JP2023089318A - Wilt disease control agent and wilt disease control method - Google Patents

Wilt disease control agent and wilt disease control method Download PDF

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JP2023089318A
JP2023089318A JP2020083556A JP2020083556A JP2023089318A JP 2023089318 A JP2023089318 A JP 2023089318A JP 2020083556 A JP2020083556 A JP 2020083556A JP 2020083556 A JP2020083556 A JP 2020083556A JP 2023089318 A JP2023089318 A JP 2023089318A
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arabinose
bacterial wilt
wilt disease
disease
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将文 清水
Masafumi Shimizu
マレク・カレド・マームード・マリアン
Khaled Mahmoud Marian Malek
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Tokai National Higher Education and Research System NUC
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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
    • A01N25/00—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
    • A01N25/02—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests containing liquids as carriers, diluents or solvents
    • 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
    • A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
    • A01N43/02—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms
    • A01N43/04—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom
    • A01N43/14—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom six-membered rings
    • A01N43/16—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom six-membered rings with oxygen as the ring hetero atom
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    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
    • A01P3/00—Fungicides

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Abstract

To provide a wilt disease control agent containing a compound capable of protecting a plant from a wilt disease, and a method for using the same.SOLUTION: A wilt disease control agent contains arabinose. Although arabinose is effective against a wilt disease in both forms of L-arabinose and D-arabinose, L-arabinose has a higher control effect. A wilt disease control method comprises pouring an arabinose aqueous solution into the soil.SELECTED DRAWING: Figure 2

Description

本発明は、植物が青枯病に罹ることを抑制する青枯病防除剤と、青枯病の防除方法に関する。 TECHNICAL FIELD The present invention relates to a bacterial wilt disease control agent for suppressing bacterial wilt disease in plants, and a method for controlling bacterial wilt disease.

青枯病は、病原細菌のラルストニア ソラナセアラム種複合体（Ｒａｌｓｔｏｎｉａ ｓｏｌａｎａｃｅａｒｕｍ ｓｐｅｃｉｅｓ ｃｏｍｐｌｅｘ）を病原とする土壌伝染性病害である。青枯病は、２００種以上の植物に発生するが、特に、トマト、ジャガイモ、ナス等のナス科植物を主な宿主とする。青枯病の病原細菌に感染した植物体は急速に萎凋し、短期間で枯死する。青枯病の被害は世界的に発生しており、防除の対策が求められている。 Bacterial wilt is a soil-borne disease caused by the pathogenic bacterium Ralstonia solanacearum species complex. Bacterial wilt occurs in more than 200 kinds of plants, and in particular, solanaceous plants such as tomatoes, potatoes, and eggplants are the main hosts. Plant bodies infected with pathogenic bacteria of bacterial wilt rapidly wither and die in a short period of time. The damage caused by bacterial wilt is occurring worldwide, and control measures are required.

青枯病を防除する一つの対策として、抵抗性台木を用いる方法が開発されており、普及している。また、抵抗性品種の開発も進められている。しかし、抵抗性台木や抵抗性品種は、突然変異病原菌の出現により抵抗性が容易に打破されることがあり、安定的な防除対策とはなっていない。そこで、抵抗性台木の利用と並行して、化学燻蒸剤を用いた土壌燻蒸消毒が行われることがある。しかしながら、青枯病に対する土壌燻蒸消毒の効果は不安定であり、消毒後も青枯病が再発する事例が多数報告されている。また、化学燻蒸剤は概して生物に対する毒性が強く、ヒトの健康や生態系、環境への悪影響が懸念されている。そのため、青枯病の病原細菌を効果的に抑制でき、且つ安全性の高い防除剤の開発が望まれている。 As one measure for controlling bacterial wilt, a method using resistant rootstocks has been developed and is in widespread use. In addition, the development of resistant cultivars is underway. However, resistant rootstocks and resistant cultivars are not a stable control measure because the resistance can be easily overcome by the emergence of mutated pathogens. Therefore, soil fumigation using a chemical fumigant is sometimes carried out in parallel with the use of resistant rootstocks. However, the effect of soil fumigation against bacterial wilt is unstable, and many cases of recurrence of bacterial wilt have been reported even after disinfection. In addition, chemical fumigants are generally highly toxic to organisms, and there are concerns about adverse effects on human health, ecosystems, and the environment. Therefore, it is desired to develop a highly safe control agent that can effectively suppress pathogenic bacteria of bacterial wilt disease.

