Composition for preventing and treating anthracnose of turpinia arguta and application of composition

Abstract

The invention discloses a composition for preventing and treating anthracnose of turpinia arguta and application of the composition, and belongs to the technical field of plant disease prevention and treatment. The composition comprises a microbial agent and a carrier, the microbial agent is prepared from bacillus mycoides, pseudomonas mendocina, pseudomonas alcaligenes and brevibacillus brevis; the carrier comprises algal polysaccharide, chitosan oligosaccharide and calcium carbonate. According to the invention, a prevention and treatment method of treating bacteria with bacteria is adopted, biocontrol bacteria with a prevention and treatment effect on turpinia montana anthracnose are screened, and through scientific and reasonable proportioning and matching of microbial carriers, reproduction of turpinia montana anthracnose germs is effectively inhibited, the morbidity and disease index of turpinia montana anthracnose are reduced, and a new experience method is provided for prevention and treatment of turpinia montana anthracnose.

Description

Technical Field

[0001] The invention relates to the technical field of plant disease prevention and control, in particular to a composition for preventing and controlling anthracnose of Cyperus rotundus and application thereof. Background Art

[0002] Plant anthracnose is a plant disease caused by fungi of the genus Colletotrichum. This type of fungus can reproduce and grow on a variety of plants. High temperature and high humidity environments are more suitable for the growth of this type of pathogen. Usually, one type of anthracnose only grows on plants of a certain family or genus. The traditional classification system of the genus Colletotrichum also uses this as the classification standard, and is divided into thousands of species such as sorghum anthracnose, flax anthracnose, and lily anthracnose. Although it has been discovered in recent years that the genus Anthrax has a wide host spectrum, and one type of anthracnose can also parasitize on plants of many different families and genera, due to the different resistance of different plants to different anthracnose, the prevention and control of anthracnose still has certain plant specificity.

[0003] Turpinia montana (Blume) Kurz. is a small tree of the genus Turpinia of the family Turpinaceae. Its roots and leaves can be used as medicine. The National Compilation of Chinese Herbal Medicines records that Turpinia montana can "promote blood circulation, dissipate blood stasis, reduce swelling and relieve pain. It can treat injuries from falls and enlarged spleen". It tastes bitter and is cold in nature. It has the effects of clearing away heat and detoxifying, relieving sore throat and reducing swelling, promoting blood circulation and relieving pain. It is mainly used to treat sore throat, sore throat, sore ulcers, and pain from falls. It has extremely high medicinal value. Turpinia montana likes a cool and humid microclimate. It can tolerate shade slightly in the seedling stage. Adult trees like sunlight and are slightly cold-resistant. It grows best in well-drained, deep and fertile soil. However, high humidity environments can easily cause infection and reproduction of anthrax, which seriously affects the yield and medicinal value of Turpinia montana.

[0004] Common anthracnose control drugs on the market include botanical fungicides and chemical preparations, but these drugs have low efficacy and poor control effects when used to control anthracnose in mountain sweet potatoes, and cannot effectively form a protective barrier on mountain sweet potatoes. Therefore, there is an urgent need to provide an efficient and safe anthracnose control drug for mountain sweet potatoes. Summary of the invention

[0005] The purpose of the present invention is to provide a composition for preventing and controlling anthracnose of mountain sweet potato and its application, so as to solve the problems existing in the above-mentioned prior art. The present invention adopts a control method of treating bacteria with bacteria, screens biocontrol bacteria with control effects on anthracnose of mountain sweet potato, and effectively inhibits the reproduction of anthracnose bacteria of mountain sweet potato through scientific and reasonable proportions and matching microbial carriers, thereby reducing the incidence and disease index of anthracnose of mountain sweet potato, and providing a new empirical method for the control of anthracnose of mountain sweet potato.

[0006] To achieve the above object, the present invention provides the following solutions:

[0007] The present invention provides a composition for preventing and treating anthracnose of Pinellia ternata, the composition comprising a microbial agent and a carrier;

[0008] The microbial agent comprises Bacillus mycoides, Pseudomonas mendocina, Pseudomonas alcaligenes and Bacillus brevis;

[0009] The carrier comprises seaweed polysaccharide, chito oligosaccharide and calcium carbonate.

