Preparation and Application of a Fungicide Containing Azoxystrobin, Oxalic Acid and Bacillus amyloliquefaciens

Abstract

The present invention discloses a composition containing azoxystrobin, oxalic acid and Bacillus amyloliquefaciens. The ratio of azoxystrobin, oxalic acid and Bacillus amyloliquefaciens in the composition is 50 - 1000 μg: 50 - 1000 μg: 1×10<supgt;5< / supgt> - 1×10<supgt;7< / supgt> cfu. The present invention also discloses the application of the composition in the preparation of a fungicide. The fungicide has a high bacteriostatic efficiency, a significant synergistic effect among the components, can reduce the usage amount of chemical pesticides, and reduce the risk of crop resistance to azoxystrobin; can effectively reduce the chemical agent residues in crops and reduce the dietary risk of agricultural products caused by the application of chemical fungicides; is environmentally friendly and can be widely used for the prevention and control of plant anthracnose and / or Fusarium wilt, which is of great significance for the development of sustainable agriculture.

Description

Technical Field

[0001] The invention relates to the technical field of pesticides, and more specifically to the preparation and application of a fungicide containing azoxystrobin, oxalic acid and Bacillus amyloliquefaciens. Background Art

[0002] Azoxystrobin, also known as Amicida and Anmicida, is a methoxyacrylate fungicide pesticide that can effectively prevent and control a variety of crop diseases caused by fungi. However, due to the abuse of azoxystrobin alone, some pathogens have developed resistance to azoxystrobin, and repeated use of azoxystrobin will also have adverse effects on soil microorganisms and soil enzymes. In order to improve the efficacy of azoxystrobin and extend its effective time, a series of chemical compound fungicides that use other chemical agents, such as fenpropimorph, pyraclostrobin and thiophanate-methyl, have appeared on the market. However, chemical compound fungicides not only cause certain pollution to the environment, but also remain in agricultural products, posing dietary risks.

[0003] Plant immune inducers are also called plant vaccines. Plant immune inducers can induce plants to produce various defense enzymes, proteins and hormones, enhance the strength of plant cell walls, and improve plant disease resistance. Plant immune inducers are considered to be an alternative way to reduce agricultural dependence on chemical pesticides, which is conducive to the development of sustainable agriculture. Plant immune inducers include oligosaccharides, lipopolysaccharides and protease inhibitors, but plant immune inducers and azoxystrobin are different types of prevention and control strategies. There are no fungicides that use plant immune inducers and azoxystrobin in combination on the market.

[0004] Biocontrol bacteria are a series of beneficial microorganisms that can prevent and control plant diseases, such as Bacillus amyloliquefaciens, Trichoderma among fungi, Bacillus subtilis among bacteria, and Streptomyces among actinomycetes. Biocontrol bacteria can change the diversity and richness of soil microorganisms, as well as the activity of enzymes and organic carbon content in the soil, thereby improving the structure of rhizosphere soil microbial communities, alleviating crop continuous cropping obstacles and reducing crop morbidity. However, biocontrol bacteria have low geographical adaptability, lower control benefits than chemical control, and poor drug resistance. The population will decline rapidly due to residual pesticides in the field or after the application of pesticides. Summary of the invention

[0005] The purpose of the present invention is to overcome the shortcomings of the prior art and provide a preparation method and application of a fungicide containing azoxystrobin, oxalic acid and Bacillus amyloliquefaciens.

[0006] The first object of the present invention is to provide a composition containing azoxystrobin, oxalic acid and Bacillus amyloliquefaciens.

[0007] The second object of the present invention is to provide a bactericide and / or a medicine.

[0008] The third object of the present invention is to provide a method for preventing and controlling soybean anthracnose.

[0009] The fourth object of the present invention is to provide a method for preparing a fungicide and / or a medicine.

[0010] In order to achieve the above object, the present invention is implemented by the following technical solutions:

[0011] A composition containing azoxystrobin, oxalic acid and Bacillus amyloliquefaciens.

[0012] Preferably, the name of the Bacillus amyloliquefaciens is JT68, and the strain was deposited in the Guangdong Provincial Institute of Microbiology Strain Collection Center on May 21, 2019, with a deposit number of GDMCC NO: 60673.

[0013] More preferably, in the composition, the ratio of azoxystrobin, oxalic acid and Bacillus amyloliquefaciens is 50-1000 μg: 50-1000 μg: 1×10 5 ~1×10 7 cfu.

[0014] More preferably, in the composition, the ratio of azoxystrobin, oxalic acid and Bacillus amyloliquefaciens is 50-500 μg: 50-500 μg: 1×10 5 ~1×10 7 cfu.

[0015] Most preferably, in the composition, the ratio of azoxystrobin, oxalic acid and Bacillus amyloliquefaciens is 50-100 μg: 50-100 μg: 1×10 5 ~1×10 7 cfu.

[0016] More preferably, in the composition, the ratio of azoxystrobin, oxalic acid and Bacillus amyloliquefaciens is 50 μg: 50 μg: 1×10 7 cfu.

[0017] The use of the composition in the preparation of fungicides and / or medicines should also be within the protection scope of the present invention.

[0018] Preferably, the bactericide and / or drug is an anthracnicide and / or a Fusarium-killing agent.

[0019] The use of the composition in the preparation of fungicides and / or drugs for preventing and controlling plant anthracnose and / or sickle wilt should also be within the protection scope of the present invention.

[0020] A bactericide and / or medicine, characterized by comprising the composition containing azoxystrobin, oxalic acid and Bacillus amyloliquefaciens.

[0021] Preferably, the name of the Bacillus amyloliquefaciens is JT68, and the strain was deposited in the Guangdong Provincial Institute of Microbiology Strain Collection Center on May 21, 2019, with a deposit number of GDMCC NO: 60673.

[0022] More preferably, in the fungicide and / or drug, the ratio of azoxystrobin, oxalic acid and Bacillus amyloliquefaciens is 50-1000 μg: 50-1000 μg: 1×10 5 ~1×10 7 cfu.

[0023] More preferably, in the fungicide and / or drug, the ratio of azoxystrobin, oxalic acid and Bacillus amyloliquefaciens is 50-500 μg: 50-500 μg: 1×10 5 ~1×10 7 cfu.

[0024] Most preferably, in the fungicide and / or drug, the ratio of azoxystrobin, oxalic acid and Bacillus amyloliquefaciens is 50-100 μg: 50-100 μg: 1×10 5 ~1×10 7 cfu.

[0025] More preferably, in the fungicide and / or drug, the ratio of azoxystrobin, oxalic acid and Bacillus amyloliquefaciens is 50 μg: 50 μg: 1×10 5 ~1×10 7 cfu.

[0026] A method for preventing and controlling soybean anthracnose comprises the following steps:

[0027] The fungicide and / or drug are used to spray the leaves of soybean seedlings.

