Weeding method for controlling cyperus rotundus by cooperating with glyphosate

Abstract

The invention provides a herbicide for preventing and treating cyperus rotundus in cooperation with glyphosate. The herbicide is prepared from bacillus velezensis fermentation liquor, phenolic acid and glyphosate. The phenolic acid comprises any one or a combination of more of p-coumaric acid, cinnamic acid, p-hydroxybenzoic acid and caffeic acid. The bacillus velezensis fermentation liquor and phenolic acid components are introduced and form a synergistic effect with the glyphosate, so that the dosage of a single dosage of the glyphosate is reduced, the control effect is guaranteed, meanwhile, the potential pressure of chemical pesticide input on the ecological environment is reduced, and the environmental compatibility of a weeding system is improved.

Description

Technical Field

[0001] This invention belongs to the field of agricultural weed control technology, specifically relating to a method and herbicide composition for controlling Cyperus rotundus by combining natural phenolic acid, Bacillus berberis fermentation broth, and glyphosate. Background Technology

[0002] Glyphosate, scientifically known as N-(phosphonomethyl)aminoacetic acid or N-(phosphonomethyl)glycine, is an organophosphorus systemic broad-spectrum non-selective herbicide. Its mechanism of action involves inhibiting enolpyruvylshikimate phosphate synthase in plants, blocking the conversion of shikimate to phenylalanine, tyrosine, and tryptophan, ultimately leading to plant death due to disruption of protein synthesis. This herbicide can be absorbed through the stems and leaves and translocated to all parts of the plant, effectively controlling more than 40 families of plants, including monocots and dicots, annuals and perennials, and herbs and shrubs. However, it exhibits poor sensitivity to some weeds and faces significant resistance issues with long-term single-use application, becoming the core bottleneck limiting its efficacy.

[0003] In mainstream planting scenarios such as orchards and farmland, glyphosate has become the core choice for farmers to control weeds due to its broad spectrum. However, its long-term and repeated application has led to the large-scale evolution of herbicide resistance in weeds. Taking citrus orchards and wheat fields, which are prone to the growth of Cyperus rotundus, as an example, in the initial stage, farmers often used 2000-4000 times diluted glyphosate for foliar spraying to control weed growth. They applied it 2-3 times a year during the key growth periods of weeds (germination period in spring, March-April, and vigorous growth period in summer, June-July). After this application pattern was carried out continuously for 3-5 years, the weed population gradually developed stable herbicide resistance. The mechanism of herbicide resistance mainly includes two aspects: First, the target enzyme in weeds—enolpyruvylshikimate phosphate synthase (EPSPS)—undergoes gene mutation (such as mutations at the Pro106Ser and Pro106Ala sites), resulting in a 60%-80% decrease in the binding affinity of glyphosate to the target, making it unable to effectively inhibit enzyme activity; second, weeds accelerate the detoxification and degradation of glyphosate in vivo by upregulating the expression levels of metabolic enzymes such as cytochrome P450 enzyme and glutathione transferase, making it difficult for the effective concentration of the herbicide to reach the action threshold. According to field monitoring data, after applying glyphosate in citrus orchards for five consecutive years in the above-mentioned conventional manner, the resistance index of the noxious weed Cyperus rotundus to glyphosate has increased from the initial 1.0 to 3.2-4.5 times, and the control efficacy of 2000-fold diluted solution against Cyperus rotundus has decreased from more than 85% initially to less than 30%, and conventional doses are no longer effective in controlling it.

[0004] Cyperus rotundus L., a perennial noxious weed belonging to the Cyperaceae family, possesses extremely strong reproductive and competitive capabilities. It has long, creeping rhizomes and tubers underground, allowing it to reproduce simultaneously through seeds, rhizomes, and tubers. During its growing season, a single Cyperus plant can produce 10-15 tubers within one month, and its population can expand 5-10 times within two months, rapidly occupying ground space and competing with orchard and farmland crops for water and fertilizer resources. It is already considered an extremely difficult weed to control. The combined effect of herbicide resistance further leads to a significant decline in the efficacy of glyphosate against Cyperus rotundus, creating a dilemma where "conventional doses are ineffective, and increased doses are inefficient."