[0010] Preferably, the microbial agent is obtained by mixing Bacillus mycoides solution, Pseudomonas mendocinosus solution, Pseudomonas alcaligenes solution and Bacillus brevis solution in a volume ratio of (50-80): (20-40): (10-20): (2-4).

[0011] Preferably, the concentrations of the Bacillus mycoides solution, Pseudomonas mendocinae solution, Pseudomonas alcaligenes solution and Bacillus brevis solution are all 10 9～12 Pieces / mL.

[0012] Preferably, the mass ratio of seaweed polysaccharide, chito oligosaccharide and calcium carbonate is (15-30):(6-10):1.

[0013] Preferably, the mass volume ratio of the carrier and the microbial agent is (10-15) g:1 mL.

[0014] The invention also provides an application of the composition in preventing and treating anthracnose of Cyperus rotundus.

[0015] The present invention also provides a method for preventing and controlling anthracnose of mountain sweet potato. After the composition is diluted into an aqueous solution of the composition, the mountain sweet potato is soaked in the solution before sowing or before cutting, or the solution is sprayed on the leaves after the mountain sweet potato shows symptoms of anthracnose.

[0016] Preferably, when the mountain fragrant round seeds are soaked before sowing or dipped in liquid before cutting, the aqueous solution of the composition is prepared by mixing the composition and water in a volume ratio of 1 to 2:10; the soaking time is 30 to 60 minutes.

[0017] Preferably, when spraying on leaves, the aqueous solution of the composition is prepared by mixing the composition and water in a volume ratio of 1 to 2:20; the spraying amount is 30 to 50 mL / plant, and the spraying frequency is 10 to 15 days / time.

[0018] The present invention discloses the following technical effects:

[0019] The present invention adopts the prevention and control principle of using bacteria to treat bacteria, selects four biocontrol bacteria, Bacillus mycoides, Pseudomonas mendocinae, Pseudomonas alcaligenes and Bacillus brevis, which have an inhibitory effect on the anthracnose pathogens of mountain fragrant roundworms to prepare a microbial agent, which can effectively inhibit the reproduction of the anthracnose pathogens of mountain fragrant roundworms and reduce the incidence rate and disease index of anthracnose of mountain fragrant roundworms. At the same time, seaweed polysaccharides, chitosan oligosaccharides and calcium carbonate are used as carriers to load the biocontrol bacteria, which not only provides necessary nutrients for the biocontrol bacteria and ensures the activity of the biocontrol bacteria, but also can increase the colonization rate of the biocontrol bacteria and further improve the prevention effect. The composition of the present invention provides a new empirical method for the prevention and control of anthracnose of mountain fragrant roundworms, and also provides a new technical idea for the prevention and control of anthracnose of other plants. DETAILED DESCRIPTION

[0020] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0021] It should be understood that the terms described in the present invention are only for describing a particular embodiment and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. The intermediate value in any stated value or stated range, and each smaller range between any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0022] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0023] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to those skilled in the art. The present invention description and examples are exemplary only.

[0024] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0025] The sources of the strains used in the embodiments of the present invention are as follows:

[0026] Bacillus mycoides, Pseudomonas mendocina, Pseudomonas alcaligenes and Bacillus brevis were purchased from Shanghai Bohu Biotechnology Co., Ltd.

[0027] Example 1

[0028] A composition for preventing and treating anthracnose of Pinellia ternata, the specific preparation process of which is as follows:

[0029] (1) Bacillus mycoides, Pseudomonas mendocina, Pseudomonas alcaligenes and Bacillus brevis were inoculated into LB medium for activation at 30°C for 36 h.

[0030] (2) Picking single colonies on the LB medium in step (1) respectively, inoculating them into LB liquid medium, and culturing them at 30° C., 200 rpm, and 24 h to obtain seed solutions of Bacillus mycoides, Pseudomonas mendocina, Pseudomonas alcaligenes, and Bacillus brevis.

