CN113767901A

CN113767901A - Application of thymol in prevention and treatment of peanut southern blight

Info

Publication number: CN113767901A
Application number: CN202110982318.4A
Authority: CN
Inventors: 何磊鸣; 周琳; 崔凯娣; 杜鹏强; 赵特; 高飞; 汪梅子; 何亚; 孙龙江
Original assignee: Henan Agricultural University
Current assignee: Henan Agricultural University
Priority date: 2021-08-25
Filing date: 2021-08-25
Publication date: 2021-12-10

Abstract

The invention discloses an application of thymol in the aspect of preventing and treating peanut southern blight, in particular to an application of plant essential oil containing thymol, such as oregano essential oil or thyme essential oil and the like in the aspect of preventing and treating peanut southern blight. The invention is found by experimental research that: the thymol and the plant essential oil containing the thymol have good inhibitory activity on the hypha growth and sclerotium germination of the southern blight, the inhibitory activity is enhanced along with the increase of the concentration of the thymol, and the thymol is obviously superior to a control medicament polyoxin. Meanwhile, the results of the potted plant control effect test and the indoor toxicity test are consistent, which shows that the thymol and the plant essential oil containing the thymol can effectively prevent and treat the peanut southern blight.

Description

Application of thymol in prevention and treatment of peanut southern blight

Technical Field

The invention belongs to the technical field of southern blight prevention and control, and particularly relates to application of thymol in prevention and control of peanut southern blight.

Background

Thymol, also known as thymol, naturally occurring in the plant oregano (A)Origanum vulgare) And thyme (Hymus mongolicus) The essential oil is also the main active component of oregano essential oil and thyme essential oil. At first, the study of Huying and the like finds that the combination of thymol and chitosan has better bacteriostatic activity on 6 bacteria such as staphylococcus aureus and the like. Later, research on thymol is also increasing in the fields of animal feed, medicine, food, and the like. The research of Manying Ying et al shows that thymol can inhibit the growth of the Bel-7402 cell of liver cancer. The research of Wangxiyu and the like finds that the addition of thymol and carvacrol in the feed can improve the intestinal microflora of weaned piglets, promote the intestinal health and promote the growth of the piglets. In the field of plant protection, researches on Zhang Jing, Lulu and the like find that thymol has better insecticidal activity on Bemisia tabaci and caenorhabditis elegans. Thymol has been less studied in plant pathogenic fungi. At present, no research report on the bioactivity and the prevention and treatment effect of thymol on southern blight bacteria is found.

Southern blight of peanut is caused by sclerotium rolfsii (Sclerotium rolfsii) Fungal soil-borne diseases caused by infection. The pathogenic bacteria have wide hosts and can infect more than 500 kinds of plants such as cereals, potatoes, beans and the like. At present, southern blight has become an important factor for restricting the yield and quality of peanuts in China. The southern blight of the peanuts mostly occurs in the adult-plant stage of the peanuts, germs mainly invade from the stem bases of the plants, hypha can expand upwards and downwards along stems, and finally stems, fruit needles, roots and pods of the plants can be damaged. Leaf wilting at the early stage of damage, covering the ground and the base of the stem with white spun silk-like hyphae, and attaching white hyphae to the root and the pod after being pulled out; in the later period, the whole plant dies, the root part and the pod become brown and rot, and even the kernel also changes color and becomes wet and rotten. The hyphae begin to gather to form sclerotium in later period, the hyphae are white small particles in initial period, gradually increase to become yellow brown, and finally become hard sclerotium in black brown or dark brown of rapeseed shape, and then the sclerotium can be irrigated and rain water flowsOr some farming operations, etc., to further increase the disease condition. At present, outbreak and disaster formation of peanut southern blight are reported in peanut planting areas of Henan, Shandong, Anhui, Liaoning, Jiangxi province and the like in China, the morbidity is generally 10% -40%, 20% -30% of yield loss of peanuts can be caused, and the yield can be reduced by more than 50% in serious cases. In 2020, southern Henan province agricultural rural hall lists the southern peanut southern blight as a category II pest and disease damage directory in southern Henan province. The research on the southern blight of the peanuts at home and abroad is less, and the prevention and control effects are limited due to the wide hosts and the long survival time of sclerotia of the peanuts because of the lack of disease-resistant varieties and agricultural measures such as deep ploughing, reasonable rotation and the like. At present, the control of the peanut southern blight mainly depends on chemical bactericides. However, the varieties of chemical bactericides registered on Chinese pesticide information network for preventing and controlling the peanut southern blight are limited, and the long-term use of the chemical bactericides can not only cause the pests to generate drug resistance, but also cause the problems of pesticide residue exceeding standard, environmental pollution, harm to human health and the like. Therefore, a new safe, efficient and pollution-free medicament is urgently needed to be found.

