience in powdery mildew research, we selected 2-amino-5-benzoylbenzimidazole, a benzimidazole derivative, for in-depth study. Benzimidazole and its derivatives are important active substances in the pesticide and pharmaceutical fields, possessing broad-spectrum biological activities such as anticancer, antibacterial, and antiparasitic effects. Benzimidazole compounds are low in toxicity and can inhibit fungal growth, leading to their increasingly widespread application. The effects of 2-amino-5-benzoylbenzimidazole on plant disease resistance have not yet been reported. This study will investigate the efficacy of 2-amino-5-benzoylbenzimidazole in controlling powdery mildew and its plant growth-promoting effects.

[0048] Example 5 This example is a test to determine the efficacy of 2-amino-5-benzoylbenzimidazole against wheat powdery mildew.

[0049] (1) 2-Amino-5-benzoylbenzimidazole Bgt Conidial germination inhibition test The prepared 2-amino-5-benzoylbenzimidazole (ABZMI, hereinafter referred to as ABZMI) solution was mixed with 2% melted sterile agar to a final concentration of 1 mg / mL. The mixture was poured into a 90×10 mm petri dish, with the same volume of sterile water used as a control instead of the fermentation supernatant. The diseased wheat leaves were gently shaken. Bgt Conidia were evenly shaken onto the solidified culture medium surface and incubated in the dark at (25±2)℃ for 24 h. Each treatment was repeated in triplicate. After the dark incubation period, the results of each treatment were observed and analyzed under an optical microscope. Bgt Number of spores germinating. Statistics. Bgt The germination rate and inhibition rate of spores were determined using the same method as in Example 3.

[0050] Results of wheat powdery mildew spore germination inhibition test as follows Figure 7 As shown, ABZMI to Bgt It has a significant inhibitory effect on spore germination. Under the same conditions, the control group Bgt Spores germinated normally, but treatment with 1 mg / mL ABZMI significantly inhibited germination. Bgt The spore germination inhibition rate was 91.34%, indicating that ABZMI has a strong antibacterial effect against wheat powdery mildew.

[0051] (2) Test on the control efficacy of wheat powdery mildew detached leaf segments When wheat reaches the one-heart, two-leaf stage, wheat leaves are harvested, with a length of 4-5 cm. The leaves are then immersed in ABZMI solutions of different concentrations (0.01 mg / mL, 0.05 mg / mL, 0.1 mg / mL, 0.5 mg / mL, 1 mg / mL), with sterile water as a control. After 12 hours of immersion, the leaves are removed and air-dried in a clean bench. Six leaves are then evenly placed in each petri dish of benzimidazole medium, with the upper leaf surface facing up and the lower leaf surface facing down. Fresh... Bgt Conidia were evenly distributed onto the leaves in each culture dish and placed in a (25±2)℃ light incubator for cultivation. Three parallel tests were conducted per group. After 7 days of cultivation, the disease index was calculated and the control efficacy was determined according to a 6-level classification method. The wheat powdery mildew grading standards are shown in Table 2, and the test results are shown in Table 3. Figure 8 As shown.

[0052] Table 2 Grading Standards for Wheat Powdery Mildew

[0053] The formula for the relationship between disease severity index and prevention and control effectiveness is as follows: Disease index = ∑[(Number of diseased leaves at each level × Corresponding level value) / (Total number of leaves surveyed × 5)] × 100 Prevention and control efficacy (%) = (Disease index of control group - Disease index of treatment group) / Disease index of control group × 100 Table 3. Control efficacy of ABZMI against detached leaf segments of wheat powdery mildew.

[0054] Note: Different lowercase letters indicate significant differences in experimental treatments. p <0.05).

[0055] like Figure 8 As shown in Table 3, compared with the control group, wheat leaves treated with gradually increasing concentrations of ABZMI showed a significant reduction in lesion area. The statistical results of the control efficacy in Table 3 indicate that ABZMI treatment can significantly (… p<0.05) Reduced the disease index of wheat powdery mildew. The disease index of wheat leaves in the control group was 54.44, while the disease index of wheat leaves treated with 1 mg / mL ABZMI was only 7.78, with a control efficacy of 86.27%, indicating that ABZMI can significantly inhibit the infection of wheat leaves by powdery mildew. In vitro leaf segment control experiments showed that ABZMI has a good control effect on wheat powdery mildew.

[0056] (3) Pot experiment to test the therapeutic effect of ABZMI on wheat powdery mildew The control experiment included seven treatments: a sterile water group (negative control), a 1 mg / mL triadimefon group (positive control), and ABZMI test groups at different concentrations (0.5 mg / mL, 1 mg / mL, 2 mg / mL, 3 mg / mL, and 4 mg / mL). Wheat seedlings were inoculated when they reached the two-heart-one-leaf stage. Bgt Conidia, shake off old leaves from the surface the day before inoculation. Bgt Spores were collected and inoculated by shaking off fresh spores. The plants were then placed in a light-controlled incubator. On days 3, 7, and 11 post-inoculation, different experimental groups were sprayed with the appropriate fungicide. Each treatment was replicated in triplicate, with at least 20 plants per pot. On day 15 post-inoculation, the severity of wheat powdery mildew was assessed using a 6-level classification system. The disease index and control efficacy for wheat powdery mildew were statistically analyzed. Results are as follows: Figure 9 As shown.