[0028] Preferably, the method comprises spraying the non-diseased leaves of soybean seedlings with the fungicide / medicine to achieve the effect of preventing and controlling soybean anthracnose.

[0029] A method for preparing a fungicide and / or a medicine comprises the following steps:

[0030] S1. mixing azoxystrobin and oxalic acid to prepare azoxystrobin-oxalic acid compound;

[0031] S2. Adding Bacillus amyloliquefaciens to the azoxystrobin-oxalic acid compound prepared in step S1, so that the ratio of azoxystrobin, oxalic acid and Bacillus amyloliquefaciens in the composition is 50-1000 μg: 50-1000 μg: 1×10 5 ~1×10 7 cfu.

[0032] Preferably, in step S1, azoxystrobin and oxalic acid are dissolved in an organic solvent to obtain azoxystrobin-oxalic acid compound.

[0033] More preferably, in step S1, azoxystrobin and oxalic acid are dissolved in acetone to prepare azoxystrobin-oxalic acid compound.

[0034] Preferably, in step S2, Bacillus amyloliquefaciens is added to the azoxystrobin-oxalic acid compound prepared in step S1, so that the concentration of Bacillus amyloliquefaciens in the composition is 1×10 5 ~1×10 7 cfu / mL.

[0035] More preferably, in step S2, Bacillus amyloliquefaciens is added to the azoxystrobin-oxalic acid compound prepared in step S1, so that the final concentration of azoxystrobin in the composition is 50-500 μg / mL.

[0036] Further more preferably, in step S2, Bacillus amyloliquefaciens is added to the azoxystrobin-oxalic acid compound prepared in step S1, so that the final concentration of azoxystrobin in the composition is 50-100 μg / mL.

[0037] Most preferably, in step S2, Bacillus amyloliquefaciens is added to the azoxystrobin-oxalic acid compound prepared in step S1, so that the final concentration of azoxystrobin in the composition is 50 μg / mL.

[0038] More preferably, in step S2, Bacillus amyloliquefaciens is added to the azoxystrobin-oxalic acid compound prepared in step S1, so that the ratio of azoxystrobin, oxalic acid and Bacillus amyloliquefaciens in the composition is 50 μg: 50 μg: 1×10 5 ~1×10 7 cfu.

[0039] The use of the bactericide and / or drug in killing anthrax and / or fusarium is also within the protection scope of the present invention.

[0040] Preferably, the anthrax bacteria is one or more of Siamese anthrax bacteria, flat-headed anthrax bacteria and Higgins anthrax bacteria.

[0041] Further preferably, the Fusarium is one or more of Fusarium solani, Fusarium oxysporum and Fusarium fusarius.

[0042] The use of the fungicide and / or drug in preventing and controlling plant anthracnose and / or sickle wilt is also within the protection scope of the present invention.

[0043] Preferably, the plant anthrax is caused by one or more of Siamese anthrax, flat-headed anthrax and Higgins anthrax.

[0044] Further preferably, the Fusarium wilt is caused by one or more of Fusarium solani, Fusarium oxysporum and Fusarium serrata.

[0045] Compared with the prior art, the present invention has the following beneficial effects:

[0046] The present invention provides a composition with high antibacterial efficiency, environmental friendliness, and significant synergistic effect by compounding two environmentally friendly and efficient agents, plant immune inducer and Bacillus amyloliquefaciens JT68 (biocontrol bacteria), with azoxystrobin. The bactericide and / or drug containing the composition provided by the present invention can reduce the risk of crop resistance to azoxystrobin, and at the same time can reduce the amount of chemical pesticides used, and improve the bactericidal efficiency of the bactericide and / or drug. The bactericide and / or drug containing the composition provided by the present invention is environmentally friendly, can be widely used in the prevention and treatment of plant anthracnose and / or sickle wilt, and is of great significance to the development of sustainable agriculture.

[0047] The three components in the composition provided by the present invention all have a synergistic effect, which overcomes the problem that azoxystrobin and a plant immune inducer interfere with each other when compounded, thereby affecting the plant immune ability, and makes up for the shortcomings of the slow effect of the plant immune inducer and the poor drug resistance of the biocontrol bacteria. At the same time, it combines the advantages of the plant immune inducer, the biocontrol bacteria and azoxystrobin, improves the disease resistance of the plant and has a significant bactericidal effect. BRIEF DESCRIPTION OF THE DRAWINGS

[0048] Figure 1 The antibacterial effect of different concentrations of myclobutanil on 6 plant pathogenic fungi.

[0049] Figure 2 The antibacterial effect of Bacillus amyloliquefaciens JT68 on 6 plant pathogenic fungi.

[0050] Figure 3 The growth of Bacillus amyloliquefaciens JT68 under single agent treatment.

[0051] Figure 4 This is the effect of myclobutanil-oxalic acid combination on the growth of six plant pathogenic fungi.

[0052] Figure 5 This is the effect of the myclobutanil-oxalic acid-Bacillus amyloliquefaciens JT68 composition on the growth of six plant pathogenic fungi.

[0053] Figure 6 The antibacterial effect of pectin oligosaccharides at different concentrations on 6 plant pathogenic fungi.

[0054] Figure 7 The antibacterial effect of different concentrations of chitosan oligosaccharides on 6 plant pathogenic fungi.

[0055] Figure 8The antibacterial effect of different concentrations of oxalic acid on 6 plant pathogenic fungi.

[0056] Fig. 9 The antibacterial effect of different concentrations of amino oligosaccharides on 6 plant pathogenic fungi.

[0057] Fig.10 This is the effect of myclobutanil-pectin oligosaccharide compound on the growth of six plant pathogenic fungi.

[0058] Fig.11 This is the effect of myclobutanil-chitosan oligosaccharide combination on the growth of six plant pathogenic fungi.

[0059] Fig.12 This is the effect of myclobutanil-amino oligosaccharide combination on the growth of six plant pathogenic fungi.

[0060] Fig.13 The greenhouse control effect of the myclobutanil-oxalic acid-Bacillus amyloliquefaciens JT68 composition on soybean anthracnose.

[0061] Fig.14 This is a schematic diagram of the classification standards for disease levels of anthracnose diseased leaves on soybean seedlings. DETAILED DESCRIPTION

[0062] The present invention is further described in detail below in conjunction with the accompanying drawings and specific examples of the specification. The examples are only used to explain the present invention and are not used to limit the scope of the present invention. The test methods used in the following examples are conventional methods unless otherwise specified; the materials and reagents used are reagents and materials that can be obtained from commercial channels unless otherwise specified.

[0063] The Bacillus amyloliquefaciens JT68 or JT68 or Bacillus amyloliquefaciens described in the examples refers to a strain with a deposit number of GDMCCNO: 60673 and a subculture strain or derivative strain thereof, and the strain was deposited in the Guangdong Provincial Institute of Microbiology Strain Collection Center on May 21, 2019.