Under the background of long creation period, high cost and the like of new pesticides, people begin to search high-activity compounds from natural products, and more efficient pesticide varieties can be developed by using the natural products as lead compounds for structural optimization and modification, for example, polyoxin, a metabolite generated by streptomyces aureofaciens, has a good prevention and treatment effect on southern blight, and pesticide registration is carried out in China. In addition, eugenol, allicin, matrine and other bactericidal substances derived from plant bodies have been registered to prevent and treat various plant diseases. The research finds that the thymol and the plant essential oil containing the thymol have strong inhibitory activity on the southern blight, have good control effect on the southern blight and have huge application prospect.

Disclosure of Invention

The invention aims to overcome the defects of the prior art and provides a new application of thymol in the aspect of peanut southern blight prevention, and the application is verified by experiments as follows: the thymol can effectively prevent and treat the peanut southern blight and has good prevention and treatment effect.

In order to achieve the purpose, the invention adopts the following technical scheme:

a new use of thymol in preventing and treating southern blight of peanut is provided.

The application can be the application of the plant essential oil containing thymol in the aspect of preventing and treating the southern blight of the peanuts.

Furthermore, thymol naturally exists in origanum vulgaris, thymus vulgaris and other plants, and is also a main active ingredient of origanum vulgaris essential oil and thymus vulgaris essential oil. Thus, the plant essential oil may be oregano essential oil or thyme essential oil, or the like.

The invention provides a mixed medicament prepared by compounding thymol or plant essential oil containing thymol and conventional auxiliary materials in the field. The mixed medicament can be suspending agent, water dispersible granule, wettable powder, granule, aqueous emulsion, missible oil or microcapsule and the like.

The essential oil of origanum vulgaris is prepared from perennial shrub or herb origanum vulgaris of origanum of Labiatae (origanum vulgaris) ((L))Origanum vulgare) The volatile oil extracted from the Chinese medicinal herb is a feed additive approved by the Ministry of agriculture. Initially, much research on oregano essential oil was focused on the field of animal nutrition and feed. Researches on the heroic character and the like find that the oregano essential oil not only has good bacteriostatic action on mould and saccharomycetes, but also has a certain antioxidation. Then, researches on the antibacterial action and the pharmacological activity of the oregano essential oil are gradually increased in the field of postharvest management of traditional Chinese medicines, foods and fruits and vegetables. Researches such as forest clearing and the like show that the origanum essential oil has the effects of enhancing the immunity of the organism, clearing heat and relieving pain and the like. In addition, the oregano essential oil can also be used as a food preservative. Zhang et al reviewed the use of oregano essential oil in the preservation of fruits and vegetables, meat products, eggs, milk and other food products. The origanum essential oil not only has good bacteriostatic action on aspergillus flavus, staphylococcus aureus, escherichia coli and the like, but also can replace a chemical bactericide for management of picked fruits and vegetables. Research on the biological activity of oregano essential oil against phytopathogenic fungi has been relatively limited. At present, no research report on the biological activity of the origanum vulgare essential oil on the southern blight florida is found.

Compared with the prior art, the invention has the following excellent effects:

1) the invention is found by experiments that: thymol has good inhibitory activity on the growth of hyphae of southern blight fungusIncreased thymol concentration, increased inhibitory activity, EC₅₀The value is 83.3 mg/L, and the inhibitory activity is obviously better than that of a control medicament polyoxin (EC)₅₀The value was 204.7 mg/L). The thymol also has a good inhibition effect on sclerotium germination of the southern blight bacterium, and the inhibition rate of the thymol on sclerotium germination is 76.67% under the treatment concentration of 100 mg/L; when the concentration of the thymol is increased to 150 mg/L and above, the inhibition rate of the thymol on sclerotium germination is up to 100%. The thymol has good treatment effect on the peanut southern blight, the control effect (62.54%) of the thymol under the dosage of 1000mg/L is obviously higher than the control effect (42.9%) of the control medicament polyoxin under the dosage of 1000mg/L, and the thymol has great application prospect in the prevention and treatment of the peanut southern blight;

