JP2007082499A - New strain of fusarium oxysporum - Google Patents
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本発明は、フザリウム属の新菌株に関し、更に詳細には、広範な植物病害防除に有効で、かつ主要作物に病原性を示さない新規微生物フザリウム・オキシスポラムＮＰＦ−９９０１菌株、ＮＰＦ−９９０５菌株及びＮＰＦ−９９１０菌株に関する。またＮＰＦ-９９０１菌株、ＮＰＦ−９９０５菌株、ＮＰＦ−９９１０菌株の少なくとも一つを含有する植物病害防除剤、並びにこれらを利用した植物病害防除方法に関する。 The present invention relates to a new strain of the genus Fusarium, and more particularly, a novel microorganism Fusarium oxysporum NPF-9901, NPF-9905, and NPF that are effective in controlling a wide range of plant diseases and that are not pathogenic to major crops. -Relates to 9910 strains. Moreover, it is related with the plant disease control agent containing at least one of NPF-9901 strain, NPF-9905 strain, NPF-9910 strain, and the plant disease control method using these.
植物病害防除法としては、輪作、太陽熱利用等の耕種的、物理的防除、化学農薬による化学的防除、病害抵抗性品種の利用、更には、天敵、弱毒ウイルス、拮抗微生物等を用いた生物的防除が挙げられる。このうち、化学農薬、特に有機合成殺菌剤の開発研究は目覚ましく発達し、より効力が高く、多数の様々な作用を有する剤が次々と開発され、更には色々な施用法が開発されたことにより、これらを用いた化学的防除法は病害防除並びに防除作業の省力化等に大きく貢献してきた。しかしながら、近年、いわゆる薬剤耐性菌の出現により、防除効果が低下するという現象が一部作物、病害で認められており、問題化してきている。また、作物の指定産地化が進むにつれて連作を余儀なくされ、その結果、化学農薬では難防除とされる土壌伝染性病害の発生も各地で深刻な問題となっている。 Plant disease control methods include cultivating and physical control such as crop rotation and solar heat, chemical control with chemical pesticides, use of disease-resistant varieties, and biological using natural enemies, attenuated viruses, antagonistic microorganisms, etc. Control is mentioned. Among these, the development research of chemical pesticides, especially organic synthetic fungicides, has developed remarkably, and more and more effective agents have been developed one after another, and various application methods have been developed. Chemical control methods using these have greatly contributed to disease control and labor saving of control work. However, in recent years, the phenomenon that the control effect is reduced due to the appearance of so-called drug-resistant bacteria has been recognized in some crops and diseases and has become a problem. In addition, as crops are designated as local production areas, continuous crops are forced, and as a result, the occurrence of soil-borne diseases, which are difficult to control with chemical pesticides, has become a serious problem.
このような背景のもと、近年、化学農薬の使用に偏った防除体系を見直し、化学農薬からより環境への安全性が高いと想定される微生物を利用した生物防除（いわゆる生物農薬あるいは微生物農薬）も提案され、一部は実用化段階に達してきつつある。 Against this backdrop, in recent years we have reviewed the control system biased towards the use of chemical pesticides, and used biocontrol using microorganisms that are expected to be safer from chemical pesticides to the environment (so-called biological pesticides or microbial pesticides). ) Has also been proposed, and some have reached the practical stage.
植物病害の生物防除に関する研究としては、弱毒ウイルスの利用、病原菌の弱病原性或るいは非病原性系統微生物の利用、拮抗微生物の利用等が試みられている。その中でも、拮抗微生物の利用に関する研究事例は多数あり、更に拮抗微生物のうちでフザリウム属菌を用いての病害防除研究についても、多数の事例はあるが、その実用化に成功した例は極めて少ない。 As research on biological control of plant diseases, use of attenuated viruses, use of weakly pathogenic or non-pathogenic microorganisms of pathogenic bacteria, use of antagonistic microorganisms, and the like have been attempted. Among them, there are many research examples on the use of antagonistic microorganisms, and there are also many examples of disease control research using Fusarium spp. Among the antagonistic microorganisms, but very few have been successfully put to practical use. .
フザリウム属菌は、一般に土壌及び植物残さに生息する糸状菌である。本属の菌は植物病原菌に拮抗することにより、病害防除活性を発現するとされている。フザリウム属菌を利用したイネ分野における病害防除活性の過去の発明としては、イネばか苗病やイネ苗立枯細菌病などに起因する病害に有効なことが開示されている（例えば、特許文献１、２参照）。また、ナスやイチゴなどの園芸作物に発生する各種の病害に有効なことが報告されている（例えば、特許文献３、４、及び非特許文献１参照）。しかしながら、１種類のフザリウム属菌で各種の植物病害を防除する実用化システムの構築に成功した例は報告されていない。
前記したように、化学農薬による病害防除は耐性菌の出現によって防除効果が低下する可能性が高く、その場合新たなる殺菌剤の開発を必要とする。また、化学農薬では難防除とされる病害防除においては、代替或るいは併用手段を講じなくてはならない。さらに、環境に対してより安全性の高い防除技術の確立も望まれている。また従来の技術では１種類のフザリウム菌により、イネや園芸作物に発生する病害を防除することは見出されておらず、このような広範囲の病害を防除する方法としては、化学農薬との併用や複数の微生物を利用して防除するなどの手段を講じなくてはならなかった。 As described above, disease control with chemical pesticides is likely to reduce the control effect due to the emergence of resistant bacteria, and in this case, it is necessary to develop a new fungicide. In addition, in the control of diseases that are difficult to control with chemical pesticides, alternative or combined measures must be taken. Furthermore, establishment of control technology with higher safety for the environment is also desired. In addition, the conventional technology has not been found to control diseases occurring in rice or horticultural crops with one type of Fusarium fungus. As a method for controlling such a wide range of diseases, combined use with chemical pesticides And measures such as controlling using multiple microorganisms had to be taken.
本発明は、このような問題を解決し、化学農薬による防除に代わる手段、あるいは併用する手段として新しい生物農薬、しかも一種の微生物により、イネや園芸作物に発生する多数の病害の防除を可能とする生物農薬を開発する目的でなされたものであり、更には環境保全等につながるものである。更に一種類の微生物のみでイネや園芸作物などに対する植物病害防除を示す優れた技術を提供するものである。 The present invention solves such a problem, and it is possible to control a large number of diseases occurring in rice and horticultural crops by a new biological pesticide as a means to replace or use in combination with a chemical pesticide, and a kind of microorganism. It was made for the purpose of developing biological pesticides, and also leads to environmental conservation. Furthermore, the present invention provides an excellent technique for controlling plant diseases against rice and horticultural crops using only one type of microorganism.
