Disinfectant for apple valsa canker pathogen and method for using the same

Abstract

To provide an effective disinfectant for apple Valsa canker pathogen that can be used in an easy manner, and a method for using the disinfectant.SOLUTION: A disinfectant for apple Valsa canker pathogen of the present invention is a mixed solution containing a culture solution including fungi of at least 14 genera, a fulvic acid solution, and zinc ions, wherein the genera include at least Streptomyces, Sulfolobus, Saccharopolyspora, Nocardia, Pseudomonas, Bacillus, Cytophaga, Candida, Cellulomonas, Clostridium, Aspergillus, Penicillium, Rhizopus, and Basidiomycetes.SELECTED DRAWING: Figure 4

Description

Technical Field

[0001] The present invention relates to a Valsa ceratosperma disinfectant solution for apples and a method for using the same, and more particularly, to a Valsa ceratosperma disinfectant solution for apples and a method for using the same, which are applied to the infected parts of Valsa ceratosperma of apple trees to disinfect Valsa ceratosperma.

Background Art

[0002] Apple canker is a plant disease in which an apple tree is infected with Valsa ceratosperma, a kind of filamentous fungus, and develops symptoms. When an apple tree is infected with Valsa ceratosperma, an affected part with a reddish-brown lesion is formed, the bark turns brown, sap leaks from the affected part, and the tip side from the affected part withers, resulting in a significant decrease in the apple production. There is no way but to cut off the part infected with Valsa ceratosperma, and when the infected part expands, the apple tree has to be cut down to prevent the spread of the infection to other apple trees.

[0003] Conventionally, as a measure for preventing the infection of apple canker, in the fall after apple harvest, the apple tree is wrapped with soil in the cultivation field and further wrapped on the outside for overwintering. However, wrapping the apple tree with soil is a very laborious task and a heavy burden on apple farmers. Moreover, this method only prevents the infection and onset of apple canker, and has almost no effect on the already developed apple canker.

[0004] Patent Document 1 describes a method for preventing infection by applying inorganic agents such as sodium carbonate and alum to apple trees. However, since Valsa ceratosperma lies dormant for nearly a year after invading an apple tree and develops apple canker after invading deep into the resin tissue, it is difficult to disinfect Valsa ceratosperma with the inorganic agents described in Patent Document 1.

Prior Art Documents

Patent Documents

[0005]

Patent Document 1

Non-Patent Literature

[0006]

Non-Patent Literature 1

Non-Patent Literature 2

Summary of the Invention

Problems to be Solved by the Invention

[0007] An object of the present invention is to provide a disinfection solution for apple rot pathogens that is effective against apple rot pathogens and can be easily applied. Another object of the present invention is to provide a method for using a disinfection solution for apple rot pathogens that disinfects and treats the affected parts of apple trees infected with apple rot pathogens, and regenerates the infected apple trees by treatment and regeneration.

Means for Solving the Problems

[0008] The disinfection solution for apple rot pathogens of the present invention is a mixed solution containing a culture solution obtained by mixed culture of bacterial flora belonging to at least the following 14 genera, zinc ions, and fulvic acid, The 14 genera are (1) Streptomyces (2) Sulfolobus (3) Saccharopolyspora (4) Nocardia (5) Pseudomonas (6) Bacillus (7) Cytophaga (8) Candida (9) Cellulomonas (10) Clostridium (11) Aspergillus (12) Penicillium (13) Rhizopus (14) Basidiomycetes and the culture solution is cultured so that the total mass of the cells of the bacterial group is 3.0 to 30.0% by mass of the total mass of the culture solution, and is filtered through a filter medium with an aperture that allows the cells of the bacterial group to pass through, and the solid components are removed. The culture solution contains 1.0 to 20.0% by mass and the fulvic acid contains 0.0001 to 0.01% by mass with respect to 100% of the total mass. It is characterized in that 0.01 to 10.0 millimoles of the zinc ions are contained with respect to 1 kg of the total mass.

