Disinfectant for apple canker and its use

Abstract

To provide an effective and easy-to-use disinfectant for apple canker bacteria, and a method for using the same. [Solution] The apple canker fungus disinfectant of the present invention is a mixed solution containing a culture solution containing fungi of at least 14 genera, a fulvic acid solution, and zinc ions, characterized in that the genera include at least the genera Streptomyces, Sulfolobus, Saccharopolyspora, Nocardia, Pseudomonas, Bacillus, Cytophaga, Candida, Cellulomonas, Clostridium, Aspergillus, Penicillium, Rhizopus, and Basidiomycota.

Description

[Technical field]

[0001] The present invention relates to a disinfectant for apple canker fungus and a method for using the same, and more particularly to a disinfectant for apple canker fungus that is applied to an infected site of an apple tree to disinfect the apple canker fungus and a method for using the same. [Background technology]

[0002] Apple canker disease is a plant disease that occurs when apple trees are infected with Valsa ceratosperma, a type of filamentous fungus. When apple trees are infected with Valsa ceratosperma, reddish-brown spots appear, the bark turns brown, sap leaks from the affected areas, and the tree dies from the tip of the affected area, resulting in a significant decrease in apple production. The only way to treat the disease is to cut off the areas infected with Valsa ceratosperma, and if the infection spreads, the tree must be cut down to prevent the infection from spreading to other apple trees.

[0003] Conventionally, as a preventive measure against apple canker, apple trees have been covered with soil from the growing area in the autumn after harvest, and then wrapped up in soil to overwinter. However, covering apple trees with mud is a laborious task that places a heavy burden on apple farmers, and this method only prevents infection and onset of apple canker, and is almost ineffective against apple canker that has already developed.

[0004] Patent Document 1 describes a method of preventing infection by applying inorganic agents such as sodium carbonate and alum to apple trees. However, since the apple canker fungus remains dormant for nearly a year after invading an apple tree and develops apple canker disease after penetrating deep into the resinous tissue, it is difficult to disinfect the apple canker fungus with inorganic agents such as those described in Patent Document 1. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] Japanese Patent Application Publication No. 8-268819 [Non-patent literature]

[0006] [Non-Patent Document 1] Norio Nakazawa and Yukio Harada, Journal of the Northern Japan Insect Control Research Institute 53:109-111(2002) [Non-Patent Document 2] "Fraction of fulvic acid by gel filtration and its chelating ability", Hidekazu Yamada, Koyo Yonebayashi, Kyosei Hattori, Shuji Morita, Abstracts of the 1972 Osaka Conference of the Japanese Society of Soil Science and Plant Nutrition, http: / / ci.nii.ac.jp / els / 110001195691.pdf?id=ART0001528728&type=pdf&lang=en&host=cinii&order_no=&ppv_type=0&lang_sw=&no=1420378856&cp= Summary of the Invention [Problem to be solved by the invention]

[0007] An object of the present invention is to provide a disinfectant solution for apple canker bacteria which is effective against apple canker bacteria and can be easily applied. Another object of the present invention is to provide a method for using an apple canker fungus disinfectant for treating and regenerating an infected apple tree by disinfecting and treating the affected part of the apple tree infected with the apple canker fungus and regenerating the infected apple tree. [Means for solving the problem]

[0008] The disinfectant solution for apple canker fungus of the present invention has at least one of the following properties: 3 A group of bacteria belonging to the genus and wood-decaying fungi A mixed solution containing a culture solution obtained by mixing the above, zinc ions, and fulvic acid, 1 3 The fungus genus is (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 ) 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 with openings that allow the bacterial cells of the bacterial group to pass through to remove solid components, The culture solution contains 1.0 to 20.0% by mass and the fulvic acid contains 0.0001 to 0.01% by mass relative to 100% total mass. The composition is characterized in that the zinc ions are contained in an amount of 0.01 to 10.0 millimoles per 1 kg of the total mass.

[0009] The present invention is characterized in that a thickener is further added to make it possible to apply the composition in a thin layer.

[0010] The method for using the Apple canker fungus disinfectant of the present invention is characterized in that a part infected with the Apple canker fungus and diseased is removed from an apple tree, and the Apple canker fungus disinfectant is applied to the remaining part and its surroundings.