2) the invention is found by experiments that: under the treatment concentrations of 1000mg/L and 500 mg/L, the inhibition rate of the oregano essential oil on the growth of the hyphae of the southern blight fungus is up to 100%, and the inhibition rate is obviously superior to that of a control medicament polyoxin (64.9% and 61.7%). The oregano essential oil has good inhibitory activity on hypha growth of southern blight bacteria, and the inhibitory activity is enhanced with the increase of essential oil concentration, and EC₅₀The value is 118.5 mg/L, and the inhibitory activity is obviously better than that of a control medicament polyoxin (EC)₅₀The value was 204.7 mg/L). The oregano essential oil also has a good inhibition effect on sclerotium germination of southern blight bacteria, and the inhibition rate of the oregano essential oil on sclerotium germination is 85% at the treatment concentration of 250 mg/L; when the concentration of the essential oil is increased to 300 mg/L, the inhibition rate of the essential oil on sclerotium germination is as high as 100%. The oregano essential oil has good prevention and treatment effect on the southern blight of peanuts, and the prevention and treatment effect of 1000mg/L of oregano essential oil on the southern blight of peanuts is 50.76%, which is equivalent to the prevention effect (42.9%) of a control medicament polyoxin under the dosage of 1000 mg/L.

Drawings

FIG. 1 is a picture showing the effect of oregano essential oil of different concentrations on controlling southern blight of peanut. From the three panels, it can be seen that southern blight has already caused peanut plant death in the control treatment. While southern blight also caused some damage to the root system of oregano essential oil treated peanut plants, it did not result in massive plant death. The oregano essential oil can inhibit the spread of pathogenic bacteria on plants, so that the bacterial strains damaged by the pathogenic bacteria can recover normal growth in the later period, thereby reducing the yield loss, and the control effect is in positive correlation with the concentration of the oregano essential oil.

Detailed Description

The technical solution of the present invention is further described in detail with reference to the following examples, but the scope of the present invention is not limited thereto.

In the following examples, thymol used was a common commercially available product, and oregano essential oil was obtained from oregano oil produced by Jiangxi Cedar Natural medicinal oil Co., Ltd. and had an oily liquid ranging from pale yellow to brownish red in appearance and had all the spicy and aromatic odor of thyme, and the main components of carvacrol were equal to or greater than 88% and thymol were equal to or greater than 2.0%.

Test methods and results

1. Virulence of thymol on growth of hyphae of southern blight fungus

The growth rate of hypha is measured by a method for inhibiting the growth rate of hypha. Dissolving 0.03 g of thymol in 0.4 mL of DMSO and Tween-80, adding 19.57 mL of sterile water to a constant volume of 20mL, and preparing into a mother solution of 1500 mg/L, wherein the volume ratio of the DMSO to the Tween-80 is 1%; then the mixture is diluted into medicine liquid with 5 series concentrations of 1300, 1100, 900, 450 and 225 mg/L by sterile water in a gradient way. Adding 1 mL of medicinal liquid with different concentrations into 9 mL of PDA culture medium to obtain 6 kinds of medicinal culture medium with final concentrations of 150, 130, 110, 90, 45, and 22.5 mg/L, and control agent polyoxin with final concentrations of 6000, 1000, 500, 100, 50, and 10mg/L, and repeating the above three times with non-medicinal culture medium containing 0.1% DMSO and 0.1% Tween-80 as control. Culturing in dark at 25 deg.C in light incubator, and measuring colony diameter by cross method after 3 d. Calculating the inhibition rate of thymol and polyoxin on the growth of the hyphae of the southern blight bacteria, and obtaining a toxicity regression equation and effective medium concentration (EC) according to a probability value analysis method₅₀) And a concentration (EC) of up to 90% inhibition₉₀）。

Hypha growth inhibition = [ (control colony diameter-cake diameter) - (treatment colony diameter-cake diameter) ]/(control colony diameter-cake diameter) × 100. The results are shown in Table 1.