本発明者らは、上記目的を達成するために各種植物の根圏、根面あるいは土壌より非常に多数のフザリウム属に属する糸状菌を分離し、これらの糸状菌について、各種作物病害に対する防除活性について検討した。その結果、ススキのクラウン内部から分離した菌株ＮＰＦ−９９０１、ＮＰＦ−９９０５、ＮＰＦ９９１０株がそれぞれ非常にすぐれた作物病害防除作用を有するという有用な新知見を得、本発明を完成するに至った。 In order to achieve the above object, the present inventors have isolated a large number of fungal fungi belonging to the genus Fusarium from the rhizosphere, root surface or soil of various plants, and these filamentous fungi have a controlling activity against various crop diseases. Was examined. As a result, useful new knowledge that the strains NPF-9901, NPF-9905, and NPF9910 isolated from the inside of the Susuki crown have excellent crop disease control action was obtained, and the present invention was completed.
このようにして新たに分離した３菌株は、後記する菌学的性質を有することから、いずれもフザリウム・オキシスポラム（Ｆｕｓａｒｉｕｍ ｏｘｙｓｐｏｒｕｍ）に属するものと認められたが、非常にすぐれた作物病害防除作用を有する点で従来既知の菌株とは明らかに区別することができるので、これらを新菌株と同定し、ＮＰＦ−９９０１株をフザリウム・オキシスポラム（Ｆｕｓａｒｉｕｍ ｏｘｙｓｐｏｒｕｍ）ＮＰＦ−９９０１と命名し、ＮＰＦ−９９０５をフザリウム・オキシスポラム（Ｆｕｓａｒｉｕｍ ｏｘｙｓｐｏｒｕｍ）ＮＰＦ−９９０５株、ＮＰＦ−９９１０株をフザリウム・オキシスポラム（Ｆｕｓａｒｉｕｍ ｏｘｙｓｐｏｒｕｍ）ＮＰＦ−９９１０と命名した。 Since the three newly isolated strains have the bacteriological properties described below, all of them were recognized as belonging to Fusarium oxysporum, but had an excellent crop disease control action. Therefore, these strains are identified as new strains, the NPF-9901 strain is named Fusarium oxysporum NPF-9901, and NPF-9905 is named Fusarium. -Oxysporum (Fusarium oxysporum) NPF-9905 strain and NPF-9910 strain were named Fusarium oxysporum (Fusarium oxysporum) NPF-9910.
これら本発明に係る新規微生物、フザリウム・オキシスポラム（Ｆｕｓａｒｉｕｍ ｏｘｙｓｐｏｒｕｍ）ＮＰＦ−９９０１菌株、フザリウム・オキシスポラム（Ｆｕｓａｒｉｕｍ ｏｘｙｓｐｏｒｕｍ）ＮＰＦ−９９０５菌株及びフザリウム・オキシスポラム（Ｆｕｓａｒｉｕｍ ｏｘｙｓｐｏｒｕｍ）ＮＰＦ−９９１０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し、各々、以下の寄託番号が付与されている。
フザリウム・オキシスポラムＮＰＦ−９９０１： ＦＥＲＭ Ｐ−２０４６９
フザリウム・オキシスポラムＮＰＦ−９９０５： ＦＥＲＭ Ｐ−２０４７０
フザリウム・オキシスポラムＮＰＦ−９９１０： ＦＥＲＭ Ｐ−２０４７１
These novel microorganisms according to the present invention, Fusarium oxysporum NPF-9901 strain, Fusarium oxysporum NPF-9905 strain and Fusarium oxysporum NPF-9910 industrial strain NPF-9910 Deposited at the Research Institute Patent Biological Depositary Center, the following deposit numbers are assigned.
Fusarium oxysporum NPF-9901: FERM P-20469
Fusarium oxysporum NPF-9905: FERM P-20470
Fusarium oxysporum NPF-9910: FERM P-20471
フザリウム、オキシスポラム（Ｆｕｓａｒｉｕｍ ｏｘｙｓｐｏｒｕｍ）ＮＰＦ−９９０１菌株、フザリウム、オキシスポラム（Ｆｕｓａｒｕｍ ｏｘｙｓｐｏｒｕｍ）ＮＰＦ−９９０５菌株及びフザリウム・オキシスポラム（Ｆｕｓａｒｉｕｍ ｏｘｙｓｐｏｒｕｍ）ＮＰＦ-９９１０菌株は、以下の性質を有する。 Fusarium, Fusarium oxysporum NPF-9901 strain, Fusarium, oxysporum NPF-9905 strain and Fusarium oxysporum NPF-9910 strain have the following properties.
（１）培地上での性質
ＰＤＡ培地（ポテトデキストロース）上及びツァペック培地（ＮａＮＯ3 ２．０ｇ、Ｋ2ＨＰＯ4 １．０ｇ、ＭｇＳＯ4・７Ｈ2Ｏ ０．５ｇ、ＫＣｌ ０．５ｇ、ＦｅＳＯ4・７Ｈ2Ｏ ０．０１ｇ、ショ糖３０ｇ、寒天１５ｇ、蒸留水１０００ｍｌ）上での菌糸伸長は早く、綿毛状の気中菌糸を僅かに生じ、培養子座は軟質である。ＰＤＡ培地上では桃色〜紫色の色素を培地中に生産する。
(1) Properties on medium PDA medium (potato dextrose) and Czapek medium (NaNO 3 2.0 g, K 2 HPO 4 1.0 g, MgSO 4 .7H 2 O 0.5 g, KCl 0.5 g, FeSO 4 (7H 2 O 0.01 g, sucrose 30 g, agar 15 g, distilled water 1000 ml) The mycelial elongation is fast, slightly producing a fluffy aerial mycelium, and the culture locus is soft. On the PDA medium, a pink to purple pigment is produced in the medium.
（２）形態的性質
小型分生胞子は、隔壁を有する菌糸から生じる隔壁のない短い小梗（ｐｈｉａｌｉｄｅ）上に擬頭状をなして形成される。形は長楕円または卵型で、０〜１隔壁、大きさは２〜４×６〜１１μｍ（平均３．５×８μｍ）である。大型分生胞子は、主にオレンジ色に着色した分生子座（Ｓｐｒｏｄｏｃｈｉａ）のモノフィアライド上に形成される。３〜５隔膜、３隔膜が主体であり、頂部は僅かにかぎ状に湾曲し、基部は踵状の形態を呈する。大きさは４〜６×４０〜５５μｍ（平均５×５０μｍ）である。厚膜胞子は菌糸上或るいは大型分生胞子上に形成され、大きさは直径７〜１２μｍである。
(2) Morphological properties Small conidia are formed in a pseudo-head shape on short phialides without septum that arise from hyphae having septa. The shape is oval or egg-shaped, 0-1 partition, and the size is 2-4 × 6-11 μm (average 3.5 × 8 μm). Large conidia are formed on monodialyrides of the Sporodia, which are mainly colored orange. 3 to 5 diaphragms and 3 diaphragms are the main components, the top is slightly crooked, and the base has a bowl-like form. The size is 4-6 × 40-55 μm (average 5 × 50 μm). Thick membrane spores are formed on mycelia or on large conidia and have a diameter of 7-12 μm.