[0064] Example 1 Antibacterial Experiment of Azoxystrobin on Plant Pathogenic Fungi

[0065] 1. Experimental Methods

[0066] Azoxystrobin was dissolved in acetone to prepare 100 mL of azoxystrobin mother solution, which was placed in a brown small-mouth bottle, labeled and set aside. Azoxystrobin mother solution was added to 4 bottles of PDA culture medium that were pre-heated and melted in a microwave oven and cooled to about 50-60° C., so that the final concentrations of azoxystrobin in the PDA culture medium were 0, 25, 50 and 100 μg / mL, respectively. The PDA culture medium was poured into a culture dish with a diameter of 9 cm while hot, and after cooling and solidification, PDA culture medium plates containing different concentrations of azoxystrobin were prepared.

[0067] Then, one mycelium block of the tested pathogenic fungus was inoculated in the center of the PDA medium plate containing different concentrations of azoxystrobin, and 3 replicates (dishes) were set for each pathogenic fungus, and the plates were placed in an incubator for dark culture at 28° C. The growth of the pathogenic fungi was observed and recorded every day. When the pathogenic fungi in the entire culture dish of the control culture dish (0 azoxystrobin concentration) were almost full, all the culture dishes containing bacteria were taken out, and the colony diameter was measured by the cross method, and the measurement data was recorded and filled in the table, and the growth of the colonies in each treatment was photographed and retained, and finally the inhibition rate was calculated.

[0068]

[0069] 2. Experimental Results

[0070] like Figure 1 As shown, the concentrations (μg / mL) of azoxystrobin in each pathogenic fungus treatment group are 0, 25, 50, and 100 from the upper left, upper right, lower left, and lower right, respectively. Figure 1 Among them, A is the Siamese anthracnose group; B is the Higgins anthracnose group; C is the flat-headed anthracnose group; D is the Fusarium solani group; E is the Cuban special type of Fusarium oxysporum group; F is the Fusarium fusilli group.

[0071] The inhibition rate is shown in Table 1. Different lowercase letters after the data of "inhibition rate" in the same column in the table indicate that the Duncan's new multiple range analysis between the treatments is significantly different (P<0.05). The plate diameter is 90 mm, so the colony diameter of the control group is 90 mm. As shown in Table 1, except for Siamese anthrax, the antibacterial effect of 50 μg / mL azoxystrobin on the other five pathogenic fungi is better than 25 and 100 μg / mL, so the subsequent experiments all use 50 μg / mL azoxystrobin as the concentration of the compound agent.

[0072] Table 1 Antibacterial effect of azoxystrobin on plant pathogenic fungi (colony diameter mm)

[0073]

[0074] Example 2 Antibacterial Experiment of Bacillus amyloliquefaciens JT68 on Plant Pathogenic Fungi

[0075] 1. Experimental Methods

[0076] The antagonistic activity of Bacillus amyloliquefaciens JT68 against six plant pathogenic fungi was determined by the confrontation culture method. The mycelial blocks (bacterial cakes) of six plant pathogenic fungi: Siamese anthracnose, Higgins anthracnose, flat-headed anthracnose, Fusarium solani, Fusarium oxysporum Cuban specialization and Fusarium fusilli were punched out with a 6mm puncher and inoculated in the center of a 9cm diameter PDA plate. Bacillus amyloliquefaciens JT68 was symmetrically inoculated 2.5cm away from the mycelial blocks of pathogenic fungi. The PDA plate without inoculation of Bacillus amyloliquefaciens JT68 was used as the control group. The plates were cultured at 28℃. When the pathogenic fungal colonies in the control group almost filled the culture dish, the antibacterial zone around Bacillus amyloliquefaciens JT68 was observed, and the radius or diameter of the antibacterial zone was measured. The growth of the colonies in each treatment was photographed and retained.

[0077] 2. Experimental Results

[0078] like Figure 2 As shown, the right side of each group of figures is the treatment group, and the upper and lower parts of each treatment group plate are all Bacillus amyloliquefaciens JT68; the left side is the PDA plate without Bacillus amyloliquefaciens JT68 as the control group. Figure 2 Among them, A is the Siamese anthracnose group; B is the Higgins anthracnose group; C is the flat-headed anthracnose group; D is the Fusarium solani group; E is the Cuban special type of Fusarium oxysporum group; F is the Fusarium fusilli group.

[0079] The inhibition rates are shown in Table 2. The results showed that although Bacillus amyloliquefaciens JT68 had a certain degree of inhibition against Siamese anthrax, Higgins anthrax, flat-headed anthrax, Fusarium solani, Fusarium oxysporum Cuban special type and Fusarium fusilli, the overall inhibition effect was not good.

[0080] Table 2 Antibacterial effect of Bacillus amyloliquefaciens JT68 on plant pathogenic fungi (diameter of inhibition zone, mm)

[0081]

[0082] Example 3 Antibacterial test of a single agent against Bacillus amyloliquefaciens JT68

[0083] 1. Experimental Methods

[0084] Drug-containing filter paper disc method: prepare drug solutions of different concentrations and soak the filter paper discs (6 mm in diameter) in them; mix 1 mL of a certain concentration of Bacillus amyloliquefaciens JT68 bacterial suspension into LB culture medium, then stick the filter paper disc soaked in the drug solution in the center of the LB culture medium, and measure the diameter of the inhibition zone around the drug-containing filter paper disc on the next day.

[0085] (1) Antibacterial test of azoxystrobin against Bacillus amyloliquefaciens JT68

[0086] The drug-containing filter paper disc method was used. Azoxystrobin solutions with concentrations of 0, 25, 50, and 100 μg / mL were prepared and used to soak filter paper discs (diameter 6 mm). The bacterial suspension of Bacillus amyloliquefaciens JT68 was mixed into 100 mL of LB solid medium that had been heated and melted and cooled to about 45°C, so that the concentration of Bacillus amyloliquefaciens JT68 in the LB solid medium was 1×10 5 ~1×10 7 cfu / mL, pour the plate, wait for the plate to cool and solidify, then stick the filter paper disc soaked with different concentrations of azoxystrobin solution in the center of the LB culture medium, and measure the diameter of the inhibition zone around the filter paper disc containing azoxystrobin the next day.

[0087] (2) Antibacterial test of pectin oligosaccharides against Bacillus amyloliquefaciens JT68

[0088] The drug-containing filter paper disc method was used. PDA culture medium containing pectin oligosaccharide with concentrations of 0, 25, 50, and 100 μg / mL was prepared for soaking filter paper discs (6 mm in diameter). The bacterial suspension of Bacillus amyloliquefaciens JT68 was mixed into 100 mL of LB solid culture medium that had been heated and melted and cooled to about 45°C, so that the concentration of Bacillus amyloliquefaciens JT68 in the LB solid culture medium was 1×10 5 ~1×10 7 cfu / mL, pour the plate, wait for the plate to cool and solidify, then stick the filter paper disc soaked with pectin oligosaccharide solution of different concentrations in the center of LB culture medium, and measure the diameter of the inhibition zone around the filter paper disc containing pectin oligosaccharide on the next day.