特許文献１には、青枯病に防除効果を示す化合物として、L体のアミノ酸であるＬ－ヒスチジン、Ｌ－アルギニン、Ｌ－リシン、Ｌ－アスパラギン酸、Ｌ－グリシン、Ｌ－フェニルアラニン、Ｌ－プロリン、Ｌ－アラニン、及びＬ－グルタミンが開示されている。特許文献２には、青枯病の防除剤として、テルペノイド関連化合物であるスクラレオールが開示されている。特許文献３には、青枯病の防除剤として、１－（４－ブロモフェニル）－６－メトキシ－１，２，３，４－テトラヒドロベータ－カルボリン及びその誘導体が開示されている。特許文献４には、ストレプトマイセス属バリダマイシン群抗生物質生産菌を培養して、バリダマイシンを生産する技術が開示されている。バリダマイシンは、青枯病防除剤として、市販されている。 Patent Document 1 discloses L-amino acids L-histidine, L-arginine, L-lysine, L-aspartic acid, L-glycine, L-phenylalanine, L- Proline, L-alanine, and L-glutamine are disclosed. Patent Document 2 discloses sclareol, which is a terpenoid-related compound, as a bacterial wilt control agent. Patent Document 3 discloses 1-(4-bromophenyl)-6-methoxy-1,2,3,4-tetrahydrobeta-carboline and derivatives thereof as a bacterial wilt control agent. Patent Document 4 discloses a technique for producing validamycin by culturing Streptomyces validamycin group antibiotic-producing bacteria. Validamycin is commercially available as a bacterial wilt disease control agent.

特許文献１に開示されているＬ体アミノ酸は、植物に根から吸収させるための好適な濃度が５ｍＭ以上である。また、特許文献２のスクラレオールや特許文献３のベータカルボリンは比較的高価な物質であり、これらを一般的な野菜の栽培に適用するためには、使用条件の最適化が必要となっている。一方、市販されている特許文献４のバリダマイシンは、トマトの生育期に使用すると、薬害を生じる恐れがあり、適用範囲が限定されている。 The L-amino acid disclosed in Patent Document 1 has a suitable concentration of 5 mM or more for absorption by plants through the roots. In addition, sclareol of Patent Document 2 and beta-carboline of Patent Document 3 are relatively expensive substances, and in order to apply them to general vegetable cultivation, it is necessary to optimize their usage conditions. On the other hand, the commercially available validamycin of Patent Document 4 may cause phytotoxicity when used during the growing season of tomatoes, and its application range is limited.

発明者らは、従来から青枯病防除法に関する研究を行っており、これまでに、青枯病に有効な根圏細菌を２株発見し、非特許文献１によって開示した。また、これらの根圏細菌を用いた青枯病防除方法を、非特許文献２に開示した。 The inventors have been conducting research on methods for controlling bacterial wilt disease, and so far have discovered two strains of rhizosphere bacteria that are effective against bacterial wilt disease, and disclosed them in Non-Patent Document 1. In addition, Non-Patent Document 2 discloses a method for controlling bacterial wilt disease using these rhizobacteria.

特許第６００７３６０号公報Japanese Patent No. 6007360 特許第５７９４５６２号公報Japanese Patent No. 5794562 特開２０１９－１５１６３１号公報JP 2019-151631 A 特開昭４７－０３９６９７号公報JP-A-47-039697

「Ｂｉｏｃｏｎｔｒｏｌ ｐｏｔｅｎｔｉａｌ ｏｆ Ｒａｌｓｔｏｎｉａ ｓｐ． ＴＣＲ１１２ ａｎｄ Ｍｉｔｓｕａｒｉａ ｓｐ． ＴＷＲ１１４ ａｇａｉｎｓｔ ｔｏｍａｔｏ ｂａｃｔｅｒｉａｌ ｗｉｌｔ．」マレク・カレド・マームード・マリアン，西岡友樹、小山博之、須賀晴久、清水将文、２０１８年、Ａｐｐｌｉｅｄ Ｓｏｉｌ Ｅｃｏｌｏｇｙ １２８号、 ７１－８０ページ``Biocontrol potential of Ralstonia sp. TCR112 and Mitsuaria sp. Applied Soil Ecology No. 128, 71 - page 80 「Ｅｎｈａｎｃｅｄ ｂｉｏｃｏｎｔｒｏｌ ｏｆ ｔｏｍａｔｏ ｂａｃｔｅｒｉａｌ ｗｉｌｔ ｕｓｉｎｇ ｔｈｅ ｃｏｍｂｉｎｅｄ ａｐｐｌｉｃａｔｉｏｎ ｏｆ Ｍｉｔｓｕａｒｉａ ｓｐ． ＴＷＲ１１４ ａｎｄ ｎｏｎｐａｔｈｏｇｅｎｉｃ Ｒａｌｓｔｏｎｉａ ｓｐ． ＴＣＲ１１２．」マレク・カレド・マームード・マリアン、森田明雄、小山博之、須賀晴久、清水将文、２０１９年、Ｊｏｕｒｎａｌ ｏｆ Ｇｅｎｅｒａｌ Ｐｌａｎｔ Ｐａｔｈｏｌｏｇｙ ８５号、 １－１３ページ"Enhanced biocontrol of tomato bacterial wilt using the combined application of Mitsuaria sp. TWR114 and nonpathogenic Ralstonia sp. TCR112." Liane, Akio Morita, Hiroyuki Koyama, Haruhisa Suga, Masafumi Shimizu, 2019, Journal of General Plant Pathology No. 85, pages 1-13.