[0011] The method of using the Apple Canker Fungus disinfectant of the present invention is characterized in that a part of an apple tree infected with the Apple Canker Fungus is removed, a wound is made in the remaining part and its surroundings parallel to the tree vein, and the Apple Canker Fungus disinfectant is then applied to the surroundings. Effect of the Invention

[0012] In the present invention, an apple tree infected with the apple canker fungus is treated by removing the infected part of the apple tree or by applying a disinfectant for the apple canker fungus to the infected part as is or by making a cut parallel to the tree veins and drying the cut part, thereby disinfecting the apple canker fungus and regenerating the infected part.

[0013] The present invention can treat apple trees infected with the apple canker fungus, thereby recovering the yield of the apple trees and preventing the spread of infection, eliminating the need to cut down apple trees infected with the apple canker fungus. Moreover, the method of using the disinfectant is simple, reducing the burden on apple farmers who have to cover infected areas with mud in the fall after apple harvest. [Brief description of the drawings]

[0014] [Figure 1] This is a photograph in lieu of a drawing showing the state of an apple tree where a part infected with apple canker has been removed. [Diagram 2] FIG. 2 is a photograph substituted for a drawing, showing the state of the apple tree 6 days after a wound was made parallel to the vein at the site infected with apple canker in the state shown in FIG. 1 and the anti-apple canker fungus disinfectant solution of Example 1 was applied. [Diagram 3] 3 is a photograph showing the condition of apple trees 36 days after application of the apple canker fungus disinfectant solution using the method of FIG. 2. [Figure 4] 3 is a photograph showing the condition one year after application of an apple canker fungus disinfectant solution using the method of FIG. 2. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0015] The disinfectant for apple canker fungus of the present invention and a method for using the same will be described below. This description is for the purpose of explaining the present invention, and is not intended to limit the technical scope of the present invention. The present invention can be implemented in various modifications without departing from the technical scope of the present invention.

[0016] Non-Patent Document 1 reports that in a test tube dual culture of a filamentous fungus isolated from an apple tree and the apple canker fungus, some of the filamentous fungus isolated from the apple tree controls the growth of the apple canker fungus. Based on this, Non-Patent Document 1 presumes that the effect of the conventional treatment of wrapping infected areas in mud in autumn after apple harvest is growth control based on the antagonism between the soil fungus in the area where the apple tree grew and the apple canker fungus. However, this growth control is only an effect in a test tube, and growth control in an actual field is unstable and not practical.

[0017] The present inventor hypothesized that the antagonistic action between soil bacteria and the apple canker fungus described in Non-Patent Document 1 is due to the action of an unknown fungal parasitic virus. In order to verify this hypothesis, the inventors investigated the possibility of a fungal parasitic virus contained in a mixture of multiple types of fungi mixed at a specified ratio and having specified properties as a whole in the fungal solution, which reduces the toxicity of the apple canker fungus and inhibits its growth.

[0018] In addition, we performed co-cultivation of a number of mixed fungi with the apple canker pathogen (Valsa ceratosperma), and found that at least 1 of 3 Bacteria belonging to the genus Bacillus and wood-decaying fungi However, the pathogenicity suppression effect of the mixture was only moderate, and was insufficient for practical use as it was. The present invention provides a practical disinfectant for apple canker bacteria by further mixing zinc ions and fulvic acid with the culture solution of the mixed bacteria.

[0019] [Bacteria used in the present invention] The mixed bacteria used in the present invention is at least one of the following: 3 A group of bacteria belonging to the genus and wood-decaying fungi The culture medium used is a mixed culture of the following bacteria: (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 ) The present invention relates to the above-mentioned 3 Bacteria belonging to the genus Bacillus and wood-decaying fungi The present invention uses a mixed bacterium containing all of the above, and further contains zinc ions and fulvic acid, thereby making it possible to treat apple canker. Here, "Basidiomycota" is not the name of a genus based on the classification of fungi, but because the Basidiomycota, a superordinate concept of Basidiomycota, is a field whose classification system is yet to be determined, "Basidiomycota" is a common name that includes the more common "mushrooms" and wood-decaying fungi. The present invention uses a mixed bacteria containing all of the bacteria belonging to the above 14 bacterial families, and further contains zinc ions and fulvic acid, making it possible to treat apple canker disease.