（３）生理学的性質
生育温度は１０〜３５℃であり、最適温度は２５〜３０℃である。ｐＨ４．０〜８．０の間で生育可能であり、最適ｐＨは６．０〜７．０である。
(3) Physiological properties The growth temperature is 10 to 35 ° C, and the optimum temperature is 25 to 30 ° C. It can grow between pH 4.0 and 8.0, and the optimum pH is 6.0 to 7.0.
本発明は、これらの新規微生物を基本的技術思想とするものである。即ち、本発明は、植物病害防除に有効で、かつ主要作物に病原性を示さない新規微生物フザリウム・オキシスポラムＮＰＦ−９９０１菌株、ＮＰＦ−９９０５菌株及びＮＰＦ−９９１０菌株である。また、ＮＰＦ−９９０１菌株、ＮＰＦ−９９０５菌株、ＮＰＦ−９９１０菌株の少なくとも一つを含有する植物病害防除剤、並びにこれらを利用した植物病害防除方法である。 The present invention uses these novel microorganisms as a basic technical idea. That is, the present invention is a novel microorganism Fusarium oxysporum NPF-9901 strain, NPF-9905 strain and NPF-9910 strain which are effective in controlling plant diseases and does not show pathogenicity in major crops. Moreover, it is a plant disease control agent containing at least one of NPF-9901 strain, NPF-9905 strain, and NPF-9910 strain, and a plant disease control method using these.
本発明に係るフザリウム・オキシスポラム（Ｆｕｓａｒｉｕｍ ｏｘｙｓｐｏｒｕｍ）ＮＰＦ−９９０１菌株（ＦＥＲＭ Ｐ−２０４６９）、ＮＰＦ−９９０５菌株（ＦＥＲＭ Ｐ−２０４７０）又はＮＰＦ−９９１０菌株（ＦＥＲＭ Ｐ−２０４７１）は、いずれも作物に対して病原性を示すことがないので（例えば、その分生胞子懸濁液に２４時間浸漬したイネ種子を播種したり、パセリなどの園芸作物の苗に株元灌注しても何らの病原性も認められなかった）、自由に防除剤の有効成分として使用することができる。本発明の防除剤に用いるＮＰＦ−９９０１菌株、ＮＰＦ−９９０５菌株又はＮＰＦ−９９１０菌株は、ふすまなどの資材培養、固形培地上での静置培養、液体培養等の公知の手段で増殖させたものを用いればよく、生存細胞が増殖するのであれば特に培地の種類、培養条件等に制限されることはない。 Fusarium oxysporum NPF-9901 strain (FERM P-20469), NPF-9905 strain (FERM P-20470) or NPF-9910 strain (FERM P-20471) according to the present invention are all for crops. (For example, seeding rice seeds soaked in the conidial spore suspension for 24 hours, or irrigating the seedlings of horticultural crops such as parsley) It was not recognized) and can be used freely as an active ingredient of a control agent. NPF-9901 strain, NPF-9905 strain or NPF-9910 strain used for the control agent of the present invention is grown by known means such as material culture such as bran, static culture on solid medium, liquid culture, etc. As long as viable cells proliferate, there are no particular restrictions on the type of culture medium, culture conditions, and the like.
本発明で用いる防除剤としては、ＮＰＦ−９９０１菌株、ＮＰＦ−９９０５菌又はＮＰＦ−９９１０菌自体のほか、その懸濁液ないし培養液、又はその処理物（濃縮物、ペースト状物、乾燥物、希釈物等）を広く包含するものである。本発明における新規微生物ＮＰＦ−９９０１菌株、ＮＰＦ−９９０５菌株又はＮＰＦ−９９１０菌株を病害防除剤として用いる場合には、各々微生物の胞子又は培養菌体を単独で用いても良いが、通常は、担体、界面活性剤、分散剤又は補助剤等を配合して常法により例えば、粉剤、粒剤、水和剤、顆粒水和剤、フロアブル剤などの形態に製剤化して使用すると更に好ましい。好適な担体としては、例えばクレー、タルク、ベントナイト、珪藻土、ホワイトカーボン、カオリン、バーミキュライト、消石灰、珪砂、硫安、尿素等の固体担体が挙げられ、界面活性剤及び分散剤としては、例えばアルキルベンゼンスルホン酸金属塩、ポリオキシエチレンアルキルアリールエーテル、アルキル硫酸ナトリウム、アルキルナフタレンスルホン酸ナトリウム、ジナフチルメタンジスルホシ酸ナトリウム、リグニン酸ナトリウム等が挙げられる。補助剤としては、例えばカルボキシメチルセルロース、ポリオキシエチレングリコール、アラビアゴム、澱粉、乳糖等が挙げられる。 As a control agent used in the present invention, in addition to NPF-9901 strain, NPF-9905 bacterium or NPF-9910 bacterium itself, a suspension or culture solution thereof, or a treated product thereof (concentrate, paste-like product, dried product, A broad range of dilutions). When the novel microorganism NPF-9901 strain, NPF-9905 strain or NPF-9910 strain in the present invention is used as a disease control agent, each spore or cultured microbial cell may be used alone, but usually a carrier It is more preferable to add a surfactant, a dispersing agent or an auxiliary agent, and formulate and use it in the form of, for example, a powder, a granule, a wettable powder, a wettable granule, or a flowable powder by a conventional method. Suitable carriers include, for example, solid carriers such as clay, talc, bentonite, diatomaceous earth, white carbon, kaolin, vermiculite, slaked lime, silica sand, ammonium sulfate, urea, etc. Examples of surfactants and dispersants include alkylbenzenesulfonic acid. Examples thereof include metal salts, polyoxyethylene alkyl aryl ethers, sodium alkyl sulfates, sodium alkyl naphthalene sulfonates, sodium dinaphthylmethane disulfoate, and sodium lignate. Examples of the adjuvant include carboxymethyl cellulose, polyoxyethylene glycol, gum arabic, starch, lactose and the like.
次に、本発明の防除剤の使用方法を述べる。まず、地上部に発生する病害の防除に用いる場合は、胞子体、培養菌体又は生菌製剤の場合とも、水等に適宜希釈した後にスプレーヤーにより植物全体に散布することにより、予防的或るいは治療的に効果を発現する。また、種子伝染性病害又は土壌伝染性病害の防除に用いる場合は、胞子体、培養菌体又は生菌製剤の場合とも、種子又は根を浸漬、噴霧、塗布或るいは粉衣処理の少なくともひとつの処理をするか、土壌に直接混和するか、水等に懸濁した後に灌注処理することにより、種子或るいは土壌中の病原菌の生育を抑制し、防除効果を発現する。 Next, a method for using the control agent of the present invention will be described. First, when used for controlling diseases that occur on the ground, in the case of spores, cultured cells, or viable cell preparations, it can be prevented or dispersed by spraying the entire plant with a sprayer after appropriate dilution in water. Or it is therapeutically effective. In addition, when used for controlling seed-borne diseases or soil-borne diseases, seeds or roots are immersed, sprayed, applied, or dressed, in the case of spores, cultured cells, or viable preparations. , Or directly mixed in the soil, or suspended in water or the like and then irrigated, thereby suppressing the growth of pathogenic bacteria in the seed or the soil and exhibiting a control effect.
