WO2013146775A1 - Bacterium belonging to genus trichoderma, agent for controlling plant pathogen, soil amendment, method for controlling plant pathogen, and method for improving soil - Google Patents
Bacterium belonging to genus trichoderma, agent for controlling plant pathogen, soil amendment, method for controlling plant pathogen, and method for improving soil Download PDFInfo
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
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- A01N63/38—Trichoderma
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
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- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/645—Fungi ; Processes using fungi
- C12R2001/885—Trichoderma
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- the present invention relates to a Trichoderma genus, a plant pathogen control agent and a soil conditioner using the same, and a plant pathogen control method and a soil improvement method.
- Natural rubber is produced by processing latex called latex, which is mainly made in the ductal cells of Hevea brasiliensis. Natural rubber is used widely and in large quantities in various applications as the main raw material for rubber products.
- plantation of Hevea brasiliensis is performed in a climatic zone suitable for the growth of Hevea brasiliensis. Such climatic zones are also suitable for the growth of pathogens that infect Hevea brasiliensis, and root rot caused by infection of Hevea brasiliensis causes serious damage to the cultivation of Hevea brasiliensis in rubber plantations. Yes.
- Root rot is a disease that causes destructive damage to the roots of Hevea brasiliensis.
- Rigidoporus microporus is known as the main pathogen of this root rot.
- Rigidoporus microporus belongs to the Basidiomycota and forms a huge fruit body at the root of infected Hevea brasiliensis.
- root rot in Hevea brasiliensis is controlled by a physical removal method by incineration of infected roots or a chemical removal method by a bactericide.
- a physical removal method by incineration of infected roots or a chemical removal method by a bactericide.
- a method for reliably controlling root rot is desired.
- Non-Patent Document 1 an attempt is made to analyze the relationship between Rigidoporus microporus genetic diversity and pathogenicity.
- Non-Patent Document 1 confirmed the correlation between geographical distribution and genetic diversity, but there was a clear correlation between pathogenicity and genetic diversity. Not confirmed. In order to apply this analysis result to the control method of root rot, further knowledge is required.
- the present invention has been made in view of the above circumstances, and is a Trichoderma sp. That can control root rot of Hevea brasiliensis, a phytopathogenic fungus control agent and a soil improver using the same, and a phytopathogenic fungus It aims at providing the control method and the soil improvement method.
- the present inventor found a Trichoderma genus having antagonistic properties against root rot of Hevea brasiliensis and completed the present invention.
- this invention provides the Trichoderma genus microbe which has the following characteristics, a phytopathogen control agent and a soil improvement agent using the same, and a phytopathogen control method.
- Trichoderma spp. That has antagonistic properties against root rot of Hevea brasiliensis.
- the Trichoderma spp. According to (1) or (2), wherein the root rot fungus is Rigidoporus microporus.
- the Trichoderma spp. According to any one of (1) to (3), which is Trichoderma spiral or Trichoderma erinaceum.
- a phytopathogenic fungus control agent comprising a Trichoderma spp.
- a method for controlling phytopathogenic fungi comprising a step of treating the phytopathogenic fungus controlling agent according to (5) with soil.
- a soil improvement method comprising a step of treating the soil improver according to (7) with soil.
- the disease of Hevea brasiliensis caused by root rot fungus can be suppressed.
- Trichoderma spp. The Trichoderma spp. Of the present invention has antagonistic properties against root rot of Hevea brasiliensis.
- Hevea brasiliensis is a Euphorbiaceae plant that produces latex (mainly polyisoprene) and has duct cells. Hevea brasiliensis is widely used as an industrial natural rubber raw material. Since the main pathogen of the root rot fungus is Rigidoporus microporus, it is preferable that the Trichoderma spp. Of the present invention have antagonistic properties to Rigidoporus microporus.
- Trichoderma genera such as Trichoderma spiral and Trichoderma erinaceum were identified.
- Trichoderma spp. Are known to have parasitic characteristics to phytopathogenic fungi (R. Wedding, Phytopathology, Vol. 22, pp. 837-845, 1932) and others including soil infectious phytopathogenic fungi It has been reported to show an antagonistic action against other microorganisms.
- the plant pathogen control agent of the present invention contains a culture, wet cell or dry cell of the Trichoderma genus of the present invention.
- PD potato dextrose
- a jar fermenter is used at a temperature of about 28 ° C. in an aerobic state.
- a method of culturing for about 4 to 4 days is mentioned.
- the plant pathogen control agent of the present invention exhibits a more effective control action against root rot fungus.
- the growth temperature is preferably 10 to 28 ° C, more preferably 15 to 28 ° C.
- the culture containing the plant pathogen control agent of the present invention is a liquid medium containing the grown Trichoderma genus of the present invention as it is.
- a culture contains medium components contained in the culture solution and secondary metabolites produced by the culture.
- the phytopathogenic fungus control agent of the present invention is preferably diluted 100 to 1000 times with water.
- the wet cells contained in the phytopathogenic fungus control agent of the present invention are those obtained by washing a culture solution collected by centrifugation, as appropriate, with pure water or the like. Unlike the culture, the wet cell does not contain a medium component or the like. However, the phytopathogenic fungus control agent of the present invention is preferably used after being diluted 100 to 1000 times with water from the viewpoint of being uniformly applied to Hevea brasiliensis.
- the dry cell contained in the plant pathogen control agent of the present invention is a product obtained by collecting a culture solution by centrifugation and converting it into a dry solid.
- Such dry solid is preferably semi-dried to a moisture content of 5 to 30% by mass.
- the drying treatment is preferably performed at a temperature that does not adversely affect the growth of the Trichoderma spp. Of the present invention.
- the plant pathogen control agent of the present invention may be one obtained by inoculating a culture of the genus Trichoderma of the present invention, a wet cell, or a dry cell into a sterilized solid medium and culturing for a certain period.
- the plant pathogen control agent of the present invention may be one obtained by adsorbing a solid culture cultured in the above-mentioned solid medium to an adsorbent.
- the adsorbent include adsorbing mineral materials such as vermiculite and zeolite; charcoal such as charcoal and activated carbon; chemically synthesized polymers and the like.
- the plant pathogen control agent of the present invention may contain an insecticide, an acaricide, a herbicide, other fungicides and the like.
- the above-mentioned plant pathogen control agent of the present invention is compost and other soil improving components. May be added.
- the compost include those composed of a medium and fermenting bacteria, and examples of the medium include a single substance or a mixture of rice bran, bran, sake lees, shochu soybean meal, and the like.
- the soil improving agent of the present invention contains compost, it can impart fertilizer effect to the soil in addition to imparting antagonistic properties against root rot fungus.
- Other soil improvement components include peat, humic acid materials, soil improvement materials such as polyvinyl alcohol materials, plant growth regulators, and the like.
- the soil conditioner of the present invention is obtained by adding the phytopathogenic fungus control agent of the present invention to compost that has been fermented
- the fermented compost is obtained by adding the phytopathogenic fungus control agent of the present invention to the compost raw material.
- the Trichoderma spp. Of the present invention may be grown.
- the plant pathogen control method of the present invention includes a step of treating the plant pathogen control agent of the present invention with soil.
- a method for controlling plant pathogens of the present invention a method of spraying a culture of Trichoderma spp. Of the present invention or a wet fungus body diluted 100 to 1000 times with water onto diseased soil; Cultivation of Trichoderma spp.
- the method etc. which spray a microbial cell on diseased soil are mentioned.
- Examples of the method for applying the plant pathogen control agent of the present invention include ground liquid spray, ground solid spray, air solution spray, and air solid spray.
- the soil improvement method of this invention has the process of processing the soil improvement agent of this invention to soil.
- the process for treating the soil includes mixing with soil, application of the plant base, application of the nursery bed, and mixing with fertilizer.
- the cultured medium of Rigidoporus microporus was punched and inoculated with a sterilized 5 mm diameter cork borer at a position 1 cm inward from the outer periphery of a 9 cm diameter petri dish containing autoclaved PDA medium.
- the cultured sample 1 was punched and inoculated with a sterilized 5 mm diameter cork borer at a position symmetrical to the position where Rigidoporus microporus was inoculated, and cultured at 28 ° C. for 5 days, 10 days, and 15 days.
- the ratio of the growth area of Sample 1 in the total growth area of Rigidoporus microporus and Sample 1 was calculated as the growth inhibition rate.
- FIG. 1 A photograph of the culture dish after 15 days of culture is shown in FIG.
- the growth area was calculated by drawing squares at intervals of 5 mm on a transparent sheet, and covering this on a petri dish and counting the squares.
- Example 2 instead of Sample 1, the sample was cultured in the same manner as in Example 1 except that Sample 2 (soil derived from compartment 2) was used. The ratio of the growth area of Sample 2 in the total growth area of Rigidoporus microporus and Sample 2 was calculated as the growth inhibition rate. The results of the growth inhibition rate for 5 days, 10 days, and 15 days are shown in FIG. A photograph of the culture petri dish after 15 days of culture is shown in FIG.