[0009] In the present invention, it is characterized in that a thickener is further added to form a form that can be applied in a thin layer.

[0010] The method of using the apple brown rot pathogen disinfectant solution of the present invention is characterized by removing the parts infected with and diseased by the apple brown rot pathogen from the apple tree, and applying the apple brown rot pathogen disinfectant solution to the remaining parts and their surroundings.

[0011] Further, the method of using the apple brown rot pathogen disinfectant solution of the present invention is characterized by removing the parts infected with and diseased by the apple brown rot pathogen from the apple tree, making wounds parallel to the tree veins on the remaining parts and their surroundings, and then applying the apple brown rot pathogen disinfectant solution to the surroundings.

Effect of the Invention

[0012] The present invention disinfects the apple rot pathogen by removing the infected part of the apple tree infected with the apple rot pathogen or the infected part, and then drying it as it is or after applying the apple rot pathogen disinfectant solution by making a scratch parallel to the tree veins on the infected part. The infected part can be regenerated to treat the apple tree infected with the apple rot pathogen.

[0013] By treating the apple tree infected with the apple rot pathogen, the present invention can recover the yield of the apple tree and prevent the spread of the infection, so there is no need to cut down the apple tree infected with the apple rot pathogen. Moreover, since the method of using the disinfectant solution is a simple operation, the burden on apple farmers who wrap the infected part with mud in the fall after the apple harvest can be reduced.

Brief Description of the Drawings

[0014]

Fig. 1

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Modes for Carrying Out the Invention

[0015] Hereinafter, the apple rot pathogen disinfectant solution of the present invention and its usage method will be described. This description is for explaining the present invention of this application, and does not limit the technical scope of the present invention of this application. The present invention of this application can be variously modified and implemented within the scope not departing from the technical scope of the present invention of this application.

[0016] Non-Patent Document 1 reports that in an in vitro confrontation culture between a filamentous fungus isolated from an apple tree and the apple rot pathogen, a part of the filamentous fungus isolated from the apple tree controls the growth of the apple rot pathogen. Furthermore, Non-Patent Document 1 presumes that the effect of the conventional treatment of wrapping the infected part with mud in the fall after apple harvesting is growth control based on the antagonistic action between the soil bacteria at the location where the apple tree grew and the apple rot pathogen. However, this growth control is only an effect in vitro, and the growth control in an actual field is unstable and not practical.

[0017] The present inventor hypothesized that the antagonistic action between the soil bacteria and the apple rot pathogen described in Non-Patent Document 1 is due to the action of an unknown mycovirus. To verify this hypothesis, the present inventor examined the possibility of reducing the toxicity and inhibiting the growth of the apple rot pathogen by a mycovirus contained in a mixed bacterium in which various types of bacteria are mixed at a predetermined ratio and the whole bacterial solution has a predetermined property.

[0018] When the co-culture of a large number of mixed bacteria and the apple rot pathogen (Valsa ceratosperma) was carried out, it was found that the pathogenicity of the apple rot pathogen could be reduced by including at least 14 bacterial genera. However, the pathogenicity inhibitory effect of the mixed bacteria was moderate and was insufficient for practical application as it was. The present invention provides a practical disinfectant for the apple rot pathogen by mixing zinc ions and fulvic acid into the culture solution of this mixed bacterium.

[0019] [Bacterial genera used in the present invention] The mixed bacterium used in the present invention uses a culture solution obtained by mixed-culturing bacterial groups belonging to at least the following 14 bacterial genera. The bacterial groups are as follows. (1) Streptomyces (2) Sulfolobus (3) Saccharopolyspora (4) Nocardia (5) Pseudomonas (6) Bacillus (7) Cytophaga (8) Candida (9) Cellulomonas (10) Clostridium (11) Aspergillus (12) Penicillium (13) Rhizopus (14) Basidiomycetes Here, "Basidiomycetes" is not the name of a genus based on the classification method of fungi. Since the taxonomic system of Basidiomycota, the higher concept of basidiomycetes, is an area where the classification system is undetermined, "Basidiomycetes" is a common name and includes more general "mushrooms" and wood-rotting fungi. The present invention can treat apple rot disease by using a mixed bacterium containing all of the bacterial groups belonging to the above 14 bacterial tribes, and further containing zinc ions and fulvic acid.