使用量としては、製剤の剤型、適用方法、適用場所、適用すべき病害の種類、所望の防除効果などに応じて使用量は適宜選定されるが、粉剤、粒剤、或るいは水で希釈する製剤の場合は、当該菌の胞子濃度が、１０2〜１０9程度、好ましくは１０4〜１０9の範囲で使用するのが望ましい。 The amount to be used is appropriately selected according to the dosage form of the preparation, application method, application place, type of disease to be applied, desired control effect, etc., but with powder, granule, or water In the case of a preparation to be diluted, it is desirable that the spore concentration of the bacterium is about 10 2 to 10 9 , preferably 10 4 to 10 9 .
本発明による新規微生物フザリウム・オキシスポラムＮＰＦ−９９０１菌株、ＮＰＦ−９９０５菌株、及びＮＰＦ−９９１０菌株の少なくとも１つを含有する防除剤は、イネ及び園芸植物に茎葉処理、植物種子又は植物根に浸漬又は粉衣処理、更には土壌に潅注又は混和処理することにより、各種作物病害に対して高い防除効果が期待でき、農業生産上有用である。また、本発明は、一種の微生物による各種の作物病害防除を可能とする生物農薬の利用であり、本発明は、省資源、省力化、環境保全等につながるものである。 A novel microorganism Fusarium oxysporum NPF-9901 strain, NPF-9905 strain, and NPF-9910 strain according to the present invention contains at least one control agent that is immersed in a foliage treatment, plant seed or plant root in rice and garden plants. High pesticidal effects against various crop diseases can be expected by powdering treatment, and further irrigation or mixing treatment on soil, which is useful for agricultural production. In addition, the present invention is the use of biological pesticides that can control various crop diseases by a kind of microorganisms, and the present invention leads to resource saving, labor saving, environmental conservation and the like.
例えば、イネ苗いもち病菌（Ｐｙｒｉｃｕｌａｒｉａ ｏｒｙｚａｅ）、イネばか苗病菌（Ｆｕｓａｒｉｕｍ ｍｏｎｉｌｉｆｏｒｍｅ）、イネもみ枯細菌病菌（Ｂｕｒｋｈｏｌｄｅｒｉａ ｇｌｕｍａｅ）、イネ褐条病菌（Ａｃｉｄｏｖｏｒａｘ ａｖｅｎａｅ）、イネ苗立枯病菌（Ｆｕｓａｒｉｕｍ ｓｐ．）といったイネ病害菌に起因する多数のイネ病害を１種類のフザリウム菌株で同時に防除することができ、本発明に係る新菌株は抗菌スペクトルが非常に広範であるという特徴を有する。したがって、病害ごとにそれぞれ別の防除剤を施用する必要がなく、省力化が達成される。農業人口の減少、農作業者の高齢化が問題となっている今日、防除回数の低減はきわめて特筆すべき著効である。 For example, rice seedling blast fungus (Pyricularia oryzae), rice shoot seedling fungus (Fusarium moniliforme), rice blast fungus (Burkholderia glumae), rice brown fungus (Acidovoraxum spp. A large number of rice diseases caused by rice pathogens can be controlled simultaneously with one type of Fusarium strain, and the new strain according to the present invention has a feature that its antibacterial spectrum is very wide. Therefore, it is not necessary to apply a different control agent for each disease, and labor saving is achieved. Today, where the decline in the agricultural population and the aging of farmers are a problem, the reduction in the number of control is a remarkable effect.
また、本発明に係る新菌株は、イネ病害を広範に防除できるだけでなく、サツマイモつる割病菌（Ｆｕｓａｒｉｕｍ ｏｘｙｓｐｏｒｕｍ）にも有効であって、各種園芸作物の病害防除にも広く使用することができる。 Moreover, the new strain according to the present invention can not only control rice diseases extensively, but also is effective against sweet potato vinespora (Fusarium oxysporum) and can be widely used for disease control of various horticultural crops.
以下、本発明を実施例により具体的に説明するが、本発明はこれらの実施例により限定されるものではない。 EXAMPLES Hereinafter, although an Example demonstrates this invention concretely, this invention is not limited by these Examples.
（実施例１：水和剤）
フザリウム・オキシスポラムＮＰＦ−９９０１菌株（ＦＥＲＭ Ｐ−２０４６９）又はフザリウム・オキシスポラムＮＰＦ−９９０５菌株（ＦＥＲＭ Ｐ−２０４７０）又はフザリウム・オキシスポラムＮＰＦ−９９１０菌株（ＦＥＲＭ Ｐ−２０４７１）をＰＤ液体培地中で培養し、得られた胞子を蒸留水に懸濁して作成したフザリウム属菌胞子懸濁液８重量部、珪藻土４０重量部、クレー５０重量部、ジナフタレンジスルホン酸ナトリウム１重量部及びリグニンスルホン酸ナトリウム１重量部を混合乾燥後、粉砕して水和剤とした。
(Example 1: wettable powder)
Culturing Fusarium oxysporum NPF-9901 strain (FERM P-20469) or Fusarium oxysporum NPF-9905 strain (FERM P-20470) or Fusarium oxysporum NPF-9910 strain (FERM P-20471); 8 parts by weight of Fusarium spore suspension prepared by suspending the obtained spores in distilled water, 40 parts by weight of diatomaceous earth, 50 parts by weight of clay, 1 part by weight of sodium dinaphthalenedisulfonate and 1 part by weight of sodium lignin sulfonate The mixture was dried and then pulverized to obtain a wettable powder.
（実施例２：粒剤）
フザリウム・オキシスポラムＮＰＦ−９９０１菌株又はフザリウム・オキシスポラムＮＰＦ−９９０５菌株又はフザリウム・オキシスポラムＮＰＦ−９９１０菌株をＰＤ液体培地中で培養し、得られた胞子を蒸留水に懸濁して作成したフザリウム属菌胞子懸濁液３０重量部、ラウリルアルコール硫酸エステルのナトリウム塩１重量部、リグニンスルホン酸ナトリウム１重量部、カルボキシメチルセルロース２重量部及びクレー９０重量部を均一に混合粉砕する。この混合物を、押出式造粒機を用いて１４〜３２メッシュの粒状に加工した後、乾燥して粒剤とした。
(Example 2: Granules)
Fusarium oxysporum NPF-9901 strain, Fusarium oxysporum NPF-9905 strain or Fusarium oxysporum NPF-9910 strain was cultured in PD liquid medium, and Fusarium spore suspension prepared by suspending the resulting spores in distilled water 30 parts by weight of a suspension, 1 part by weight of sodium salt of lauryl alcohol sulfate, 1 part by weight of sodium lignin sulfonate, 2 parts by weight of carboxymethyl cellulose and 90 parts by weight of clay are mixed and ground uniformly. This mixture was processed into granules of 14 to 32 mesh using an extrusion granulator and then dried to obtain granules.