- ⁇ Comparative example 1> instead of Sample 1, the cells were cultured in the same manner as in Example 1 except that Sample 3 (soil derived from compartment 3) was used. The ratio of the growth area of Sample 3 to the total growth area of Rigidoporus microporus and Sample 3 was calculated as a growth inhibition rate. The results of the growth inhibition rate for 5 days, 10 days, and 15 days are shown in FIG. A photograph of the culture petri dish after 15 days of culture is shown in FIG.
- Trichoderma spp. which can control root rot of Hevea brasiliensis, plant pathogen control agent and soil improver using the same, and plant pathogen control method and soil improvement method.
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Abstract
This bacterium belonging to the genus Trichoderma is antagonistic to the bacterium that causes white root rot disease in Hevea brasiliensis. This agent for controlling a plant pathogen comprises the cultured product, wet biomass, or dry biomass of this bacterium belonging to the genus Trichoderma. This soil amendment comprises the cultured product, wet biomass, or dry biomass of this bacterium belonging to the genus Trichoderma. This method for controlling a plant pathogen comprises a step for treating soil using this agent for controlling a plant pathogen. This method for improving soil comprises a step for treating soil using this soil amendment.
Description
本発明は、Ｔｒｉｃｈｏｄｅｒｍａ属菌、これを用いた植物病原菌防除剤及び土壌改良剤、並びに、植物病原菌防除方法及び土壌改良方法に関する。
本願は、２０１２年３月２６日に、日本に出願された特願２０１２－０７０２２５号に基づき優先権を主張し、その内容をここに援用する。 The present invention relates to a Trichoderma genus, a plant pathogen control agent and a soil conditioner using the same, and a plant pathogen control method and a soil improvement method.
This application claims priority based on Japanese Patent Application No. 2012-070225 filed in Japan on March 26, 2012, the contents of which are incorporated herein by reference.
本願は、２０１２年３月２６日に、日本に出願された特願２０１２－０７０２２５号に基づき優先権を主張し、その内容をここに援用する。 The present invention relates to a Trichoderma genus, a plant pathogen control agent and a soil conditioner using the same, and a plant pathogen control method and a soil improvement method.
This application claims priority based on Japanese Patent Application No. 2012-070225 filed in Japan on March 26, 2012, the contents of which are incorporated herein by reference.
天然ゴムは、主にＨｅｖｅａ ｂｒａｓｉｌｉｅｎｓｉｓの乳管細胞内で造られているラテックスという乳液を加工することにより製造される。天然ゴムは、ゴム製品の主原料として、様々な用途において幅広くかつ大量に用いられている。
このラテックス採取を目的として、Ｈｅｖｅａ ｂｒａｓｉｌｉｅｎｓｉｓの生育に適した気候区において、Ｈｅｖｅａ ｂｒａｓｉｌｉｅｎｓｉｓの植林が行われている。かかる気候区は、Ｈｅｖｅａ ｂｒａｓｉｌｉｅｎｓｉｓに感染する病原菌の生育においても適しており、Ｈｅｖｅａ ｂｒａｓｉｌｉｅｎｓｉｓに病原菌が感染することによりもたらされる根白腐れ病が、ゴム農園におけるＨｅｖｅａ ｂｒａｓｉｌｉｅｎｓｉｓの栽培に深刻な被害を与えている。 Natural rubber is produced by processing latex called latex, which is mainly made in the ductal cells of Hevea brasiliensis. Natural rubber is used widely and in large quantities in various applications as the main raw material for rubber products.
For the purpose of collecting latex, plantation of Hevea brasiliensis is performed in a climatic zone suitable for the growth of Hevea brasiliensis. Such climatic zones are also suitable for the growth of pathogens that infect Hevea brasiliensis, and root rot caused by infection of Hevea brasiliensis causes serious damage to the cultivation of Hevea brasiliensis in rubber plantations. Yes.
このラテックス採取を目的として、Ｈｅｖｅａ ｂｒａｓｉｌｉｅｎｓｉｓの生育に適した気候区において、Ｈｅｖｅａ ｂｒａｓｉｌｉｅｎｓｉｓの植林が行われている。かかる気候区は、Ｈｅｖｅａ ｂｒａｓｉｌｉｅｎｓｉｓに感染する病原菌の生育においても適しており、Ｈｅｖｅａ ｂｒａｓｉｌｉｅｎｓｉｓに病原菌が感染することによりもたらされる根白腐れ病が、ゴム農園におけるＨｅｖｅａ ｂｒａｓｉｌｉｅｎｓｉｓの栽培に深刻な被害を与えている。 Natural rubber is produced by processing latex called latex, which is mainly made in the ductal cells of Hevea brasiliensis. Natural rubber is used widely and in large quantities in various applications as the main raw material for rubber products.
For the purpose of collecting latex, plantation of Hevea brasiliensis is performed in a climatic zone suitable for the growth of Hevea brasiliensis. Such climatic zones are also suitable for the growth of pathogens that infect Hevea brasiliensis, and root rot caused by infection of Hevea brasiliensis causes serious damage to the cultivation of Hevea brasiliensis in rubber plantations. Yes.
根白腐れ病は、Ｈｅｖｅａ ｂｒａｓｉｌｉｅｎｓｉｓの根に破壊的なダメージを与える病気である。この根白腐れ病の主な病原菌として、Ｒｉｇｉｄｏｐｏｒｕｓ ｍｉｃｒｏｐｏｒｕｓが知られている。
Ｒｉｇｉｄｏｐｏｒｕｓ ｍｉｃｒｏｐｏｒｕｓは、担子菌門に属し、感染したＨｅｖｅａ ｂｒａｓｉｌｉｅｎｓｉｓの根に巨大な子実体を形成する。 Root rot is a disease that causes destructive damage to the roots of Hevea brasiliensis. Rigidoporus microporus is known as the main pathogen of this root rot.
Rigidoporus microporus belongs to the Basidiomycota and forms a huge fruit body at the root of infected Hevea brasiliensis.
Ｒｉｇｉｄｏｐｏｒｕｓ ｍｉｃｒｏｐｏｒｕｓは、担子菌門に属し、感染したＨｅｖｅａ ｂｒａｓｉｌｉｅｎｓｉｓの根に巨大な子実体を形成する。 Root rot is a disease that causes destructive damage to the roots of Hevea brasiliensis. Rigidoporus microporus is known as the main pathogen of this root rot.
Rigidoporus microporus belongs to the Basidiomycota and forms a huge fruit body at the root of infected Hevea brasiliensis.
従来、Ｈｅｖｅａ ｂｒａｓｉｌｉｅｎｓｉｓにおける根白腐れ病は、感染した根の焼却による物理的除去方法、又は殺菌剤による化学的除去方法により制御されている。
しかしながら、これらの方法では、根白腐れ病にかかったＨｅｖｅａ ｂｒａｓｉｌｉｅｎｓｉｓを回復させるのに手遅れの場合が多々あり、確実に根白腐れ病を制御する方法が望まれている。 Conventionally, root rot in Hevea brasiliensis is controlled by a physical removal method by incineration of infected roots or a chemical removal method by a bactericide.
However, in these methods, there are many cases in which it is too late to recover Hevea brasiliensis suffering from root rot, and a method for reliably controlling root rot is desired.
しかしながら、これらの方法では、根白腐れ病にかかったＨｅｖｅａ ｂｒａｓｉｌｉｅｎｓｉｓを回復させるのに手遅れの場合が多々あり、確実に根白腐れ病を制御する方法が望まれている。 Conventionally, root rot in Hevea brasiliensis is controlled by a physical removal method by incineration of infected roots or a chemical removal method by a bactericide.
However, in these methods, there are many cases in which it is too late to recover Hevea brasiliensis suffering from root rot, and a method for reliably controlling root rot is desired.
これに対して、根白腐れ病の制御系の改良発展には、病原菌であるＲｉｇｉｄｏｐｏｒｕｓ ｍｉｃｒｏｐｏｒｕｓの遺伝的多様性を理解することが重要である。例えば非特許文献１において、Ｒｉｇｉｄｏｐｏｒｕｓ ｍｉｃｒｏｐｏｒｕｓの遺伝的多様性と病原性の関係を解析する試みがなされている。
In contrast, it is important to understand the genetic diversity of Rigidoporus microporus, the pathogen, in order to improve and develop the control system for root rot. For example, in Non-Patent Document 1, an attempt is made to analyze the relationship between Rigidoporus microporus genetic diversity and pathogenicity.