[0020] [Bacterial species used in the present invention] In the present invention, as the mixed bacterium containing the bacterial groups belonging to the above 14 genera, for example, a mixed bacterium containing the following 36 bacterial species can be selected. (1) Streptomyces Streptomyces griseus (streptomycin-producing bacterium) (2) Sulfolobus Sulfolobus acidocaldarius ldarius This bacterium is a thermophilic acidophilic archaeon. (3) Saccharopolyspora Saccharopolyspora erythraea is a bacterium that produces erythromycin. (4) Nocardia Nocardia asteroides is an infectious bacterium that causes severe lung infections in immunocompromised hosts. (5) Pseudomonas Pseudomonas fluorescens is a common Gram-negative bacillus that produces a variety of secondary metabolites. Pseudomonas mendocina is involved in toluene degradation. Pseudomonas alcaligenes is capable of degrading polycyclic aromatic hydrocarbons. Pseudomonas veronii is capable of degrading a variety of aromatic organic compounds. Pseudomonas pseudoalkaligenens is capable of metabolizing rare substrates and is particularly effective at decomposing cyanide compounds. Pseudomonas glumae is a bacterial pathogen of rice blast and also has antibacterial activity against several important plant pathogenic bacteria. Pseudomonas gladioli is a pathogen of both humans and plants and has antagonistic interactions with other microorganisms. Pseudomonas glumae This bacterium is also the bacterial pathogen of rice bakanae disease. It also has antibacterial activity against several important plant pathogenic bacteria. Pseudomonas gladioli This bacterium is a pathogen of both humans and plants and has antagonistic interactions with other microorganisms. Pseudomonas glumae Pseudomonas avenae This bacterium causes brown streak symptoms in infected susceptible plants. Pseudomonas putida This bacterium can decompose organic solvents such as toluene. (6) Genus Bacillus Bacillus thuringiensis ) This bacterium produces insecticidal components and is approved as a biopesticide. Bacillus subtilis (7) Genus Cytophaga Cytophaga hutchinsonii This bacterium exhibits gliding motility and is an aerobic cellulose-decomposing bacterium found in soil environments. It is. (8) Genus Candida Candida etchellsii This bacterium is added as a post-ripening yeast to the brewing of soy sauce and miso. Candida versatilis This bacterium is added as a post-ripening yeast to the brewing of soy sauce and miso. Candida stellata This bacterium is involved in wine brewing. (9) Genus Cellulomonas Cellulomonas flavigena This bacterium is a cellulose-decomposing bacterium isolated from manure. (10) Genus Clostridium Clostridium thermocellum umu This bacterium is a cellulase. Clostridium acetobut obutyricum) This bacterium is a bacterium that produces acetone, butanol, and ethanol from starch. It is. Clostridium acetobutyri cum) This bacterium is a bacterium that produces acetone, butanol, and ethanol from starch. It is. Clostridium acetobut yricum) This bacterium is a bacterium used as an intestinal regulator (Bacillus mesentericus). Clostridium ljungdahl ii) This bacterium is a bacterium used in ethanol production. (11) Aspergillus Aspergillus nigar This bacterium is a bacterium used in the production of organic acids such as Aspergillus niger, citric acid, and amylase. It is. Aspergillus oryzae This bacterium is used to make Aspergillus oryzae, soy sauce, miso, sake, shochu, and fermented liquor. It is utilized. Aspergillus penicillo ides) This bacterium is used in the production of fermented foods such as dried bonito and raw ham. Aspergillus brasillen sis) This bacterium is the most common bacterial species in Aspergillus and is widely present in the soil. Aspergillus terreus This bacterium is used to produce organic acids and enzymes. Aspergillus sojae This bacterium is a soy sauce koji mold. (12) Penicillium Penicillium chrysogenum This bacterium is a penicillin-producing bacterium. Penicillium camemberti This bacterium is the blue mold for Camembert cheese. Penicillium glabrum This bacterium is a plant pathogen that infects strawberries. Penicillium digitatum This bacterium is the bacterium that causes post-harvest decay of fruits. (13) Rhizopus Rhizopus oryzae This bacterium produces various enzymes and is used in fields such as the food industry and the pharmaceutical industry. Yes. (14) Basidiomycetes White rot fungus This bacterium decomposes cellulose.