植物栽培の現場では、安全且つ容易に使用できる青枯病の防除剤が求められている。本発明は係る現状に鑑みてなされたものであって、新規な青枯病防除剤と、青枯病の防除方法とを提供することを目的としている。 In the field of plant cultivation, there is a demand for a bacterial wilt control agent that can be used safely and easily. The present invention has been made in view of the current situation, and aims to provide a novel bacterial wilt disease control agent and a method for controlling bacterial wilt disease.

本発明の青枯病防除剤は、アラビノースを含有していることを特徴とする。 The bacterial wilt disease control agent of the present invention is characterized by containing arabinose.

発明者は、青枯病の防除剤として使用できる可能性のある物質を種々検討した結果、アラビノースが青枯病の防除効果を有することを見いだして、本発明をなすに至った。 As a result of various investigations of substances that can be used as agents for controlling bacterial wilt, the inventors have found that arabinose has an effect of controlling bacterial wilt, and have completed the present invention.

本発明の青枯病防除剤は、アラビノースがＬ－アラビノースであることが好ましい。すなわち、本発明の青枯病防除剤は、Ｌ－アラビノースを含むことを特徴とする。 In the bacterial wilt disease control agent of the present invention, arabinose is preferably L-arabinose. That is, the bacterial wilt disease control agent of the present invention is characterized by containing L-arabinose.

本発明はまた、青枯病の防除方法を提供する。本発明の青枯病の防除方法は、アラビノース水溶液を土壌に潅注することを特徴とする。 The present invention also provides a method for controlling bacterial wilt disease. The method for controlling bacterial wilt of the present invention is characterized by irrigating the soil with an aqueous solution of arabinose.

本発明の青枯病防除剤は、アラビノースを含有していることを特徴とする。アラビノースは、従来から食品添加物として用いられる五炭糖であり、その安全性が確認されている。従って、水耕栽培や土壌栽培に適用しても、環境に悪影響を及ぼすことがない。 The bacterial wilt disease control agent of the present invention is characterized by containing arabinose. Arabinose is a pentose conventionally used as a food additive, and its safety has been confirmed. Therefore, even if it is applied to hydroponics or soil cultivation, it does not adversely affect the environment.

アラビノースは、従来知られている青枯病防除剤よりも安価であることから、農業生産者が安心して使用できる。また、非常に簡易な方法で使用することができる。 Since arabinose is cheaper than conventionally known bacterial wilt disease control agents, agricultural producers can use it with confidence. Also, it can be used in a very simple way.

図１は、アラビノースの添加と他の物質の添加による青枯病の防除効果の違いをシードリング試験により比較した結果を示す図である。FIG. 1 is a diagram showing the results of comparing the difference in the control effect against bacterial wilt caused by the addition of arabinose and the addition of other substances by a seeding test. 図２は、トマトにおけるＬ－アラビノースとＤ－アラビノースの青枯病抑制効果の比較結果を示す図である。FIG. 2 is a diagram showing the results of comparison of the bacterial wilt disease inhibitory effects of L-arabinose and D-arabinose in tomato. 図３は、トマトにおけるＬ－アラビノースの土壌潅注濃度と、青枯病抑制効果の関係を示す図である。FIG. 3 is a diagram showing the relationship between soil irrigation concentration of L-arabinose in tomato and bacterial wilt disease inhibitory effect. 図４は、トマトにおけるＬ－アラビノースとＬ－ヒスチジンの青枯病抑制効果の比較結果を示す図である。FIG. 4 is a diagram showing the results of comparison of the bacterial wilt disease inhibitory effects of L-arabinose and L-histidine in tomato. 図５は、Ｌ－アラビノースを潅注したトマトの根圏と茎内の青枯病菌の生菌密度抑制効果を示す図である。FIG. 5 is a diagram showing the effect of suppressing the viable cell density of R. wilt in the rhizosphere and stem of tomatoes irrigated with L-arabinose. 図６は、ポット試験の対照区におけるトマトの青枯病の発病状態を示す図面代用写真である。FIG. 6 is a drawing-substituting photograph showing the onset state of tomato bacterial wilt disease in the control plot of the pot test. 図７は、ポット試験のＬ－アラビノースを潅注したトマトの青枯病の発病状態を示す図面代用写真である。FIG. 7 is a drawing-substituting photograph showing the disease state of bacterial wilt disease in tomatoes irrigated with L-arabinose in a pot test. 図８は、ポット試験のＤ－アラビノースを潅注したトマトの青枯病の発病状態を示す図面代用写真である。FIG. 8 is a photograph substituting for a drawing showing the disease state of bacterial wilt disease in tomatoes irrigated with D-arabinose in a pot test. 図９は、ポット試験のＬ－ヒスチジンを潅注したトマトの青枯病の発病状態を示す図面代用写真である。FIG. 9 is a drawing-substituting photograph showing the disease state of bacterial wilt disease in tomatoes irrigated with L-histidine in a pot test.

