JP4301920B2 - Control agent and control method for cruciferous plant diseases - Google Patents
Control agent and control method for cruciferous plant diseases Download PDFInfo
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Description
本発明は、学名バリオボラックス属(Variovorax)に属する細菌を用いたアブラナ科植物病害の防除剤および防除方法に関する。 The present invention relates to a control agent and a control method for cruciferous plant diseases using bacteria belonging to the genus Variovorax.
アブラナ科植物病害の主要な土壌病害である根こぶ病は、ハクサイ、キャベツ、カリフラワー、ブロッコリーなどアブラナ科植物300種以上の植物に発生する土壌糸状菌病害である。根こぶ病は、これらアブラナ科作物の安定した生産に支障をきたしている。病徴としては、植付け後20日位から根にこぶが生成し、初期から感染した場合にはハクサイやキャベツでは結球せず、後半以降に感染した場合でも収穫物が大きくならず、全く収穫が得られないことも見られる。 Clubroot disease, which is a major soil disease of cruciferous plant diseases, is a soil filamentous fungal disease that occurs in more than 300 species of cruciferous plants such as Chinese cabbage, cabbage, cauliflower, broccoli. Clubroot disease hinders the stable production of these cruciferous crops. As a symptom, knots are generated on the roots around 20 days after planting, and when infected from the beginning, no cabbage is formed with Chinese cabbage or cabbage. It can be seen that it cannot be obtained.
現在、アブラナ科植物の根こぶ病害の防除には、土壌消毒剤としていくつかの化学薬剤が使用されている。しかし、これらの化学薬剤には、環境上の問題や使用者及び近隣住民への安全性の問題、さらに近年の消費者の減農薬・無農薬指向に合致しないというような問題がある。更には、病原菌の菌密度が高い場合には、効果が劣ることも稀ではない。 Currently, several chemical agents are used as soil disinfectants for the control of clubroot diseases of cruciferous plants. However, these chemical agents have environmental problems, safety problems for users and neighboring residents, and problems that do not meet the recent trend of reducing pesticides and pesticides by consumers. Furthermore, when the density of pathogenic bacteria is high, it is not uncommon for the effect to be inferior.
そこで、この病害の防除には、防除効果が高く、水質汚染などの環境汚染及び安全性を満足する防除剤の開発が望まれている。 Therefore, for the control of this disease, it is desired to develop a control agent that has a high control effect and satisfies environmental pollution such as water pollution and safety.
これまでにアブラナ科植物根こぶ病に対しての生物防除法は、糸状菌(非特許文献1)やバチルス属細菌(特許文献1)がある。また本願に関連する技術として、特許文献2において、シュードモナス属ベトナミエンシス(Pseudomonas vietnamiensis)に属する微生物が、ハクサイ等のアブラナ科植物の黄化病(バーティシリウム病害)に防除効果を有することが開示されている。
本発明の課題は、アブラナ科植物病害であるハクサイ根こぶ病、キャベツ根こぶ病等のアブラナ科根こぶ病に対して防除効果が高く、環境汚染のない防除剤および防除方法を提供することである。 An object of the present invention is to provide a control agent and a control method that have a high control effect on Brassicaceae clubroot diseases such as Chinese cabbage clubroot disease and cabbage root clubroot disease that are cruciferous plant diseases, and have no environmental pollution. is there.
本発明者らは、上記課題を解決するために鋭意検討を行った結果、ハクサイ等アブラナ科植物の根こぶ病に対して高い防除効果を有するバリオボラックス属細菌CGF4526株(Variovorax sp. CGF4526)を見出し、本細菌を有効成分とする防除剤を、アブラナ科植物に接触させることで、根こぶ病害の防除を特に効果的に行えることを見出し、本発明を完成した。 As a result of intensive studies to solve the above-mentioned problems, the present inventors have found that the Variophorax sp. CGF4526 strain (Variovorax sp. CGF4526) has a high control effect on clubroot of cruciferous plants such as Chinese cabbage. The present inventors have found that a control agent containing the present bacterium as an active ingredient can be brought into contact with a cruciferous plant to effectively control root-knot diseases.