[0021] [Production of culture solution] <Zinc ion> Among filamentous fungi related to cytoplasmic incompatibility, even if they are co-cultured, the transmission (transfer) of mycophagous viruses usually does not occur. However, in the presence of zinc ions, when these are co-cultured, the virus migrates (transmits) from the filamentous fungus infected with the mycophagous virus to the filamentous fungus not infected with the mycophagous virus. For this reason, zinc ions are used. When zinc ions and fulvic acid are added to the culture solution of this mixed fungus and co-cultured with the apple rot pathogen, and the growth inhibitory effect of this mixed fungus is measured, the mixed solution exhibits a strong and practical inhibitory effect on the apple rot pathogen.

[0022] <Fulvic acid> Fulvic acid is a substance that gives a yellow to orange-yellow color to the acidic supernatant when extracted with an inorganic acid after extracting and removing humic acid from soil or coal with a dilute alkali. It is an amorphous acidic substance soluble in water and ethanol, does not have a single chemical structural formula, and neither its composition nor molecular weight is constant. The composition and molecular weight vary depending on the raw material and collection conditions (see, for example, Non-Patent Document 2). Fulvic acid has a strong affinity with zinc ions and acts to assist the uptake of zinc ions into the living body. It is preferable to use a purified fulvic acid solution produced by the method described in Non-Patent Document 2.

[0023] The content of the fulvic acid solution is preferably 0.0001 to 0.01 when the mass of the entire apple rot pathogen disinfectant solution is 100% by mass. If the amount of the fulvic acid solution is less than 0.0001% by mass, a sufficient defensive effect against the apple rot pathogen cannot be obtained. On the other hand, even if an amount of fulvic acid exceeding 0.01% by mass is added, it becomes expensive, and the defensive effect against the apple rot pathogen does not increase, which is not economically preferable.

[0024] <Thickener> The apple rot pathogen disinfectant of the present invention can be made into a form that can be applied in the form of a coating film having a thickness by adding a thickener. The operation of wrapping the apple tree with soil in the cultivation field in the fall after apple harvesting is a very laborious operation for apple farmers. According to the present invention, however, this wrapping operation can be replaced with the spraying operation of the apple rot pathogen disinfectant with a thickener added. As the thickener, various ones including gum arabic, xanthan gum, pectin, carrageenan, gum arabic, etc. can be used, and any one can be used as long as it meets the purpose of the present invention.

[0025] [Production of culture solution] [Pre-culture] For the bacteria of at least 36 bacterial species belonging to at least 14 genera of the present invention, each bacterium is, for example, 1×10 5 cfu / cm 3 At the same concentration, the same amount is mixed with the mother agent of the pre-culture and water, heated to an appropriate temperature, and pre-cultured while adding a small amount of the mother agent to the medium little by little. The mother agent used for the pre-culture for starting fermentation is not particularly limited as long as it can fix and carry fungi on its surface or internal structure. Specifically, for example, barks, sawdust, charcoal powder, coffee grounds, rice husks, bokashi obtained from rice bran, etc. can be preferably used, and one kind or a mixture of two or more kinds thereof can be used.