[0031] (3) The seed liquid of Bacillus mycoides, Pseudomonas mendocinae, Pseudomonas alcaligenes and Bacillus brevis were inoculated into the fermentation medium (5.0 g of peptone, 3.0 g of beef extract, 5.0 g of NaCl, 1.0 L of distilled water, pH 7.0) at 30°C and 200 rpm until the concentration of the fermentation medium reached 10 10 / mL, and the fermentation broths of Bacillus mycoides, Pseudomonas mendocina, Pseudomonas alcaligenes and Bacillus brevis were obtained.

[0032] (4) The fermentation broths of Bacillus mycoides, Pseudomonas mendocina, Pseudomonas alcaligenes and Bacillus brevis are mixed in a volume ratio of 70:30:15:3 to obtain a microbial agent; seaweed polysaccharide, chitosan oligosaccharide and calcium carbonate are mixed in a mass ratio of 20:8:1 to obtain a carrier. The carrier and the microbial agent are evenly mixed in a ratio of 15 g:1 mL, so that the microbial agent is evenly attached to the carrier, thereby obtaining a composition for preventing and controlling anthracnose of mountain fragrant roundworms.

[0033] Example 2

[0034] A composition for preventing and treating anthracnose of Pinellia ternata, the specific preparation process of which is as follows:

[0035] (1) Bacillus mycoides, Pseudomonas mendocina, Pseudomonas alcaligenes and Bacillus brevis were inoculated into LB medium for activation at 30°C for 48 h.

[0036] (2) Picking single colonies on the LB medium in step (1) respectively, inoculating them into LB liquid medium, and culturing them at 30° C., 200 rpm, and 24 h to obtain seed solutions of Bacillus mycoides, Pseudomonas mendocina, Pseudomonas alcaligenes, and Bacillus brevis.

[0037] (3) The seed liquid of Bacillus mycoides, Pseudomonas mendocinae, Pseudomonas alcaligenes and Bacillus brevis were inoculated into the fermentation medium (5.0 g of peptone, 3.0 g of beef extract, 5.0 g of NaCl, 1.0 L of distilled water, pH 7.0) at 30°C and 200 rpm until the concentration of the fermentation medium reached 10 9 / mL, and the fermentation broths of Bacillus mycoides, Pseudomonas mendocina, Pseudomonas alcaligenes and Bacillus brevis were obtained.

[0038] (4) The fermentation broths of Bacillus mycoides, Pseudomonas mendocina, Pseudomonas alcaligenes and Bacillus brevis are mixed in a volume ratio of 40:10:5:2 to obtain a microbial agent; seaweed polysaccharide, chitosan oligosaccharide and calcium carbonate are mixed in a mass ratio of 15:10:1 to obtain a carrier. The carrier and the microbial agent are evenly mixed in a ratio of 10 g:1 mL, so that the microbial agent is evenly attached to the carrier, thereby obtaining a composition for preventing and controlling anthracnose of mountain fragrant roundworms.

[0039] Example 3

[0040] A composition for preventing and treating anthracnose of Pinellia ternata, the specific preparation process of which is as follows:

[0041] (1) Bacillus mycoides, Pseudomonas mendocina, Pseudomonas alcaligenes and Bacillus brevis were inoculated into LB medium for activation at 30°C for 36 h.

[0042] (2) Picking single colonies on the LB medium in step (1) respectively, inoculating them into LB liquid medium, and culturing them at 30° C., 200 rpm, and 24 h to obtain seed solutions of Bacillus mycoides, Pseudomonas mendocina, Pseudomonas alcaligenes, and Bacillus brevis.

[0043] (3) The seed liquid of Bacillus mycoides, Pseudomonas mendocinae, Pseudomonas alcaligenes and Bacillus brevis were inoculated into the fermentation medium (5.0 g of peptone, 3.0 g of beef extract, 5.0 g of NaCl, 1.0 L of distilled water, pH 7.0) at 30°C and 200 rpm until the concentration of the fermentation medium reached 10 12 / mL, and the fermentation broths of Bacillus mycoides, Pseudomonas mendocina, Pseudomonas alcaligenes and Bacillus brevis were obtained.