なお、本出願人は、アブラナ科植物の根こぶ病に対して高い防除効果を有するアシドボラックス属デラフィールデイ(Acidovorax delafieldii)に属する微生物を見いだし、既に特許出願をしている(特願2002−158565)。また、同じくアブラナ科植物の根こぶ病に対して高い防除効果を有するシュードモナス属ベトナミエンシス(Pseudomonas vietnamiensis)に属する細菌を見出し、特許出願している(特願2003−171251)。 In addition, the present applicant has found a microorganism belonging to the genus Acidovorax delafieldii having a high control effect against clubroot of cruciferous plants, and has already filed a patent application (Japanese Patent Application 2002). -158565). In addition, a bacterium belonging to the genus Pseudomonas vietnamiensis having a high control effect against clubroot of a cruciferous plant has been found and a patent application has been filed (Japanese Patent Application No. 2003-171251).
本発明に係るバリオボラックス属細菌CGF4526株(Variovorax sp. CGF4526)は、アブラナ科植物の根こぶ病害を効果的に防除するのみならず、保存安定性にも優れ、植物の生長を促進する作用も併せ持ち、農薬としての高い実用性を有している。 Varioborax sp. Strain CGF4526 (Variovorax sp. CGF4526) according to the present invention not only effectively controls root-knot disease of cruciferous plants, but also has excellent storage stability and promotes plant growth. In addition, it has high utility as an agrochemical.
すなわち本発明は、バリオボラックス属(Variovorax)菌株並びにその生菌を有効成分として含有することを特徴とするアブラナ科植物根こぶ病の防除剤並びにそれを用いたアブラナ科植物根こぶ病害の防除方法である。 That is, the present invention provides a control agent for Brassicaceae plant root-knot disease, comprising the Variovorax genus (Variovorax) strain and its living bacteria as active ingredients, and control of Brassicaceae plant root-knot disease using the same. Is the method.
本発明の微生物は、アブラナ科植物根こぶ病害に対して高い防除効果を有するバリオボラックス属細菌CGF4526(Variovorax sp. CGF4526)菌株である。 The microorganism of the present invention is a Variovorax sp. CGF4526 strain having a high control effect against cruciferous root-knot disease.
バリオボラックス属細菌CGF4526(Variovorax sp. CGF4526)は、イネや野菜から分離収集した約7000菌株の細菌から、糸状菌に対する抗菌活性、さらにポット栽培ハクサイによる病害防除検定試験による選抜の結果得られた菌株である。 Varioborax sp. CGF4526 (Variovorax sp. CGF4526) was obtained from approximately 7000 bacterial strains isolated and collected from rice and vegetables, as well as antibacterial activity against filamentous fungi, and further selected by disease control tests using pot-cultivated Chinese cabbage It is a strain.
バリオボラックス属細菌CGF4526(Variovorax sp. CGF4526)は、光学顕微鏡および電子顕微鏡での形態観察の結果、細胞の大きさは、1〜3μmの桿菌であり、細胞の多形性はなく、いずれも運動性を有していた。グラム反応は、陰性で、内胞子は形成しなかった。 Variovorax sp. CGF4526 (Variovorax sp. CGF4526) is a gonococcal 1 to 3 μm cell as a result of morphological observation with an optical microscope and an electron microscope. It had motility. Gram reaction was negative and no endospore was formed.
その他の細菌学的性質について、以下に示す。
1.培養的性質
CGF4526の栄養寒天培地における生育状態を以下に示す。観察は、30℃、3日間培養後に行った。
Other bacteriological properties are shown below.
1. Culture properties
The growth state of CGF4526 on nutrient agar is shown below. Observation was performed after culturing at 30 ° C. for 3 days.
CGF4526のコロニー形態は、クリーム色、円形、全縁滑らか、低凸状、光沢ありである。
2.一般的性質
The colony morphology of CGF4526 is cream, round, smooth all edges, low convex and glossy.
2. General properties
3.16s−rRNA分析
16s−rRNAの塩基配列1530bpを決定した。解析は得られた16s−rRNAの塩基配列を用いて相同性検索を行った結果、相同率の上位はバリオボラックス・パラドキサス(Variovorax paradoxus)であった。
3.16s-rRNA Analysis The base sequence 1530 bp of 16s-rRNA was determined. As a result of the analysis of homology using the obtained 16s-rRNA base sequence, the highest homology was Variovorax paradoxus.