しかしながら、非特許文献１に記載された解析結果からは、地理的分布と遺伝的多様性との間に相関が確認されたが、病原性と遺伝的多様性の間には明確な相関性が確認されない。かかる解析結果を根白腐れ病の制御方法に応用するには、更なる知見を必要としている。
However, the analysis results described in Non-Patent Document 1 confirmed the correlation between geographical distribution and genetic diversity, but there was a clear correlation between pathogenicity and genetic diversity. Not confirmed. In order to apply this analysis result to the control method of root rot, further knowledge is required.
本発明は、上記事情に鑑みてなされたものであって、Ｈｅｖｅａ ｂｒａｓｉｌｉｅｎｓｉｓの根白腐れ病を制御することができるＴｒｉｃｈｏｄｅｒｍａ属菌、これを用いた植物病原菌防除剤及び土壌改良剤、並びに、植物病原菌防除方法及び土壌改良方法を提供することを課題とする。
The present invention has been made in view of the above circumstances, and is a Trichoderma sp. That can control root rot of Hevea brasiliensis, a phytopathogenic fungus control agent and a soil improver using the same, and a phytopathogenic fungus It aims at providing the control method and the soil improvement method.
本発明者は、上記課題を解決すべく鋭意研究した結果、Ｈｅｖｅａ ｂｒａｓｉｌｉｅｎｓｉｓの根白腐れ病菌に対して拮抗性を有するＴｒｉｃｈｏｄｅｒｍａ属菌を見出し、本発明を完成させた。
As a result of intensive research aimed at solving the above-mentioned problems, the present inventor found a Trichoderma genus having antagonistic properties against root rot of Hevea brasiliensis and completed the present invention.
すなわち、本発明は、下記の特徴を有するＴｒｉｃｈｏｄｅｒｍａ属菌、これを用いた植物病原菌防除剤及び土壌改良剤、並びに、植物病原菌防除方法を提供する。
（１）Ｈｅｖｅａ ｂｒａｓｉｌｉｅｎｓｉｓの根白腐れ病菌に対して拮抗性を有するＴｒｉｃｈｏｄｅｒｍａ属菌。
（２）生物農薬として用いられる（１）に記載のＴｒｉｃｈｏｄｅｒｍａ属菌。
（３）前記根白腐れ病菌は、Ｒｉｇｉｄｏｐｏｒｕｓ ｍｉｃｒｏｐｏｒｕｓである（１）または（２）に記載のＴｒｉｃｈｏｄｅｒｍａ属菌。
（４）Ｔｒｉｃｈｏｄｅｒｍａ ｓｐｉｒａｌｅ又はＴｒｉｃｈｏｄｅｒｍａ ｅｒｉｎａｃｅｕｍである（１）～（３）のいずれかに記載のＴｒｉｃｈｏｄｅｒｍａ属菌。
（５）（１）～（４）のいずれかに記載のＴｒｉｃｈｏｄｅｒｍａ属菌の培養物、湿菌体、又は乾燥菌体を含有する植物病原菌防除剤。
（６）（５）に記載の植物病原菌防除剤を土壌に処理する工程を有する植物病原菌防除方法。
（７）（１）～（４）のいずれかに記載のＴｒｉｃｈｏｄｅｒｍａ属菌の培養物、湿菌体、又は乾燥菌体を含有する土壌改良剤。
（８）（７）に記載の土壌改良剤を土壌に処理する工程を有する土壌改良方法。 That is, this invention provides the Trichoderma genus microbe which has the following characteristics, a phytopathogen control agent and a soil improvement agent using the same, and a phytopathogen control method.
(1) Trichoderma spp. That has antagonistic properties against root rot of Hevea brasiliensis.
(2) The Trichoderma genus microbe as described in (1) used as a biopesticide.
(3) The Trichoderma spp. According to (1) or (2), wherein the root rot fungus is Rigidoporus microporus.
(4) The Trichoderma spp. According to any one of (1) to (3), which is Trichoderma spiral or Trichoderma erinaceum.
(5) A phytopathogenic fungus control agent comprising a Trichoderma spp. Culture, wet cell or dry cell according to any one of (1) to (4).
(6) A method for controlling phytopathogenic fungi, comprising a step of treating the phytopathogenic fungus controlling agent according to (5) with soil.
(7) A soil conditioner containing a Trichoderma spp. Culture, wet cell or dry cell according to any one of (1) to (4).
(8) A soil improvement method comprising a step of treating the soil improver according to (7) with soil.
（１）Ｈｅｖｅａ ｂｒａｓｉｌｉｅｎｓｉｓの根白腐れ病菌に対して拮抗性を有するＴｒｉｃｈｏｄｅｒｍａ属菌。
（２）生物農薬として用いられる（１）に記載のＴｒｉｃｈｏｄｅｒｍａ属菌。
（３）前記根白腐れ病菌は、Ｒｉｇｉｄｏｐｏｒｕｓ ｍｉｃｒｏｐｏｒｕｓである（１）または（２）に記載のＴｒｉｃｈｏｄｅｒｍａ属菌。
（４）Ｔｒｉｃｈｏｄｅｒｍａ ｓｐｉｒａｌｅ又はＴｒｉｃｈｏｄｅｒｍａ ｅｒｉｎａｃｅｕｍである（１）～（３）のいずれかに記載のＴｒｉｃｈｏｄｅｒｍａ属菌。
（５）（１）～（４）のいずれかに記載のＴｒｉｃｈｏｄｅｒｍａ属菌の培養物、湿菌体、又は乾燥菌体を含有する植物病原菌防除剤。
（６）（５）に記載の植物病原菌防除剤を土壌に処理する工程を有する植物病原菌防除方法。
（７）（１）～（４）のいずれかに記載のＴｒｉｃｈｏｄｅｒｍａ属菌の培養物、湿菌体、又は乾燥菌体を含有する土壌改良剤。
（８）（７）に記載の土壌改良剤を土壌に処理する工程を有する土壌改良方法。 That is, this invention provides the Trichoderma genus microbe which has the following characteristics, a phytopathogen control agent and a soil improvement agent using the same, and a phytopathogen control method.
(1) Trichoderma spp. That has antagonistic properties against root rot of Hevea brasiliensis.
(2) The Trichoderma genus microbe as described in (1) used as a biopesticide.
(3) The Trichoderma spp. According to (1) or (2), wherein the root rot fungus is Rigidoporus microporus.
(4) The Trichoderma spp. According to any one of (1) to (3), which is Trichoderma spiral or Trichoderma erinaceum.
(5) A phytopathogenic fungus control agent comprising a Trichoderma spp. Culture, wet cell or dry cell according to any one of (1) to (4).
(6) A method for controlling phytopathogenic fungi, comprising a step of treating the phytopathogenic fungus controlling agent according to (5) with soil.
(7) A soil conditioner containing a Trichoderma spp. Culture, wet cell or dry cell according to any one of (1) to (4).
(8) A soil improvement method comprising a step of treating the soil improver according to (7) with soil.
本発明によれば、根白腐れ病菌によるＨｅｖｅａ ｂｒａｓｉｌｉｅｎｓｉｓの病害を抑制することができる。
According to the present invention, the disease of Hevea brasiliensis caused by root rot fungus can be suppressed.
［Ｔｒｉｃｈｏｄｅｒｍａ属菌］
本発明のＴｒｉｃｈｏｄｅｒｍａ属菌は、Ｈｅｖｅａ ｂｒａｓｉｌｉｅｎｓｉｓの根白腐れ病菌に対して拮抗性を有する。
Ｈｅｖｅａ ｂｒａｓｉｌｉｅｎｓｉｓは、ラテックス（主にポリイソプレン）を産生し、乳管細胞を有するトウダイグサ科の植物である。Ｈｅｖｅａ ｂｒａｓｉｌｉｅｎｓｉｓは、工業用天然ゴム原料として汎用されている。
根白腐れ病菌の主な病原菌は、Ｒｉｇｉｄｏｐｏｒｕｓ ｍｉｃｒｏｐｏｒｕｓであるため、本発明のＴｒｉｃｈｏｄｅｒｍａ属菌は、Ｒｉｇｉｄｏｐｏｒｕｓ ｍｉｃｒｏｐｏｒｕｓに対して拮抗性を有することが好ましい。 [Trichoderma spp.]
The Trichoderma spp. Of the present invention has antagonistic properties against root rot of Hevea brasiliensis.
Hevea brasiliensis is a Euphorbiaceae plant that produces latex (mainly polyisoprene) and has duct cells. Hevea brasiliensis is widely used as an industrial natural rubber raw material.
Since the main pathogen of the root rot fungus is Rigidoporus microporus, it is preferable that the Trichoderma spp. Of the present invention have antagonistic properties to Rigidoporus microporus.