以下、本発明に係る青枯病防除剤の組成と、青枯病防除剤を用いた青枯病防除方法を実施するための形態について、図面を参照しつつ詳細に説明する。 BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the composition of the bacterial wilt disease control agent according to the present invention and the mode for carrying out the bacterial wilt disease control method using the bacterial wilt disease control agent will be described in detail with reference to the drawings.

図１は、本発明の青枯病防除剤の主な成分であるアラビノースの添加と、他の物質の添加による青枯病の防除効果の違いをシードリング試験により比較した結果を示す図である。 FIG. 1 is a diagram showing the results of comparing the difference in the control effect against bacterial wilt disease by adding arabinose, which is the main component of the bacterial wilt disease control agent of the present invention, and by adding other substances, by a seeding test. .

今回のシードリング試験では、濃度を１０ｍＭに調整したアラビノース溶液を用いた。また、アラビノースと効果を比較する比較試験として、トリプトファン、スレオニン、ヒスチジン、グリシン、グルタミン、グルタミン酸、アスパラギン酸、アスパラギン、Ｂ―アラニン、Ｌ―アラニン、コハク酸、ピルビン酸、２―オキソグルタル酸、マロン酸、グリコール酸、ガラクツロン酸、フマル酸、マンニトール、フルクトース、イノシン、キシロース、スクロース、マンノース、グルコース、ガラクトースの、計２５種類の栄養成分を、アラビノースと同一濃度の溶液に調整して試験に用いた。更に、対照区として、栄養成分の溶液の代わりに滅菌水を用いて、試験を行った。 In this seeding test, an arabinose solution adjusted to a concentration of 10 mM was used. In addition, tryptophan, threonine, histidine, glycine, glutamine, glutamic acid, aspartic acid, asparagine, B-alanine, L-alanine, succinic acid, pyruvic acid, 2-oxoglutarate, malonic acid, as a comparative test comparing the effects with arabinose. , glycolic acid, galacturonic acid, fumaric acid, mannitol, fructose, inosine, xylose, sucrose, mannose, glucose, and galactose. Furthermore, as a control group, a test was conducted using sterilized water instead of the nutrient solution.

シードリング試験の方法を説明する。直径２．５ｃｍ、高さ１０ｃｍの植物試験管に、滅菌したバーミキュライト３．４ｇを入れ、そこに青枯病菌の細胞懸濁液（約８×１０５細胞／ｍＬ）２ｍＬを潅注した。この植物試験管に、濃度を１０ｍＭに調整したアラビノース溶液１ｍＬを潅注した。 The seedling test method will be explained. A 2.5 cm diameter, 10 cm high plant test tube was filled with 3.4 g of sterilized vermiculite and irrigated with 2 mL of a cell suspension of R. solanacearum (approximately 8×10 5 cells/mL). The plant tube was irrigated with 1 mL of arabinose solution adjusted to a concentration of 10 mM.

比較試験として、バーミキュライトと青枯病菌の細胞懸濁液を入れたそれぞれの植物試験管に、１０ｍＭの栄養成分水溶液１ｍＬを、１種類ずつ潅注した。対照区として、同様の植物試験管に、滅菌水１ｍＬを潅注したものを用いた。 As a comparative test, 1 mL of 10 mM nutrient solution was irrigated into each plant test tube containing vermiculite and bacterial wilt cell suspensions. As a control, a similar plant test tube irrigated with 1 mL of sterile water was used.