以上の細菌学的性質により、CGF4526は、運動性を有するグラム陰性の桿菌で、カタラーゼ活性陽性、オキシダーゼ活性陽性、内胞子を形成しないことにより、シュードモナスグループに属する細菌であり、資化性からシュードモナスグループIIに属する細菌に分類された。また、16s−rRNA解析から、バリオボラックス属細菌に属することがわかった。なお、バリオボラックス属の中では、バリオボラックス・パラドキサス(Variovorax paradoxus)と考えられるものの、断定するには至っていないために、バリオボラックス・エスピー(Variovorax sp.)とした。 Due to the above bacteriological properties, CGF4526 is a gram-negative gonococci with motility and is a bacterium belonging to the Pseudomonas group by being positive for catalase activity, positive for oxidase activity and not forming endospores. It was classified into bacteria belonging to Group II. Moreover, it was found from 16s-rRNA analysis that it belongs to the genus Varioborax. In the genus Varioborax, it is considered as Variovorax paradoxus, but since it has not yet been determined, it was named Variovorax sp.
本発明の該当菌株は、独立行政法人産業技術総合研究所特許生物寄託センターに寄託され、以下の受託番号を得ている。
The corresponding strain of the present invention has been deposited with National Institute of Advanced Industrial Science and Technology, International Patent Organism Depositary, has obtained the following entrusted number.
バリオボラックス属細菌CGF4526(Variovorax sp. CGF4526):FERM BP-10160
Variovorax sp. CGF4526: FERM BP-10160
本発明におけるアブラナ科植物根こぶ病害の防除剤または防除方法を用いれば、アブラナ科植物根こぶ病の病害に対して発病を強く抑制することができ、現在使用されている化学薬剤と同等以上の効果を奏する。また、本発明の防除剤の使用は既存の化学薬剤のように農薬による環境汚染を引き起こすことはない。さらに、本発明の防除剤は市場において安定な状態で流通させることができる。 If the control agent or the control method of the Brassicaceae plant clubroot disease in the present invention, the onset can be strongly suppressed against the Brassicaceae plant clubroot disease, which is equal to or better than the currently used chemical agents. There is an effect. In addition, the use of the control agent of the present invention does not cause environmental pollution by agricultural chemicals unlike existing chemical agents. Furthermore, the control agent of the present invention can be distributed in a stable state on the market.
次に、これらの菌の培養および防除剤としての製剤は、慣用の手法で行うことができるが、以下に例をもって説明する。ここで使用する培地は菌が増殖するものであれば特に限定するものではない。生育に可能な炭素源、窒素源、無機物を適当に含有している培地であれば、天然培地、合成培地のいずれも用いることができる。培地としてはブイヨン培地、キングB培地、PS培地、PDB培地などが例示できる。以上のような培地で15〜42℃好ましくは28℃〜35℃で12〜48時間培養し増殖させたのちに遠心分離機もしくは膜濃縮機により濃縮して集菌を行い培地成分を取り除く。この操作により菌体の濃度は通常1〜50×1010cfu/ml程度に濃縮される。本発明の防除剤は、培養後の生菌をそのまま使用しても良いが、一般には農薬として使用可能な固体または液体の製剤として使用される。したがって、培養した湿菌体に糖類とグルタミン酸ナトリウム、リン酸ナトリウム緩衝液からなる保護剤を加え、真空乾燥するものである。 Next, the culture of these bacteria and the preparation as a control agent can be carried out by a conventional method, and will be described below with examples. The medium used here is not particularly limited as long as the bacteria grow. Any natural or synthetic medium can be used as long as it is a medium appropriately containing a carbon source, a nitrogen source and an inorganic substance capable of growing. Examples of the medium include bouillon medium, King B medium, PS medium, and PDB medium. After culturing and growing in the above medium at 15 to 42 ° C., preferably 28 to 35 ° C. for 12 to 48 hours, the cells are concentrated by centrifugation or a membrane concentrator to remove the medium components. By this operation, the concentration of the bacterial cells is usually concentrated to about 1 to 50 × 10 10 cfu / ml. As the control agent of the present invention, viable bacteria after culturing may be used as they are, but generally they are used as solid or liquid preparations usable as agricultural chemicals. Therefore, a protective agent comprising a saccharide, sodium glutamate, and a sodium phosphate buffer is added to the cultured wet cells and vacuum dried.
真空乾燥する前に保護剤と混合した菌体を予備凍結し、凍結したまま真空乾燥することが菌の生存率を維持するためには好ましい。なお、保護剤は水溶液の状態で菌体と混合してもよく、固体のまま混合してもよい。 In order to maintain the viability of the bacteria, it is preferable to pre-freeze the microbial cells mixed with the protective agent before vacuum drying and then vacuum-dry them while they are frozen. In addition, a protective agent may be mixed with a microbial cell in the state of aqueous solution, and may be mixed with solid.