本発明のＴｒｉｃｈｏｄｅｒｍａ属菌は、Ｈｅｖｅａ ｂｒａｓｉｌｉｅｎｓｉｓの根白腐れ病菌に対して拮抗性を有する。
Ｈｅｖｅａ ｂｒａｓｉｌｉｅｎｓｉｓは、ラテックス（主にポリイソプレン）を産生し、乳管細胞を有するトウダイグサ科の植物である。Ｈｅｖｅａ ｂｒａｓｉｌｉｅｎｓｉｓは、工業用天然ゴム原料として汎用されている。
根白腐れ病菌の主な病原菌は、Ｒｉｇｉｄｏｐｏｒｕｓ ｍｉｃｒｏｐｏｒｕｓであるため、本発明のＴｒｉｃｈｏｄｅｒｍａ属菌は、Ｒｉｇｉｄｏｐｏｒｕｓ ｍｉｃｒｏｐｏｒｕｓに対して拮抗性を有することが好ましい。 [Trichoderma spp.]
The Trichoderma spp. Of the present invention has antagonistic properties against root rot of Hevea brasiliensis.
Hevea brasiliensis is a Euphorbiaceae plant that produces latex (mainly polyisoprene) and has duct cells. Hevea brasiliensis is widely used as an industrial natural rubber raw material.
Since the main pathogen of the root rot fungus is Rigidoporus microporus, it is preferable that the Trichoderma spp. Of the present invention have antagonistic properties to Rigidoporus microporus.
根白腐れ病菌に対して拮抗性を有する菌をスクリーニングする方法として、対峙培養法により拮抗の有無を定性測定する方法が挙げられる。
後述する実施例において、対峙培養法によりＲｉｇｉｄｏｐｏｒｕｓ ｍｉｃｒｏｐｏｒｕｓに拮抗性を有する菌をスクリーニングしたところ、Ｔｒｉｃｈｏｄｅｒｍａ ｓｐｉｒａｌｅやＴｒｉｃｈｏｄｅｒｍａ ｅｒｉｎａｃｅｕｍといったＴｒｉｃｈｏｄｅｒｍａ属菌が同定された。
Ｔｒｉｃｈｏｄｅｒｍａ属菌は、植物病原菌に寄生する特徴があることが知られており（Ｒ．Ｗｅｉｎｄｉｎｇ、Ｐｈｙｔｏｐａｔｈｏｌｏｇｙ、第２２巻、第８３７～８４５頁、１９３２年）、土壌伝染性植物病原菌をはじめとする他の微生物に対して拮抗的な作用を示すことが報告されている。 As a method for screening a bacterium having antagonistic properties against root rot fungus, a method of qualitatively measuring the presence or absence of antagonism by an anti-culturing method can be mentioned.
In the examples described later, when bacteria having an antagonistic property to Rigidoporus microporus were screened by the antipodal culture method, Trichoderma genera such as Trichoderma spiral and Trichoderma erinaceum were identified.
Trichoderma spp. Are known to have parasitic characteristics to phytopathogenic fungi (R. Wedding, Phytopathology, Vol. 22, pp. 837-845, 1932) and others including soil infectious phytopathogenic fungi It has been reported to show an antagonistic action against other microorganisms.
後述する実施例において、対峙培養法によりＲｉｇｉｄｏｐｏｒｕｓ ｍｉｃｒｏｐｏｒｕｓに拮抗性を有する菌をスクリーニングしたところ、Ｔｒｉｃｈｏｄｅｒｍａ ｓｐｉｒａｌｅやＴｒｉｃｈｏｄｅｒｍａ ｅｒｉｎａｃｅｕｍといったＴｒｉｃｈｏｄｅｒｍａ属菌が同定された。
Ｔｒｉｃｈｏｄｅｒｍａ属菌は、植物病原菌に寄生する特徴があることが知られており（Ｒ．Ｗｅｉｎｄｉｎｇ、Ｐｈｙｔｏｐａｔｈｏｌｏｇｙ、第２２巻、第８３７～８４５頁、１９３２年）、土壌伝染性植物病原菌をはじめとする他の微生物に対して拮抗的な作用を示すことが報告されている。 As a method for screening a bacterium having antagonistic properties against root rot fungus, a method of qualitatively measuring the presence or absence of antagonism by an anti-culturing method can be mentioned.
In the examples described later, when bacteria having an antagonistic property to Rigidoporus microporus were screened by the antipodal culture method, Trichoderma genera such as Trichoderma spiral and Trichoderma erinaceum were identified.
Trichoderma spp. Are known to have parasitic characteristics to phytopathogenic fungi (R. Wedding, Phytopathology, Vol. 22, pp. 837-845, 1932) and others including soil infectious phytopathogenic fungi It has been reported to show an antagonistic action against other microorganisms.
従来、根白腐れ病菌の病害防除には、化学物質である農薬が用いられていたが、自然環境の保護に対する要求が高まっており、毒性の低い植物病原性防除剤が注目されている。
従って、根白腐れ病に対して拮抗性を有する本発明のＴｒｉｃｈｏｄｅｒｍａ属菌は、生物農薬として用いられることが好ましい。 Conventionally, pesticides, which are chemical substances, have been used to control root rot fungi. However, demands for protection of the natural environment are increasing, and phytopathogenic control agents with low toxicity are drawing attention.
Therefore, the Trichoderma spp. Of the present invention having antagonistic properties against root rot is preferably used as a biopesticide.
従って、根白腐れ病に対して拮抗性を有する本発明のＴｒｉｃｈｏｄｅｒｍａ属菌は、生物農薬として用いられることが好ましい。 Conventionally, pesticides, which are chemical substances, have been used to control root rot fungi. However, demands for protection of the natural environment are increasing, and phytopathogenic control agents with low toxicity are drawing attention.
Therefore, the Trichoderma spp. Of the present invention having antagonistic properties against root rot is preferably used as a biopesticide.
［植物病原菌防除剤］
本発明の植物病原菌防除剤は、本発明のＴｒｉｃｈｏｄｅｒｍａ属菌の培養物、湿菌体、又は乾燥菌体を含有する。
本発明のＴｒｉｃｈｏｄｅｒｍａ属菌の培養方法としては、ＰＤ（ポテト・デキストロース）ブロースに、前記Ｔｒｉｃｈｏｄｅｒｍａ属菌を接種して、約２８℃の温度でジャーファーメンターを用いて、好気的状態で、２日～４日程度培養する方法が挙げられる。係る培養方法によってＴｒｉｃｈｏｄｅｒｍａ属菌の数を増殖させることにより、本発明の植物病原菌防除剤は、根白腐れ病菌に対してより効果的な防除作用を示す。なお、菌の生存率をあげるためには、生育温度は１０～２８℃が好ましく、１５～２８℃がより好ましい。 [Plant pathogen control agent]
The plant pathogen control agent of the present invention contains a culture, wet cell or dry cell of the Trichoderma genus of the present invention.
As a method for culturing the genus Trichoderma of the present invention, PD (potato dextrose) broth is inoculated with the Trichoderma genus, and a jar fermenter is used at a temperature of about 28 ° C. in an aerobic state. A method of culturing for about 4 to 4 days is mentioned. By growing the number of Trichoderma spp. By such a culture method, the plant pathogen control agent of the present invention exhibits a more effective control action against root rot fungus. In order to increase the survival rate of the bacteria, the growth temperature is preferably 10 to 28 ° C, more preferably 15 to 28 ° C.
本発明の植物病原菌防除剤は、本発明のＴｒｉｃｈｏｄｅｒｍａ属菌の培養物、湿菌体、又は乾燥菌体を含有する。
本発明のＴｒｉｃｈｏｄｅｒｍａ属菌の培養方法としては、ＰＤ（ポテト・デキストロース）ブロースに、前記Ｔｒｉｃｈｏｄｅｒｍａ属菌を接種して、約２８℃の温度でジャーファーメンターを用いて、好気的状態で、２日～４日程度培養する方法が挙げられる。係る培養方法によってＴｒｉｃｈｏｄｅｒｍａ属菌の数を増殖させることにより、本発明の植物病原菌防除剤は、根白腐れ病菌に対してより効果的な防除作用を示す。なお、菌の生存率をあげるためには、生育温度は１０～２８℃が好ましく、１５～２８℃がより好ましい。 [Plant pathogen control agent]
The plant pathogen control agent of the present invention contains a culture, wet cell or dry cell of the Trichoderma genus of the present invention.
As a method for culturing the genus Trichoderma of the present invention, PD (potato dextrose) broth is inoculated with the Trichoderma genus, and a jar fermenter is used at a temperature of about 28 ° C. in an aerobic state. A method of culturing for about 4 to 4 days is mentioned. By growing the number of Trichoderma spp. By such a culture method, the plant pathogen control agent of the present invention exhibits a more effective control action against root rot fungus. In order to increase the survival rate of the bacteria, the growth temperature is preferably 10 to 28 ° C, more preferably 15 to 28 ° C.
