JP2019038788A - Microbial-containing control material and control method for potato scab - Google Patents
Microbial-containing control material and control method for potato scab Download PDFInfo
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本発明は、ジャガイモそうか病菌に対する防除作用を有する微生物等に関する。 The present invention relates to a microorganism or the like having a control action against potato scab fungus.
本発明はまた、当該微生物を用いてジャガイモの種子、種イモ、葉及び栽培土壌の少なくとも一つを処理することを含むジャガイモそうか病の防除方法に関する。 The present invention also relates to a method for controlling potato scab comprising treating at least one of potato seeds, seed potatoes, leaves and cultivated soil using the microorganism.
本発明はさらに、上記処理と、他の薬剤、例えば栽培土壌への土壌消毒剤の処理の組み合わせによる防除体系におけるジャガイモそうか病防除方法に関する。 The invention further relates to a method for controlling potato scab in a control system by a combination of the above treatments and the treatment of other agents, such as soil disinfectants on cultivated soil.
ジャガイモそうか病は世界的に問題となっている難防除土壌病害であり、その対策は非常に重要な課題となっている。日本国内のジャガイモ生産高は2014年度の統計で245.6万t（農林水産省、統計資料）、その約8割の191.6万tが北海道で、次いで長崎県（10.5万t）、鹿児島県（9.4万t）、茨城県（4.2万t）の順に生産高が多い。そうか病に罹病したジャガイモは食用、食品加工用としての価値を著しく落とし、被害がひどい場合には澱粉の含有量や品質の低下を起こす。そうか病は連作障害であり、集約的な生産体制では有効な対策が難しいのが現状である。病原菌は罹病イモの植え付けにより広がり、被害植物とともに土壌中で越冬し、汚染土壌の飛散等によって伝染する。若い塊茎ほど罹病しやすく、皮目・気孔・傷口から侵入するため発症の確認ができず、栽培期間中の対策が困難である。 Potato scab is a difficult problem for controlling soil diseases worldwide, and its control is a very important issue. Potato production in Japan was 2.456,000 tons (Ministry of Agriculture, Forestry and Fisheries, statistical data) in statistics of 2014, and about 19.1 million tons of about 80% is Hokkaido, followed by Nagasaki prefecture (105,000 tons), Kagoshima prefecture (94,000 t), Ibaraki prefecture (42,000 t) in the order of production volume. Potatoes that are affected by scab significantly reduce their value for food and food processing, and if the damage is severe, they cause deterioration in starch content and quality. Scab disease is a disorder of continuous cropping, and effective measures are difficult at present in an intensive production system. The pathogen spreads by planting diseased potatoes, and overwinters in the soil with the affected plants, and is transmitted by scattering of the contaminated soil. Younger tubers are more susceptible to disease, and they can not be confirmed as they develop from skin, pores, and wounds, making it difficult to take measures during the cultivation period.
ジャガイモそうか病の原因菌はストレプトマイセス属菌という放線菌の仲間で、高温、乾燥の土壌条件下でよく繁殖し、有機物に富む中性〜アルカリ性の土壌を好み、作物がなくとも土壌中の有機物を栄養源として極めて長期間生存する。日本国内では３種類のそうか病菌が分布し、ジャガイモの主要な産地である北海道、長崎県、鹿児島県で大きな問題となっている。そうか病の症状は、病原菌が生産するタキストミンという物質（毒素）に対して抵抗するためにかさぶた状の組織を形成するため生ずる。 The causative agent of potato scab is an actinomycete member of the genus Streptomyces, which propagates well under hot and dry soil conditions and prefers neutral to alkaline soils that are rich in organic matter, even in the soil without crops Survive for a very long time using the organic matter of In Japan, three kinds of scab fungus are distributed, and it is a major problem in Hokkaido, Nagasaki and Kagoshima prefectures, which are major producing areas of potato. The symptoms of scab are caused by the formation of scaly tissue to resist the substance (toxin) called tachytomin produced by the pathogen.