[0044] (4) The fermentation broths of Bacillus mycoides, Pseudomonas mendocina, Pseudomonas alcaligenes and Bacillus brevis are mixed in a volume ratio of 25:20:10:1 to obtain a microbial agent; seaweed polysaccharide, chitosan oligosaccharide and calcium carbonate are mixed in a mass ratio of 30:6:1 to obtain a carrier. The carrier and the microbial agent are evenly mixed in a ratio of 12 g:1 mL, so that the microbial agent is evenly attached to the carrier, thereby obtaining a composition for preventing and controlling anthracnose of mountain fragrant roundworms.

[0045] Comparative Example 1

[0046] The only difference between this comparative example and Example 1 is that only Bacillus mycoides is used to prepare the microbial agent, and the concentration of the microbial agent is the same as that in Example 1.

[0047] Comparative Example 2

[0048] The only difference between this comparative example and Example 1 is that only Pseudomonas mendocinae is used to prepare the microbial agent, and the concentration of the microbial agent is the same as that in Example 1.

[0049] Comparative Example 3

[0050] The only difference between this comparative example and Example 1 is that only Pseudomonas alcaligenes is used to prepare the microbial agent, and the concentration of the microbial agent is the same as that in Example 1.

[0051] Comparative Example 4

[0052] The only difference between this comparative example and Example 1 is that seaweed polysaccharide is omitted.

[0053] Comparative Example 5

[0054] The difference between this comparative example and Example 1 is that the volume ratio of the Bacillus mycoides bacterial solution, the Pseudomonas mendocinosus bacterial solution, the Pseudomonas alcaligenes bacterial solution and the Bacillus brevis bacterial solution is adjusted to 1:1:1:1.

[0055] Test Example 1

[0056] In April 2023, the plots where anthrax had occurred were selected as experimental fields and randomly divided into 9 experimental plots, each with an area of ​​20m 2, respectively, the experimental group (Examples 1 to 3, Comparative Examples 1 to 5) and the control group. The compositions of Examples 1 to 3 and Comparative Examples 1 to 5 were mixed with water in a volume ratio of 1:10 to prepare a composition aqueous solution. Before sowing the mountain fragrant round, the seeds were soaked in the above-mentioned composition aqueous solution for 1 hour, and the control group was soaked in sterile water for 1 hour. Then the soaked mountain fragrant round seeds were sown in the corresponding plots respectively. The management measures after sowing are shown in "DB36 / T 1606-2022 Technical Regulations for Cultivation and Management of Mountain Fragrant Round". In the 5th month after sowing, the incidence rate of each plot was counted (incidence rate % = diseased plants / total number of plants × 100), and the control rate was calculated (control rate % = (control group incidence rate-experimental group incidence rate) / control group incidence rate). The results are shown in Table 1.

[0057] Table 1 Disease incidence and control rate in each experimental plot

[0058]

[0059]

[0060] As can be seen from Table 1, Bacillus mycoides, Pseudomonas mendocinae, and Pseudomonas alcaligenes all have control effects on anthracnose of mountain sweet circles, but the effect is not as good as the combination, and the control effect of the combination needs to be based on a specific ratio to be exerted. In addition, the type of raw materials of the bacterial agent carrier also has a certain influence on the control effect. In addition to being able to load strains, seaweed polysaccharides can increase the viscosity of the aqueous solution of the composition, improve the colonization rate of the strains, and enhance the control effect.

[0061] Test Example 2

[0062] In July 2023, the field where anthrax occurred was selected and randomly divided into 11 experimental plots, each with an area of ​​20m 2 , respectively, experimental group (Examples 1 to 3, comparative examples 1 to 5), blank control group and positive control group (positive control groups 1 and 2).

[0063] The compositions of Examples 1 to 3 and Comparative Examples 1 to 5 were mixed with water in a volume ratio of 1:20 to prepare a composition aqueous solution, and the corresponding group of composition aqueous solutions were sprayed on the leaves of the corresponding group of mountain fragrant round plants. The spraying amount was calculated as 40 mL / plant, and the spraying frequency was 10 days / time. The blank control group was sprayed with sterile water at the same spraying amount. Positive control group 1 used the plant-derived fungicide preparation of Example 1 of patented technology CN102365939A, and used it in the manner described in Example 7 thereof; positive control group 2 used the composite microbial agent of Example 3 of patented technology CN114868773A, and used it in the manner described in Example 6 thereof.