[0089] (3) Antibacterial test of chitosan oligosaccharide on Bacillus amyloliquefaciens JT68

[0090] The drug-containing filter paper disc method was used. PDA medium containing chitosan oligosaccharide with chitosan oligosaccharide concentrations of 0, 25, 50, and 100 μg / mL was prepared and used to soak filter paper discs (diameter 6 mm). The bacterial suspension of Bacillus amyloliquefaciens JT68 was mixed into 100 mL LB solid medium that had been heated and melted and cooled to about 45°C, so that the concentration of Bacillus amyloliquefaciens JT68 in the LB solid medium was 1×10 5 ~1×10 7 cfu / mL, pour the plate, wait for the plate to cool and solidify, then stick the filter paper disc soaked with different concentrations of chitosan oligosaccharide solution in the center of LB culture medium, and measure the diameter of the inhibition zone around the filter paper disc containing chitosan oligosaccharide the next day.

[0091] (4) Antibacterial test of oxalic acid on Bacillus amyloliquefaciens JT68

[0092] The drug-containing filter paper disc method was used. Prepared oxalic acid-containing PDA medium with oxalic acid concentrations of 0, 25, 50, and 100 μg / mL, respectively, and used to soak the filter paper disc (diameter 6 mm). Mixed the bacterial suspension of Bacillus amyloliquefaciens JT68 into 100 mL of LB solid medium that had been heated and melted and cooled to about 45°C, the concentration of Bacillus amyloliquefaciens JT68 in the LB solid medium was 1×10 5 ~1×10 7 cfu / mL, pour the plate, wait for the plate to cool and solidify, then stick the filter paper disc soaked with different concentrations of oxalic acid solution in the center of LB culture medium, and measure the diameter of the inhibition zone around the filter paper disc containing oxalic acid the next day.

[0093] (5) Antibacterial test of amino oligosaccharides against Bacillus amyloliquefaciens JT68

[0094] The drug-containing filter paper disc method was used. PDA culture medium containing amino oligosaccharides with concentrations of 0, 25, 50, and 100 μg / mL of amino oligosaccharides was prepared for soaking filter paper discs (6 mm in diameter). The bacterial suspension of Bacillus amyloliquefaciens JT68 was mixed into 100 mL of LB solid culture medium that had been heated and melted and cooled to about 45°C, so that the concentration of Bacillus amyloliquefaciens JT68 in the LB solid culture medium was 1×10 5 ~1×10 7 cfu / mL, pour the plate, wait for the plate to cool and solidify, then stick the filter paper disc soaked with different concentrations of amino oligosaccharide solution in the center of LB culture medium, and measure the diameter of the inhibition zone around the filter paper disc containing amino oligosaccharide the next day.

[0095] 2. Experimental Results

[0096] like Figure 3 As shown, the concentrations (μg / mL) of the drugs in each group are 0, 25, 50, and 100 from the upper left, upper right, lower left, and lower right, respectively. A is the azoxystrobin treatment group, B is the chitosan oligosaccharide treatment group, C is the pectin oligosaccharide treatment group, D is the oxalic acid treatment group, and E is the amino oligosaccharide treatment group.

[0097] Depend on Figure 3 It can be seen that myclobutanil and the other four plant immune inducers (chitosan oligosaccharides, pectin oligosaccharides, oxalic acid and amino oligosaccharides) have no antibacterial effect on Bacillus amyloliquefaciens JT68 at different concentrations.

[0098] Example 4 Antibacterial experiment of azoxystrobin and oxalic acid combination on plant pathogenic fungi

[0099] 1. Experimental Methods

[0100] First, prepare azoxystrobin mother solution and oxalic acid mother solution of the same concentration; mix the azoxystrobin mother solution and the oxalic acid mother solution at a mass ratio of 1:1 to prepare azoxystrobin and oxalic acid compound. Add the azoxystrobin and oxalic acid compound to two bottles of PDA culture medium that are pre-heated and melted and cooled to about 50-60°C, respectively, so that the final concentrations of the azoxystrobin and oxalic acid compound in the PDA culture medium are 0 and 50 μg / mL, respectively, pour the drug-containing PDA culture medium into a culture dish with a diameter of 9 cm while hot, and prepare PDA culture medium plates containing different concentrations of azoxystrobin and oxalic acid compound after cooling and solidification.

[0101] Then, a mycelium block of the tested pathogenic fungus was inoculated in the center of the PDA culture medium plate containing different concentrations of azoxystrobin and oxalic acid compound, and 3 replicates (dishes) were set for each pathogenic fungus, and the plate was placed in an incubator for dark culture at 28° C. The growth of the pathogenic fungi was observed and recorded every day. When the pathogenic fungi in the entire culture dish in the control culture dish (the concentration of azoxystrobin and oxalic acid compound was 0) were almost full, all the culture dishes containing bacteria were taken out, and the colony diameter was measured by the cross method, and the measurement data was recorded and filled in the table, and the growth of the colonies in each treatment was photographed and retained, and finally the inhibition rate was calculated.

[0102] 2. Experimental Results

[0103] like Figure 4 As shown, for each pathogenic fungus treatment group, from left to right, the concentrations (μg / mL) of the combination of azoxystrobin and oxalic acid were 0 and 50, respectively. Figure 4 Among them, A is the Siamese anthracnose group; B is the Higgins anthracnose group; C is the flat-headed anthracnose group; D is the Fusarium solani group; E is the Cuban special type of Fusarium oxysporum group; F is the Fusarium fusilli group.

[0104] The inhibition rate is shown in Table 3. The plate diameter is 90 mm, so the colony diameter of the control group is 90 mm. As shown in Table 3, the combination of azoxystrobin and oxalic acid has an inhibitory effect on the six plant pathogenic fungi, and except for Siamese anthracnose, the combination of azoxystrobin and oxalic acid shows a significant synergistic effect on the other five plant pathogenic fungi.

[0105] Table 3 Antibacterial effect of azoxystrobin and oxalic acid complex on plant pathogenic fungi (colony diameter mm)

[0106]

[0107] Example 5 Antibacterial experiment of the compound of azoxystrobin, oxalic acid and Bacillus amyloliquefaciens JT68 on plant pathogenic fungi

[0108] 1. Experimental Methods

[0109] First, prepare azoxystrobin mother solution and oxalic acid mother solution with the same concentration; mix the azoxystrobin mother solution and oxalic acid mother solution at a mass ratio of 1:1 to prepare azoxystrobin-oxalic acid composite agent. Add the azoxystrobin-oxalic acid composite agent to a PDA culture medium that has been preheated and melted and cooled to about 50 to 60°C, and then add a bacterial solution of Bacillus amyloliquefaciens JT68, so that the concentration ratio of azoxystrobin, oxalic acid and Bacillus amyloliquefaciens JT68 in the PDA culture medium is 50 μg / mL: 50 μg / mL: 1×10 5 ~1×10 7 cfu / mL, pour the drug-containing PDA medium into a culture dish with a diameter of 9 cm while it is still hot, and after cooling and solidification, prepare a PDA medium plate containing azoxystrobin, oxalic acid and Bacillus amyloliquefaciens JT68 compound.