TABLE 1 virulence of thymol against floral southern blight hyphae

As is clear from Table 1, thymol has a good inhibitory activity on the hyphal growth of southern blight bacterium, and the inhibitory activity is increased with the increase of thymol concentration, EC₅₀The value is 83.3 mg/L, and the inhibitory activity is obviously better than that of a control medicament polyoxin (EC)₅₀The value was 204.7 mg/L).

Inhibition of sclerotium germination of southern blight fungus by thymol

Dissolving 0.06 g of thymol in 1 mL of DMSO and Tween-80, adding 18.94 mL of sterile water, diluting to 20mL, and preparing into mother liquor of 3000 mg/L for standby, wherein the volume ratio of the DMSO to the Tween-80 is 3% and 2%, respectively. Then the solution is diluted into 5 liquid medicines with the concentration series of 2500, 2000, 1500, 1000 and 500 mg/L in a gradient way. Adding 1 mL of the medicinal liquid into 9 mL of PDA culture medium, and making into 6 series of medicinal culture medium with final concentration of 300, 250, 200, 150, 100, and 50 mg/L. The sclerotium with the same shape and size is selected respectively and placed on the drug-containing culture medium, 20 sclerotium are placed in each treatment, drug-free culture medium containing 0.3% DMSO and 0.2% Tween-80 is used as a control, and each concentration is repeated three times. And after 2 d of culture on a culture medium containing medicine, observing the germination condition of sclerotium, and when the hypha generated by the sclerotium is more than or equal to 2 mm, determining the sclerotium to germinate. Sclerotium germination inhibition rate = (control sclerotium germination number-treated sclerotium germination number)/(control sclerotium germination number) × 100. The results are shown in Table 2.

TABLE 2 inhibitory Effect of thymol on sclerotia germination of southern blight fungus

The results in table 2 show that: the thymol also has a good inhibition effect on sclerotium germination of the southern blight bacteria. Under the treatment concentration of 100 mg/L, the inhibition rate of thymol on sclerotium germination is 76.67%; when the concentration of the thymol is increased to 150 mg/L and above, the inhibition rate of the thymol on sclerotium germination is up to 100%.

Potted plant control effect of thymol on peanut southern blight

A spraying method is adopted to test the potted plant control effect of thymol on the peanut southern blight.

The test peanut variety is Yuhua 9326, peanut seeds with full grains and consistent sizes are selected and placed in a basin, one third of the seeds are submerged by adding sterile water, and the peanut seeds are placed in an incubator for half a day. Mixing matrix soil and vermiculite according to a volume ratio of 3:1, adjusting the water holding capacity of the soil by using sterile water to 70%, placing the soil in a pot, and sowing the germinated seeds with radicles facing downwards.

Artificially inoculating germs: after one week of sowing, selecting healthy, growing and consistent peanut plants, inoculating 1 fungus cake on the stem base of each peanut plant, and burying with soil. When white hyphae appear on peanut plants and soil, spraying the root stems and the surrounding soil of the peanut plants with 500 mg/L thymol and 1000mg/L polyoxin liquid medicines respectively, wherein 10 mL of liquid medicine is sprayed on each plant. Sterile water containing 0.1% DMSO and 0.1% Tween-80 was used as a control. Each treatment was 15 strains, replicated three times. After 3 days of administration, the disease condition was identified according to the following grading criteria:

level 0: no disease spots;

level 1: the lesion is smaller than 1/3 pedicles;

and 3, level: the disease spots are larger than 2/3 stems, and a small amount of hypha is on the ground;

and 5, stage: the disease spots are larger than 2/3 stems, and a large amount of hypha is on the ground;

and 7, stage: the disease spot is larger than 2/3 stems, the ground has a large amount of hyphae, and the overground part of the disease spot has wilting;

and 9, stage: the plants died.

According to the disease index = (each stage diseased plant x relative stage value)/(total plant number x 9) × 100,

The prevention and treatment effect (%) is = (the index of illness state treated by contrast-the index of illness state treated by medicament)/the index of illness state treated by contrast is multiplied by 100, and the index of illness state and prevention effect of different treatments are calculated. The results are shown in Table 3.

TABLE 3 potted control of peanut southern blight by thymol

Note: data in the table are mean ± sem. The different letter representations in the same column are tested by the least significant difference methodP <The difference at the 0.05 level was significant.