本発明の植物病原菌防除剤が含有する培養物とは、増殖させた本発明のＴｒｉｃｈｏｄｅｒｍａ属菌を含む液体培地そのままのものである。かかる培養物には、培養液中に含まれる培地成分や培養によって生成した二次代謝産物が含まれている。これらがＨｅｖｅａ ｂｒａｓｉｌｉｅｎｓｉｓに与える影響を防止するため、本発明の植物病原菌防除剤は、水で１００～１０００倍に希釈して用いられることが好ましい。
The culture containing the plant pathogen control agent of the present invention is a liquid medium containing the grown Trichoderma genus of the present invention as it is. Such a culture contains medium components contained in the culture solution and secondary metabolites produced by the culture. In order to prevent the effects of these on Hevea brasiliensis, the phytopathogenic fungus control agent of the present invention is preferably diluted 100 to 1000 times with water.
本発明の植物病原菌防除剤が含有する湿菌体とは、培養液を遠心分離により集菌したものを、適宜、純水等で洗浄したものである。湿菌体は、培養物と異なり、培地成分等を含んでいない。しかし、本発明の植物病原菌防除剤は、Ｈｅｖｅａ ｂｒａｓｉｌｉｅｎｓｉｓに均一に散布する観点から、やはり水で１００～１０００倍に希釈して用いられることが好ましい。
The wet cells contained in the phytopathogenic fungus control agent of the present invention are those obtained by washing a culture solution collected by centrifugation, as appropriate, with pure water or the like. Unlike the culture, the wet cell does not contain a medium component or the like. However, the phytopathogenic fungus control agent of the present invention is preferably used after being diluted 100 to 1000 times with water from the viewpoint of being uniformly applied to Hevea brasiliensis.
本発明の植物病原菌防除剤が含有する乾燥菌体とは、培養液を遠心分離により集菌したものを、乾燥固形物にしたものである。かかる乾燥固形物は、５～３０質量％の水分含量になるまで半乾燥されたものが好ましい。乾燥処理は、本発明のＴｒｉｃｈｏｄｅｒｍａ属菌の生育に悪影響を与えない温度下で行われることが好ましい。
The dry cell contained in the plant pathogen control agent of the present invention is a product obtained by collecting a culture solution by centrifugation and converting it into a dry solid. Such dry solid is preferably semi-dried to a moisture content of 5 to 30% by mass. The drying treatment is preferably performed at a temperature that does not adversely affect the growth of the Trichoderma spp. Of the present invention.
また、本発明の植物病原菌防除剤は、本発明のＴｒｉｃｈｏｄｅｒｍａ属菌の培養物、湿菌体、又は乾燥菌体を滅菌した固体培地に接種して一定期間培養したものであってもよい。
Further, the plant pathogen control agent of the present invention may be one obtained by inoculating a culture of the genus Trichoderma of the present invention, a wet cell, or a dry cell into a sterilized solid medium and culturing for a certain period.
また、本発明の植物病原菌防除剤は、上述した固体培地に培養した固体培養物を吸着剤に吸着させたものであってもよい。吸着剤としては、バーミキュライト、ゼオライト等の吸着性鉱物材料；木炭、活性炭素等の炭類；化学合成されたポリマー等を挙げることができる。
Further, the plant pathogen control agent of the present invention may be one obtained by adsorbing a solid culture cultured in the above-mentioned solid medium to an adsorbent. Examples of the adsorbent include adsorbing mineral materials such as vermiculite and zeolite; charcoal such as charcoal and activated carbon; chemically synthesized polymers and the like.
また、本発明の植物病原菌防除剤は、殺虫剤、殺ダニ剤、除草剤、他の殺菌剤等を含んでいてもよい。
Moreover, the plant pathogen control agent of the present invention may contain an insecticide, an acaricide, a herbicide, other fungicides and the like.
［土壌改良剤］
本発明の土壌改良剤は、本発明のＴｒｉｃｈｏｄｅｒｍａ属菌の培養物、湿菌体、又は乾燥菌体を含有するものであれば、上述した本発明の植物病原菌防除剤に堆肥やその他土壌改良成分を添加したものであってもよい。
堆肥としては、例えば、培地と発酵菌からなるものが挙げられ、培地としては、米糠、フスマ、酒粕、焼酎粕大豆粕等の単独物又は混合物が挙げられる。
本発明の土壌改良剤が、堆肥を含有する場合には、根白腐れ病菌に対して拮抗性を付与することに加えて、土壌に肥料効果を付与することができる。
また、その他土壌改良成分として、泥炭、腐植酸資材、ポリビニルアルコール系資材等の土壌改良資材及び植物生長調節剤等が挙げられる。
本発明の土壌改良剤は、本発明の植物病原菌防除剤を、発酵の完了した堆肥に加えたものであっても、堆肥の原料に、本発明の植物病原菌防除剤を加えて、堆肥の発酵と同時に本発明のＴｒｉｃｈｏｄｅｒｍａ属菌の増殖を行わせたものであってもよい。 [Soil conditioner]
As long as the soil conditioner of the present invention contains the culture, wet cells, or dried cells of the genus Trichoderma of the present invention, the above-mentioned plant pathogen control agent of the present invention is compost and other soil improving components. May be added.
Examples of the compost include those composed of a medium and fermenting bacteria, and examples of the medium include a single substance or a mixture of rice bran, bran, sake lees, shochu soybean meal, and the like.
When the soil improving agent of the present invention contains compost, it can impart fertilizer effect to the soil in addition to imparting antagonistic properties against root rot fungus.
Other soil improvement components include peat, humic acid materials, soil improvement materials such as polyvinyl alcohol materials, plant growth regulators, and the like.
Even if the soil conditioner of the present invention is obtained by adding the phytopathogenic fungus control agent of the present invention to compost that has been fermented, the fermented compost is obtained by adding the phytopathogenic fungus control agent of the present invention to the compost raw material. Simultaneously, the Trichoderma spp. Of the present invention may be grown.
本発明の土壌改良剤は、本発明のＴｒｉｃｈｏｄｅｒｍａ属菌の培養物、湿菌体、又は乾燥菌体を含有するものであれば、上述した本発明の植物病原菌防除剤に堆肥やその他土壌改良成分を添加したものであってもよい。
堆肥としては、例えば、培地と発酵菌からなるものが挙げられ、培地としては、米糠、フスマ、酒粕、焼酎粕大豆粕等の単独物又は混合物が挙げられる。
本発明の土壌改良剤が、堆肥を含有する場合には、根白腐れ病菌に対して拮抗性を付与することに加えて、土壌に肥料効果を付与することができる。
また、その他土壌改良成分として、泥炭、腐植酸資材、ポリビニルアルコール系資材等の土壌改良資材及び植物生長調節剤等が挙げられる。
本発明の土壌改良剤は、本発明の植物病原菌防除剤を、発酵の完了した堆肥に加えたものであっても、堆肥の原料に、本発明の植物病原菌防除剤を加えて、堆肥の発酵と同時に本発明のＴｒｉｃｈｏｄｅｒｍａ属菌の増殖を行わせたものであってもよい。 [Soil conditioner]
As long as the soil conditioner of the present invention contains the culture, wet cells, or dried cells of the genus Trichoderma of the present invention, the above-mentioned plant pathogen control agent of the present invention is compost and other soil improving components. May be added.
Examples of the compost include those composed of a medium and fermenting bacteria, and examples of the medium include a single substance or a mixture of rice bran, bran, sake lees, shochu soybean meal, and the like.
When the soil improving agent of the present invention contains compost, it can impart fertilizer effect to the soil in addition to imparting antagonistic properties against root rot fungus.
Other soil improvement components include peat, humic acid materials, soil improvement materials such as polyvinyl alcohol materials, plant growth regulators, and the like.
Even if the soil conditioner of the present invention is obtained by adding the phytopathogenic fungus control agent of the present invention to compost that has been fermented, the fermented compost is obtained by adding the phytopathogenic fungus control agent of the present invention to the compost raw material. Simultaneously, the Trichoderma spp. Of the present invention may be grown.
［植物病原菌防除方法］
本発明の植物病原菌防除方法は、本発明の植物病原菌防除剤を土壌に処理する工程を有する。本発明の植物病原菌防除方法としては、本発明のＴｒｉｃｈｏｄｅｒｍａ属菌の培養物又は湿菌体を水で１００～１０００倍に希釈したものを病害土壌に散布する方法；本発明のＴｒｉｃｈｏｄｅｒｍａ属菌の培養物又は湿菌体を固体培地に接種した固体培養物を病害土壌に施す方法；上記固体培養物を吸着材に吸着させてなる防除材を病害土壌に施す方法；本発明のＴｒｉｃｈｏｄｅｒｍａ属菌の乾燥菌体を病害土壌に散布する方法等が挙げられる。
本発明の植物病原菌防除剤の施用方法としては、地上液体散布、地上固形散布、空中液剤散布、空中固形散布等が挙げられる。 [Plant pathogen control method]
The plant pathogen control method of the present invention includes a step of treating the plant pathogen control agent of the present invention with soil. As a method for controlling plant pathogens of the present invention, a method of spraying a culture of Trichoderma spp. Of the present invention or a wet fungus body diluted 100 to 1000 times with water onto diseased soil; Cultivation of Trichoderma spp. Of the present invention A method of applying a solid culture obtained by inoculating a solid medium with a product or wet cells to a diseased soil; a method of applying a control material obtained by adsorbing the solid culture to an adsorbent to the diseased soil; drying the Trichoderma spp. Of the present invention The method etc. which spray a microbial cell on diseased soil are mentioned.