本病害の対策としては、硫酸アンモニウム、硫酸第1鉄、及び硫黄華等の酸性資材の施用による土壌酸性化、並びに化学農薬による種イモの消毒の他、土壌消毒等が知られている。しかし、土壌消毒を行った圃場では、土壌中の微生物層が貧弱となるため、汚染されたイモを植えた場合は、逆にそうか病の発生率が増加することが知られている。また、別の対策としては、くん蒸剤（クロルピクリン剤等）による土壌消毒や種イモ消毒が行われているが、環境への影響が懸念される。さらにまた、ｐＨを低く調整することによる対策も行われているが、発生の激しい圃場では効果が低いことと、収量が低下してしまうことが問題となっている。最近では、太陽熱消毒、還元消毒の取組も始まっているが、大規模面積で栽培されている圃場では実施が難しい。 As measures against this disease, soil acidification by application of acidic materials such as ammonium sulfate, ferrous sulfate and sulfur blue, and soil disinfection etc. in addition to disinfection of seed potato by chemical pesticides are known. However, in a field where soil disinfection has been carried out, the microbial layer in the soil is poor, and it is known that the incidence of scab on the contrary increases when planted with contaminated potatoes. In addition, as another measures, soil disinfection and seed potato disinfection with fumigant (chlorpicrin agent etc.) are carried out, but there is concern about environmental impact. Furthermore, although the countermeasure by adjusting pH low is also performed, it is a problem that an effect is low and the yield falls in the field where the outbreak is severe. Recently, solar heat disinfection and reduction disinfection efforts have also begun, but implementation is difficult in fields grown on large-scale areas.
上記の通り、本病害の防除法としては、種イモ及び土壌の化学農薬による消毒が一般的であるが、近年は農薬の使用規制が厳しくなっている。また、農薬を使用しない安心かつ安全な農作物を消費者が求める傾向にあり、環境負荷の点からも農薬の使用回数の低減が望ましいことから、農薬に代わってジャガイモそうか病を防除できる資材が求められている。化学農薬を使用しない防除方法として太陽熱消毒、有用微生物の利用（非特許文献1）、及び輪作等も行なわれているが、そうか病を防除するためのより有効な技術の開発が必要とされている。 As described above, as a method for controlling this disease, disinfection of seed potatoes and soil with chemical pesticides is generally used, but in recent years, the use regulations of pesticides have become strict. In addition, consumers tend to demand safer and safer crops that do not use pesticides, and it is desirable to reduce the number of times pesticides are used from the point of environmental load, so materials that can control potato scab can be substituted for pesticides. It has been demanded. Solar heat disinfection, utilization of useful microorganisms (Non-Patent Document 1), and rotation are also conducted as control methods that do not use chemical pesticides, but development of more effective techniques for controlling scab disease is required. ing.
本発明は、ジャガイモそうか病を防除及び／又は予防に有効な資材を提供することを課題とする。また、本資材を用い、減農薬による環境保全型農業が可能となるような、ジャガイモそうか病の生物的防除方法を提供することを課題とする。 An object of the present invention is to provide a material effective for controlling and / or preventing potato scab. Another object of the present invention is to provide a method for biological control of potato scab that enables environmental conservation agriculture with reduced pesticides using this material.
本発明者らは、上記課題を解決するため鋭意研究を行い、ジャガイモ栽培圃場より根圏土壌又はジャガイモ表皮に存在する微生物群からジャガイモそうか菌を抑制する微生物の検索を行い、ジャガイモそうか菌に対して高い防除作用（例えば、抑制能）を有する微生物を見出し、本発明を完成するに至った。本発明者は、特に、ストレプトマイセス・パナシラディシス(Streptomyces panaciradicis)PSA-107（受託番号：NITE P-02407）、バチルス・メガテリウム(Bacillus megaterium) PSB-139（受託番号：NITE P-02408）、バチルス・ベレゼンシス（Bacillus velezensis）NA5-3（受託番号：NITE P-02482）、バチルス・アリアブハッタイ（Bacillus aryabhattai）NA6（受託番号：NITE P-02483）、タラロマイセス・フラバス（Talaromyces flavus）PSF-234（受託番号：NITE P-02426）、及びタラロマイセス・ピノフィラス（Talaromyces pinophilus）PSF-332（受託番号：NITE P-02427）からなる群から選択される微生物がジャガイモそうか病の防除に有効であることを見出した。 The present inventors conducted intensive research to solve the above-mentioned problems, and searched for microorganisms that suppress potato scabies from microorganisms present in the rhizosphere soil or potato skin from a potato cultivation field, We have found a microorganism having a high control activity (for example, the ability to suppress) against In particular, the present inventors have identified Streptomyces panaciradicis PSA-107 (Accession number: NITE P-02407), Bacillus megaterium PSB-139 (Accession number: NITE P-02408), Bacillus.・ Bellezensis (Bacillus velezensis) NA5-3 (Accession No .: NITE P-02482), Bacillus ariabhattai (Bacillus aryabhattai) NA6 (Accession No .: NITE P-02483), Talaromyces flavus (Talaromyces flavus) PSF-234 ( The microorganism selected from the group consisting of Accession No .: NITE P-02426) and Talaromyces pinophilus PSF-332 (Accession No .: NITE P-02427) is effective for controlling potato scab I found it.