（実施例３：イネ苗いもち病防除効果試験）
フザリウム・オキシスポラムＮＰＦ−９９０１菌、ＮＰＦ−９９０５菌及びＮＰＦ−９９１０菌をＰＤ液体培地中で培養し、得られた胞子を蒸留水に懸濁し、胞子懸濁液を調製した。水温１５℃で浸種処理を行ったイネいもち病菌罹病籾（品種：コシヒカリ）３３ｇを、この胞子懸濁液或るいは実施例１で作製したＮＰＦ−９９０１菌株の水和剤懸濁液に浴比１：２（籾：懸濁液）で、３２℃、２４時間催芽時浸漬処理を行った後、１処理あたり３３ｇを均等に３区に分けて育苗培土に播種した。３日間の加温処理（３２℃）後、ガラス温室にて１８日間育苗した後に、全苗について発病の有無を調査し、数１により発病苗率を算出した。試験規模は１区８．５×１３．５ｃｍ（１１５ｃｍ2；プラスチックパック）で３連制で行い、結果を表１に示した。
(Example 3: Rice seedling blast control effect test)
Fusarium oxysporum NPF-9901, NPF-9905, and NPF-9910 were cultured in PD liquid medium, and the resulting spores were suspended in distilled water to prepare a spore suspension. Bath ratio of 33 g of rice blast fungus (variety: Koshihikari) that had been soaked at a water temperature of 15 ° C. was added to the spore suspension or the wettable powder suspension of NPF-9901 strain prepared in Example 1. After immersing treatment at germination for 24 hours at 32 ° C. with 1: 2 (spear: suspension), 33 g per treatment was equally divided into three sections and sown on the seedling culture soil. After 3 days of heating treatment (32 ° C.), the seedlings were grown in a glass greenhouse for 18 days, and then the presence or absence of disease was investigated for all the seedlings. The test scale was 8.5 × 13.5 cm (115 cm 2 ; plastic pack) in 1 section, and the results were shown in Table 1.
（比較例１：市販フザリウム・オキシスポラム製剤（マルカライト水和剤：エーザイ生科研社製）のイネ苗いもち病防除効果試験）
実施例３と同様に、水温１５℃で浸種処理を行ったイネいもち病菌罹病籾（品種：コシヒカリ）３３ｇをマルカライト水和剤の１０倍希釈液に浴比１：２（籾：希釈液）で３２℃、２４時間催芽時浸漬処理を行った後、１処理あたり３３ｇを均等に３区に分けて育苗培土に播種した。３日間の加温処理（３２℃）後、ガラス温室にて１８日間育苗した後に、全苗について発病の有無を調査し、数１により発病苗率を算出した。試験規模は１区８．５×１３．５ｃｍ（１１５ｃｍ2；プラスチックパック）で３連制で行い、結果を表１に示した。
(Comparative example 1: Rice seedling blast control effect test of commercial fusarium oxysporum preparation (Marcalite wettable powder: Eisai Seikagaku Co., Ltd.))
In the same manner as in Example 3, 33 g of rice blast fungus (variety: Koshihikari) subjected to seeding treatment at a water temperature of 15 ° C. in a 10-fold diluted solution of marcalite wettable powder was 1: 2 (pox: diluted solution). After performing the soaking treatment at 32 ° C. for 24 hours at 33 ° C., 33 g per treatment was equally divided into three sections and sown on the seedling culture soil. After 3 days of heating treatment (32 ° C.), the seedlings were grown in a glass greenhouse for 18 days, and then the presence or absence of disease was investigated for all the seedlings. The test scale was 8.5 × 13.5 cm (115 cm 2 ; plastic pack) in 1 section, and the results were shown in Table 1.
結果は表１に示す通り、ＮＰＦ−９９０１菌、ＮＰＦ−９９０５菌、及びＮＰＦ−９９１０菌とも、イネ苗いもち病に対して明らかな発病抑制効果を示した。またＮＰＦ−９９０１菌株の分生胞子を用いて調製した水和剤製剤においても同様の発病抑制効果が認められた。一方、比較例で試験を行った市販フザリウム・オキシスポラム製剤（マルカライト水和剤）ではほとんど発病抑制効果が認められず、本発明の３菌株の有用性が明らかとなった。 As a result, as shown in Table 1, NPF-9901, NPF-9905, and NPF-9910 showed a clear disease-inhibiting effect against rice seedling blast. Moreover, the same disease-inhibiting effect was recognized also in the wettable powder formulation prepared using the conidia of NPF-9901 strain. On the other hand, the commercially available fusarium oxysporum preparation (Marcalite wettable powder) tested in the comparative example showed almost no disease suppression effect, and the usefulness of the three strains of the present invention became clear.
（実施例４：イネばか苗病防除効果試験）
フザリウム・オキシスポラムＮＰＦ−９９０１菌、ＮＰＦ−９９０５菌及びＮＰＦ−９９１０菌をＰＤ液体培地中で培養し、得られた胞子を蒸留水に懸濁し、胞子懸濁液を調製した。水温１５℃で浸種処理を行ったイネばか苗病菌罹病籾（品種：アカモチ）３３ｇを、この胞子懸濁液或るいは実施例１で作製したＮＰＦ−９９０１菌株の水和剤懸濁液に浴比１：２（籾：懸濁液）で３２℃、２４時間催芽時浸漬処理を行った後、１処理あたり３３ｇを均等に３区に分けて育苗培土に播種し、覆土した。３日間の加温処理（３２℃）後、ガラス温室にて１８日間育苗した後に、全苗について発病の有無を調査し、数１により発病苗率を算出した。試験規模は１区８．５×１３．５ｃｍ（１１５ｃｍ2；プラスチックパック）で３連制で行い、結果を表２に示した。
(Example 4: Rice blast disease control effect test)
Fusarium oxysporum NPF-9901, NPF-9905, and NPF-9910 were cultured in PD liquid medium, and the resulting spores were suspended in distilled water to prepare a spore suspension. Bath 33 g of rice seedling fungus diseased rice cultivar (variety: Akamochi) that had been soaked at a water temperature of 15 ° C. in a spore suspension or a wettable powder suspension of NPF-9901 strain prepared in Example 1. After immersing in germination at a ratio of 1: 2 (spear: suspension) at 32 ° C. for 24 hours, 33 g per treatment was equally divided into three sections and seeded in a seedling culture soil and covered with soil. After 3 days of heating treatment (32 ° C.), the seedlings were grown in a glass greenhouse for 18 days, and then the presence or absence of disease was investigated for all the seedlings. The test scale was 8.5 × 13.5 cm (115 cm 2 ; plastic pack) in 1 section, and the results were shown in Table 2.