As can be seen from the above table 3, thymol has a good treatment effect on the peanut southern blight, and the control effect (62.54%) of thymol at the dose of 1000mg/L is significantly higher than the control effect (42.9%) of polyoxin at the dose of 1000mg/L of a control medicament, which indicates that thymol has a great application prospect in the prevention and treatment of the peanut southern blight.

Inhibition effect of oregano essential oil on growth of hyphae of peanut southern blight bacteria

The inhibitory activity of the oregano essential oil on the hypha growth of sclerotium rolfsii is measured by adopting a hypha growth rate inhibition method. 0.1 g of oregano essential oil is dissolved in 0.2 mL of DMSO and Tween-80, 9.7 mL of sterile water is added to the solution to reach a constant volume of 10 mL, and a mother solution of 10000 mg/L is prepared, wherein the volume ratio of the DMSO to the Tween-80 is 1%. Then diluting with sterile water to obtain medicinal liquid of 5000 mg/L. Adding 1 mL of two different concentrations of medicinal liquid into 9 mL of PDA culture medium to obtain medicinal culture medium with final concentrations of 1000mg/L and 500 mg/L respectively. The final concentrations of the control agent, polyoxin, were 1000mg/L and 500 mg/L. Control was performed in drug-free medium containing 0.1% DMSO and 0.1% Tween-80, and repeated three times per concentration. Culturing in dark at 25 deg.C in light incubator, and measuring colony diameter by cross method after 3 d. The inhibition ratio of oregano essential oil was calculated according to the inhibition ratio of hypha growth = [ (control colony diameter-cake diameter) - (treated colony diameter-cake diameter) ]/(control colony diameter-cake diameter) × 100. The results are shown in Table 4.

TABLE 4 inhibition ratio of oregano essential oil of different concentrations on the growth of hyphae of southern blight fungus

As can be seen from Table 4, the inhibition rate of the oregano essential oil on the growth of the hyphae of the southern blight fungus is as high as 100% under the treatment concentration of 1000mg/L and 500 mg/L, and the inhibition rate is obviously better than that of the control medicament polyoxin (64.9% and 61.7%).

Toxicity of oregano essential oil on growth of hyphae of peanut southern blight bacteria

The growth rate of hypha is measured by a method for inhibiting the growth rate of hypha. 0.1 g of oregano essential oil is dissolved in 0.4 mL of DMSO and Tween-80, 19.5 mL of sterile water is added to the solution to reach a constant volume of 20mL, and a mother solution of 5000 mg/L is prepared, wherein the volume ratio of the DMSO to the Tween-80 is 1%. Then the mixture is diluted into liquid medicine with 6 series concentrations of 2500, 1250, 625, 312.5, 156.25 and 78.125 mg/L by sterile water gradient. 1 mL of the liquid medicine with different concentrations is respectively added into 9 mL of PDA culture medium to prepare 7 kinds of drug-containing culture medium with final concentrations of 500, 250, 125, 62.5, 31.25, 15.625 and 7.8125 mg/L, the final concentrations of the control drug polyoxin are 6000, 1000, 500, 100, 50 and 10mg/L, and drug-free culture medium containing 0.1% DMSO and 0.1% Tween-80 is used as a control, and each concentration is repeated three times. Culturing in dark at 25 deg.C in light incubator, and measuring colony diameter by cross method after 3 d. Calculating the inhibition rate of thymol and polyoxin on the growth of the hyphae of the southern blight bacteria, and obtaining a toxicity regression equation and effective medium concentration (EC) according to a probability value analysis method₅₀) And a concentration (EC) of up to 90% inhibition₉₀）。

Hypha growth inhibition = [ (control colony diameter-cake diameter) - (treatment colony diameter-cake diameter) ]/(control colony diameter-cake diameter) × 100. The results are shown in Table 5.

TABLE 5 toxicity of Oregano essential oil on hyphae of southern blight bacterium

As can be seen from Table 5, the oregano essential oil has good inhibitory activity on the hypha growth of southern blight, and the inhibitory activity is enhanced with the increase of the concentration of the essential oil, EC₅₀The value is 118.5 mg/L, and the inhibitory activity is obviously better than that of a control medicament polyoxin (EC)₅₀The value was 204.7 mg/L).