本発明の菌体の固定化は、保護剤としてサッカロース、フルクトース、グルコースおよびソルビトールの一種または二種以上からなる糖類を用いることで行い、菌体と混合し、真空乾燥もしくは凍結真空などの方法で乾燥することによって行うことができる。 The microbial cells of the present invention are immobilized by using a saccharide consisting of one or more of saccharose, fructose, glucose and sorbitol as a protective agent, mixed with the microbial cells, and dried by a method such as vacuum drying or freezing vacuum. This can be done by drying.
ここで培養したバリオボラックス属細菌CGF4526株は、しかるべき担体と混合し、粉剤もしくは粒剤とすることができる。この場合の担体には、タルク、炭酸カルシウム、ケイソウ土等の鉱物性粉末や、ピートモス、さらには、ポリビニルアルコールなどの高分子化合物、ザンサンゴムやアルギン酸などの天然高分子化合物などがある。菌体の濃度は、105cfu/g以上、好ましくは107cfu/g以上とする。水和剤の場合には、109cfu/g以上、好ましくは1010cfu/g以上とする。 The cultivated Varioborax sp. Strain CGF4526 can be mixed with an appropriate carrier to form a powder or granule. Carriers in this case include mineral powders such as talc, calcium carbonate, and diatomaceous earth, peat moss, polymer compounds such as polyvinyl alcohol, and natural polymer compounds such as xanthan gum and alginic acid. The concentration of the bacterial cells is 10 5 cfu / g or more, preferably 10 7 cfu / g or more. In the case of a wettable powder, it is 10 9 cfu / g or more, preferably 10 10 cfu / g or more.
一般にハクサイ、キャベツ、カリフラワー、ブロッコリーなどアブラナ科植物の生産は育苗トレイに播種し、3〜5週間育苗した後に、畑に定植を行う。本発明の防除剤は、薬害の問題もなく使用できる。本発明の防除剤は、播種時の育苗培土に混合したり、播種前後の種子にかん注することができる。さらに定植前の苗の根をその希釈液に浸漬したり、育苗中または定植直前の苗にかん注する方法も可能である。播種時及び定植時の両方の処理を組み合わせると根への本菌の定着がより効率的になるため好ましい。 In general, the production of cruciferous plants such as Chinese cabbage, cabbage, cauliflower, broccoli is sown in a seedling tray, and after 3 to 5 weeks of seedling, they are planted in a field. The control agent of the present invention can be used without any problem of chemical damage. The control agent of the present invention can be mixed with seedling culture soil at the time of sowing or can be poured into seeds before and after sowing. Furthermore, it is possible to immerse the seedling roots before planting in the diluted solution, or pour them into seedlings during or just before planting. It is preferable to combine the treatments at the time of sowing and planting because the establishment of the fungus at the root becomes more efficient.
本発明の防除剤を上記方法で使用する場合、育苗培土への混合処理の場合は、土壌1L当りに1g以上混合し、均一になるように撹拌する。培土中の菌濃度は106cfu/g以上、好ましくは107cfu/g以上になるように調整する。 When using the control agent of this invention by the said method, in the case of the mixing process to raising seedling culture soil, 1g or more is mixed per 1L of soil, and it stirs so that it may become uniform. The bacterial concentration in the culture medium is adjusted to be 10 6 cfu / g or more, preferably 10 7 cfu / g or more.
また、育苗された苗を播種前後もしくは定植前に本防除剤希釈液に浸漬処理する場合の希釈液の菌濃度は107cfu/ml以上、好ましくは108cfu/ml以上になるように調整する。 In addition, when soaking seedlings before and after sowing or before planting, adjust the bacterial concentration in the diluted solution to 10 7 cfu / ml or more, preferably 10 8 cfu / ml or more. To do.