したがって、本発明は、以下の態様を包含する。
（１）ストレプトマイセス・パナシラディシス（Streptomyces panaciradicis） PSA-107株（受託番号：NITE P-02407）、又はジャガイモそうか病菌に対し防除作用を有するその派生株。
（２）バチルス・メガテリウム（Bacillus megaterium） PSB-139株（受託番号：NITE P-02408）、バチルス・ベレゼンシス（Bacillus velezensis）NA5-3株（受託番号：NITE P-02482）、及びバチルス・アリアブハッタイ（Bacillus aryabhattai）NA6株（受託番号：NITE P-02483）からなる群から選択される菌株、又はジャガイモそうか病菌に対し防除作用を有するその派生株。
（３）タラロマイセス・フラバス（Talaromyces flavus）PSF-234株（受託番号：NITE P-02426）、若しくはタラロマイセス・ピノフィラス（Talaromyces pinophilus）PSF-332株（受託番号：NITE P-02427）、又はジャガイモそうか病菌に対し防除作用を有するその派生株。
（４）（１）〜（３）のいずれかに記載の株からなる群から選択される少なくとも１種の菌株を含むことを特徴とする、ジャガイモそうか病防除資材。
（５）菌株が、ストレプトマイセス・パナシラディシス（Streptomyces panaciradicis） PSA-107株（受託番号：NITE P-02407）、バチルス・メガテリウム（Bacillus megaterium） PSB-139株（受託番号：NITE P-02408）、バチルス・ベレゼンシス（Bacillus velezensis）NA5-3株（受託番号：NITE P-02482）、バチルス・アリアブハッタイ（Bacillus aryabhattai）NA6株（受託番号：NITE P-02483）、タラロマイセス・フラバス（Talaromyces flavus）PSF-234株（受託番号：NITE P-02426）、及びタラロマイセス・ピノフィラス（Talaromyces pinophilus）PSF-332株（受託番号：NITE P-02427）からなる群から選択される少なくとも１種の菌株であることを特徴とする、（４）記載のジャガイモそうか病防除資材。
（６）（１）〜（３）のいずれかに記載の株からなる群から選択される少なくとも１種の菌株を用いて、ジャガイモの種子、種イモ、葉及び栽培土壌の少なくとも一つを処理することを含むことを特徴とする、ジャガイモそうか病の防除方法。
（７）菌株が、ストレプトマイセス・パナシラディシス（Streptomyces panaciradicis） PSA-107株（受託番号：NITE P-02407）、バチルス・メガテリウム（Bacillus megaterium） PSB-139株（受託番号：NITE P-02408）、バチルス・ベレゼンシス（Bacillus velezensis）NA5-3株（受託番号：NITE P-02482）、バチルス・アリアブハッタイ（Bacillus aryabhattai）NA6株（受託番号：NITE P-02483）、タラロマイセス・フラバス（Talaromyces flavus）PSF-234株（受託番号：NITE P-02426）、及びタラロマイセス・ピノフィラス（Talaromyces pinophilus）PSF-332株（受託番号：NITE P-02427）からなる群から選択される少なくとも１種の菌株であることを特徴とする、（６）記載の方法。
（８）（４）又は（５）記載のジャガイモそうか病防除資材を用いて、ジャガイモの種子、種イモ、葉及び栽培土壌の少なくとも一つを処理することを含むことを特徴とする、ジャガイモそうか病の防除方法。
Therefore, the present invention includes the following aspects.