[0026] [Main culture] When the temperature of the pre-culture solution rises to 45°C to 55°C, fermentation bases are sequentially added in place of the mother agent. In this example, organic waste discharged from an aquatic product processing factory was used as the fermentation base, but the fermentation base to be used is not limited to this as long as it can be used in the present invention. As time passes during the culture, the input amount of the fermentation base material is gradually increased, the temperature is raised to 60 to 70°C, and appropriate fermentation is maintained by adjusting the input amount of the organic waste so as to maintain this temperature. Then, the organic waste input by fermentation almost completely disappears, and the mass of the bacterial cells gradually increases.

[0027] The culturing treatment is preferably continued until the total mass of the bacterial cells reaches 3 to 30% by mass based on the mass of the culture solution. If the total mass of the bacterial cells is less than 3% by mass based on the mass of the culture solution, a sufficient protective effect against the apple rot pathogen cannot be obtained. On the other hand, it is often difficult to culture so that it exceeds 30% by mass, and since the bacteria enter the stationary phase, the reaction rate of fermentation may decrease. Moreover, since the activity of the fungi may change, it is preferable to terminate the culture.

[0028] After the completion of this culturing, filtration is performed using a filter medium with an opening size through which the bacterial cells pass to remove fermentation residues, generated precipitates, dust, etc., to obtain a culture solution. Here, water-soluble heavy metal components containing one or more of cadmium, mercury, and arsenic contained in the organic waste change to water-insoluble solids and precipitate during the culturing. Therefore, when the solids are removed by filtration, at least a part of the water-soluble heavy metal components becomes water-insoluble and is removed.

[0029] [Examples] <Example 1> Each of the bacteria of 36 bacterial species belonging to the above 14 genera of bacteria was mixed in the same amount at a concentration of 1×10 5 cfu / cm 3 and pre-cultured, and a culture solution of mixed bacteria was produced by subjecting the pre-culture solution to main culturing. The bacteria of the 36 bacterial species used are as follows. Streptomycin-producing bacteria: Sulfurobolus acidocaldarius: Saccharopolyspora erythraea: Nocardia asteroides: Pseudomonas fluorescens: Pseudomonas mendocina: Pseudomonas alcaligenes: Pseudomonas veronii: Pseudomonas pseudoalcaligenes: Pseudomonas gruma: Pseudomonas gladioli: Pseudomonas avenae: Pseudomonas putida: Bacillus thuringiensis: Bacillus subtilis: Cytophaga hutchinsonii: Candida etchellsii: Candida versatilis: Candida stellata: Cellulomonas flavigena: Clostridium thermocellum: Clostridium acetobutylicum: Clostridium butyricum: Clostridium ljungdahlii: Aspergillus niger: Aspergillus oryzae: Aspergillus penicilloides: Aspergillus brasiliensis: Aspergillus terreus: Aspergillus sojae: Penicillium chrysogenum: Penicillium camemberti: Penicillium glabrum: Penicillium digitalitatum: Rhizopus oryzae: White rot fungus

[0030] Next, 287.5 mg (1.00 mmol) of zinc sulfate (heptahydrate) and 1.0 g of fulvic acid were added to 2.0 kg of the prepared culture solution, and further water was added to make 100 kg to obtain the apple ring rot pathogen disinfectant of Example 1.

[0031] <Example 2> 1438 mg (5.00 mmol) of zinc sulfate (heptahydrate), 5.0 g of fulvic acid, and 1.2 kg of gum arabic as a thickener were added to 20.0 kg of the culture solution, and further water was added to make 100 kg to obtain the apple ring rot pathogen disinfectant of Example 2. The apple ring rot pathogen disinfectant of Example 2 was thickened compared to Example 1, and when applied to the infected part of ring rot, it formed a thin layer at the infected part and could be surely applied.

[0032] <Example 3> Example of use of apple ring rot pathogen disinfectant 1 Apple rot often shows moist lesion areas with sap at the infected sites from April to June. When the disease appears on a branch, the branch should be removed; when it appears on the trunk, the knobby bark should be removed and incinerated. The spraying solution of the apple rot pathogen disinfectant of Example 1 was evenly applied to the remaining part after removal and its surrounding area.