次に、バリオボラックス属細菌CGF4526の選抜について詳しく記載する。CGF4526は、イネ、野菜および雑草などから分離・収集した細菌の中から糸状菌であるトマト萎凋病菌(Fusarium oxysporum)およびハクサイ黄化病菌(Verticillium daliae)に対する抗菌活性、さらにハクサイ苗を用いた黄化病および根こぶ病ポット検定試験による選抜の結果、ハクサイ黄化病およびアブラナ科植物根こぶ病に対し、防除効果を有することを見出し、選抜された。具体的には、圃場から採取した植物の根を水道水で洗浄した後、根を細かく裁断し、滅菌水に入れ、ミキサーで潰した。その潰した液を、適当に希釈し、ブイヨン寒天培地に塗布し、培養を行った。そこで出現したコロニーを単離、保存し、供試菌株とした。 Next, selection of the Varioborax genus CGF4526 will be described in detail. CGF4526 is an antibacterial activity against fungi, Fusarium oxysporum and Verticillium daliae, among fungi isolated and collected from rice, vegetables and weeds, and yellowing using Chinese cabbage seedlings As a result of selection by the pot test of rot and clubroot, it was found that it has a controlling effect against Chinese cabbage yellowing disease and cruciferous plant clubroot. Specifically, after the plant roots collected from the field were washed with tap water, the roots were finely cut, put into sterilized water, and crushed with a mixer. The crushed liquid was appropriately diluted, applied to a bouillon agar medium, and cultured. The colonies that appeared there were isolated and stored, and used as test strains.
トマト萎凋病菌(Fusarium oxysporum)およびハクサイ黄化病菌(Verticillium daliae)に対する抗菌活性の測定は、トマト萎凋病菌(Fusarium oxysporum)およびハクサイ黄化病菌(Verticillium daliae)と供試菌株をPDA培地(ポテトデキストロース培地)上で対峙培養を行うことにより測定した。培養は、25℃で1週間または3週間行った。 Antibacterial activity against tomato wilt fungus (Fusarium oxysporum) and Chinese cabbage yellow rot fungus (Verticillium daliae) was measured using PDA medium (potato dextrose medium) with Fusarium oxysporum and tomato wilt fungus (Verticillium daliae). ) Measured by performing counterculture on The culture was performed at 25 ° C. for 1 week or 3 weeks.
さらに、対峙培養で得られた抗菌活性を有する菌株について、ハクサイ黄化病及びハクサイ根こぶ病に対して防除試験を行った。方法は、供試菌株の108cfu/ml希釈液にハクサイ苗の根を植付け時に24時間浸漬処理を行い、汚染土壌に植え付けた後、4〜5週間目に発病調査を行った。その結果、根こぶ病を強く抑制することがわかった。 Furthermore, a control test was conducted on Chinese cabbage yellowing disease and Chinese cabbage root-knot disease with respect to the strain having antibacterial activity obtained by anti-cultivation. In the method, the roots of Chinese cabbage seedlings were immersed in a diluted solution of 10 8 cfu / ml of the test strain for 24 hours and planted in contaminated soil. As a result, it was found that the clubroot was strongly suppressed.
次に実施例を示すが、本発明は以下の実施例によって限定されるものではない。なお、実施例に用いた培地の組成を次に示す。
ブイヨン培地:肉エキス 3g、ペプトン 10g、NaCl 15g、水1L、pH7.0
PDA培地(ポテトデキストロース培地):ポテト滲出液200g、ブドウ糖20g、水1L、pH5.6)。
EXAMPLES Next, examples will be shown, but the present invention is not limited to the following examples. The composition of the medium used in the examples is shown below.
Bouillon medium: meat extract 3g, peptone 10g, NaCl 15g, water 1L, pH7.0
PDA medium (potato dextrose medium): potato exudate 200 g, glucose 20 g, water 1 L, pH 5.6).
〔実施例1〕 ハクサイ根こぶ病に対する防除試験
バリオボラックス属細菌CGF4526株をブイヨン液体培地で24時間培養し、得られた菌体を遠心により分離し、供試菌の懸濁液を調整した。この懸濁液に、ハクサイ苗(品種:新理想めぐみ)を浸漬し、懸濁液に浸漬したまま24時間置いた。その後、汚染土壌に植え付けた。汚染土壌は、ハクサイ根こぶ菌に罹病したハクサイ根(根こぶ付き)をホモジナイズし混和した畑土壌を使用した。33日間後に発病の有無を調査し、防除効果の判定を行った。その結果を表2に示す。
[Example 1] Control test against Chinese cabbage root-knot disease Varioborax sp. CGF4526 strain was cultured in bouillon liquid medium for 24 hours, and the resulting cells were separated by centrifugation to prepare a suspension of the test bacteria. . In this suspension, Chinese cabbage seedling (variety: New ideal Megumi) was immersed and left in the suspension for 24 hours. Then planted in contaminated soil. As the contaminated soil, field soil was used in which Chinese cabbage roots (with root knots) affected by Chinese cabbage root koji were homogenized and mixed. After 33 days, the presence or absence of the disease was investigated and the control effect was determined. The results are shown in Table 2.