(1) Streptomyces panaciradicis PSA-107 strain (Accession No .: NITE P-02407), or a derivative thereof having a control activity against potato scab fungus.
(2) Bacillus megaterium (Bacillus megaterium) PSB-139 strain (Accession No .: NITE P-02408), Bacillus verezensis (Bacillus velezensis) NA5-3 strain (Accession No .: NITE P-02482), and Bacillus ariab. A strain selected from the group consisting of Thai (Bacillus aryabhattai) strain NA6 (Accession No .: NITE P-02483), or a derivative thereof having a controlling action against potato scab.
(3) Talaromyces flavus (Talaromyces flavus) PSF-234 (Accession No .: NITE P-02426), or Talaromyces pinophilus PSF-332 (Accession No .: NITE P-02427), or potato scum The derivative which has control action with respect to disease bacteria.
(4) A potato scab controlling material comprising at least one strain selected from the group consisting of the strains according to any of (1) to (3).
(5) The strain is Streptomyces panacradicis PSA-107 (Accession No .: NITE P-02407), Bacillus megaterium PSB-139 (Accession No .: NITE P-02408), Bacillus velezensis strain NA5-3 (Accession No .: NITE P-02482), Bacillus ariab hattai strain NA6 (Accession No .: NITE P-02483), Talaromyces flavus (PSA) PSF At least one strain selected from the group consisting of -234 strain (Accession number: NITE P-02426), and Talaromyces pinophilus PSF-332 strain (Accession number: NITE P-02427) The potato scab control material according to (4), which is characterized by
(6) Using at least one strain selected from the group consisting of strains according to any of (1) to (3), at least one of potato seeds, seed potatoes, leaves and cultivated soil is treated A method for controlling potato scab, comprising:
(7) The strain is Streptomyces panacradicis PSA-107 (Accession No .: NITE P-02407), Bacillus megaterium PSB-139 (Accession No .: NITE P-02408), Bacillus velezensis strain NA5-3 (Accession No .: NITE P-02482), Bacillus ariab hattai strain NA6 (Accession No .: NITE P-02483), Talaromyces flavus (PSA) PSF At least one strain selected from the group consisting of -234 strain (Accession number: NITE P-02426), and Talaromyces pinophilus PSF-332 strain (Accession number: NITE P-02427) (6) the method according to (6).
(8) A potato comprising: treating at least one of potato seeds, seed potatoes, leaves and cultivated soil, using the potato scab control material according to (4) or (5). How to control scab.
本発明は、ジャガイモそうか菌に対して高い防除作用を有する微生物を特徴としており、この微生物によってジャガイモそうか菌を防除することが可能である。 The present invention is characterized by a microorganism having a high controlling action on potato scaly fungus, which is capable of controlling potato scaly fungus.
以下、本発明について詳細に説明する。
本発明は、ジャガイモそうか病菌に対する防除作用（例えば、抑制能）を有する新規微生物を提供する。
Hereinafter, the present invention will be described in detail.
The present invention provides a novel microorganism having a control action (for example, the ability to suppress) against potato scab.