[0033] <Example 4> Example of using the apple rot pathogen disinfectant 2 Apple rot often shows moist lesion areas with sap at the infected sites from April to June. When the disease appears on a branch, the branch should be removed; when it appears on the trunk, the knobby bark should be removed and incinerated. Without cutting the tree veins, cuts were made at intervals of 6 - 10 mm parallel to the tree veins at the infected sites, and the spraying solution of the apple rot pathogen disinfectant of Example 1 was evenly applied to the remaining part after removal and its surrounding area.

[0034] <Example 5> To produce the apple rot pathogen disinfectant of Example 5, xanthan gum was added as a thickener to Example 1 to thicken it to an amount where the whole disinfectant forms a thin - layer coating state. The disinfectant of Example 5 was thickened compared to Example 1, and when this disinfectant was applied to the infected sites of apple rot, it could be surely applied in a thin - layer form.

[0035] <Comparative Example 1> Except for removing the streptomycin - producing bacteria of the genus Streptomyces from the 36 bacterial species used in Example 1, the disinfectant of Comparative Example 1 was produced in the same way as Example 1 and applied to the sites infected with apple rot. As a result, apple rot could not be cured.

[0036] <Advantages of the Invention> The treatment effects when the apple rot pathogen disinfectant of Example 1 was applied by the method of Example 1 are shown in Figures 1 - 4. Figure 1 is a photo - substitute drawing showing the apple infected site before treatment, and the infected site is wet with exuded liquid. Figure 2 is a photo substitute for a drawing 6 days after applying the apple ring rot pathogen disinfectant solution of Example 1 to the infected area at a rate of 100 milliliters per square meter. The seepage of the solution has stopped and the infected area has dried out. Figure 3 is a photo substitute for a drawing 36 days after applying the apple ring rot pathogen disinfectant solution. Further drying has progressed and the area that was the infected area has hardened. Figure 4 is a photo substitute for a drawing 1 year after applying the apple ring rot pathogen disinfectant solution. It has become so clean that it is hard to believe it was the infected area, and young buds are emerging from the site where the apple ring rot disease occurred.

Claims

1. A mixed liquid containing a culture solution of a mixed culture of bacteria belonging to at least the following 14 genera, zinc ions, and fulvic acid, The 14 genera are: (1) Streptomyces (2) Sulfolobus (3) Saccharopolyspora (4) Nocardia (5) Pseudomonas (6) Bacillus (7) Cytophaga (Cytophaga) (8) Candida (9) Cellulomonas (10) Clostridium (11) Aspergillus (12) Penicillium (13) Rhizopus (14) Basidiomycota genus and the culture medium is cultured so that the total mass of the bacterial cells of the bacterial group is 3.0 to 30.0% by mass of the total mass of the culture medium, and is filtered through a filter material having an opening that allows the bacterial cells of the bacterial group to pass through to remove solid components; The culture solution is contained in an amount of 1.0 to 20.0% by mass and the fulvic acid is contained in an amount of 0.0001 to 0.01% by mass relative to 100% of the total mass; 1. A disinfectant solution for apple canker fungus, comprising 0.01 to 10.0 millimoles of zinc ions per 1 kg of total weight.

2. 2. The disinfectant solution for apple canker sore according to claim 1, further comprising a thickener added thereto so that the disinfectant solution can be applied in a thin layer.

3. A method for using the disinfectant solution for apple canker fungus, which comprises removing a part of an apple tree infected with apple canker fungus and applying the disinfectant solution for apple canker fungus according to claim 1 or 2 to the remaining part and its surroundings.

4. This method for using the disinfectant against apple canker fungus comprises removing a part of an apple tree infected with the apple canker fungus and injuring the remaining part and its surroundings parallel to the tree veins, and then applying the disinfectant against apple canker fungus according to claim 1 or 2 to the remaining part and its surroundings.

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