それぞれの植物試験管に、トマト種子（品種：ポンテローザ）１０粒を播種し、２８℃・１２時間日長の人工気象器内で１週間培養した。培養後、トマト苗を回収し、発病度を３段階の発病指数に数値化して評価した。評価の基準は、以下の通りである。
０：無病徴。
１：胚軸の一部に壊死症状が認められる。
２：完全に枯死又は苗の大部分に壊死症状が認められる。 10 tomato seeds (cultivar: Ponterosa) were sown in each plant test tube and cultured for 1 week in an artificial climate chamber at 28°C and a day length of 12 hours. After culturing, the tomato seedlings were collected, and the degree of disease incidence was quantified into a three-stage disease incidence index and evaluated. The evaluation criteria are as follows.
0: No symptoms.
1: Necrotic symptoms are observed in part of the hypocotyl.
2: Completely dead or necrotic symptoms observed in most of the seedlings.

この発病指数を基に、発病度（以下、発病株率とも言う）を以下の式で算出した。
発病度（％）
＝Σ（発病指数×各指数に分類された苗の数）÷（調査苗数×２）×１００％ Based on this disease incidence index, the degree of disease incidence (hereinafter also referred to as diseased strain rate) was calculated by the following formula.
Disease severity (%)
= Σ (disease index x number of seedlings classified into each index) ÷ (number of seedlings surveyed x 2) x 100%

図１に示すとおり、アラビノースを潅注した場合のトマト苗の発病度が、６０％であるのに対して、比較試験に用いたすべての物質では、９０％以上の発病度となった。この結果から、アラビノースが、青枯病防除剤としての作用効果を有していることが確認された。 As shown in FIG. 1, the disease incidence of tomato seedlings irrigated with arabinose was 60%, whereas all the substances used in the comparison test resulted in a disease incidence of 90% or more. From these results, it was confirmed that arabinose has an action and effect as a bacterial wilt disease control agent.

また、アラビノースの水溶液を土壌に潅注することが、青枯病の防除方法として有効であることが確認された。 It was also confirmed that irrigation of the soil with an aqueous solution of arabinose is effective as a method for controlling bacterial wilt disease.

図２は、アラビノースの光学異性体であるＬ－アラビノースとＤ－アラビノースの青枯病防除効果の違いを比較した結果を示す図である。この試験では、ポット試験により、より長期間の防除効果を検証している。ポット試験では、濃度を０．５重量％に調整したＬ－アラビノース溶液とＤ－アラビノース溶液を用いた。また、対照区として、滅菌水を用いて、試験を行った。 FIG. 2 is a diagram showing the results of comparing the difference in the bacterial wilt disease-controlling effects of L-arabinose and D-arabinose, which are optical isomers of arabinose. In this test, the pot test verifies the control effect over a longer period of time. In the pot test, L-arabinose solution and D-arabinose solution adjusted to 0.5% by weight were used. Also, as a control group, a test was conducted using sterilized water.

ポット試験の方法を説明する。直径９ｃｍのポットに育苗培土を詰め、４葉期のトマト苗（品種：ポンテローザ）を植え付けた。一つのポットに、Ｌ－アラビノース０．５重量％水溶液を６０ｍＬ潅注した。他のポットに、Ｄ－アラビノース０．５重量％水溶液を６０ｍＬ潅注した。さらに、対照区として、同様のポットに６０ｍＬの滅菌水を潅注した。１日後、それぞれのポットに、青枯病菌の細胞懸濁液を、育苗培土１ｇあたり約１０７細胞となるように潅注接種し、３０℃のガラス温室内で２週間栽培した。この間、トマト苗における発病の有無を毎日調査した。 The pot test method will be explained. A pot with a diameter of 9 cm was filled with nursery soil, and four-leaf stage tomato seedlings (cultivar: Ponterosa) were planted. One pot was irrigated with 60 mL of 0.5% by weight L-arabinose aqueous solution. Another pot was irrigated with 60 mL of 0.5% by weight D-arabinose in water. Furthermore, as a control, a similar pot was irrigated with 60 mL of sterilized water. One day later, each pot was irrigated with a cell suspension of R. solanacearum to give about 10 7 cells per 1 g of seedling culture medium, and cultivated in a glass greenhouse at 30° C. for 2 weeks. During this period, the presence or absence of disease on the tomato seedlings was checked every day.

図２に示したとおり、Ｌ－アラビノースとＤ－アラビノースは、いずれも、青枯病の防除効果を有している。しかしながら、育苗１４日目の発病度は、Ｄ－アラビノースが６０％であるのに対し、Ｌ－アラビノースは４０％であり、Ｌ－アラビノースの方が、青枯病の防除効果がより高いことが確認された。 As shown in FIG. 2, both L-arabinose and D-arabinose have the effect of controlling bacterial wilt disease. However, the degree of disease on the 14th day of raising seedlings was 60% for D-arabinose and 40% for L-arabinose, indicating that L-arabinose is more effective in controlling bacterial wilt disease. confirmed.