１．微生物
本発明の微生物としては、ジャガイモそうか病菌に対する防除作用（例えば、抑制能）を有する微生物が用いられる。放線菌（Streptomyces属）、細菌（Bacillus属）、糸状菌（Talaromyces属）等、ジャガイモそうか病菌に対し防除作用（例えば、抑制能）を有する微生物であれば、本発明の微生物として用いることができる。このうち、ストレプトマイセス・パナシラディシス(Streptomyces panaciradicis)PSA-107（受託番号：NITE P-02407）、バチルス・メガテリウム(Bacillus megaterium) PSB-139（受託番号：NITE P-02408）、バチルス・ベレゼンシス（Bacillus velezensis）NA5-3（受託番号：NITE P-02482）、バチルス・アリアブハッタイ（Bacillus aryabhattai）NA6（受託番号：NITE P-02483）、タラロマイセス・フラバス（Talaromyces flavus）PSF-234（受託番号：NITE P-02426）、及びタラロマイセス・ピノフィラス（Talaromyces pinophilus）PSF-332（受託番号：NITE P-02427）からなる群から選択される菌株、又はその派生株が、ジャガイモそうか病菌に対する防除作用（例えば、抑制能）が高く有効に用いられる。
1. Microorganism As the microorganism of the present invention, a microorganism having a control action (for example, suppressing ability) against potato scab fungus is used. Microorganisms having a control action (for example, inhibition ability) against potato scab, such as actinomycetes (Streptomyces genus), bacteria (Bacillus genus), filamentous fungi (Talaromyces genus), etc., may be used as the microorganism of the present invention it can. Among these, Streptomyces panaciradicis (Streptomyces panaciradicis) PSA-107 (Accession No .: NITE P-02407), Bacillus megaterium (Bacillus megaterium) PSB-139 (Accession No .: NITE P-02408), Bacillus berezensis (Bacillus) velezensis) NA5-3 (Accession No .: NITE P-02482), Bacillus aryabhattai NA6 (Accession No .: NITE P-02483), Talaromyces flavus (Talaromyces flavus) PSF-234 (Accession No .: NITE) P-02426), and a strain selected from the group consisting of Talaromyces pinophilus PSF-332 (Accession No .: NITE P-02427), or a derivative thereof has a control activity against potato scab (eg, Control ability) is used effectively.
＜有用微生物の分離＞
圃場から健全なじゃがいもを掘り取り、水道水で十分に洗浄して、塊茎等に付着した泥等を洗い落したのち、これらの部位を摩砕して得られた試料を寒天培地に塗布してコロニーを発生させた。次いで、各コロニーを単離、培養してＤＮＡを抽出した。
<Separation of useful microorganisms>
After digging healthy potatoes from the field and thoroughly washing with tap water to wash away mud etc. adhering to tubers etc., a sample obtained by grinding these sites is applied to agar medium A colony was generated. Then, each colony was isolated and cultured to extract DNA.
以下に菌種の同定について記載する。
DNA抽出はDSPD法（Ikeda et al., 2004, Microbes Environ 19, 301-309）に従って行い、またPCR条件は以下の通りである。
The identification of bacterial species is described below.
DNA extraction is performed according to the DSPD method (Ikeda et al., 2004, Microbes Environ 19, 301-309), and the PCR conditions are as follows.
プライマーについて、細菌・放線菌の同定に使用したプライマーは、ユニバーサルプライマーの27Fと1525Rである。プライマーのヌクレオチド配列（5’→3’）は以下のとおりである。
27F：AGAGTTTGATCMTGGCTCAG（配列番号１）
1525R：AAGGAGGTGWTCCARCC（配列番号２）
Regarding primers, the primers used for identification of bacteria and actinomycetes are universal primers 27F and 1525R. The nucleotide sequence (5 'to 3') of the primer is as follows.
27F: AGAGTTTGATCMTGGCTCAG (SEQ ID NO: 1)
1525R: AAGGAGGTGWTCCRCC (SEQ ID NO: 2)
糸状菌の同定に使用したプライマーは、ユニバーサルプライマーのITS1-FとITS4である。プライマーのヌクレオチド配列（5’→3’）は以下のとおりである。
ITS1-F：CTTGGTCATTTAGAGGAAGTAA（配列番号３）
ITS4：TCCTCCGCTTATTGATATGC（配列番号４）
PCR反応液を表１に、反応条件を表２にそれぞれ示した。
The primers used to identify filamentous fungi are the universal primers ITS1-F and ITS4. The nucleotide sequence (5 'to 3') of the primer is as follows.
ITS1-F: CTTGGTCATTTAGAGGA AGTAA (SEQ ID NO: 3)
ITS4: TCCTCCGCTTATTGATATGC (SEQ ID NO: 4)
The PCR reaction solution is shown in Table 1, and the reaction conditions are shown in Table 2.