Examples of the method for applying the plant pathogen control agent of the present invention include ground liquid spray, ground solid spray, air solution spray, and air solid spray.
本発明の植物病原菌防除方法は、本発明の植物病原菌防除剤を土壌に処理する工程を有する。本発明の植物病原菌防除方法としては、本発明のＴｒｉｃｈｏｄｅｒｍａ属菌の培養物又は湿菌体を水で１００～１０００倍に希釈したものを病害土壌に散布する方法；本発明のＴｒｉｃｈｏｄｅｒｍａ属菌の培養物又は湿菌体を固体培地に接種した固体培養物を病害土壌に施す方法；上記固体培養物を吸着材に吸着させてなる防除材を病害土壌に施す方法；本発明のＴｒｉｃｈｏｄｅｒｍａ属菌の乾燥菌体を病害土壌に散布する方法等が挙げられる。
本発明の植物病原菌防除剤の施用方法としては、地上液体散布、地上固形散布、空中液剤散布、空中固形散布等が挙げられる。 [Plant pathogen control method]
The plant pathogen control method of the present invention includes a step of treating the plant pathogen control agent of the present invention with soil. As a method for controlling plant pathogens of the present invention, a method of spraying a culture of Trichoderma spp. Of the present invention or a wet fungus body diluted 100 to 1000 times with water onto diseased soil; Cultivation of Trichoderma spp. Of the present invention A method of applying a solid culture obtained by inoculating a solid medium with a product or wet cells to a diseased soil; a method of applying a control material obtained by adsorbing the solid culture to an adsorbent to the diseased soil; drying the Trichoderma spp. Of the present invention The method etc. which spray a microbial cell on diseased soil are mentioned.
Examples of the method for applying the plant pathogen control agent of the present invention include ground liquid spray, ground solid spray, air solution spray, and air solid spray.
［土壌改良方法］
本発明の土壌改良方法は、本発明の土壌改良剤を土壌に処理する工程を有する。土壌に処理する工程は、具体的には土壌混和、株元施用、苗床施用及び肥料に混ぜて施す、等などがある。かかる工程を有することにより、根白腐れ病菌の病害を抑制できることに加えて、病害土壌に肥料効果を付与することができる。 [Soil improvement method]
The soil improvement method of this invention has the process of processing the soil improvement agent of this invention to soil. Specifically, the process for treating the soil includes mixing with soil, application of the plant base, application of the nursery bed, and mixing with fertilizer. By having this process, in addition to being able to suppress the disease of root rot fungus, it is possible to impart a fertilizer effect to the diseased soil.
本発明の土壌改良方法は、本発明の土壌改良剤を土壌に処理する工程を有する。土壌に処理する工程は、具体的には土壌混和、株元施用、苗床施用及び肥料に混ぜて施す、等などがある。かかる工程を有することにより、根白腐れ病菌の病害を抑制できることに加えて、病害土壌に肥料効果を付与することができる。 [Soil improvement method]
The soil improvement method of this invention has the process of processing the soil improvement agent of this invention to soil. Specifically, the process for treating the soil includes mixing with soil, application of the plant base, application of the nursery bed, and mixing with fertilizer. By having this process, in addition to being able to suppress the disease of root rot fungus, it is possible to impart a fertilizer effect to the diseased soil.
次に実施例を示して本発明をさらに詳細に説明するが、本発明は以下の実施例に限定されるものではない。
Next, the present invention will be described in more detail with reference to examples. However, the present invention is not limited to the following examples.
［対峙培養法によるＲｉｇｉｄｏｐｏｒｕｓ ｍｉｃｒｏｐｏｒｕｓに対する拮抗性試験］
インドネシアにおけるブリヂストン社のゴム農園から採取された土壌を用いて、以下の拮抗性試験を行った。尚、前記ゴム農園において、土壌採取は、それぞれ場所（区画１～４）を変えて行った。
〈実施例１〉
ＰＤＡ培地（ポテト・デキストロース・アガー培地）に、Ｒｉｇｉｄｏｐｏｒｕｓ ｍｉｃｒｏｐｏｒｕｓ、又はＳａｍｐｌｅ１（区画１由来の土壌）を、２８℃で５日間、それぞれ培養した。 [Antagonism test for Rigidoporus microporus by anti-culture method]
The following antagonistic tests were conducted using soil collected from Bridgestone Rubber Plantation in Indonesia. In the rubber plantation, soil sampling was performed at different locations (sections 1 to 4).
<Example 1>
Rigidoporus microporus or Sample 1 (soil from compartment 1) was cultured in PDA medium (potato dextrose agar medium) at 28 ° C. for 5 days.
インドネシアにおけるブリヂストン社のゴム農園から採取された土壌を用いて、以下の拮抗性試験を行った。尚、前記ゴム農園において、土壌採取は、それぞれ場所（区画１～４）を変えて行った。
〈実施例１〉
ＰＤＡ培地（ポテト・デキストロース・アガー培地）に、Ｒｉｇｉｄｏｐｏｒｕｓ ｍｉｃｒｏｐｏｒｕｓ、又はＳａｍｐｌｅ１（区画１由来の土壌）を、２８℃で５日間、それぞれ培養した。 [Antagonism test for Rigidoporus microporus by anti-culture method]
The following antagonistic tests were conducted using soil collected from Bridgestone Rubber Plantation in Indonesia. In the rubber plantation, soil sampling was performed at different locations (sections 1 to 4).
<Example 1>
Rigidoporus microporus or Sample 1 (soil from compartment 1) was cultured in PDA medium (potato dextrose agar medium) at 28 ° C. for 5 days.
オートクレーブ滅菌したＰＤＡ培地を分注した直径９ｃｍのシャーレの外周から１ｃｍ内側の位置に、培養したＲｉｇｉｄｏｐｏｒｕｓ ｍｉｃｒｏｐｏｒｕｓを滅菌した直径５ｍｍのコルクボーラーで培地ごと打ち抜いて接種した。
次いで、Ｒｉｇｉｄｏｐｏｒｕｓ ｍｉｃｒｏｐｏｒｕｓを接種した位置と対称の位置に、培養したＳａｍｐｌｅ１を滅菌した直径５ｍｍのコルクボーラーで培地ごと打ち抜いて接種し、２８℃で５日間、１０日間、１５日対峙培養した。Ｒｉｇｉｄｏｐｏｒｕｓ ｍｉｃｒｏｐｏｒｕｓ及びＳａｍｐｌｅ１の生育総面積中のＳａｍｐｌｅ１の生育面積の割合を、生育阻止率として算出した。５日間、１０日間、１５日の生育阻止率の結果を図１に示す。１５日間培養後の培養シャーレの写真を図２の（ａ）に示す。
尚、生育面積は、透明なシートに５ｍｍ間隔のマス目をひき、これをシャーレ上にかぶせてマス目を数えることにより計算した。 The cultured medium of Rigidoporus microporus was punched and inoculated with a sterilized 5 mm diameter cork borer at a position 1 cm inward from the outer periphery of a 9 cm diameter petri dish containing autoclaved PDA medium.
Next, the cultured sample 1 was punched and inoculated with a sterilized 5 mm diameter cork borer at a position symmetrical to the position where Rigidoporus microporus was inoculated, and cultured at 28 ° C. for 5 days, 10 days, and 15 days. The ratio of the growth area of Sample 1 in the total growth area of Rigidoporus microporus and Sample 1 was calculated as the growth inhibition rate. The results of the growth inhibition rate for 5 days, 10 days, and 15 days are shown in FIG. A photograph of the culture dish after 15 days of culture is shown in FIG.
In addition, the growth area was calculated by drawing squares at intervals of 5 mm on a transparent sheet, and covering this on a petri dish and counting the squares.
次いで、Ｒｉｇｉｄｏｐｏｒｕｓ ｍｉｃｒｏｐｏｒｕｓを接種した位置と対称の位置に、培養したＳａｍｐｌｅ１を滅菌した直径５ｍｍのコルクボーラーで培地ごと打ち抜いて接種し、２８℃で５日間、１０日間、１５日対峙培養した。Ｒｉｇｉｄｏｐｏｒｕｓ ｍｉｃｒｏｐｏｒｕｓ及びＳａｍｐｌｅ１の生育総面積中のＳａｍｐｌｅ１の生育面積の割合を、生育阻止率として算出した。５日間、１０日間、１５日の生育阻止率の結果を図１に示す。１５日間培養後の培養シャーレの写真を図２の（ａ）に示す。
尚、生育面積は、透明なシートに５ｍｍ間隔のマス目をひき、これをシャーレ上にかぶせてマス目を数えることにより計算した。 The cultured medium of Rigidoporus microporus was punched and inoculated with a sterilized 5 mm diameter cork borer at a position 1 cm inward from the outer periphery of a 9 cm diameter petri dish containing autoclaved PDA medium.