[0110] A PDA culture medium plate without azoxystrobin, oxalic acid and the compound agent of Bacillus amyloliquefaciens JT68 was prepared as a control culture dish.

[0111] Inoculate a mycelium block of the tested pathogenic fungus in the center of each PDA medium plate, set up 3 replicates (dishes) for each pathogenic fungus, and place them in an incubator for dark culture at 28°C. Observe and record the growth of pathogenic fungi every day. When the pathogenic fungi in the entire culture dish of the control culture dish are almost full, take out all the culture dishes containing bacteria, measure the colony diameter by the cross method, record the measurement data, fill in the table, take pictures of the growth of the colonies in each treatment, retain them, and finally calculate the inhibition rate.

[0112] 2. Experimental Results

[0113] like Figure 5 As shown, for each pathogenic fungus treatment group from left to right, the PDA medium plate containing azoxystrobin, oxalic acid and Bacillus amyloliquefaciens JT68 compound is on the left, and the control culture dish is on the right. Figure 5 Among them, A is the Siamese anthracnose group; B is the Higgins anthracnose group; C is the flat-headed anthracnose group; D is the Fusarium solani group; E is the Cuban special type of Fusarium oxysporum group; F is the Fusarium fusilli group.

[0114] The antibacterial rate is shown in Table 4. The plate diameter is 90 mm, so the colony diameter of the control group is 90 mm. As shown in Table 4, the compound of azoxystrobin, oxalic acid and Bacillus amyloliquefaciens JT68 has an antibacterial effect on the six plant pathogenic fungi, and the compound of azoxystrobin, oxalic acid and Bacillus amyloliquefaciens JT68 shows a significant synergistic effect on the six plant pathogenic fungi.

[0115] Table 4 Antibacterial effect of azoxystrobin-oxalic acid-Bacillus amyloliquefaciens JT68 compound on plant pathogenic fungi (colony diameter mm)

[0116]

[0117] Comparative Example 1 Antibacterial Experiment of Pectin Oligosaccharide on Plant Pathogenic Fungi

[0118] 1. Experimental Methods

[0119] Pectin oligosaccharides were added to 4 bottles of PDA culture medium that were preheated and melted in a microwave oven and cooled to about 50-60°C, so that the final concentrations of pectin oligosaccharides in the PDA culture medium were 0, 25, 50 and 100 μg / mL, respectively. The PDA culture medium containing pectin oligosaccharides of different concentrations was poured into a culture dish with a diameter of 9 cm while hot, and after cooling and solidification, PDA culture medium plates containing pectin oligosaccharides of different concentrations were prepared.

[0120] Then, a mycelium block of the tested pathogenic fungus was inoculated in the center of each PDA medium plate, and 3 replicates (dishes) were set for each pathogenic fungus, and they were placed in an incubator for dark culture at 28°C. The growth of pathogenic fungi was observed and recorded every day. When the pathogenic fungi in the entire culture dish of the control culture dish (pectin oligosaccharide concentration was 0) were almost full, all the culture dishes containing bacteria were taken out, and the colony diameter was measured by the cross method. The measurement data was recorded and filled in the table, and the growth of the colonies in each treatment was photographed and retained, and finally the inhibition rate was calculated.

[0121] 2. Experimental Results

[0122] like Figure 6 As shown, the concentrations (μg / mL) of pectin oligosaccharides in each pathogenic fungus treatment group are 0, 25, 50, and 100 from the upper left, upper right, lower left, and lower right, respectively. Figure 6 Among them, A is the Siamese anthracnose group; B is the Higgins anthracnose group; C is the flat-headed anthracnose group; D is the Fusarium solani group; E is the Cuban special type of Fusarium oxysporum group; F is the Fusarium fusilli group.

[0123] The antibacterial rate is shown in Table 5. The plate diameter is 90 mm, so the colony diameter of the control group or the group without antibacterial effect is 90 mm. As shown in Table 5, pectin oligosaccharide alone has no antibacterial effect on the six plant pathogenic fungi.

[0124] Table 5 Antibacterial effect of pectin oligosaccharides on plant pathogenic fungi (colony diameter mm)

[0125]

[0126] Comparative Example 2 Antibacterial Experiment of Chitosan Oligosaccharide on Plant Pathogenic Fungi

[0127] 1. Experimental Methods

[0128] Chitosan oligosaccharide was added to 4 bottles of PDA culture medium which were preheated in a microwave oven to melt and cooled to about 50-60°C, so that the final concentrations of chitosan oligosaccharide in the PDA culture medium were 0, 25, 50 and 100 μg / mL, respectively. The PDA culture medium containing different concentrations of chitosan oligosaccharide was poured into a culture dish with a diameter of 9 cm while hot, and after cooling and solidification, PDA culture medium plates containing different concentrations of chitosan oligosaccharide were prepared.

[0129] Then, a mycelium block of the tested pathogenic fungus was inoculated in the center of the PDA medium plate containing different concentrations of chitosan oligosaccharides, and 3 replicates (dishes) were set for each pathogenic fungus, and the plates were placed in an incubator for dark culture at 28° C. The growth of the pathogenic fungi was observed and recorded every day. When the pathogenic fungi in the entire culture dish in the control culture dish (chitosan oligosaccharide concentration was 0) were almost full, all the culture dishes containing bacteria were taken out, and the colony diameter was measured by the cross method, and the measurement data was recorded and filled in the table, and the growth of the colonies in each treatment was photographed and retained, and finally the inhibition rate was calculated.

[0130] 2. Experimental Results

[0131] like Figure 7 As shown, the concentrations of chitosan oligosaccharides (μg / mL) in each pathogenic fungus treatment group are 0, 25, 50, and 100 from the upper left, upper right, lower left, and lower right, respectively. Figure 7 Among them, A is the Siamese anthracnose group; B is the Higgins anthracnose group; C is the flat-headed anthracnose group; D is the Fusarium solani group; E is the Cuban special type of Fusarium oxysporum group; F is the Fusarium fusilli group.

[0132] The inhibition rate is shown in Table 6. The plate diameter is 90 mm, so the colony diameters of the control group and the group without antibacterial effect are both 90 mm. As shown in Table 6, chitosan oligosaccharide alone has only a weak antibacterial effect on Higgins anthracnose, but has no antibacterial effect on the other five plant pathogenic fungi.