（比較例２：市販フザリウム・オキシスポラム製剤（マルカライト水和剤：エーザイ生科研社製）のイネばか苗病防除効果試験）
実施例４と同様に、水温１５℃で浸種処理を行ったイネばか苗病菌罹病籾（品種：アカモチ）３３ｇをマルカライト水和剤の１０倍希釈液に浴比１：２（籾：希釈液）で、３２℃、２４時間催芽時浸漬処理を行った後、１処理あたり３３ｇを均等に３区に分けて育苗培土に播種し、覆土した。３日間の加温処理（３２℃）後、ガラス温室にて１８日間育苗した後に、全苗について発病の有無を調査し、数１により発病苗率を算出した。試験規模は１区８．５×１３．５ｃｍ（１１５ｃｍ2；プラスチックパック）で３連制で行い、結果を表２に示した。
(Comparative Example 2: Rice Fusarium Oxysporum Formulation (Marcalite wettable powder: Eisai Seikagaku Co., Ltd.) rice sapling disease control effect test)
In the same manner as in Example 4, 33 g of rice sapling afflicted rice cultivar (variety: Akamochi) that had been subjected to seeding treatment at a water temperature of 15 ° C. in a 10-fold diluted solution of marcalite wettable powder, 1: 2 (pox: diluted solution) ) At 32 ° C. for 24 hours, after immersing, 33 g per treatment was evenly divided into 3 sections and seeded on the seedling culture soil and covered with soil. After 3 days of heating treatment (32 ° C.), the seedlings were grown in a glass greenhouse for 18 days, and then the presence or absence of disease was investigated for all the seedlings. The test scale was 8.5 × 13.5 cm (115 cm 2 ; plastic pack) in 1 section, and the results were shown in Table 2.
結果は表２に示す通り、ＮＰＦ−９９０１菌、ＮＰＦ−９９０５菌、及びＮＰＦ−９９１０菌とも、イネばか苗病に対して明らかな発病抑制効果を示した。またＮＰＦ−９９０１菌株の分生胞子を用いて調製した水和剤製剤においても同様の発病抑制効果が認められた。一方、比較例で試験を行った市販フザリウム・オキシスポラム（マルカライト水和剤）ではほとんど発病抑制効果が認められず、本発明の３菌株の有用性が明らかとなった。 As a result, as shown in Table 2, NPF-9901 bacteria, NPF-9905 bacteria, and NPF-9910 bacteria showed a clear disease-inhibiting effect against rice sapling seedling disease. Moreover, the same disease-inhibiting effect was recognized also in the wettable powder formulation prepared using the conidia of NPF-9901 strain. On the other hand, the commercially available fusarium oxysporum (Marcalite wettable powder) tested in the comparative example showed almost no disease-inhibiting effect, and the usefulness of the three strains of the present invention became clear.
（実施例５：イネもみ枯細菌病防除効果試験）
フザリウム・オキシスポラムＮＰＦ−９９０１菌、ＮＰＦ−９９０５菌及びＮＰＦ−９９１０菌をＰＤ液体培地中で培養し、得られた胞子を蒸留水に懸濁し、胞子懸濁液を調製した。水温１５℃で浸種処理を行ったイネもみ枯細菌病菌罹病籾（品種：コシヒカリ）３３ｇを、この胞子懸濁液或るいは実施例１で作製したＮＰＦ−９９０１菌株の水和剤懸濁液に浴比１：２（籾：懸濁液）で３２℃、２４時間催芽時浸漬処理を行った後、１処理あたり３３ｇを均等に３区に分けて育苗培土に播種し、覆土した。３日間の加温処理（３２℃）後、ガラス温室にて１８日間育苗した後に、全苗について発病の有無を調査し、数１により発病苗率を算出した。試験規模は１区８．５×１３．５ｃｍ（１１５ｃｍ2；プラスチックパック）で３連制で行い、結果を表３に示した。
(Example 5: Rice blast bacterial disease control effect test)
Fusarium oxysporum NPF-9901, NPF-9905, and NPF-9910 were cultured in PD liquid medium, and the resulting spores were suspended in distilled water to prepare a spore suspension. 33 g of rice blast fungus diseased varieties (variety: Koshihikari) that had been soaked at a water temperature of 15 ° C. were added to this spore suspension or a wettable powder suspension of NPF-9901 strain prepared in Example 1. After immersing in germination for 24 hours at 32 ° C. with a bath ratio of 1: 2 (koji: suspension), 33 g per treatment was equally divided into three sections and seeded in a seedling culture soil and covered with soil. After 3 days of heating treatment (32 ° C.), the seedlings were grown in a glass greenhouse for 18 days, and then the presence or absence of disease was investigated for all the seedlings. The test scale was 8.5 × 13.5 cm (115 cm 2 ; plastic pack) in 1 section, and the results were shown in Table 3.
（比較例３：市販フザリウム・オキシスポラム製剤（マルカライト水和剤：エーザイ生科研社製）のイネもみ枯細菌病防除効果試験）
実施例５と同様に、水温１５℃で浸種処理を行ったイネもみ枯細菌病菌罹病籾（品種：コシヒカリ）３３ｇをマルカライト水和剤の１０倍希釈液に浴比１：２（籾：希釈液）で、３２℃、２４時間催芽時浸漬処理を行った後、１処理あたり３３ｇを均等に３区に分けて育苗培土に播種し、覆土した。３日間の加温処理（３２℃）後、ガラス温室にて１８日間育苗した後に、全苗について発病の有無を調査し、数１により発病苗率を算出した。試験規模は１区８．５×１３．５ｃｍ（１１５ｃｍ2：プラスチックパック）で３連制で行い、結果を表３に示した。
(Comparative Example 3: Rice Fusarium Bacterial Disease Control Effect Test of Commercial Fusarium Oxysporum Formulation (Marcalite Hydrating Agent: Eisai Seikagaku Co., Ltd.))
In the same manner as in Example 5, 33 g of rice bran bacterial disease-affected rice bran (variety: Koshihikari) that had been subjected to seeding treatment at a water temperature of 15 ° C. in a 10-fold diluted solution of marcalite wettable powder was used at a bath ratio of 1: 2 (籾: diluted). Liquid), and after immersing treatment at 32 ° C. for 24 hours, 33 g per treatment was equally divided into three sections and sown in a seedling culture soil and covered with soil. After 3 days of heating treatment (32 ° C.), the seedlings were grown in a glass greenhouse for 18 days, and then the presence or absence of disease was investigated for all the seedlings. The test scale was 8.5 × 13.5 cm (115 cm 2 : plastic pack) in 1 section, and the results were shown in Table 3.