細菌・放線菌については、得られた16S ribosomal RNA遺伝子の増幅産物を次世代シーケンサーで解析した。用いたプライマーは27F(Lane et al. 1991 16S rRNA用ユニバーサルプライマー)である。 For bacteria and actinomycetes, the amplified product of the obtained 16S ribosomal RNA gene was analyzed by a next-generation sequencer. The primer used is 27F (Lane et al. 1991 universal primer for 16S rRNA).
糸状菌については、得られた18S rRNA遺伝子の増幅産物を次世代シーケンサーで解析した。用いたプライマーは、上記ITS1-Fである。 For filamentous fungi, the amplification product of the obtained 18S rRNA gene was analyzed by a next-generation sequencer. The primer used is the above ITS1-F.
いずれも、用いた分析機器は、Applied Biosystems 3730xl DNA Analyzer（ライフテクノロジーズジャパン社）である。 All analytical instruments used were Applied Biosystems 3730xl DNA Analyzer (Life Technologies Japan).
得られた配列データは、波形を確認したのちNCBI (National Center for Biotechnology Information)が提供するBLASTを用いて相同性検索を行った。結果を表３に示す。 The obtained sequence data were subjected to homology search using BLAST provided by NCBI (National Center for Biotechnology Information) after confirming the waveform. The results are shown in Table 3.
Streptomyces panaciradicis PSA-107の形態的、培地上の特徴、生理学的特徴等を以下に示す。 The morphological, medium characteristics, physiological characteristics and the like of Streptomyces panacradicis PSA-107 are shown below.
形態的特徴について、イースト・麦芽寒天培地(ISP No.2)、オートミール寒天培地(ISP No.3)、スターチ無機塩寒天培地(ISP No.4)で良好な生育を見せ、豊富な菌糸を形成する。 Regarding morphological characteristics, yeast-malt agar medium (ISP No. 2), oatmeal agar medium (ISP No. 3), and starch mineral salt agar medium (ISP No. 4) show good growth and form abundant mycelium. Do.
コロニーの形態について、イースト・麦芽寒天培地平板培養で黄茶色のコロニーを形成し、オートミール寒天培地平板培養で灰緑色のコロニーを形成し、スターチ無機塩寒天培地平板培養で灰緑色のコロニーを形成し、グリセリン・アスパラギン寒天培地(ISP No.5)平板培養で白色のコロニーを形成する。
細胞の形態について、胞子形成菌糸は直状又は直曲状である。
Regarding colony morphology, yellow-brown colonies are formed by yeast-malt agar medium plate culture, gray-green colonies are formed by oatmeal agar medium plate culture, and gray-green colonies are formed by starch mineral salt agar medium culture. White colonies are formed on glycerol-asparagine agar (ISP No. 5) plating.
With respect to the morphology of cells, sporulating hyphae are straight or straight.
生育温度について、R2A寒天培地、YPMG寒天培地を用いて培養し、10〜40℃まで生育し、最適生育温度はR2A寒天培地では30〜40℃、YPMG寒天培地では35℃である。 The growth temperature is cultured using R2A agar medium and YPMG agar medium and grown to 10-40 ° C. The optimum growth temperature is 30-40 ° C. for R2A agar medium and 35 ° C. for YPMG agar medium.
耐塩性について、7％NaCl含有イースト・麦芽寒天培地では正常に生育するが、8％では明らかな生育阻害を受ける。 With regard to salt tolerance, the yeast-malt agar medium containing 7% of NaCl grows normally, but 8% undergoes obvious growth inhibition.
炭素源の利用性試験では、Pridham-Gottlieb寒天培地(ISP No.9)を使用した結果、glucose、D-fructose、D-xylose、L-arabinose、D-raffinose、sucrose、inositolに対して資化性があり、L-rhamnose、D- mannitolに対して資化性がなかった。 In the availability test of carbon source, as a result of using Pridham-Gottlieb agar medium (ISP No. 9), utilization for glucose, D-fructose, D-xylose, L-arabinose, D-raffinose, sucrose and inositol It was sexual and was not assimilated to L-rhamnose and D-mannitol.