[0133] Table 6 Antibacterial effect of chitosan oligosaccharide on plant pathogenic fungi (colony diameter mm)

[0134]

[0135] Comparative Example 3 Antibacterial Experiment of Oxalic Acid on Plant Pathogenic Fungi

[0136] 1. Experimental Methods

[0137] Oxalic acid was added to four bottles of PDA culture medium that had been preheated in a microwave oven to melt and then cooled to about 50-60°C, so that the final concentrations of oxalic acid in the PDA culture medium were 0, 25, 50 and 100 μg / mL, respectively. The PDA culture medium containing different concentrations of oxalic acid was poured into a culture dish with a diameter of 9 cm while hot, and after cooling and solidification, PDA culture medium plates containing different concentrations of oxalic acid were prepared.

[0138] Then, a mycelium block of the tested pathogenic fungus was inoculated in the center of the PDA medium plate with different concentrations of oxalic acid, and 3 replicates (dishes) were set for each pathogenic fungus, and the plates were placed in an incubator and cultured in the dark at 28° C. The growth of the pathogenic fungi was observed and recorded every day. When the pathogenic fungi in the entire culture dish of the control culture dish (oxalic acid concentration was 0) were almost full, all the culture dishes containing bacteria were taken out, and the colony diameter was measured by the cross method, and the measurement data was recorded and filled in the table, and the growth of the colonies in each treatment was photographed and retained, and finally the inhibition rate was calculated.

[0139] 2. Experimental Results

[0140] like Figure 8 As shown, the concentrations of oxalic acid (μg / mL) in each pathogenic fungus treatment group are 0, 25, 50, and 100 from the upper left, upper right, lower left, and lower right, respectively. Figure 8 Among them, A is the Siamese anthracnose group; B is the Higgins anthracnose group; C is the flat-headed anthracnose group; D is the Fusarium solani group; E is the Cuban special type of Fusarium oxysporum group; F is the Fusarium fusilli group.

[0141] The inhibition rate is shown in Table 7. The plate diameter is 90 mm, so the colony diameters of the control group and the group without antibacterial effect are both 90 mm. As shown in Table 7, oxalic acid alone has no antibacterial effect on the six plant pathogenic fungi.

[0142] Table 7 Antibacterial effect of oxalic acid on plant pathogenic fungi (colony diameter mm)

[0143]

[0144] Comparative Example 4 Antibacterial Experiment of Amino Oligosaccharides on Plant Pathogenic Fungi

[0145] 1. Experimental Methods

[0146] To the PDA culture medium that was preheated in a microwave oven to melt and cooled to about 50-60°C, amino oligosaccharides were added respectively to make the final concentrations of amino oligosaccharides in the PDA culture medium 0, 25, 50 and 100 μg / mL, respectively. The PDA culture medium containing different concentrations of amino oligosaccharides was poured into a culture dish with a diameter of 9 cm while hot, and after cooling and solidification, PDA culture medium plates containing different concentrations of amino oligosaccharides were prepared.

[0147] Then, a mycelium block of the tested pathogenic fungus was inoculated in the center of the PDA culture medium plate containing different concentrations of amino oligosaccharides, and 3 replicates (dishes) were set for each pathogenic fungus, and the plates were placed in an incubator for dark culture at 28° C. The growth of the pathogenic fungi was observed and recorded every day. When the pathogenic fungi in the entire culture dish of the control culture dish (amino oligosaccharide concentration was 0) were almost full, all the culture dishes containing bacteria were taken out, and the colony diameter was measured by the cross method, and the measurement data was recorded and filled in the table, and the growth of the colonies in each treatment was photographed and retained, and finally the inhibition rate was calculated.

[0148] 2. Experimental Results

[0149] like Fig. 9 As shown, the concentrations of amino oligosaccharides (μg / mL) in each pathogenic fungus treatment group are 0, 25, 50, and 100, respectively, from the upper left, upper right, lower left, and lower right. Fig. 9 Among them, A is the Siamese anthracnose group; B is the Higgins anthracnose group; C is the flat-headed anthracnose group; D is the Fusarium solani group; E is the Cuban special type of Fusarium oxysporum group; F is the Fusarium fusilli group.

[0150] The antibacterial rate is shown in Table 8. The plate diameter is 90 mm, so the colony diameters of the control group and the group without antibacterial effect are both 90 mm. As shown in Table 8, amino oligosaccharides alone have no antibacterial effect on the six plant pathogenic fungi.

[0151] Table 8 Antibacterial effect of amino oligosaccharides on plant pathogenic fungi (colony diameter mm)

[0152]

[0153] Comparative Example 5 Antibacterial experiment of azoxystrobin and pectin oligosaccharide compound on plant pathogenic fungi

[0154] 1. Experimental Methods

[0155] First, prepare azoxystrobin mother solution and pectin oligosaccharide mother solution of the same concentration; mix the azoxystrobin mother solution and pectin oligosaccharide mother solution at a mass ratio of 1:1 to prepare azoxystrobin and pectin oligosaccharide compound. Add the azoxystrobin and pectin oligosaccharide compound to the PDA culture medium that has been preheated and melted and cooled to about 50-60°C, respectively, so that the final concentrations of the azoxystrobin-pectin oligosaccharide compound in the PDA culture medium are 0 and 50 μg / mL, respectively, pour the PDA culture medium into a culture dish with a diameter of 9 cm while hot, and prepare PDA culture medium plates containing different concentrations of azoxystrobin and pectin oligosaccharide compound after cooling and solidification.

[0156] Then, a mycelium block of the tested pathogenic fungus was inoculated in the center of each PDA medium plate, and 3 replicates (dishes) were set for each pathogenic fungus, and they were placed in an incubator for dark culture at 28°C. The growth of the pathogenic fungi was observed and recorded every day. When the pathogenic fungi in the control culture dish (the concentration of the azoxystrobin and pectin oligosaccharide compound was 0) were almost full of the entire culture dish, all the culture dishes containing bacteria were taken out, and the colony diameter was measured by the cross method, and the measurement data was recorded and filled in the table, and the growth of the colonies in each treatment was photographed and retained, and finally the inhibition rate was calculated.

[0157] 2. Experimental Results

[0158] like Fig.10 As shown, for each pathogenic fungus treatment group, from left to right, the concentrations (μg / mL) of the combination of azoxystrobin and pectin oligosaccharides were 0 and 50, respectively. Fig.10 Among them, A is the Siamese anthracnose group; B is the Higgins anthracnose group; C is the flat-headed anthracnose group; D is the Fusarium solani group; E is the Cuban special type of Fusarium oxysporum group; F is the Fusarium fusilli group.

[0159] The antibacterial rate is shown in Table 9. The plate diameter is 90 mm, so the colony diameter of the control group is 90 mm. As shown in Table 9, the combination of azoxystrobin and pectin oligosaccharide has an antibacterial effect on the six plant pathogenic fungi.