[0005] Microbial pesticides have great potential for development in pest and disease control due to their advantages such as safety for humans and the environment, low incidence of resistance, long-term controllability, and high modifiability. Among these advantages, the carrier of the microbial strain is crucial for the production of microbial inoculants and determines their effectiveness. While microbial pesticides have many advantages, they also have disadvantages such as slow control effects, narrow control range, and poor stability. Therefore, utilizing carriers to carry microorganisms to fully utilize their active ingredients and achieve optimal control effects, as well as addressing the specific problems of microbial pesticides to enable wider application of microbial herbicides in the field, are important current research directions.

[0006] Plant allelopathic effects are a chemical ecology phenomenon in which plants directly or indirectly influence the growth and development of neighboring plants by releasing certain chemical substances they produce. Phenolic acids are the main allelochemicals produced by higher plants. Compared to glyphosate, phenolic acids are relatively safe. As natural secondary metabolites of plants, they are easily degraded by microorganisms in the natural environment, causing less disturbance to the ecosystem. Coumaric acid, cinnamic acid, caffeic acid, and p-hydroxybenzoic acid are widely distributed phenolic acids in nature, possessing various biological activities such as insect resistance, antibacterial activity, and weed suppression, and are the main allelochemicals of many plants.

[0007] Microbial herbicides are often not accepted by farmers and cannot be effectively promoted when used in the field due to their slow weed-controlling effect. According to the national pesticide reduction policy, research could be conducted on combining microbial herbicides with chemical herbicides. This would reduce the amount of chemical herbicides used while also addressing the slow-acting problem of microbial herbicides. Currently, research in this area is relatively scarce, but it represents another important direction for implementing herbicide reduction measures.

[0008] Therefore, by developing a glyphosate-co-phenol-based Bacillus baicalensis fermentation broth for the control of Cyperus rotundus, it is possible to effectively control Cyperus rotundus that is antagonistic to glyphosate, reduce the amount of glyphosate used, and keep the cost of medication within a reasonable range. Summary of the Invention

[0009] This invention introduces Bacillus berberis fermentation broth and phenolic acid components, combining biological herbicides with chemical herbicides to form a synergistic effect with glyphosate. This reduces the amount of glyphosate used alone, ensures the control effect, and at the same time controls Cyperus rotundus, which is antagonistic to glyphosate. This reduces the potential pressure on the ecological environment from chemical pesticide input and improves the environmental compatibility of the weeding system.

[0010] The Bacillus berberis fermentation broth was prepared by using LB liquid medium with strain BV-J11, with an inoculum size of 2-5%, and cultured in a shaker at 25-30℃ and 180-200rpm for two days to obtain the Bacillus berberis fermentation broth. The obtained fermentation broth was centrifuged to obtain the supernatant. The strain preservation number is M 20231502.

[0011] In the early stages of this invention, Bacillus berberis was cultured in LB liquid medium at an inoculation rate of 2-5% in a shaker at 25-30°C and 180-200 rpm for two days to obtain a Bacillus berberis fermentation broth. The fermentation broth was then centrifuged to obtain the supernatant. The strain preservation number was M 20231502. It showed a significant synergistic herbicidal effect with glyphosate and was therefore identified as a compound component.

[0012] The selection of p-coumaric acid, cinnamic acid, p-hydroxybenzoic acid, and caffeic acid as synergistic ingredients was based on a comprehensive consideration of multiple factors: From the perspective of their commonality in nature, they are widely found in the rhizosphere exudates and decomposition residues of various crops such as wheat, rice, and soybeans, as well as weeds, making them readily available; through preliminary activity screening experiments (using Cyperus rotundus as the test weed and setting different phenolic acid gradient concentrations), it was found that these phenolic acids could produce a significant synergistic herbicidal effect with glyphosate at relatively low concentrations (8.21 - 36.04 mg / L), exhibiting strong allelopathic activity, and were therefore determined to be the compound ingredients.

[0013] To achieve the above objectives, the technical solution of the present invention is as follows: A herbicide that synergistically controls Cyperus rotundus with glyphosate comprises Bacillus berberis fermentation broth, phenolic acid, and glyphosate.