結果は表３に示す通り、ＮＰＦ−９９０１菌、ＮＰＦ−９９０５菌、及びＮＰＦ−９９１０菌とも、イネもみ枯細菌病に対して明らかな発病抑制効果を示した。またＮＰＦ−９９０１菌株の分生胞子を用いて調製した水和剤製剤においても同様の発病抑制効果が認められた。一方、比較例で試験を行った市販フザリウム・オキシスポラム製剤（マルカライト水和剤）ではほとんど発病抑制効果が認められず、本発明の３菌株の有用性が明らかとなった。 As a result, as shown in Table 3, NPF-9901 bacteria, NPF-9905 bacteria, and NPF-9910 bacteria all showed a clear pathogenesis-inhibiting effect against the rice wilt bacterial disease. Moreover, the same disease-inhibiting effect was recognized also in the wettable powder formulation prepared using the conidia of NPF-9901 strain. On the other hand, the commercially available fusarium oxysporum preparation (Marcalite wettable powder) tested in the comparative example showed almost no disease suppression effect, and the usefulness of the three strains of the present invention became clear.
（実施例６：イネ褐条病防除効果試験）
フザリウム・オキシスポラムＮＰＦ−９９０１菌、ＮＰＦ−９９０５菌及びＮＰＦ−９９１０菌をＰＤ液体培地中で培養し、得られた胞子を蒸留水に懸濁し、胞子懸濁液を調製した。水温１５℃で浸種処理を行ったイネ褐条病菌罹病籾（品種：コシヒカリ）３３ｇを、この胞子懸濁液或るいは実施例１で作製したＮＰＦ−９９０１菌株の水和剤懸濁液に浴比１：２（籾：懸濁液）で、３２℃、２４時間催芽時浸漬処理を行った後、１処理あたり３３ｇを均等に３区に分けて育苗培土に播種し、覆土した。３日間の加温処理（３２℃）後、ガラス温室にて１８日間育苗した後に、全苗について発病の有無を調査し、数１により発病苗率を算出した。試験規模は１区８．５×１３．５ｃｍ（１１５ｃｍ2：プラスチックパック）で３連制で行い、結果を表４に示した。
(Example 6: Rice brown stripe disease control effect test)
Fusarium oxysporum NPF-9901, NPF-9905, and NPF-9910 were cultured in PD liquid medium, and the resulting spores were suspended in distilled water to prepare a spore suspension. 33 g of rice brown streak disease afflicted rice (variety: Koshihikari) that had been soaked at a water temperature of 15 ° C. was bathed in this spore suspension or a wettable powder suspension of NPF-9901 strain prepared in Example 1. After immersing treatment at germination for 24 hours at 32 ° C. at a ratio of 1: 2 (koji: suspension), 33 g per treatment was equally divided into three sections and seeded in a seedling culture soil and covered with soil. After 3 days of heating treatment (32 ° C.), the seedlings were grown in a glass greenhouse for 18 days, and then the presence or absence of disease was investigated for all the seedlings. The test scale was 8.5 × 13.5 cm (115 cm 2 : plastic pack) in 1 section, and the results were shown in Table 4.
（比較例４：市販フザリウム・オキシスポラム製剤（マルカライト水和剤：エーザイ生科研社製）のイネ褐条病防除効果試験）
実施例６と同様に、水温１５℃で浸種処理を行ったイネ褐条病菌罹病籾（品種：コシヒカリ）３３ｇをマルカライト水和剤の１０倍希釈液に浴比１：２（籾：希釈液）で、３２℃、２４時間催芽時浸漬処理を行った後、１処理あたり３３ｇを均等に３区に分けて育苗培土に播種し、覆土した。３日間の加温処理（３２℃）後、ガラス温室にて１８日間育苗した後に、全苗について発病の有無を調査し、数１により発病苗率を算出した。試験規模は１区８．５×１３．５ｃｍ（１１５ｃｍ2：プラスチックパック）で３連制で行い、結果を表４に示した。
(Comparative Example 4: Rice brown stripe disease control effect test of commercially available Fusarium oxysporum preparation (Marcalite wettable powder: manufactured by Eisai Seikaken))
In the same manner as in Example 6, 33 g of rice brown streak disease-affected rice bran (variety: Koshihikari) that had been soaked at a water temperature of 15 ° C. in a 10-fold diluted solution of marcalite wettable powder, 1: 2 (pox: diluted solution) ) At 32 ° C. for 24 hours, after immersing, 33 g per treatment was evenly divided into 3 sections and seeded on the seedling culture soil and covered with soil. After 3 days of heating treatment (32 ° C.), the seedlings were grown in a glass greenhouse for 18 days, and then the presence or absence of disease was investigated for all the seedlings. The test scale was 8.5 × 13.5 cm (115 cm 2 : plastic pack) in 1 section, and the results were shown in Table 4.
結果は表４に示すとおり、ＮＰＦ−９９０１菌、ＮＰＦ−９９０５菌、及びＮＰＦ−９９１０菌とも、イネ褐条病に対して明らかな発病抑制効果を示した。またＮＰＦ−９９０１菌株の分生胞子を用いて調製した水和剤製剤においても同様の発病抑制効果が認められた。一方、比較例で試験を行った市販フザリウム・オキシスポラム製剤（マルカライト水和剤）ではほとんど発病抑制効果が認められず、本発明の３菌株の有用性が明らかとなった。 As a result, as shown in Table 4, NPF-9901 bacteria, NPF-9905 bacteria, and NPF-9910 bacteria showed a clear pathogenesis-inhibiting effect against rice brown stripe disease. Moreover, the same disease-inhibiting effect was recognized also in the wettable powder formulation prepared using the conidia of NPF-9901 strain. On the other hand, the commercially available fusarium oxysporum preparation (Marcalite wettable powder) tested in the comparative example showed almost no disease suppression effect, and the usefulness of the three strains of the present invention became clear.