[0160] Table 9 Antibacterial effect of azoxystrobin and pectin oligosaccharide compound on plant pathogenic fungi (colony diameter mm)

[0161]

[0162] Comparative Example 6 Antibacterial Experiment of Azoxystrobin and Chitosan Oligosaccharide Compound on Plant Pathogenic Fungi

[0163] 1. Experimental Methods

[0164] First, prepare azoxystrobin mother solution and chitosan oligosaccharide mother solution of the same concentration; mix the azoxystrobin mother solution and chitosan oligosaccharide mother solution at a mass ratio of 1:1 to prepare azoxystrobin and chitosan oligosaccharide compound. Add the azoxystrobin and chitosan oligosaccharide compound to a PDA culture medium that has been preheated and melted and cooled to about 50-60°C, respectively, so that the final concentrations of the azoxystrobin and chitosan oligosaccharide compound in the PDA culture medium are 0 and 50 μg / mL, respectively, pour the PDA culture medium into a culture dish with a diameter of 9 cm while hot, and prepare PDA culture medium plates containing different concentrations of azoxystrobin and chitosan oligosaccharide compound after cooling and solidification.

[0165] Then, a mycelium block of the tested pathogenic fungus was inoculated in the center of each PDA medium plate, and 3 replicates (dishes) were set for each pathogenic fungus, and they were placed in an incubator for dark culture at 28°C. The growth of the pathogenic fungi was observed and recorded every day. When the pathogenic fungi in the entire culture dish of the control culture dish (the concentration of the azoxystrobin and chitosan oligosaccharide compound was 0) were almost full, all the culture dishes containing bacteria were taken out, and the colony diameter was measured by the cross method, and the measurement data was recorded and filled in the table, and the growth of the colonies in each treatment was photographed and retained, and finally the inhibition rate was calculated.

[0166] 2. Experimental Results

[0167] like Fig.11 As shown, for each pathogenic fungus treatment group, from left to right, the concentrations (μg / mL) of the combination of azoxystrobin and chitosan oligosaccharide were 0 and 50, respectively. Fig.11 Among them, A is the Siamese anthracnose group; B is the Higgins anthracnose group; C is the flat-headed anthracnose group; D is the Fusarium solani group; E is the Cuban special type of Fusarium oxysporum group; F is the Fusarium fusilli group.

[0168] The antibacterial rate is shown in Table 10. The plate diameter is 90 mm, so the colony diameters of the control group and the group without antibacterial effect are both 90 mm. As shown in Table 10, the combination of azoxystrobin and chitosan oligosaccharide has an antibacterial effect on the five plant pathogenic fungi except Siamese anthracnose.

[0169] Table 10 Antibacterial effect of azoxystrobin and chitosan oligosaccharide compound on plant pathogenic fungi (colony diameter mm)

[0170]

[0171] Comparative Example 7 Antibacterial Experiment of Azoxystrobin and Amino Acid Oligosaccharide Compound on Plant Pathogenic Fungi

[0172] 1. Experimental Methods

[0173] First, prepare azoxystrobin mother solution and amino acid oligosaccharide mother solution of the same concentration; mix the azoxystrobin mother solution and amino acid oligosaccharide mother solution at a mass ratio of 1:1 to prepare azoxystrobin and amino acid oligosaccharide compound. Add the azoxystrobin and amino acid oligosaccharide compound to the PDA culture medium that has been preheated and melted and cooled to about 50-60°C, respectively, so that the final concentrations of the azoxystrobin and amino acid oligosaccharide compound in the PDA culture medium are 0 and 50 μg / mL, respectively, pour the PDA culture medium into a culture dish with a diameter of 9 cm while hot, and prepare PDA culture medium plates containing different concentrations of azoxystrobin and amino acid oligosaccharide compound after cooling and solidification.

[0174] Then, a mycelium block of the tested pathogenic fungus was inoculated in the center of each PDA medium plate, and 3 replicates (dishes) were set for each pathogenic fungus, and they were placed in an incubator for dark culture at 28°C. The growth of the pathogenic fungi was observed and recorded every day. When the pathogenic fungi in the entire culture dish of the control culture dish (the concentration of the azoxystrobin and amino acid oligosaccharide compound was 0) were almost full, all the culture dishes containing bacteria were taken out, and the colony diameter was measured by the cross method, and the measurement data was recorded and filled in the table, and the growth of the colonies in each treatment was photographed and retained, and finally the inhibition rate was calculated.

[0175] 2. Experimental Results

[0176] like Fig.12 As shown, for each pathogenic fungus treatment group, from left to right, the concentrations (μg / mL) of the combination of azoxystrobin and amino acid oligosaccharides were 0 and 50, respectively. Fig.12 Among them, A is the Siamese anthracnose group; B is the Higgins anthracnose group; C is the flat-headed anthracnose group; D is the Fusarium solani group; E is the Cuban special type of Fusarium oxysporum group; F is the Fusarium fusilli group.

[0177] The inhibition rate is shown in Table 11. The plate diameter is 90 mm, so the colony diameter of the control group is 90 mm. As shown in Table 11, the combination of azoxystrobin and amino acid oligosaccharide has an inhibitory effect on 6 plant pathogenic fungi, but except for Siamese anthracnose and Higgins anthracnose, the inhibitory effect of the combination of azoxystrobin and amino acid oligosaccharide on the other 4 plant pathogenic fungi is not as good as that of the combination of azoxystrobin and oxalic acid.

[0178] Table 11 Antibacterial effect of azoxystrobin and amino oligosaccharides on plant pathogenic fungi (colony diameter mm)

[0179]

[0180] Example 6: Greenhouse control test of soybean anthracnose

[0181] 1. Experimental Methods

[0182] Soybean seeds are selected from healthy soybean seeds that are full and free of insects. They are first disinfected in 75% alcohol for 30 seconds, then disinfected in 5% sodium hypochlorite solution for 5 minutes, and finally rinsed with sterile water for 3 times. They are then placed in a nutrient pot filled with vermiculite that has been sterilized at 121°C in an iron barrel by autoclaving for 30 minutes, and germinated at 25°C in a constant temperature incubator, where the temperature is set at 18-20°C under dark conditions. When more than 95% of the soybean seeds can germinate successfully, they are transplanted into a nutrient pot filled with a sterilized substrate (peat nutrient soil and vermiculite are mixed at a mass ratio of 3:1 at 121°C and then placed in an iron barrel by autoclaving for 30 minutes).

[0183] The inoculum (spores) was a spore suspension of Colletotrichum sojae. The concentration of the spore suspension of Colletotrichum sojae was set to 1.0×10 6 / mL; 10mL of spore suspension was evenly sprayed on the front and back sides of the leaves of soybean seedlings with two leaves and one heart (the first pair of true leaves were fully expanded).