[0014] The phenolic acids include any one or a combination of p-coumaric acid, cinnamic acid, p-hydroxybenzoic acid, and caffeic acid.

[0015] The Bacillus velezensis fermentation broth described herein is strain BV-J11, deposited at the China Center for Type Culture Collection (CCC) with accession number M 20231502, classified as Bacillus velezensis BV-J11, deposited on August 17, 2023, at Wuhan University, Wuhan, China.

[0016] The glyphosate concentration is 1000-4000 mg / L, and is any value within the range of 1000-4000 mg / L.

[0017] The concentration of phenolic acid was controlled at 8-40 mg / L after being mixed with the Bacillus vesiculosus fermentation broth, and could be any value within the range of 8-40 mg / L.

[0018] As a preferred embodiment, the glyphosate concentration is 2000 mg / L.

[0019] The phenolic acid mentioned is p-coumaric acid. After p-coumaric acid is mixed with Bacillus belye fermentation broth, the concentration is controlled to be 8-10 mg / L, and it is any value within 8-10 mg / L. The phenolic acid mentioned is cinnamic acid. After mixing cinnamic acid with the Bacillus vesiculosus fermentation broth, the concentration is controlled to be 28-30 mg / L, and it is any value within 28-30 mg / L. The phenolic acid mentioned is p-hydroxybenzoic acid. After p-hydroxybenzoic acid is mixed with the fermentation broth of Bacillus belye, the concentration is controlled to be 12-15 mg / L, and it is any value within 12-15 mg / L. The phenolic acid mentioned is caffeic acid. After mixing caffeic acid with Bacillus belye fermentation broth, the concentration is controlled to be 36-38 mg / L, and any value within 36-38 mg / L.

[0020] This invention also provides a method for compounding herbicides that synergistically control glyphosate-resistant Cyperus rotundus, comprising the following steps: (1) Dilute the 30% glyphosate stock solution with water to prepare a glyphosate aqueous solution; (2) Phenolic acid was dissolved in Bacillus vesiculosus fermentation broth and mixed to prepare a mixed solution; (3) The glyphosate aqueous solution from step (1) is mixed with the mixed solution from step (2) to obtain a glyphosate synergistic herbicide for controlling Cyperus rotundus.

[0021] The present invention also provides a herbicide for controlling Cyperus rotundus, which is antagonistic to glyphosate, comprising the herbicide described above.

[0022] This invention also provides a method for controlling Cyperus rotundus that is antagonistic to glyphosate, wherein the herbicide is sprayed onto the leaves of Cyperus rotundus at a rate of 100-150 mL / acre to achieve control.

[0023] The application method involves diluting the herbicidal composition and then spraying it evenly onto the stems and leaves of the weeds before planting the crops.

[0024] The herbicide is in the form of an aqueous solution.

[0025] The glyphosate and its salt are: N-phosphonomethylglycine (CAS# 46544), and its salt is isopropylamine salt.

[0026] The phenolic acids mentioned are: trans-4-hydroxycinnamic acid (CAS# 501-98-4, hereinafter referred to as "p-coumaric acid"), 3-phenyl-2-acrylic acid (CAS# 621-82-9, hereinafter referred to as "cinnamic acid"), p-hydroxybenzoic acid (CAS# 99-96-7, hereinafter referred to as "p-hydroxybenzoic acid"), and caffeic acid (CAS# 331-39-5, hereinafter referred to as "caffeic acid").

[0027] The aforementioned herbicides were used to control Cyperus rotundus, a perennial sedge that exhibits significant antagonism against glyphosate, in both cultivated and non-cultivated land.

[0028] Beneficial technical effects of the present invention: This invention introduces Bacillus berberis fermentation broth and phenolic acid components, which form a synergistic effect with glyphosate. While reducing the amount of glyphosate used and ensuring the control effect, it also reduces the potential pressure on the ecological environment from chemical pesticide input and improves the environmental compatibility of the weed control system.