Inhibition effect of oregano essential oil on germination of sclerotium of sclerotium of sclerotium of sclerotium of sclerotium of sclerotium of sclerotium of ingredient of ingredient of ingredient origanum of ingredient of

Dissolving 0.1 g of oregano essential oil in 1 mL of DMSO and Tween-80, adding 18.9 mL of sterile water to reach a constant volume of 20mL, and preparing into mother liquor of 5000 mg/L for later use, wherein the volume ratio of the DMSO to the Tween-80 is 3% and 2%, respectively. Then the solution is diluted into liquid medicines with 6 series concentrations of 3000, 2500, 2000, 1500, 1000 and 500 mg/L in a gradient way. Adding 1 mL of the medicinal liquid into 9 mL of PDA culture medium, and making into 6 series of medicinal culture medium with final concentration of 300, 250, 200, 150, 100, and 50 mg/L. The sclerotium with the same shape and size is selected respectively and placed on the drug-containing culture medium, 20 sclerotium are placed in each treatment, drug-free culture medium containing 0.3% DMSO and 0.2% Tween-80 is used as a control, and each concentration is repeated three times. And (4) observing the germination condition of sclerotia after 2 days on the culture medium containing the medicine, and germinating when the hypha generated by the sclerotia is more than or equal to 2 mm. Sclerotium germination inhibition rate = (control sclerotium germination number-treated sclerotium germination number)/(control sclerotium germination number) × 100. The results are shown in Table 6.

TABLE 6 inhibitory Effect of oregano essential oil on sclerotium germination of southern blight fungus

The results in Table 6 show that the oregano essential oil also has a good inhibition effect on sclerotium germination of southern blight bacteria. Under the treatment concentration of 250 mg/L, the inhibition rate of the oregano essential oil on sclerotium germination is 85%; when the concentration of the essential oil is increased to 300 mg/L, the germination of sclerotium can be completely inhibited.

Potted plant control effect of oregano essential oil on peanut southern blight

The test peanut variety is Yuhua 9326, peanut seeds with full grains and consistent sizes are selected and placed in a basin, one third of the seeds are submerged by adding sterile water, and the peanut seeds are placed in an incubator for half a day. Then mixing the matrix soil and vermiculite according to the volume ratio of 3:1, adding sterile water to adjust the soil to 70% humidity, placing the soil in a pot, and sowing the germinated seeds with radicles facing downwards.

Artificially inoculating germs: after one week of sowing, plants with the same health, growth vigor and size are selected, 1 fungus cake is inoculated on the base of the stem of each plant, and the plants are buried in soil. When white hypha appears on peanut plants and soil, uniformly spraying the root stems and the surrounding soil of the plants with 500 mg/L oregano essential oil and 1000mg/L polyoxin respectively, and spraying 10 mL of pesticide liquid on each plant. Sterile water containing 0.1% DMSO and 0.1% Tween-80 was used as a control. Each treatment of 15 plants was repeated three times. 3 d after the application, the disease condition is identified according to the following grading standards:

level 0: no disease spots;

level 1: the lesion is smaller than 1/3 pedicles;

and 9, stage: the plants died.

The prevention and treatment effect (%) is = (the index of illness state treated by contrast-the index of illness state treated by medicament)/the index of illness state treated by contrast is multiplied by 100, and the index of illness state and prevention effect of different treatments are calculated. The results are shown in Table 7 and FIG. 1.

TABLE 7 preventive and therapeutic effects of oregano essential oil on southern blight of peanut

Note: data in the table are mean ± sem. The different letter representations in the same column are tested by the least significant difference methodP<The difference at the 0.05 level was significant.

As can be seen from Table 7 and FIG. 1, the oregano essential oil has good control effect on peanut southern blight. The control effect of 1000mg/L oregano essential oil on peanut southern blight is 50.76%, which is equivalent to the control effect (42.9%) of control medicament polyoxin under the dosage of 1000 mg/L.

Claims

1. Application of thymol in preventing and treating peanut southern blight is provided.

2. Use according to claim 1, wherein the plant essential oil containing thymol is used for controlling peanut southern blight.

3. Use according to claim 2, wherein the plant essential oil is oregano essential oil or thyme essential oil.

4. The use as claimed in claim 1, wherein thymol or the plant essential oil containing thymol is compounded with adjuvants conventional in the art to prepare a mixed medicament.

5. The use according to claim 4, wherein the mixed medicament is a suspension, a water dispersible granule, a wettable powder, a granule, an emulsion in water, an emulsifiable concentrate or a microcapsule.