[0184] Treatment group A was sprayed with 10 mL of soybean anthracnose spore suspension and then 10 mL of distilled water as a positive control group;

[0185] After spraying 10 mL of the spore suspension of soybean anthracnose, treatment group B was sprayed with 10 mL of azoxystrobin-oxalic acid-Bacillus amyloliquefaciens JT68 composition, and the preparation method was as follows: firstly preparing azoxystrobin mother solution and oxalic acid mother solution with the same concentration; mixing the azoxystrobin mother solution and the oxalic acid mother solution at a concentration ratio of 1:1 to prepare azoxystrobin-oxalic acid compound; adding Bacillus amyloliquefaciens to the azoxystrobin-oxalic acid compound, so that the ratio of azoxystrobin, oxalic acid and Bacillus amyloliquefaciens in the azoxystrobin-oxalic acid-Bacillus amyloliquefaciens JT68 composition was 50 μg / mL:50 μg / mL:1×10 5 ~1×10 7 cfu / mL;

[0186] Treatment group C was first sprayed with 10 mL of azoxystrobin-oxalic acid-Bacillus amyloliquefaciens JT68 composition (prepared in the same way as treatment group B), and then sprayed with 10 mL of soybean anthracnose spore suspension;

[0187] Treatment group D was a negative control group which was not sprayed with the spore suspension of soybean anthracnose and the azoxystrobin-oxalic acid-Bacillus amyloliquefaciens JT68 composition.

[0188] Each treatment was repeated 3 times, for a total of 9 strains. The disease was investigated 7 days after inoculation.

[0189] According to the symptoms on soybean leaves 7 days after inoculation with soybean anthracnose, soybean anthracnose was graded according to the leaf spot rate. The disease levels of soybean anthracnose leaves at the seedling stage are shown in Table 1, which are divided into 6 levels: 0, 1, 3, 5, 7, and 9. The disease index was calculated according to the disease level (Li Yue, 2022).

[0190]

[0191] Table 1 Symptom identification and lesion classification standards for soybean anthracnose resistance

[0192]

[0193] The disease index of soybean was calculated and divided into 6 levels according to the evaluation standard of soybean resistance to anthracnose: high resistance (HR): DI=0, disease resistance (R): 0<DI<10, moderate resistance (MR): 10≤DI<20, moderate sensitivity (MS): 20≤DI<40, susceptible (S): 40≤DI<60, highly susceptible (HS): DI≥60 (Li Yue, 2022).

[0194]

[0195] The classification standard of disease level of anthracnose diseased leaves in soybean seedling stage is as follows Fig.14 shown.

[0196] 2. Experimental Results

[0197] The experimental results are as follows Fig.13 The diseased leaf level and disease index of soybean anthracnose in different treatment groups are shown in Table 12. It can be seen from Table 12 that the treatment group sprayed with the composition of azoxystrobin-oxalic acid-Bacillus amyloliquefaciens JT68 after the occurrence of lesions has significantly reduced the proportion of diseased leaves, lower diseased leaf level and lower disease index compared with the positive control group sprayed with a spore suspension of soybean anthracnose, indicating that the composition of azoxystrobin-oxalic acid-Bacillus amyloliquefaciens JT68 has significant antibacterial efficiency against soybean anthracnose.

[0198] Compared with the treatment group sprayed with azoxystrobin-oxalic acid-Bacillus amyloliquefaciens JT68 composition first and then sprayed with the spore suspension of soybean anthracnose, the disease index of the experimental group was lower than that of the treatment group sprayed with azoxystrobin-oxalic acid-Bacillus amyloliquefaciens JT68 composition after the disease appeared. The proportion of diseased leaves was smaller and the disease index was lower. This shows that the azoxystrobin-oxalic acid-Bacillus amyloliquefaciens JT68 composition not only has the significant antibacterial properties of azoxystrobin, but also retains the protective effects of plant immune inducers and biocontrol bacteria on plants, and has a significant ability to prevent and control crop diseases.

[0199] Table 12 Diseased leaf grade and disease index of soybean anthracnose under different treatments

[0200]

[0201] The above embodiments are preferred implementation modes of the present invention, but the implementation modes of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention should be equivalent replacement methods and are included in the protection scope of the present invention.

Claims

1. Use of a composition in the preparation of a fungicide and / or a drug, characterized in that, the composition contains azoxystrobin, oxalic acid and Bacillus amyloliquefaciens JT68, and the Bacillus amyloliquefaciens JT68 was deposited at the Strain Preservation Center of Guangdong Institute of Microbiology on May 21, 2019, with the deposit number GDMCC NO: 60673; the fungicide and / or the drug is an anthracnose-killing agent and / or a fusarium-killing agent; the anthracnose-causing bacteria are one or more of Colletotrichum siamense, Colletotrichum truncatum and Colletotrichum higginsianum; the fusarium are one or more of Fusarium solani, Fusarium oxysporum f. sp. cubense and Fusarium falciforme; The ratio of azoxystrobin, oxalic acid and Bacillus amyloliquefaciens JT68 is 50 - 1000 µg: 50 - 1000 µg: 1×10 5 - 1×10 7 cfu.

2. Use of a composition in the preparation of a drug for preventing and treating plant anthracnose and / or fusarium wilt, characterized in that, the composition contains azoxystrobin, oxalic acid and Bacillus amyloliquefaciens JT68, and the Bacillus amyloliquefaciens JT68 was deposited at the Strain Preservation Center of Guangdong Institute of Microbiology on May 21, 2019, with the deposit number GDMCC NO: 60673; the plant anthracnose is caused by one or more of Colletotrichum siamense, Colletotrichum truncatum and Colletotrichum higginsianum; the fusarium wilt is caused by one or more of Fusarium solani, Fusarium oxysporum f. sp. cubense and Fusarium falciforme; The ratio of azoxystrobin, oxalic acid and Bacillus amyloliquefaciens JT68 is 50 - 1000 µg: 50 - 1000 µg: 1×10 5 - 1×10 7 cfu.

3. A fungicide and / or a drug, characterized in that, it contains azoxystrobin, oxalic acid and Bacillus amyloliquefaciens JT68, and the Bacillus amyloliquefaciens JT68 was deposited at the Strain Preservation Center of Guangdong Institute of Microbiology on May 21, 2019, with the deposit number GDMCC NO: 60673; the fungicide and / or the drug is an anthracnose-killing agent and / or a fusarium-killing agent; the anthracnose-causing bacteria are one or more of Colletotrichum siamense, Colletotrichum truncatum and Colletotrichum higginsianum; the fusarium are one or more of Fusarium solani, Fusarium oxysporum f. sp. cubense and Fusarium falciforme; The ratio of azoxystrobin, oxalic acid and Bacillus amyloliquefaciens JT68 is 50 - 1000 µg: 50 - 1000 µg: 1×10 5 - 1×10 7 cfu.

4. A method for preventing and treating soybean anthracnose, characterized in that, spraying the leaves of soybean seedlings with the fungicide / or drug described in claim 3.

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