以下に、Ｌ－アラビノースを青枯病防除剤として用いる場合の、好適な濃度の評価結果を示す。図３は、０．１重量％、０．２５重量％、０．５重量％の３水準の濃度のＬ－アラビノース水溶液を用いて行った、ポット試験の１４日目の結果である。ここでは、各試験の結果について、統計解析の一種であるチューキー検定を行い、Ｌ－アラビノースの濃度の違いによって発病度に有意差があるか否かを判定した。図中、異なるアルファベットを付与した試験結果の間には５％水準で有意差があることを示している。 Below, evaluation results of suitable concentration when L-arabinose is used as a bacterial wilt disease control agent are shown. FIG. 3 shows the results on the 14th day of a pot test using L-arabinose aqueous solutions with three concentrations of 0.1% by weight, 0.25% by weight, and 0.5% by weight. Tukey's test, which is a kind of statistical analysis, was performed on the results of each test to determine whether or not there was a significant difference in the severity of disease depending on the concentration of L-arabinose. The figure shows that there is a significant difference at the 5% level between the test results assigned different alphabets.

試験の結果、Ｌ－アラビノースは、０．１重量％の水溶液を潅注した場合でも、青枯病の防除効果が得られていることが確認された。また、潅注する水溶液の濃度にほぼ比例して、青枯病を防除することができ、０．２５重量％の水溶液を潅注することで、ポット試験を行った一群のトマト苗において、青枯病を防除できていることが確認された。 As a result of the test, it was confirmed that L-arabinose was effective in controlling bacterial wilt disease even when irrigated with a 0.1% by weight aqueous solution. In addition, bacterial wilt can be controlled approximately in proportion to the concentration of the aqueous solution to be irrigated. was confirmed to have been prevented.

以上のことから、本発明の青枯病の防除方法は、Ｌ－アラビノースを０．１重量％以上含む水溶液を土壌に潅注することが好ましいことが明らかとなった。さらに、より好ましくは、０．５重量％のＬ－アラビノースを含む水溶液を土壌に潅注することで、一層確実に青枯病を防除できることが明らかとなった。 From the above, it was clarified that in the method for controlling bacterial wilt of the present invention, it is preferable to irrigate the soil with an aqueous solution containing 0.1% by weight or more of L-arabinose. Furthermore, it was found that bacterial wilt can be more reliably controlled by irrigating the soil with an aqueous solution containing 0.5% by weight of L-arabinose, more preferably.

アラビノースは、食品添加物に用いられる糖類であり、土壌に高濃度で潅注しても、環境に悪影響を及ぼすことはない。従って、1重量％以上飽和濃度以下の水溶液で使用しても問題はない。しかしながら、より安価で効率よく青枯病の防除効果を得るには、０．１～１．０重量％、より好ましくは０．２５～０．５重量％の水溶液を潅注することが最適である。 Arabinose is a sugar used in food additives, and can be irrigated in soil at high concentrations without adverse environmental effects. Therefore, there is no problem even if it is used in an aqueous solution of 1% by weight or more and the saturated concentration or less. However, irrigation with an aqueous solution of 0.1 to 1.0% by weight, more preferably 0.25 to 0.5% by weight, is optimal in order to obtain the effect of controlling bacterial wilt disease more inexpensively and efficiently. .

図４に、青枯病防除剤として知られているＬ－ヒスチジンとＬ－アラビノースの効果を比較した図を示す。図４は、０．５重量％のＬ－アラビノース水溶液と、１０ｍＭのＬ－ヒスチジン水溶液とを用い、同時期に行ったポット試験の結果である。この試験により、Ｌ―ヒスチジンを潅注することで、青枯病の発病が抑制できることが改めて確認された。しかしながら、Ｌ－ヒスチジンを潅注した一群のトマトは、Ｌ－アラビノースを潅注した一群のトマトと比較して、発病する時期が早く、６日目から病変が確認される。また発病株率も高く、１４日目では発病株率が８０％に達した。一方、Ｌ－アラビノースを潅注したトマトは、１０日目から病変が確認されたものの、１４日経過後の発病度は４０％であった。このことから、Ｌ－アラビノースは、Ｌ－ヒスチジンと同等以上の青枯病の防除効果を有することが確認された。 FIG. 4 shows a diagram comparing the effects of L-histidine and L-arabinose, which are known as bacterial wilt control agents. FIG. 4 shows the results of a pot test conducted at the same time using a 0.5% by weight L-arabinose aqueous solution and a 10 mM L-histidine aqueous solution. This test confirmed anew that the onset of bacterial wilt disease can be suppressed by irrigating with L-histidine. However, the group of tomatoes irrigated with L-histidine develops disease earlier than the group of tomatoes irrigated with L-arabinose, and lesions are confirmed from day 6 onwards. In addition, the rate of diseased strains was high, reaching 80% on the 14th day. On the other hand, in the tomatoes irrigated with L-arabinose, although lesions were confirmed from the 10th day, the degree of disease development after 14 days was 40%. From this, it was confirmed that L-arabinose has an effect of controlling bacterial wilt disease equal to or greater than that of L-histidine.