[0029] Compared with single-agent formulations, it can achieve better control effects with reduced dosage of the corresponding single agent, and shows a significant synergistic effect in controlling Cyperus rotundus, which is antagonistic to glyphosate. Detailed Implementation

[0030] To make the objectives, technical solutions, and advantages of this invention clearer and more concise, the invention is described using the following specific embodiments, but the invention is by no means limited to these examples. The following descriptions are merely preferred embodiments of the invention and are used only to describe the invention; they should not be construed as limiting the scope of the invention. It should be noted that any modifications, equivalent substitutions, and improvements made within the spirit and principles of this invention should be included within the scope of protection of this invention.

[0031] All examples were prepared according to the following steps: 1. Component A A glyphosate aqueous solution was prepared by diluting a 30% mass fraction glyphosate stock solution (Sigma-Aldrich) with deionized water.

[0032] 2. Component B Bacillus belysin was cultured in LB liquid medium at an inoculation rate of 2% for two days in a shaker at 30°C and 180 rpm to obtain the Bacillus belysin fermentation broth. The fermentation broth was then centrifuged to obtain the supernatant.

[0033] 3. Component C Cinnamic acid was dissolved in the supernatant of Bacillus vesiculosus to prepare a mixed solution.

[0034] 4. Component D p-hydroxybenzoic acid was dissolved in Bacillus berberis supernatant to prepare a mixed solution.

[0035] 5. Component E Caffeic acid was dissolved in the supernatant of Bacillus vesiculosus to prepare a mixed solution.

[0036] 5. Component F The coumaric acid was dissolved in the supernatant of Bacillus vesiculosus to prepare a mixed solution.

[0037] Example 1: Toxicity test of each component on wheat This embodiment belongs to the field of agricultural crop toxicity technology, specifically involving a comparative study of the effects of various components and different concentrations of component A on wheat.

[0038] 1. Experimental Materials Wheat materials: About 10-14 days after sowing, select wheat plants with a height of about 15 cm, 2 fully unfolded true leaves and 1 emerging heart leaf as experimental subjects. Plant one plant per pot, and 5 parallel groups per group.

[0039] Reagent A: Dilute 30% glyphosate stock solution (Sigma-Aldrich) with deionized water to prepare 4000 mg / L (0.4% concentration) and 2000 mg / L (0.2% concentration) glyphosate aqueous solutions. Reagent C: Component C was prepared into a 29.63 mg / L solution using Bacillus de Bellesia fermentation broth.

[0040] Reagent D: Component D was prepared into a 13.81 mg / L solution using Bacillus de Bellesia fermentation broth.

[0041] Reagent E: Prepare a 36.04 mg / L solution of component E using Bacillus de Bellesia fermentation broth.

[0042] Reagent F: Prepare a solution of component F at 8.21 mg / L using Bacillus de Bellesia fermentation broth.

[0043] 2. Experimental Methods Single-agent treatment group A1: The leaves were sprayed with an equal amount of 4000 mg / L reagent A.

[0044] Single-agent treatment group A2: The leaves were sprayed with an equal amount of 2000 mg / L reagent A.

[0045] Single-agent treatment group C: The leaves were sprayed with 29.63 mg / L reagent C.

[0046] Single-agent treatment group D: The leaves were sprayed with 13.81 mg / L reagent C.

[0047] Single-agent treatment group E: The leaves were sprayed with 36.04 mg / L reagent C.

[0048] Single-agent treatment group F: The leaves were sprayed with 8.21 mg / L reagent C.

[0049] 3. Experimental Procedure Starting from the day after treatment begins, observe and record the growth of wheat at the same time every day for 18 consecutive days, focusing on leaf color, morphology, and presence of wilting, and calculate the wilting rate per wheat plant. The wilting rate per plant is calculated using the following formula: wilting rate (%) = (number of wilted leaves / total number of leaves) × 100%.

[0050] The results are shown in Table 1:

[0051] Table of wheat wilting rate due to interaction of various components with glyphosate The results showed that glyphosate (reagent A) was significantly toxic to crops (wheat), characterized by early onset, continuous escalation of damage, and an extremely high final wilting rate (67.78%-74.67%); while the reagents to be verified (C, D, E, F) were basically non-toxic to crops (wheat), characterized by late onset, rapid cessation of toxicity, and an extremely low final wilting rate (6.67%-10.00%), and did not affect the normal growth of crops.