（実施例７：イネ苗立枯病防除効果試験）
フザリウム・オキシスポラムＮＰＦ−９９０１菌、ＮＰＦ−９９０５菌及びＮＰＦ−９９１０菌をＰＤ液体培地中で培養し、得られた胞子を蒸留水に懸濁し、胞子懸濁液を調製した。健全種籾（品種：コシヒカリ）３３ｇを、浴比１：２（籾：水）で１５℃５日間浸種を行った。浸種終了後、３２℃で２４時間催芽を行い、１処理あたり３３ｇを均等に３区に分けて、イネ苗立枯病菌（Ｆｕｓａｒｉｕｍ ｓｐ．）を含む育苗培土に播種し、この播種面に胞子懸濁液或るいは実施例１で作製したＮＰＦ−９９０１菌株の水和剤懸濁液を各４０ｍｌ灌注処理し、覆土した。対照化学薬剤のダコレート水和剤（クミアイ化学社製）は４００倍希釈液を４０ｍｌ処理した。３日間の加温処理（３２℃）後、ガラス温度にて１８日間育苗した後に、全苗について発病の有無を調査し、数１により発病苗率を算出した。試験規模は１区９．０×１２．５ｃｍ（１１２．５ｃｍ2；プラスチックパック）で３連制で行い、結果を表５に示した。
(Example 7: Rice seedling blight control effect test)
Fusarium oxysporum NPF-9901, NPF-9905, and NPF-9910 were cultured in PD liquid medium, and the resulting spores were suspended in distilled water to prepare a spore suspension. 33 g of healthy seed pods (variety: Koshihikari) were soaked at 15 ° C. for 5 days at a bath ratio of 1: 2 (potato: water). After completion of the sowing, germination was carried out at 32 ° C. for 24 hours, and 33 g per treatment was equally divided into three sections and sown on a seedling culture soil containing rice seedling-resistant fungus (Fusarium sp.). Each 40 ml of the turbid liquid or the wettable powder suspension of NPF-9901 strain prepared in Example 1 was irrigated and covered with soil. A control chemical agent, dacolate wettable powder (Kumiai Chemical Co., Ltd.) was treated with 40 ml of a 400-fold diluted solution. After 3 days of heating treatment (32 ° C.), the seedlings were grown for 18 days at a glass temperature, and then all seedlings were examined for the presence or absence of disease. The test scale was 9.0 × 12.5 cm (112.5 cm 2 ; plastic pack) in 1 section, and the results were shown in Table 5.
結果は表５に示す通り、ＮＰＦ−９９０１菌、ＮＰＦ−９９０５菌、及びＮＰＦ−９９１０菌とも、イネ苗立枯病に対して明らかな発病抑制効果を示した。またＮＰＦ−９９０１菌株の分生胞子を用いて調製した水和剤製剤においても同様の発病抑制効果が認められた。またその効果は市販されている化学農薬と同等の効果であり、本発明の３菌株の有用性が明らかとなった。 As a result, as shown in Table 5, NPF-9901 bacteria, NPF-9905 bacteria, and NPF-9910 bacteria all showed a clear disease-suppressing effect against rice seedling blight. Moreover, the same disease-inhibiting effect was recognized also in the wettable powder formulation prepared using the conidia of NPF-9901 strain. Moreover, the effect is equivalent to a commercially available chemical pesticide, and the usefulness of the three strains of the present invention has been clarified.
（実施例８：サツマイモつる割病防除効果試験）
フザリウム・オキシスポラムＮＰＦ−９９０１菌、ＮＰＦ−９９０５菌及びＮＰＦ−９９１０菌をＰＤ液体培地中で培養し、得られた胞子を蒸留水に懸濁し、胞子懸濁液を調製した。この胞子懸濁液或るいは実施例１で作製したＮＰＦ−９９０１菌株の水和剤懸濁液に、サツマイモ健全苗（品種：ベニコマチ）を１晩浸漬処理した。処理苗は予め作成しておいたサツマイモつる割病汚染土壌を詰めた１／５０００アールワグネルポットに定植した。１試験区につき１０本の苗を供試し、定植２２日後に全苗の発病程度を表６の基準によって調査し、数２により発病度を算出した。結果を表７に示した。
(Example 8: Sweet potato vine split disease control effect test)
Fusarium oxysporum NPF-9901, NPF-9905, and NPF-9910 were cultured in PD liquid medium, and the resulting spores were suspended in distilled water to prepare a spore suspension. In this spore suspension or a wettable powder suspension of NPF-9901 produced in Example 1, sweet potato seedlings (variety: Benikomachi) were immersed overnight. The treated seedlings were planted in a 1/5000 Earl Wagner pot filled with previously prepared sweet potato vine split disease contaminated soil. Ten seedlings were tested per test plot, and after 22 days of planting, the severity of all seedlings was examined according to the criteria shown in Table 6, and the disease severity was calculated from Equation 2. The results are shown in Table 7.
（比較例５：市販フザリウム・オキシスポラム製剤（マルカライト水和剤：エーザイ生科研社製）のサツマイモつる割病防除効果試験）
実施例８と同様に、サツマイモ健全苗（品種：ベニコマチ）をマルカライト水和剤の１０倍希釈液に１晩浸漬処理した。処理苗は予め作成しておいたサツマイモつる割病汚染土壌を詰めた１／５０００アールワグネルポットに定植した。１試験区につき１０本の苗を供試し、定植２２日後に全苗の発病程度を表６の基準によって調査し、数２により発病度を算出した。結果を表７に示した。
(Comparative example 5: Sweet potato vine split disease control effect test of commercially available fusarium oxysporum preparation (Marcalite wettable powder: Eisai Seikagaku Co., Ltd.))
As in Example 8, healthy sweet potato seedlings (variety: Benikomachi) were immersed in a 10-fold diluted solution of Marcalite wettable powder overnight. The treated seedlings were planted in a 1/5000 Earl Wagner pot filled with previously prepared sweet potato vine split disease contaminated soil. Ten seedlings were tested per test plot, and after 22 days of planting, the severity of all seedlings was examined according to the criteria shown in Table 6, and the disease severity was calculated from Equation 2. The results are shown in Table 7.
結果は表７に示す通り、ＮＰＦ−９９０１菌、ＮＰＦ−９９０５菌、及びＮＰＦ−９９１０菌とも、サツマイモつる割病に対して明らかな発病抑制効果を示した。またＮＰＦ−９９０１菌株の分生胞子を用いて調製した水和剤製剤においても同様の発病抑制効果が認められた。また、比較例で試験を行った市販フザリウム・オキシスポラム製剤（マルカライト水和剤）でも高い発病抑制効果が認められ、本発明の３菌株の有用性は市販のフザリウム・オキシスポラム製剤と同等のであることが明らかとなった。 As a result, as shown in Table 7, NPF-9901 bacteria, NPF-9905 bacteria, and NPF-9910 bacteria showed a clear disease-inhibiting effect against sweet potato vine split disease. Moreover, the same disease-inhibiting effect was recognized also in the wettable powder formulation prepared using the conidia of NPF-9901 strain. In addition, the commercially available fusarium oxysporum formulation (Marcalite wettable powder) tested in the comparative example also shows a high disease-inhibiting effect, and the usefulness of the three strains of the present invention is equivalent to the commercially available fusarium oxysporum formulation. Became clear.
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| CN106834129A (en) * | 2016-09-28 | 2017-06-13 | 福建省农业科学院作物研究所 | The non-pathogenic mutation bacterial strain of Fusarium oxysporum sweet potato specialized form with biological and ecological methods to prevent plant disease, pests, and erosion function |
| CN107034143A (en) * | 2016-09-28 | 2017-08-11 | 福建省农业科学院作物研究所 | Prevent and treat the non-pathogenic mutation bacterial strain of Fusarium oxysporum sweet potato specialized form of stem rot of sweet potato |
| CN110558337A (en) * | 2019-07-19 | 2019-12-13 | 湖南科技学院 | biocontrol preparation for preventing and treating rice blast and preparation method thereof |
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