Next, the cultured sample 1 was punched and inoculated with a sterilized 5 mm diameter cork borer at a position symmetrical to the position where Rigidoporus microporus was inoculated, and cultured at 28 ° C. for 5 days, 10 days, and 15 days. The ratio of the growth area of Sample 1 in the total growth area of Rigidoporus microporus and Sample 1 was calculated as the growth inhibition rate. The results of the growth inhibition rate for 5 days, 10 days, and 15 days are shown in FIG. A photograph of the culture dish after 15 days of culture is shown in FIG.
In addition, the growth area was calculated by drawing squares at intervals of 5 mm on a transparent sheet, and covering this on a petri dish and counting the squares.
〈実施例２〉
Ｓａｍｐｌｅ１に代えて、Ｓａｍｐｌｅ２（区画２由来の土壌）を用いた以外は、実施例１と同様の方法で対峙培養した。Ｒｉｇｉｄｏｐｏｒｕｓ ｍｉｃｒｏｐｏｒｕｓ及びＳａｍｐｌｅ２の生育総面積中のＳａｍｐｌｅ２の生育面積の割合を、生育阻止率として算出した。５日間、１０日間、１５日の生育阻止率の結果を図１に示す。１５日間培養後の培養シャーレの写真を図２の（ｂ）に示す。 <Example 2>
Instead of Sample 1, the sample was cultured in the same manner as in Example 1 except that Sample 2 (soil derived from compartment 2) was used. The ratio of the growth area of Sample 2 in the total growth area of Rigidoporus microporus and Sample 2 was calculated as the growth inhibition rate. The results of the growth inhibition rate for 5 days, 10 days, and 15 days are shown in FIG. A photograph of the culture petri dish after 15 days of culture is shown in FIG.
Ｓａｍｐｌｅ１に代えて、Ｓａｍｐｌｅ２（区画２由来の土壌）を用いた以外は、実施例１と同様の方法で対峙培養した。Ｒｉｇｉｄｏｐｏｒｕｓ ｍｉｃｒｏｐｏｒｕｓ及びＳａｍｐｌｅ２の生育総面積中のＳａｍｐｌｅ２の生育面積の割合を、生育阻止率として算出した。５日間、１０日間、１５日の生育阻止率の結果を図１に示す。１５日間培養後の培養シャーレの写真を図２の（ｂ）に示す。 <Example 2>
Instead of Sample 1, the sample was cultured in the same manner as in Example 1 except that Sample 2 (soil derived from compartment 2) was used. The ratio of the growth area of Sample 2 in the total growth area of Rigidoporus microporus and Sample 2 was calculated as the growth inhibition rate. The results of the growth inhibition rate for 5 days, 10 days, and 15 days are shown in FIG. A photograph of the culture petri dish after 15 days of culture is shown in FIG.
〈比較例１〉
Ｓａｍｐｌｅ１に代えて、Ｓａｍｐｌｅ３（区画３由来の土壌）を用いた以外は、実施例１と同様の方法で対峙培養した。Ｒｉｇｉｄｏｐｏｒｕｓ ｍｉｃｒｏｐｏｒｕｓ及びＳａｍｐｌｅ３の生育総面積中のＳａｍｐｌｅ３の生育面積の割合を、生育阻止率として算出した。５日間、１０日間、１５日の生育阻止率の結果を図１に示す。１５日間培養後の培養シャーレの写真を図２の（ｃ）に示す。 <Comparative example 1>
Instead of Sample 1, the cells were cultured in the same manner as in Example 1 except that Sample 3 (soil derived from compartment 3) was used. The ratio of the growth area of Sample 3 to the total growth area of Rigidoporus microporus and Sample 3 was calculated as a growth inhibition rate. The results of the growth inhibition rate for 5 days, 10 days, and 15 days are shown in FIG. A photograph of the culture petri dish after 15 days of culture is shown in FIG.
Ｓａｍｐｌｅ１に代えて、Ｓａｍｐｌｅ３（区画３由来の土壌）を用いた以外は、実施例１と同様の方法で対峙培養した。Ｒｉｇｉｄｏｐｏｒｕｓ ｍｉｃｒｏｐｏｒｕｓ及びＳａｍｐｌｅ３の生育総面積中のＳａｍｐｌｅ３の生育面積の割合を、生育阻止率として算出した。５日間、１０日間、１５日の生育阻止率の結果を図１に示す。１５日間培養後の培養シャーレの写真を図２の（ｃ）に示す。 <Comparative example 1>
Instead of Sample 1, the cells were cultured in the same manner as in Example 1 except that Sample 3 (soil derived from compartment 3) was used. The ratio of the growth area of Sample 3 to the total growth area of Rigidoporus microporus and Sample 3 was calculated as a growth inhibition rate. The results of the growth inhibition rate for 5 days, 10 days, and 15 days are shown in FIG. A photograph of the culture petri dish after 15 days of culture is shown in FIG.
〈比較例２〉
Ｓａｍｐｌｅ１に代えて、Ｓａｍｐｌｅ４（区画４由来の土壌）を用いた以外は、実施例１と同様の方法で対峙培養した。Ｒｉｇｉｄｏｐｏｒｕｓ ｍｉｃｒｏｐｏｒｕｓ及びＳａｍｐｌｅ４の生育総面積中のＳａｍｐｌｅ４の生育面積の割合を、生育阻止率として算出した。５日間、１０日間、１５日の生育阻止率の結果を図１に示す。１５日間培養後の培養シャーレの写真を図２の（ｄ）に示す。 <Comparative example 2>
Instead of Sample 1, counterculture was performed in the same manner as in Example 1 except that Sample 4 (soil derived from compartment 4) was used. The ratio of the growth area of Sample 4 to the total growth area of Rigidoporus microporus and Sample 4 was calculated as a growth inhibition rate. The results of the growth inhibition rate for 5 days, 10 days, and 15 days are shown in FIG. A photograph of the culture petri dish after 15 days of culture is shown in FIG.
Ｓａｍｐｌｅ１に代えて、Ｓａｍｐｌｅ４（区画４由来の土壌）を用いた以外は、実施例１と同様の方法で対峙培養した。Ｒｉｇｉｄｏｐｏｒｕｓ ｍｉｃｒｏｐｏｒｕｓ及びＳａｍｐｌｅ４の生育総面積中のＳａｍｐｌｅ４の生育面積の割合を、生育阻止率として算出した。５日間、１０日間、１５日の生育阻止率の結果を図１に示す。１５日間培養後の培養シャーレの写真を図２の（ｄ）に示す。 <Comparative example 2>
Instead of Sample 1, counterculture was performed in the same manner as in Example 1 except that Sample 4 (soil derived from compartment 4) was used. The ratio of the growth area of Sample 4 to the total growth area of Rigidoporus microporus and Sample 4 was calculated as a growth inhibition rate. The results of the growth inhibition rate for 5 days, 10 days, and 15 days are shown in FIG. A photograph of the culture petri dish after 15 days of culture is shown in FIG.
図１に示されるように、Ｓａｍｐｌｅ１及び２由来の土壌から培養された菌（実施例１及び２）は、対峙培養５日後から８５％以上の生育阻止率を示すことが確認された。一方、Ｓａｍｐｌｅ３及び４由来の土壌から培養された菌（比較例１及び２）は、対峙培養の期間を経るにつれて生育阻止率が低下し、対峙培養１５日後には、生育阻止率が６５％を下回ることが確認された。
As shown in FIG. 1, it was confirmed that the bacteria cultured in the soils derived from Samples 1 and 2 (Examples 1 and 2) exhibited a growth inhibition rate of 85% or more after 5 days of the counterculture. On the other hand, the bacteria (Comparative Examples 1 and 2) cultivated from the soils derived from Samples 3 and 4 have a growth inhibition rate that decreases as they pass through the period of anti-cultivation. It was confirmed that it was below.