[0052] Example 2: Pot experiment on the weed control effect of C synergistically with A on Cyperus rotundus This embodiment belongs to the field of agricultural weed control technology, specifically involving a comparative study on the weeding effects of component C and different concentrations of component A on Cyperus rotundus weed.

[0053] 1. Experimental Materials Weed materials: Cyperus rotundus plants with uniform growth, 5-6 leaves, and a height of 15 cm were selected as experimental subjects. One plant was planted in each pot, and there were 5 parallel groups in each group.

[0054] Reagent A: Same as Reagent A described in Example 1.

[0055] Reagent C: Component C was prepared into a 29.63 mg / L solution using Bacillus berberis fermentation broth.

[0056] 2. Experimental Methods Single-agent treatment group A1: The leaves were sprayed with an equal amount of 4000 mg / L reagent A.

[0057] Single-agent treatment group A2: The leaves were sprayed with an equal amount of 2000 mg / L reagent A.

[0058] Single-agent treatment group C: The leaves were sprayed with 9.71 mg / L reagent C.

[0059] Synergistic treatment group A+C: Spray the leaves with a mixture of 2000 mg / L reagent A2 and 9.71 mg / L reagent C at a ratio of 1:1.

[0060] 3. Experimental Procedure Starting from the day after treatment begins, observe and record the growth of Cyperus rotundus at the same time every day for 18 consecutive days, focusing on leaf color, morphology, and presence of wilting, and calculate the wilting rate for each plant. The wilting rate per plant is calculated using the following formula: wilting rate (%) = (number of wilted leaves / total number of leaves) × 100%.

[0061] The results are shown in Table 2:

[0062] Table 3: Synergistic Effect of Mixed Solution C on Glyphosate on Wilting Rate of Cyperus rotundus

[0063] Synergistic effect of compound solution C with glyphosate and cyperus rotundus synergistic index table The results showed that treatment with 2000 mg / L glyphosate alone was significantly less effective than the 4000 mg / L glyphosate treatment group. However, there was no significant difference in efficacy between the 29.63 mg / L compound solution C and the 2000 mg / L glyphosate synergistic treatment group compared to the 4000 mg / L glyphosate single-agent treatment. Furthermore, the 29.63 mg / L compound solution C alone did not achieve a lethal effect on *Cyperus rotundus*. This demonstrates that at this concentration, the synergistic use of compound solution C with low-concentration glyphosate can significantly enhance the herbicidal effect, achieving the level of action of high-concentration glyphosate single-agent treatment.

[0064] In summary, the data in the table above shows that the Cyperus rotundus herbicide prepared by this invention retains its weed-control effect while reducing the amount of glyphosate used. It effectively controls the noxious weed Cyperus rotundus and is harmless to crops.

[0065] Weed control effect: This invention solves the problem of the harmful weed Cyperus rotundus. Since a single application can control the harm of Cyperus rotundus, it reduces the cost of herbicide use and weakens the herbicide resistance of weeds.

[0066] Example 3: Pot experiment on the weed control effect of D synergistically with A on Cyperus rotundus This embodiment belongs to the field of agricultural weed control technology, specifically involving a comparative study on the weeding effects of component D and different concentrations of component A on Cyperus rotundus weed.

[0067] 1. Experimental Materials Weed materials: Cyperus rotundus plants with uniform growth, 5-6 leaves, and a height of 15 cm were selected as experimental subjects. One plant was planted in each pot, and there were 5 parallel groups in each group.

[0068] Reagent A: Same as Reagent A described in Example 1.

[0069] Reagent D: Prepare a 13.81 mg / L solution of component D using Bacillus belye fermentation broth.

[0070] 2. Experimental Methods Single-agent treatment group A1: The leaves were sprayed with an equal amount of 4000 mg / L reagent A.

[0071] Single-agent treatment group A2: The leaves were sprayed with an equal amount of 2000 mg / L reagent A.

[0072] Single-agent treatment group D: The leaves were sprayed with 13.81 mg / L reagent D.

[0073] Synergistic treatment group A+D: A foliar spray of 2000 mg / L reagent A and 13.81 mg / L reagent D at a 1:1 ratio was applied.