図６～９に、ポット試験を行った一群のトマトの苗の図面代用写真を示す。図６は、対照区として滅菌水を潅注した後、青枯病菌を植菌して１４日経過したトマトの苗である。ほぼ全数が激しい萎凋症状を呈している。図７は、Ｌ－アラビノースを潅注した後、青枯病菌を植菌して１４日経過したトマトの苗である。図８は、Ｄ－アラビノースを潅注した後、青枯病菌を植菌して１４日経過したトマトの苗である。図９は、Ｌ－ヒスチジンを潅注した後、青枯病菌を植菌して１４日経過したトマトの苗である。Ｌ－アラビノースの青枯病防除効果が最も高いことは、試験に供したトマトの状態から確認されている。 Figures 6 to 9 show photographs in place of drawings of a group of tomato seedlings subjected to the pot test. FIG. 6 shows tomato seedlings irrigated with sterilized water as a control and then inoculated with the bacterial wilt fungus for 14 days. Almost all of them show severe atrophic symptoms. FIG. 7 shows tomato seedlings 14 days after irrigation with L-arabinose and inoculation with bacterial wilt. FIG. 8 shows tomato seedlings irrigated with D-arabinose and then inoculated with R. wilt for 14 days. FIG. 9 shows tomato seedlings 14 days after irrigation with L-histidine and inoculation with bacterial wilt. It has been confirmed from the conditions of the tomatoes subjected to the test that L-arabinose has the highest bacterial wilt control effect.

Ｌ－アラビノースが植物に対してどのように作用しているかを試験により確認した。図５は、０．５重量％のＬ－アラビノース水溶液を潅注したトマトの根圏の青枯病菌の生菌密度抑制効果と、０．５重量％のＬ－アラビノース水溶液を潅注したトマトの茎内の青枯病菌の生菌密度抑制効果を示す図である。なお、対照区は、滅菌水を潅注して行っている。図５において、記号ＮＤは、「検出されず（Ｎｏt Ｄｅｔｅｃｔｅｄ）」との結果を示しており、記号ｎ．ｓ．は、「スチューデントのｔ検定において有意差なし」との結果を示している。 Tests were conducted to confirm how L-arabinose acts on plants. FIG. 5 shows the effect of suppressing the viable bacterial density of the rhizosphere of tomatoes irrigated with a 0.5% by weight L-arabinose aqueous solution, and the effect in the tomato stems irrigated with a 0.5% by weight L-arabinose aqueous solution. is a diagram showing the effect of suppressing the viable cell density of R. wilt. The control group was irrigated with sterilized water. In FIG. 5, the symbol ND indicates a result of "Not Detected", and the symbol n. s. indicates the result of "no significant difference in Student's t-test".

青枯病菌の根圏土壌の生菌密度測定の方法は、以下の通りである。ポット試験の方法によって、０．５重量％のＬ－アラビノース水溶液を潅注処理したトマト苗のうち、青枯病菌を接種して３日後および７日後に、無症状のトマト苗をポットから抜き取り、根に強固に付着した土壌を回収した。土壌を滅菌水で段階希釈し、得られた希釈液１００μＬを青枯病菌用の半選択培地（Ｍｏｄｉｆｉｅｄ ｓｅｍｉ－ｓｅｌｅｃｔｉｖｅ ｍｅｄｉｕｍ Ｓｏｕｔｈ Ａｆｒｉｃａ）に塗抹した後、３０℃で３日間培養した。培地上に現れた青枯病菌のコロニー数から、根圏土壌中および茎内の青枯病菌の生菌密度（Ｌｏｇ ＣＦＵ／ｇ）を推定した。 The method for measuring the density of viable bacteria in the rhizosphere soil of R. wilt is as follows. By the pot test method, among the tomato seedlings irrigated with 0.5% by weight of L-arabinose aqueous solution, 3 days and 7 days after inoculation with the bacterial wilt fungus, asymptomatic tomato seedlings were removed from the pot, Soil that adhered firmly to the surface was collected. The soil was serially diluted with sterilized water, and 100 μL of the obtained diluted solution was smeared on a semi-selective medium for bacterial wilt (Modified semi-selective medium South Africa) and cultured at 30° C. for 3 days. Viable cell density (Log CFU/g) of R. wilt in the rhizosphere soil and in the stem was estimated from the number of R. solanacearum colonies that appeared on the medium.