[0020] [Bacteria species used in the present invention] In the present invention, the above 1 3 Bacteria belonging to the genus Bacillus and wood-decaying fungi As a mixed bacteria containing a group of bacteria belonging to the above, for example, a mixed bacteria containing the following 36 species of bacteria can be selected. (1) Streptomyces Streptomycin-producing bacteria (Streptomyces griseus) (2) Sulfolobus Sulfolobus acidocalis ldarius) This bacterium is a thermophilic acid-tolerant archaea. (3) Saccharopolyspora Saccharopolyspora erythraea ythraea This bacterium is an erythromycin producer. (4) Nocal J Genus Nocardia Nocal J A. Asteroides (Nocardia asteroides) This bacterium causes severe pulmonary infections in immunocompromised hosts. (5) Pseudomonas Pseudomonas fluorescens n.s. This bacterium is a common Gram-negative bacillus that produces a variety of secondary metabolites. Pseudomonas mendocina This bacterium is involved in toluene degradation. Pseudomonas alucarigenes es) This bacterium decomposes polycyclic aromatic hydrocarbons. Pseudomonas veronii This bacterium degrades a variety of aromatic organic compounds. Pseudomonas pseudoalcaligenes alkaligenens) The bacterium is able to metabolize unusual substrates, in particular cyanide compounds. Pseudomonas glumae This bacterium is the causative agent of rice grain rot. It also has antibacterial properties against several important plant pathogenic bacteria. Has antibacterial activity. Pseudomonas gladioli This bacterium is a human and plant pathogen and interacts antagonistically with other microorganisms. Pseudomonas avenae The fungus causes brown striping symptoms on infected susceptible plants. Pseudomonas putida This bacterium can decompose organic solvents such as toluene. (6) Bacillus Bacillus thuringiensis The fungus produces insecticidal compounds and is licensed as a biological control agent. Bacillus subtilis (7)Cytophaga Cytophaga hutchinsonii This bacterium exhibits gliding motility and is an aerobic cellulolytic bacterium found in soil environments. (8) Candida Candida etchellsii This fungus is added as a post-ripening yeast in the brewing of soy sauce and miso. Candida versatilis This fungus is added as a post-ripening yeast in the brewing of soy sauce and miso. Candida stellata This fungus is involved in wine making. (9) Cellulominas Cellulomonas flavigena This bacterium is a cellulolytic bacterium isolated from compost. (10) Clostridium Clostridium thermocellum umu) This fungus is a cellulolytic enzyme. Clostridium acetobutylicum t y ricum) This fungus converts acetone into starch. hmm, This is a bacterium that produces butanol and ethanol. Clostridium butyricum ) This bacterium is used as an intestinal regulator (Miyairi bacteria). Clostridium ljungdahl ii) This bacterium is used in ethanol production. (11) Aspergillus Aspergillus nigar This fungus is black koji mold, a microorganism that produces organic acids such as citric acid and amylase. It is a fungus. Aspergillus oryzae This fungus is Japanese koji mold fungus. It is. To make soy sauce, miso, sake, shochu, brewed alcohol, etc. It is used for this purpose. Aspergillus penicilloidas ides) This fungus is used to produce fermented foods such as bonito flakes and prosciutto. Aspergillus Brush R Aspergillus brasillensis sis) This fungus is the most common species of Aspergillus and is widespread in soil. Aspergillus terreus The bacteria is used to produce organic acids and enzymes. Soy sauce koji mold (Aspergillus sojae) This bacterium is a soy sauce koji bacterium. (12) Penicillium Penicillium chrysogenum This bacterium is a penicillin producer. Penicillium camenberti This fungus is the same mold that is used to make Camembert cheese. Penicillium glabrum This fungus is a plant pathogen that infects strawberries. Penicillium digitatum This bacterium is a cause of post-harvest decay of fruit. (13) Rhizopus Rhizopus oryzae This bacterium produces many kinds of enzymes and is used in the food and pharmaceutical industries. There are 。

[0021] [Production of culture medium] <Zinc ion> Normally, fungal parasitic viruses do not spread (transfer) between filamentous fungi that are incompatible with each other when they are co-cultured, but in the presence of zinc ions, the virus migrates (transmits) from a filamentous fungus infected with a fungal parasitic virus to a filamentous fungus that is not infected with the virus when they are co-cultured. Zinc ions are used for this purpose. Zinc ions and fulvic acid were added to the culture solution of this mixed fungus, which was then co-cultured with the apple canker fungus. The growth inhibitory effect of this mixed fungus was measured, and the mixture showed a strong and practical inhibitory effect on the apple canker fungus.