[0074] 3. Experimental Procedure Starting from the day after treatment begins, observe and record the growth of Cyperus rotundus at the same time every day for 18 consecutive days, focusing on leaf color, morphology, and presence of wilting, and calculate the wilting rate for each plant. The wilting rate per plant is calculated using the following formula: wilting rate (%) = (number of wilted leaves / total number of leaves) × 100%.

[0075] The results are shown in Table 4:

[0076] Table of the synergistic effect of compound solution D and glyphosate on the wilting rate of Cyperus rotundus Table 5:

[0077] Synergistic effect of compound solution D with glyphosate and cyperus rotundus synergistic index table The results showed that treatment with 2000 mg / L glyphosate alone was significantly less effective than the 4000 mg / L glyphosate treatment group. However, there was no significant difference in efficacy between the 13.81 mg / L compound solution D and the 2000 mg / L glyphosate synergistic treatment group compared to the 4000 mg / L glyphosate single-agent treatment. Furthermore, 13.81 mg / L compound solution D alone did not achieve a lethal effect on Cyperus rotundus. This demonstrates that at this concentration, the synergistic use of compound solution D with low-concentration glyphosate can significantly enhance the herbicidal effect, achieving the level of action of high-concentration glyphosate single-agent treatment.

[0078] In summary, the data in the table above shows that the Cyperus rotundus herbicide prepared by this invention retains its weed-control effect while reducing the amount of glyphosate used. It effectively controls the noxious weed Cyperus rotundus and is harmless to crops.

[0079] Weed control effect: This invention solves the problem of the harm caused by the noxious weed Cyperus rotundus. Since a single application can control the harm caused by Cyperus rotundus, it reduces the cost of herbicide use, weakens the herbicide resistance of weeds, and reduces the damage to the soil.

[0080] Example 4: Pot experiment on the weed control effect of E synergistically with A on Cyperus rotundus This embodiment belongs to the field of agricultural weed control technology, specifically involving a comparative study on the weeding effects of component E and different concentrations of component A on Cyperus rotundus weed.

[0081] 1. Experimental Materials Weed materials: Cyperus rotundus plants with uniform growth, 5-6 leaves, and a height of 15 cm were selected as experimental subjects. One plant was planted in each pot, and there were 5 parallel groups in each group.

[0082] Reagent A: Same as Reagent A described in Example 1.

[0083] Reagent E: Prepare a 36.04 mg / L solution of component E using Bacillus belye fermentation broth.

[0084] 2. Experimental Methods Single-agent treatment group A1: The leaves were sprayed with an equal amount of 4000 mg / L reagent A.

[0085] Single-agent treatment group A2: The leaves were sprayed with an equal amount of 2000 mg / L reagent A.

[0086] Single-agent treatment group E: The leaves were sprayed with 36.04 mg / L reagent E.

[0087] Synergistic treatment group A+E: A foliar spray of 2000 mg / L reagent A and 36.04 mg / L reagent E at a 1:1 ratio was applied.

[0088] 3. Experimental Procedure Starting from the day after treatment begins, observe and record the growth of Cyperus rotundus at the same time every day for 18 consecutive days, focusing on leaf color, morphology, and presence of wilting, and calculate the wilting rate for each plant. The wilting rate per plant is calculated using the following formula: wilting rate (%) = (number of wilted leaves / total number of leaves) × 100%.

[0089] The results are shown in Table 6:

[0090] Table of the synergistic effect of compound solution E and glyphosate on the wilting rate of Cyperus rotundus Table 7:

[0091] Synergistic effect of compound solution E on glyphosate synergistic index of Cyperus rotundus The results showed that treatment with 2000 mg / L glyphosate alone was significantly less effective than the 4000 mg / L glyphosate treatment group. However, there was no significant difference in efficacy between the 36.04 mg / L compound solution E and the 2000 mg / L glyphosate synergistic treatment group compared to the 4000 mg / L glyphosate single-agent treatment. Furthermore, the 36.04 mg / L compound solution E alone did not achieve a lethal effect on *Cyperus rotundus*. This demonstrates that at this concentration, the synergistic use of compound solution E with low-concentration glyphosate can significantly enhance the herbicidal effect, achieving the level of action of high-concentration glyphosate single-agent treatment.