[0057] Depend on Figure 9 It can be seen that, compared with the sterile water treatment (negative control), the disease index of wheat gradually decreased and the control effect gradually increased with the increase of ABZMI concentration, indicating that ABZMI has a significant therapeutic effect on wheat powdery mildew. Specifically, the disease index of wheat in the control group was 77.78, while in the 4 mg / mL ABZMI treatment group, the disease index of wheat decreased to 16.67, with a control effect of 78.56%, which was not significantly different from the positive control triadimefon. p >0.05). The results showed that ABZMI could reduce the infectivity of powdery mildew on leaves, thereby alleviating the occurrence of wheat powdery mildew, and the higher the concentration of ABZMI, the more significant the therapeutic effect. p <0.05).

[0058] (4) Pot experiment to test the preventive effect of ABZMI on wheat powdery mildew. The preventative trial consisted of seven treatments: a sterile water group (negative control), a 1 mg / mL triadimefon group (positive control), and ABZMI test groups at different concentrations (0.5 mg / mL, 1 mg / mL, 2 mg / mL, 3 mg / mL, and 4 mg / mL). When wheat seedlings reached the two-heart-one-leaf stage, the seedlings were sprayed with the appropriate treatment for each test group once daily for three consecutive days, followed by inoculation. Bgt Conidia were used, with three replicates per treatment, each containing at least 20 seedlings. On day 15 post-inoculation, the disease severity of wheat seedlings was assessed using a six-level classification system, and the disease index and control efficacy were statistically analyzed. Results are as follows: Figure 10 As shown.

[0059] Depend on Figure 10 It was found that compared with the sterile water treatment (negative control), the disease index of wheat in the ABZMI treatment group was reduced, indicating that ABZMI has a significant preventive effect against wheat powdery mildew. The wheat powdery mildew disease index was inversely proportional to the ABZMI concentration; the control effect of wheat powdery mildew was directly proportional to the ABZMI concentration. In the preventive treatment group of wheat, the disease index of the control reached 73.33, while after treatment with 4 mg / mL ABZMI, the wheat disease index decreased to 10, with a control effect of 86.34%, which was significantly different from the positive control triadimefon (83.31%) (p<0.05). This indicates that ABZMI treatment of wheat leaves has a protective effect on wheat, significantly reducing the infectivity of wheat powdery mildew fungus, and ABZMI has a preventive effect against wheat powdery mildew (p<0.05), and the preventive effect is superior to that of the chemical fungicide triadimefon.

[0060] Example 6 This example is an experiment to determine the growth-promoting effect of 2-amino-5-benzoylbenzimidazole on wheat seedlings.

[0061] Wheat seeds were surface-sterilized with 1% (w / v) sodium hypochlorite solution for 5 min, followed by rinsing three times with sterile water. The sterilized wheat seeds were then soaked for 12 h in different concentrations of ABZMI treatment solutions (0.01 mg / mL, 0.05 mg / mL, 0.1 mg / mL, 0.5 mg / mL, 1 mg / mL), with sterile water treatment serving as a control. After rinsing the treated wheat seeds with sterile water and drying the filter paper, they were placed in glass petri dishes with a layer of moistened filter paper at the bottom and cultured in a plant light incubator at (25±2)℃, (50±5)% relative humidity, and a light:dark (L:D) ratio of 14 / 10 h. Each group had three replicates. After 10 days of culture, the root length, plant height, dry weight, and other growth parameters of the wheat seedlings were measured. The results are shown in Table 4. Figure 11 .

[0062] Table 4 Effects of different treatments on wheat growth

[0063] Note: Different lowercase letters indicate significant differences in experimental treatments. p <0.05).

[0064] Depend on Figure 11It can be seen that, compared with sterile water, low concentrations of ABZMI promote wheat growth to some extent. Table 4 shows that, compared with the control group treated with water, wheat treated with 0.05 mg / mL ABZMI showed a 15% increase in fresh weight and a 12.73% increase in dry weight. High concentrations of ABZMI treatment inhibited wheat growth; after treatment with higher concentrations of ABZMI, the root length and plant height of wheat plants decreased compared to the treatment group. It is speculated that ABZMI may mimic or interfere with the auxin signaling pathway. At low concentrations, it activates auxin response factors, promoting cell elongation and division, manifested as increased root length and plant height; however, at high concentrations, it may over-activate or feedback-inhibit this pathway, leading to growth inhibition.

[0065] As described above, the basic principles, main features, and advantages of the present invention have been well described. The above embodiments and specifications are merely descriptions of preferred embodiments of the present invention, and the present invention is not limited to the above embodiments. Various changes and improvements made to the technical solutions of the present invention by those skilled in the art without departing from the spirit and scope of the present invention should fall within the protection scope defined by the present invention.

Claims

Application of 1,2-amino-5-benzoylbenzimidazole in the control of wheat powdery mildew.

2. In the application according to claim 1, the application concentration of the 2-amino-5-benzoylbenzimidazole is 0.01 mg / mL to 4.0 mg / mL.

3. In the application according to claim 2, the application concentration of the 2-amino-5-benzoylbenzimidazole is 4.0 mg / mL. The application of 4,2-amino-5-benzoylbenzimidazole in promoting wheat growth, characterized in that, The final concentration of the 2-amino-5-benzoylbenzimidazole is 0.01 g / mL to 0.1 mg / mL.

5. The application of 2-amino-5-benzoylbenzimidazole according to claim 4 in promoting wheat growth, characterized in that, The final concentration of the 2-amino-5-benzoylbenzimidazole is 0.05 mg / mL.

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