[0022] <Fulvic acid> Fulvic acid is a substance that gives a yellow or orange-yellow color to the acidic supernatant when extracted with an inorganic acid after humic acid has been extracted and removed from soil or coal with a dilute alkali, and is an amorphous acidic substance that is soluble in water and ethanol, does not have a single chemical structural formula, and its composition and molecular weight are not fixed; the composition and molecular weight change depending on the raw material and collection conditions (see, for example, Non-Patent Document 2). Fulvic acid has a strong affinity for zinc ions and acts to assist the uptake of zinc ions into the body. It is preferable to use a purified fulvic acid solution produced by the method described in Non-Patent Document 2 as the fulvic acid.

[0023] The content of the fulvic acid solution is preferably 0.0001 to 0.01 when the mass of the entire disinfectant solution for apple canker is 100% by mass. If the amount of the fulvic acid solution is less than 0.0001% by mass, sufficient protective effect against apple canker is not obtained, while adding fulvic acid in an amount exceeding 0.01% by mass is expensive but does not increase protective effect against apple canker, which is economically undesirable.

[0024] <Thickener> The disinfectant against apple canker of the present invention can be made into a form that can be applied in the form of a thick coating film by adding a thickener. The work of covering apple trees with soil from the cultivation area in autumn after apple harvest is a hard work for apple farmers, but according to the present invention, this mud covering work can be replaced with the work of spraying the disinfectant against apple canker to which a thickener has been added. As the thickening agent, various types including gum arabic, xanthan gum, pectin, carrageenan, gum arabic, etc. can be used, and any can be used as long as it meets the objectives of the present invention.

[0025] [Production of culture medium] (preculture) The at least one 3 A group of bacteria belonging to the genus and wood-decaying fungi At least 36 species of bacteria belong to the same family, each of which is, for example, 1 × 10 5 cfu / cm 3 The same concentration and amount of the pre-culture base material and water are mixed and heated to a moderate temperature, and the base material is added in small amounts to the medium for pre-culture. The base material used for pre-culture to initiate fermentation is not particularly limited as long as it can fix and support fungi on its surface or in its internal structure.Specifically, for example, bark, sawdust, charcoal powder, coffee grounds, rice husks, and bokashi obtained from rice bran can be suitably used, and these can be used alone or in combination of two or more kinds.

[0026] (main culture) When the temperature of the preculture solution rises to 45° C. to 55° C., the fermentation base is gradually added in place of the base material. In this example, organic waste discharged from a fishery product processing plant was used as the fermentation base, but the fermentation base to be used is not limited to this as long as it is usable in the present invention. As the cultivation time progresses, the amount of fermentation substrate is gradually increased, the temperature is raised to 60-70°C, and the amount of organic waste added is adjusted to maintain this temperature, thereby maintaining proper fermentation. The organic waste added is then almost completely consumed by fermentation, and the mass of the bacteria gradually increases.

[0027] The culture treatment is preferably continued until the total mass of the fungus becomes 3 to 30% by mass relative to the mass of the culture solution. If the total mass of the fungus becomes less than 3% by mass relative to the mass of the culture solution, sufficient protective effect against the apple canker fungus cannot be obtained, while it is often difficult to culture the fungus to a mass of more than 30% by mass, and the fermentation reaction rate may decrease as the bacteria enter a resting period, and furthermore, the activity of the fungus may change, so it is preferable to terminate the culture.

[0028] After the main cultivation is completed, the culture liquid is obtained by filtering using a filter medium with openings large enough for the bacteria to pass through, and removing the fermentation residue, the precipitate that has formed, garbage, etc. Here, water-soluble heavy metal components, including one or more of cadmium, mercury, and arsenic, contained in the organic waste, are converted to water-insoluble solids and precipitate during cultivation, so that by filtering and removing the solids, at least a portion of the water-soluble heavy metal components are made water-insoluble and removed.