Bacillus megaterium PSB-139の形態的、培地上の特徴、生理学的特徴等を以下に示す。
細胞形態について、グラム陽性桿菌であり、サイズ1〜2μm程度である。
コロニーの色は、クリーム色である。
The morphological, culture characteristics, physiological characteristics and the like of Bacillus megaterium PSB-139 are shown below.
The cell morphology is gram-positive bacilli and is about 1 to 2 μm in size.
The color of the colony is cream.
生育温度について、R2A寒天培地、YPMG寒天培地を用いて培養し、10〜40℃まで生育し、最適生育温度はR2A寒天培地では30〜40℃、YPMG寒天培地では35℃である。 The growth temperature is cultured using R2A agar medium and YPMG agar medium and grown to 10-40 ° C. The optimum growth temperature is 30-40 ° C. for R2A agar medium and 35 ° C. for YPMG agar medium.
耐塩性について、9％NaCl含有イースト・麦芽寒天培地では正常に生育するが、10％では生育阻害を受ける。 As for salt tolerance, it grows normally in a yeast-malt agar medium containing 9% NaCl but suffers growth inhibition at 10%.
微生物分類・検定同定システムによる資化性調査では、BIOLOG GP2 MICROPLATE (Biolog, Inc. Cat.No. 1014)を使用して調査した結果は、以下のとおりである。 The results of investigation using BIOLOG GP2 MICROPLATE (Biolog, Inc. Cat. No. 1014) in the assimilability research by the microorganism classification and test identification system are as follows.
β-cyclodextrin、D-fructose、α-D-glucose、maltose、maltotriose、D-mannitol、D-mannose、D-melibiose、palatinose、D-psicose、sucrose、D-trehalose、D-xylose、glycerolに対して資化性があった。 against β-cyclodextrin, D-fructose, α-D-glucose, maltose, maltotriose, D-mannitol, D-mannose, D-melibiose, palatinose, D-psicose, sucrose, D-trehalose, D-xylose, glycerol It was assimilable.
dextrin、glycogen、L-arabinose、arbutin、D-cellobiose、α-D-lactose、lactulose、β-methyl-D-galactoside、β-methyl-D-glucoside、D-raffinose、D-ribose、salicin、stachyose、D-tagatose、acetic acid、 2'-deoxy adenosine、adenosine-5'-monophosphate、thymidine-5'-monophosphate、D-fructose-6-phosphateに対して資化性がなかった。 dextrin, glycogen, L-arabinose, arbutin, D-cellobiose, α-D-lactose, lactulose, β-methyl-D-galactoside, β-methyl-D-glucoside, D-raffinose, D-ribose, salicin, stachyose, There was no assimilability for D-tagatose, acetic acid, 2'-deoxy adenosine, adenosine-5'-monophosphate, thymidine-5'-monophosphate, and D-fructose-6-phosphate.
Bacillus velezensis NA5-3の形態的、培地上の特徴、生理学的特徴等を以下に示す。
細胞形態について、グラム陽性桿菌であり、サイズは1〜2μm程度である。
コロニーの色は、クリーム色である。
The morphological, culture characteristics, physiological characteristics and the like of Bacillus velezensis NA5-3 are shown below.
The cell morphology is gram-positive bacilli, and the size is about 1 to 2 μm.
The color of the colony is cream.
生育温度について、R2A寒天培地、YPMG寒天培地を用いて培養し、15〜40℃まで生育し、最適生育温度はR2A寒天培地、YPMG寒天培地ともに30℃〜40℃であった。 The growth temperature was cultured using R2A agar medium and YPMG agar medium and grown up to 15-40 ° C. The optimum growth temperature was 30 ° C.-40 ° C. for both R2A agar medium and YPMG agar medium.
耐塩性について、7％NaCl含有イースト・麦芽寒天培地では正常に生育するが、8％では明らかな生育阻害を受ける。 With regard to salt tolerance, the yeast-malt agar medium containing 7% of NaCl grows normally, but 8% undergoes obvious growth inhibition.