[0064] The test was terminated 30 days after the first spraying. At this time, the experimental group and the blank control group were sprayed 3 times, the positive control group 1 was sprayed 6 times, and the positive control group 2 was sprayed 3 times. The disease index of each plot before and after the start of the test was counted, and the control effect was calculated (control effect % = (blank control group disease index - experimental group / positive control group disease index) / blank control group disease index × 100). The results are shown in Table 2.

[0065] Table 2 The control effect of each test plot

[0066] Group Effectiveness% Blank control group - Example 1 91.62 Example 2 90.77 Example 3 89.10 Comparative Example 1 65.71 Comparative Example 2 46.95 Comparative Example 3 52.11 Comparative Example 4 60.28 Comparative Example 5 78.94 Positive control group 1 56.32 Positive control group 2 49.16

[0067] As can be seen from Table 2, for the mountain osmanthus plants that have already developed anthracnose, Bacillus mycoides, Pseudomonas mendocinae, Pseudomonas alcaligenes and Bacillus brevis have control effects when used alone, but their effects are lower than those when used in combination, and the control effect of the combination is based on the specific ratio of the four bacteria, which is consistent with the results of Test Example 1. The effect is more obvious when seaweed polysaccharides are used for foliar spraying. In the absence of seaweed polysaccharides, the strains are easily lost with water, resulting in a decrease in the control effect.

[0068] In addition, according to the results of Examples 1 to 3 and positive control groups 1 and 2, it can be seen that the anthracnose control method is indeed plant-specific, and the drugs used to control grape anthracnose (positive control group 1) and strawberry anthracnose (positive control group 2) are less effective in controlling anthracnose of Campestris ovata.

[0069] The embodiments described above are only descriptions of the preferred modes of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all fall within the protection scope determined by the claims of the present invention.

Claims

1. A composition for preventing and treating anthracnose of Psoralea corylifolia, characterized in that: The composition comprises a microbial agent and a carrier; The microbial agent comprises Bacillus mycoides, Pseudomonas mendocina, Pseudomonas alcaligenes and Bacillus brevis; The carrier comprises seaweed polysaccharide, chito oligosaccharide and calcium carbonate.

2. The composition according to claim 1, characterized in that The microbial agent is obtained by mixing bacillus mycoides solution, pseudomonas mendocinae solution, pseudomonas alcaligenes solution and bacillus brevis solution in a volume ratio of (50-80): (20-40): (10-20): (2-4).

3. The composition according to claim 2, characterized in that The concentrations of the Bacillus mycoides solution, Pseudomonas mendocinae solution, Pseudomonas alcaligenes solution and Bacillus brevis solution were all 10 9～12 Pieces / mL.

4. The composition according to claim 1, characterized in that The mass ratio of the seaweed polysaccharide, chito oligosaccharide and calcium carbonate is (15-30):(6-10):

1.

5. The composition according to claim 1, characterized in that The mass volume ratio of the carrier and the microbial agent is (10-15) g:1 mL.

6. Use of the composition according to any one of claims 1 to 5 in preventing and controlling anthracnose of Cyperus rotundus.

7. A method for preventing and treating anthracnose of Psoralea corylifolia, characterized in that: After diluting the composition according to any one of claims 1 to 5 into an aqueous solution of the composition, the composition can be used for soaking seeds of the safflower before sowing or dipping before cutting, or sprayed on the leaves after the safflower shows symptoms of anthracnose.

8. The method according to claim 7, characterized in that When the mountain fragrant round is used for soaking seeds before sowing or dipping in liquid before cutting, the composition aqueous solution is prepared by mixing the composition and water in a volume ratio of 1 to 2:10; the soaking time is 30 to 60 minutes.

9. The method according to claim 7, characterized in that: When spraying on leaves, the composition aqueous solution is prepared by mixing the composition and water in a volume ratio of 1 to 2:20; the spraying amount is 30 to 50 mL / plant, and the spraying frequency is 10 to 15 days / time.

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