[0092] The data in the table above shows that the Cyperus rotundus herbicide prepared by this invention retains its weed-control effect while reducing the amount of glyphosate used. It effectively controls the noxious weed Cyperus rotundus and is harmless to crops.

[0093] Weed control effect: This invention solves the problem of the harmful weed Cyperus rotundus. Since a single application can control the harm of Cyperus rotundus, it reduces the cost of herbicide use and weakens the herbicide resistance of weeds.

[0094] Example 5: Pot experiment on the weed control effect of F synergistic with A on Cyperus rotundus This embodiment belongs to the field of agricultural weed control technology, specifically involving a comparative study on the weeding effects of component F and component A at different concentrations on Cyperus rotundus weed.

[0095] 1. Experimental Materials Weed materials: Cyperus rotundus plants with uniform growth, 5-6 leaves, and a height of 15 cm were selected as experimental subjects. One plant was planted in each pot, and there were 5 parallel groups in each group.

[0096] Reagent A: Dilute 30% glyphosate stock solution (Sigma-Aldrich) with deionized water to prepare 4000 mg / L (0.4% concentration) and 2000 mg / L (0.2% concentration) glyphosate aqueous solutions. Reagent B: Prepare a solution of component B at 8.21 mg / L using Bacillus de Bellesia fermentation broth.

[0097] 2. Experimental Methods Single-agent treatment group A1: The leaves were sprayed with an equal amount of 4000 mg / L reagent A.

[0098] Single-agent treatment group A2: The leaves were sprayed with an equal amount of 2000 mg / L reagent A.

[0099] Single-agent treatment group F: The leaves were sprayed with 8.21 mg / L reagent B.

[0100] Synergistic treatment group A+F: 2000 mg / L reagent A was mixed with 8.21 mg / L reagent F at a ratio of 1:1 and sprayed onto the leaves.

[0101] 3. Experimental Procedure Starting from the day after treatment begins, observe and record the growth of Cyperus rotundus at the same time every day for 18 consecutive days, focusing on leaf color, morphology, and presence of wilting, and calculate the wilting rate for each plant. The wilting rate per plant is calculated using the following formula: wilting rate (%) = (number of wilted leaves / total number of leaves) × 100%.

[0102] The results are shown in Table 8:

[0103] Table of the synergistic effect of compound solution F and glyphosate on the wilting rate of Cyperus rotundus Table 9:

[0104] Synergistic effect of compound solution F with glyphosate and cyperus rotundus synergistic index table The results showed that treatment with 2000 mg / L glyphosate alone was significantly less effective than the 4000 mg / L glyphosate treatment group. However, there was no significant difference in efficacy between the 8.21 mg / L compound solution B and the 2000 mg / L glyphosate synergistic treatment group compared to the 4000 mg / L glyphosate single-agent treatment. Furthermore, 8.21 mg / L compound solution B alone did not achieve a lethal effect on *Cyperus rotundus*. This demonstrates that at this concentration, the synergistic use of compound solution B with low-concentration glyphosate can significantly enhance the herbicidal effect, achieving the level of action of high-concentration glyphosate single-agent treatment.

[0105] The data in the table above shows that the Cyperus rotundus herbicide prepared by this invention retains its weed-control effect while reducing the amount of glyphosate used. It effectively controls the noxious weed Cyperus rotundus and is harmless to crops.

[0106] Weed control effect: This invention solves the problem of the harm caused by the noxious weed Cyperus rotundus. Since a single application can control the harm caused by Cyperus rotundus, it reduces the cost of herbicide use, weakens the herbicide resistance of weeds, and reduces the damage to the soil.

[0107] In summary, comparing the synergistic effects of the four compound solutions with glyphosate (see table above): Compound solution F achieved a 100% wilting rate on day 15, demonstrating better efficacy than 4000 mg / L glyphosate alone; Compound solutions C, D, and E achieved 100% wilting rates on day 17, respectively, all exceeding the efficacy of high-concentration glyphosate. Among these, p-coumaric acid showed the fastest synergistic onset of action, while p-hydroxybenzoic acid exhibited the best final control efficacy. The appropriate type of phenolic acid solution can be selected based on the specific application scenario (e.g., weed density, control cycle).