青枯病菌の茎内の生菌密度測定の方法は、以下の通りである。ポット試験の方法によって、０．５重量％のＬ－アラビノース水溶液を潅注処理したトマト苗のうち、青枯病菌を接種して３日後および７日後に、無症状のトマト苗をポットから抜き取り、茎を回収し、乳棒と乳鉢で磨砕した。茎の磨砕物を滅菌水で段階希釈し、得られた希釈液１００μＬを青枯病菌用の半選択培地（Ｍｏｄｉｆｉｅｄ ｓｅｍｉ－ｓｅｌｅｃｔｉｖｅ ｍｅｄｉｕｍ Ｓｏｕｔｈ Ａｆｒｉｃａ）に塗抹した後、３０℃で３日間培養した。培地上に現れた青枯病菌のコロニー数から、根圏土壌中および茎内の青枯病菌の生菌密度（Ｌｏｇ ＣＦＵ／ｇ）を推定した。 The method for measuring the density of viable bacteria in the stem of R. wilt is as follows. Of the tomato seedlings irrigated with 0.5% by weight of L-arabinose aqueous solution by the pot test method, 3 days and 7 days after inoculation with the bacterial wilt fungus, asymptomatic tomato seedlings were removed from the pot, was collected and ground with a pestle and mortar. The homogenate of the stem was serially diluted with sterilized water, and 100 μL of the obtained diluted solution was smeared on a semi-selective medium for Saccharomyces cerevisiae (Modified semi-selective medium South Africa) and cultured at 30° C. for 3 days. Viable cell density (Log CFU/g) of R. wilt in the rhizosphere soil and in the stem was estimated from the number of R. solanacearum colonies that appeared on the medium.

試験の結果、Ｌ－アラビノース水溶液を潅注処理した土壌であっても、青枯病菌は、接種後３日後に増殖が始まっていることが確認された。しかしながら、Ｌ－アラビノースを潅注した場合、潅注しなかった場合と比較して、土壌中の青枯病菌の生菌密度には一定の低減効果があり、Ｌ－アラビノースが土中の青枯病菌を防除する効果があることが確認された。 As a result of the test, it was confirmed that even in the soil irrigated with the L-arabinose aqueous solution, the bacterial wilt fungus began to proliferate 3 days after the inoculation. However, when L-arabinose was irrigated, there was a certain reduction effect on the viable cell density of R. wilt in the soil compared to when it was not irrigated. It was confirmed that there is an effect to control.

一方で、Ｌ－アラビノース水溶液を潅注処理したトマトの茎内部では、青枯病菌の接種後７日後であっても、対照区と比較して、生菌密度が極めて低かった。このことは、Ｌ－アラビノースが、土壌に対する作用よりも、植物に対する作用によって青枯病の発症を抑制する効果があることを示している。 On the other hand, in the tomato stems irrigated with the L-arabinose aqueous solution, even 7 days after inoculation with the bacterial wilt, the viable cell density was extremely low compared to the control. This indicates that L-arabinose has the effect of suppressing the onset of bacterial wilt disease by acting on plants rather than acting on soil.

以上の試験結果から、アラビノースが青枯病の防除効果を有していることが明らかとなった。特にＬ－アラビノースは、青枯病の防除効果が高いことが確認された。また、本発明の青枯病防除方法は、アラビノース水溶液を土壌に潅注することで高い効果が得られることが明らかとなった。 From the above test results, it was clarified that arabinose has an effect of controlling bacterial wilt disease. In particular, L-arabinose was confirmed to be highly effective in controlling bacterial wilt disease. Moreover, it was found that the bacterial wilt disease control method of the present invention is highly effective by irrigating the soil with an aqueous solution of arabinose.

本発明の青枯病防除剤と青枯病防除方法は、青枯病菌によって引き起こされる様々な作物の病変を抑止することができるため、農薬の代替品や土壌改良剤として利用することが可能である。 Since the bacterial wilt disease control agent and the bacterial wilt disease control method of the present invention can suppress the lesions of various crops caused by the bacterial wilt disease fungus, it can be used as a substitute for agricultural chemicals and a soil conditioner. be.

Claims (3)

アラビノースを含有することを特徴とする青枯病防除剤。 A bacterial wilt disease control agent containing arabinose. アラビノースがＬ－アラビノースであることを特徴とする請求項１記載の青枯病防除剤。 2. The agent for controlling bacterial wilt disease according to claim 1, wherein the arabinose is L-arabinose. アラビノース水溶液を土壌に潅注することを特徴とする青枯病の防除方法。 A method for controlling bacterial wilt, which comprises irrigating soil with an aqueous arabinose solution.
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