[0029] [Example] <Example 1> The 13 bacterial genera and 36 species of wood-rotting fungi were each mixed at 1 × 10 5 cfu / cm 3 The 36 species of bacteria used were as follows: Streptomycin-producing bacteria: Sulfolobus acidocallidarius: Saccharopolyspora erythraea: Nocardia asteroides: Pseudomonas florescens: Pseudomonas mendocina: Pseudomonas alcaligenes: Pseudomonas veronii: Pseudomonas pseudoalcaligenes: Pseudomonas glumae: Pseudomonas gladioli: Pseudomonas avenae: Pseudomonas putida: Bacillus thuringiensis: Bacillus subtilis: Cytophaga hutchinsoni: Candida esselcii: Candida versatilis: Candida stellata: Cellulomonas flavigena: Clostridium thermocellum: Clostridium acetobutylicum: Clostridium butyricum: Clostridium ljundalii: Aspergillus niger: Aspergillus oryzae: Aspergillus Penicilloidas:Aspergillus brassiensis:Aspergillus terreus:Soy sauce koji mold:Penicillium chrysogenum:Penicillium camenbergii:Penicillium glabram:Penicillium digitatum:Rhizopus oryzae: white decay fungi

[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 produced culture solution, and water was further added to make up 100 kg, to obtain the apple canker fungus 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 water was further added to make 100 kg, to obtain the apple canker fungus disinfectant of Example 2. The apple canker fungus disinfectant of Example 2 was thickened compared to Example 1, and when applied to an area infected with ring canker fungus, it formed a thin layer and could be applied reliably to the infected area.

[0032] Example 3: Example 1 of use of disinfectant for apple rot fungus In apple canker, moist lesions with sap are often found at the infected area in April to June. If the disease occurs on a branch, the branch is removed, and if the disease occurs on the trunk, the knobby bark is removed and incinerated, and the Apple canker fungus disinfectant solution of Example 1 is evenly applied to the remaining part and its surroundings.

[0033] Example 4: Example 2 of use of disinfectant for apple rot fungus In apple canker, sap-soaked lesions are often found at the infected area in April to June. If the disease occurs on a branch, the branch is removed, and if the disease occurs on the trunk, the knobby bark is removed and incinerated. The infected area is cut at intervals of 6 to 10 mm parallel to the veins so as not to cut the veins, and the Apple canker fungus disinfectant solution of Example 1 is sprayed on the infected area, and the remaining part and its surroundings are evenly coated with the Apple canker fungus disinfectant solution.

[0034] <Example 5> In Example 5, a disinfectant against apple canker fungus was produced by adding xanthan gum as a thickener in an amount such that the entire disinfectant would form a thin layer when applied, as compared to Example 1. The disinfectant in Example 5 was thickened compared to Example 1, and when this disinfectant was applied to an area infected with apple canker fungus, it was reliably applied to the infected area in the form of a thin layer.

[0035] <Comparative Example 1> A disinfectant solution of Comparative Example 1 was prepared in the same manner as in Example 1, except that streptomycin-producing bacteria of the genus Streptomyces were excluded from the 36 bacterial species used in Example 1. The disinfectant solution was applied to areas infected with apple canker disease, but the apple canker disease could not be cured.

[0036] <Effects of the Invention> The therapeutic effect when the disinfectant solution against apple canker fungus of Example 1 was applied by the method of Example 1 is shown in Figs. Figure 1 is a photograph showing the infected area of ​​an apple before treatment; the infected area is wet and exuding fluid. Figure 2 is a photograph taken 6 days after the application of 100 milliliters per square meter of the disinfectant solution for apple canker fungus of Example 1 to the infected area. The liquid has stopped seeping out and the infected area has become dry. Figure 3 is a photo taken 36 days after the application of the apple canker fungus disinfectant. The area that was infected has become hard due to further drying. Figure 4 is a photograph used in place of a drawing, taken one year after the application of the apple canker fungus disinfectant. The area has become so clean that it is hard to believe it was once infected, and young buds are emerging from the scars where the apple canker fungus had developed.

Claims

1. A mixed solution containing a culture solution obtained by co-culturing at least the following 13 bacterial genera and wood-rotting fungi, zinc ions, and fulvic acid, The 13 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 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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