[0108] This invention utilizes phenolic acids such as p-coumaric acid, cinnamic acid, p-hydroxybenzoic acid, and caffeic acid, along with Bacillus berberis fermentation broth, as synergistic components. The concentration of phenolic acids is controlled between 8.21 and 36.04 mg / L during formulation. This herbicide is then combined with glyphosate to prepare a herbicide for Cyperus rotundus. As demonstrated in the above examples, this herbicide retains its weed-controlling effect while reducing the amount of glyphosate required, effectively controlling the noxious weed Cyperus rotundus without harming crops.

[0109] In terms of weed control efficacy: This invention utilizes the synergistic effect of Bacillus baicalensis fermentation broth and glyphosate to control the damage caused by Cyperus rotundus with a single application. While reducing weed control costs, the compound formulation delays the development of herbicide resistance in weeds (avoiding the resistance problems caused by long-term use of glyphosate alone), and reduces soil pollution by leveraging the natural properties of biological herbicides, thus solving the dual challenges of controlling noxious weeds and protecting the ecology.

Claims

1. A herbicide that synergistically controls Cyperus rotundus with glyphosate, characterized in that, Including Bacillus vesiculosus fermentation broth, phenolic acid, and glyphosate.

2. The herbicide for synergistic control of Cyperus rotundus with glyphosate according to claim 1, characterized in that, The phenolic acids include any one or a combination of p-coumaric acid, cinnamic acid, p-hydroxybenzoic acid, and caffeic acid.

3. The herbicide for synergistic control of Cyperus rotundus with glyphosate according to claim 1, characterized in that, The Bacillus berberis fermentation broth was prepared by using LB liquid medium with strain BV-J11, with an inoculum size of 2-5%, and cultured in a shaker at 25-30℃ and 180-200rpm for two days to obtain the Bacillus berberis fermentation broth. The obtained fermentation broth was centrifuged to obtain the supernatant. The strain preservation number is M 20231502.

4. The herbicide for synergistic control of Cyperus rotundus by glyphosate according to claim 1, characterized in that, The concentration of glyphosate is 1000-4000 mg / L; The concentration of phenolic acid was controlled at 8-40 mg / L after being mixed with the Bacillus vesiculosus fermentation broth.

5. The herbicide for synergistic control of Cyperus rotundus with glyphosate according to claim 4, characterized in that, The concentration of glyphosate is 1000-2000 mg / L; The phenolic acid mentioned is coumaric acid, and the concentration of coumaric acid is controlled at 8-10 mg / L after being mixed with the fermentation broth of Bacillus belye.

6. The herbicide for synergistic control of Cyperus rotundus with glyphosate according to claim 5, characterized in that, The phenolic acid mentioned can also be cinnamic acid. After mixing cinnamic acid with Bacillus vesiculosus fermentation broth, the concentration is controlled to be 28-30 mg / L.

7. The herbicide for synergistic control of Cyperus rotundus with glyphosate according to claim 5, characterized in that, The phenolic acid mentioned can also be p-hydroxybenzoic acid. After mixing p-hydroxybenzoic acid with the Bacillus vesiculosus fermentation broth, the concentration is controlled to be 12-15 mg / L.

8. The herbicide for synergistic control of Cyperus rotundus with glyphosate according to claim 5, characterized in that, The phenolic acid can also be caffeic acid. After mixing caffeic acid with Bacillus vesiculosus fermentation broth, the concentration is controlled to be 36-38 mg / L.

9. The method for compounding a herbicide according to any one of claims 1-8 that synergistically controls Cyperus rotundus with glyphosate, characterized in that, Includes the following steps: (1) Dilute the 30% glyphosate stock solution with water to prepare a glyphosate aqueous solution; (2) Phenolic acid was dissolved in Bacillus vesiculosus fermentation broth and mixed to prepare a mixed solution; (3) The glyphosate aqueous solution from step (1) is mixed with the mixed solution from step (2) to obtain a glyphosate synergistic herbicide for controlling Cyperus rotundus.

10. A herbicide for controlling Cyperus rotundus, characterized in that, Includes the herbicide according to any one of claims 1-8.

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