バリオボラックス属細菌CGF4526株の懸濁液の菌濃度は、1×108cfu/mlで行った。その結果、バリオボラックス属細菌CGF4526に高い防除効果が認められた。ハクサイ根こぶ病の検定は根部の根こぶの生成程度から発病度を算出し、評価した。
根部発病指数0;健全、1;根の10%以下に根こぶの付着を認める、2;根の10%から50%に根こぶの付着を認める、3;根の50%以上に根こぶの付着を認める。
発病度=100×{Σ(指数の値)×(各指数に該当する個体数)}÷{3×(供試株数)}。
The cell concentration of the suspension of the Varioborax sp. Strain CGF4526 was 1 × 10 8 cfu / ml. As a result, a high control effect was observed in the varioborax genus CGF4526. In the test of Chinese cabbage root-knot disease, the disease severity was calculated from the degree of root-knot formation at the root and evaluated.
Root disease index 0; healthy, 1; 10% or less of the roots show attachment of the humps 2; 10% to 50% of the roots show attachment of the humps; 3; 50% or more of the roots Adhesion is recognized.
Incidence of disease = 100 × {Σ (the value of the exponent) × (number body number corresponding to each index)} ÷ {3 × (test試株number)}.
〔実施例2〕 ブロッコリー根こぶ病に対する防除試験
バリオボラックス属細菌CGF4526株をブイヨン液体培地で24時間培養し、得られた菌体を遠心により分離し、供試菌の懸濁液を調整した。この懸濁液に、ブロッコリー苗(品種:緑帝)を浸漬し、懸濁液に浸漬したまま24時間置いた。その後、ハクサイ根こぶ病の罹病根の破砕液を混和した汚染土壌を詰めた10.5cmのポリポットに移植した。約30日間後に発病の有無を調査し、防除効果の判定を行った。その結果を表3に示す。バリオボラックス属細菌CGF4526株の懸濁液の菌濃度は、108cfu/mlで行った。その結果、バリオボラックス属細菌CGF4526に高い防除効果が認められた。
[Example 2] Control test against broccoli root-knot disease Varioborax sp. CGF4526 strain was cultured in bouillon liquid medium for 24 hours, and the resulting cells were separated by centrifugation to prepare a suspension of the test bacteria. . Broccoli seedlings (variety: Green Emperor) were immersed in this suspension and left for 24 hours while being immersed in the suspension. Then, it transplanted to the 10.5cm polypot filled with the contaminated soil which mixed the crushing liquid of the root of a cabbage root-knot disease. About 30 days later, the presence or absence of disease was investigated, and the control effect was determined. The results are shown in Table 3. The cell concentration of the suspension of the Varioborax sp. Strain CGF4526 was 10 8 cfu / ml. As a result, a high control effect was observed in the varioborax genus CGF4526.
〔実施例3〕 ハクサイ根こぶ病に対する防除試験(圃場試験)
バリオボラックス属細菌CGF4526株をブイヨン液体培地で24時間培養し、得られた菌体を遠心により分離し、播種土壌に108cfu/gの菌濃度に混和した。ハクサイ種子(品種;新理想めぐみ)を播種し、3週間育苗した。同様に培養したバリオボラックス属細菌CGF4526株菌の懸濁液(108cfu/ml)を調整し、ハクサイ苗を浸漬し、懸濁液に浸漬したまま24時間置いた。その後、ハクサイ根こぶ病の罹病根の破砕液を混和した畑に定植した。約65日間後に発病の有無を調査し、防除効果の判定を行った。その結果を表4に示す。その結果、バリオボラックス属細菌CGF4526に高い防除効果が認められた。
[Example 3] Control test against Chinese cabbage root-knot disease (field test)
Varioborax sp. Strain CGF4526 was cultured in a broth liquid medium for 24 hours, and the resulting cells were separated by centrifugation, and mixed with the seed soil at a bacterial concentration of 10 8 cfu / g. Chinese cabbage seeds (variety: New ideal Megumi) were sown and grown for 3 weeks. In the same manner, a suspension (10 8 cfu / ml) of the cultivated varioborax bacterium CGF4526 was prepared, and the Chinese cabbage seedling was immersed and left in the suspension for 24 hours. After that, the plants were planted in a field mixed with the crushed solution of root-knot disease. After about 65 days, the presence or absence of disease was investigated and the control effect was determined. The results are shown in Table 4. As a result, a high control effect was observed in the varioborax genus CGF4526.
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