Ｓａｍｐｌｅ１～４から単離された菌株の分生胞子及び分生胞子茎の形態的特性を顕微鏡で観察した。さらに、それぞれの菌株からＰｒｅｐＭａｎ Ｕｌｔｒａ Ｒｅａｇｅｎｔ（Ａｐｐｌｉｅｄ Ｂｉｏｓｙｓｔｅｍｓ社）を用いてＤＮＡ抽出を行い、抽出ＤＮＡを鋳型として、リボソーマルＤＮＡのＩｎｔｅｒｎａｌ Ｔｒａｎｓｃｒｉｂｅｄ Ｓｐａｃｅｒ保存領域中の非特異的プライマー配列（ＩＴＳ１: ５’－ＴＣＣＧＴＡＧＧＴＧＡＡＣＣＴＧＣＧＧ－３’、ＩＴＳ４：５’－ＴＣＣＴＣＣＧＣＴＴＡＴＴＧＡＴＡＴＧＣ－３’）を用いてＰＣＲでＤＮＡ断片を得た。得られたＤＮＡ断片について、ＡＢＩ ｐｒｉｓｍ ３１３０ ｇｅｎｅｔｉｃ ａｎａｌｙｚｅｒ（Ａｐｐｌｉｅｄ Ｂｉｏｓｙｓｔｅｍｓ社）を用いてシークエンシングし、ＤＮＡ配列を解析した。Ｎａｔｉｏｎａｌ Ｃｅｎｔｅｒ ｆｏｒ Ｂｉｏｔｅｃｈｎｏｌｏｇｙ Ｉｎｆｏｒｍａｔｉｏｎ（ＮＣＢＩ）のＢｌａｓｔ検索で登録データとの相同性を比較した結果、Ｓａｍｐｌｅ１は、Ｔｒｉｃｈｏｄｅｒｍａ ｓｐｉｒａｌｅと確認され、Ｓａｍｐｌｅ２は、Ｔｒｉｃｈｏｄｅｒｍａ ｅｒｉｎａｃｅｕｍと確認され、Ｓａｍｐｌｅ３～４は、Ｔｒｉｃｈｏｄｅｒｍａ属菌でないことが確認された。各Ｓａｍｐｌｅの同定結果を表１に示す。
尚、表１のＩＮＨＩＮＢＩＴＩＯＮ欄は、図１と同様、対峙培養１５日後の生育阻害率を示します。＋－は５０％～６０％の阻害率を示し、＋は６０％～７０％の阻害率を示し、＋＋は７０％～８０％の阻害率を示し、＋＋＋は９０％～１００％の阻害率を示す。 The morphological characteristics of conidia and conidia of the strains isolated from Samples 1 to 4 were observed under a microscope. Furthermore, DNA extraction was performed from each strain using PrepMan Ultra Reagent (Applied Biosystems), and using the extracted DNA as a template, a non-specific primer sequence (ITS1: 5′-TCCGCTGGTGGATGGATGGATGGATGG 3 ′, ITS4: 5′-TCCTCCCGCTTTATTAGATCGC-3 ′), a DNA fragment was obtained by PCR. The obtained DNA fragment was sequenced using ABI prism 3130 genetic analyzer (Applied Biosystems), and the DNA sequence was analyzed. As a result of comparing the homology with the registration data in the Blast search of National Center for Biotechnology Information (NCBI), Sample 1 was confirmed to be Trichoderma spiral, Sample 2 was confirmed to be Trichoderma erinaceum, It was confirmed. The identification results of each sample are shown in Table 1.
In addition, the INHINBITION column in Table 1 shows the growth inhibition rate after 15 days of culture in the same manner as in FIG. ++ indicates an inhibition rate of 50% to 60%, + indicates an inhibition rate of 60% to 70%, ++ indicates an inhibition rate of 70% to 80%, and ++ indicates an inhibition rate of 90% to 100% Indicates.
尚、表１のＩＮＨＩＮＢＩＴＩＯＮ欄は、図１と同様、対峙培養１５日後の生育阻害率を示します。＋－は５０％～６０％の阻害率を示し、＋は６０％～７０％の阻害率を示し、＋＋は７０％～８０％の阻害率を示し、＋＋＋は９０％～１００％の阻害率を示す。 The morphological characteristics of conidia and conidia of the strains isolated from Samples 1 to 4 were observed under a microscope. Furthermore, DNA extraction was performed from each strain using PrepMan Ultra Reagent (Applied Biosystems), and using the extracted DNA as a template, a non-specific primer sequence (ITS1: 5′-TCCGCTGGTGGATGGATGGATGGATGG 3 ′, ITS4: 5′-TCCTCCCGCTTTATTAGATCGC-3 ′), a DNA fragment was obtained by PCR. The obtained DNA fragment was sequenced using ABI prism 3130 genetic analyzer (Applied Biosystems), and the DNA sequence was analyzed. As a result of comparing the homology with the registration data in the Blast search of National Center for Biotechnology Information (NCBI), Sample 1 was confirmed to be Trichoderma spiral, Sample 2 was confirmed to be Trichoderma erinaceum, It was confirmed. The identification results of each sample are shown in Table 1.
In addition, the INHINBITION column in Table 1 shows the growth inhibition rate after 15 days of culture in the same manner as in FIG. ++ indicates an inhibition rate of 50% to 60%, + indicates an inhibition rate of 60% to 70%, ++ indicates an inhibition rate of 70% to 80%, and ++ indicates an inhibition rate of 90% to 100% Indicates.
以上の結果から、本発明によれば、根白腐れ病菌によるＨｅｖｅａ ｂｒａｓｉｌｉｅｎｓｉｓの病害を抑制することができることが明らかである。
From the above results, it is clear that according to the present invention, the disease of Hevea brasiliensis caused by root white rot fungus can be suppressed.
Ｈｅｖｅａ ｂｒａｓｉｌｉｅｎｓｉｓの根白腐れ病を制御することができるＴｒｉｃｈｏｄｅｒｍａ属菌、これを用いた植物病原菌防除剤及び土壌改良剤、並びに、植物病原菌防除方法及び土壌改良方法を提供する。
Provided are Trichoderma spp. Which can control root rot of Hevea brasiliensis, plant pathogen control agent and soil improver using the same, and plant pathogen control method and soil improvement method.
Provided are Trichoderma spp. Which can control root rot of Hevea brasiliensis, plant pathogen control agent and soil improver using the same, and plant pathogen control method and soil improvement method.
Claims (8)
- Ｈｅｖｅａ ｂｒａｓｉｌｉｅｎｓｉｓの根白腐れ病菌に対して拮抗性を有するＴｒｉｃｈｏｄｅｒｍａ属菌。 Trichoderma spp. That have antagonistic properties against root rot of Hevea brasiliensis.
- 生物農薬として用いられる請求項１に記載のＴｒｉｃｈｏｄｅｒｍａ属菌。 The Trichoderma genus microbe of Claim 1 used as a biological pesticide.
- 前記根白腐れ病菌は、Ｒｉｇｉｄｏｐｏｒｕｓ ｍｉｃｒｏｐｏｒｕｓである請求項１又は２に記載のＴｒｉｃｈｏｄｅｒｍａ属菌。 The Trichoderma spp. According to claim 1 or 2, wherein the root rot fungus is Rigidoporus microporus.
- Ｔｒｉｃｈｏｄｅｒｍａ ｓｐｉｒａｌｅ又はＴｒｉｃｈｏｄｅｒｍａ ｅｒｉｎａｃｅｕｍである請求項１～３のいずれか一項に記載のＴｒｉｃｈｏｄｅｒｍａ属菌。 The Trichoderma spp. According to any one of claims 1 to 3, which is Trichoderma spiral or Trichoderma erinaceum.
- 請求項１～４のいずれか一項に記載のＴｒｉｃｈｏｄｅｒｍａ属菌の培養物、湿菌体、又は乾燥菌体を含有する植物病原菌防除剤。 A phytopathogenic fungus control agent comprising a culture, wet cell or dry cell of Trichoderma spp. According to any one of claims 1 to 4.
- 請求項５に記載の植物病原菌防除剤を土壌に処理する工程を有する植物病原菌防除方法。 A plant pathogen control method comprising a step of treating the plant pathogen control agent according to claim 5 with soil.
- 請求項１～４のいずれか一項に記載のＴｒｉｃｈｏｄｅｒｍａ属菌の培養物、湿菌体、又は乾燥菌体を含有する土壌改良剤。 A soil conditioner containing a culture, wet cell or dry cell of Trichoderma spp. According to any one of claims 1 to 4.
- 請求項７に記載の土壌改良剤を土壌に処理する工程を有する土壌改良方法。
The soil improvement method which has the process of processing the soil improvement agent of Claim 7 to soil.
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| CN114258924A (en) * | 2021-12-30 | 2022-04-01 | 新疆农业科学院微生物应用研究所(中国新疆—亚美尼亚生物工程研究开发中心) | Application of trichoderma hedgehog in preparation of medicine for preventing and treating pear black spot |
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Non-Patent Citations (2)
| Title |
|---|
| HASHIM I ET AL.: "Effects of Integrating Trichoderma and Fungicides on Control of White Root Disease of Hevea Rubber", J NAT RUBBER RES, vol. 12, no. 1, 1997, pages 43 - 57 * |
| MARIA,E. ET AL.: "The potency of fungal antagonists to combat root rot in industrial Acacia mangium plantation", PHYTOPATHOLOGY, vol. 101, no. 6, 2011, pages S114 * |
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