微生物分類・検定同定システムによる資化性調査では、BIOLOG GP2 MICROPLATE (Biolog, Inc. Cat.No. 1014)を使用して調査した結果は、以下の通りである。
dextrin、arbutin、D-cellobiose、D-fructose、α-D-glucose、maltose、maltotriose、D-mannitol、salicin、sucrose、D-trehalose、glycerolに対して資化性があった。
β-cyclodextrin、glycogen、L-arabinose、α-D-lactose、lactulose、D-mannose、D-melibiose、β-methyl-D-galactoside、β-methyl-D-glucoside、palatinoce、D-psicose、D-raffinose、D-ribose、stachyose、D-tagatose、D-xylose、acetic acid、 2'-deoxy adenosine、adenosine-5'-monophosphate、thymidine-5'-monophosphate、D-fructose-6-phosphateに対して資化性がなかった。
The results of investigation using BIOLOG GP2 MICROPLATE (Biolog, Inc. Cat. No. 1014) in the assimilability research by the microorganism classification and test identification system are as follows.
Dextrin, arbutin, D-cellobiose, D-fructose, α-D-glucose, maltose, maltotriose, D-mannitol, salicin, sucrose, D-trehalose, and glycerol were assimilable.
β-cyclodextrin, glycogen, L-arabinose, α-D-lactose, lactose, D-mannose, D-melibiose, β-methyl-D-galactoside, β-methyl-D-glucoside, palatinoce, D-psicose, D-psicose Useful for raffinose, D-ribose, stachyose, D-tagatose, D-xylose, acetic acid, 2'-deoxy adenosine, adenosine-5'-monophosphate, thymidine-5'-monophosphate, D-fructose-6-phosphate There was no
Bacillus aryabhattai NA6の形態的、培地上の特徴、生理学的特徴等を以下に示す。
細胞形態について、グラム陽性桿菌であり、サイズは1〜2μm程度である。
コロニーの色は、クリーム色である。
The morphological, culture characteristics, physiological characteristics, etc. of Bacillus aryabhattai NA6 are shown below.
The cell morphology is gram-positive bacilli, and the size is about 1 to 2 μm.
The color of the colony is cream.
生育温度について、R2A寒天培地、YPMG寒天培地を用いて培養し、10〜40℃まで生育し、最適生育温度はR2A寒天培地、YPMG寒天培地ともに25℃〜30℃であった。 The growth temperature was cultured using R2A agar medium and YPMG agar medium and grown to 10-40 ° C. The optimum growth temperature was 25 ° C.-30 ° C. for both R2A agar medium and YPMG agar medium.
耐塩性について、8％NaCl含有イースト・麦芽寒天培地では正常に生育するが、9％では明らかな生育阻害を受ける。 With regard to salt tolerance, the yeast-malt agar medium containing 8% NaCl grows normally, but 9% undergoes obvious growth inhibition.
微生物分類・検定同定システムによる資化性調査では、BIOLOG GP2 MICROPLATE (Biolog, Inc. Cat.No. 1014)を使用して調査した結果は、以下の通りである。
dextrin、D-fructose、α-D-glucose、maltose、maltotriose、D-mannitol、D-melibiose、D-psicose、D-ribose、sucrose、D-trehalose、glycerolに対して資化性があった。
β-cyclodextrin、glycogen、L-arabinose、arbutin、D-cellobiose、α-D-lactose、lactulose、D-mannose、β-methyl-D-galactoside、β-methyl-D-glucoside、palatinoce、D-raffinose、salicin、stachyose、D-tagatose、D-xylose、acetic acid、 2'-deoxy adenosine、adenosine-5'-monophosphate、thymidine-5'-monophosphate、D-fructose-6-phosphateに対して資化性がなかった。
The results of investigation using BIOLOG GP2 MICROPLATE (Biolog, Inc. Cat. No. 1014) in the assimilability research by the microorganism classification and test identification system are as follows.
Dextrin, D-fructose, α-D-glucose, maltose, maltotriose, D-mannitol, D-melibiose, D-psicose, D-ribose, sucrose, D-trehalose, and glycerol were assimilated.
β-cyclodextrin, glycogen, L-arabinose, arbutin, D-cellobiose, α-D-lactose, lactulose, D-mannose, β-methyl-D-galactoside, β-methyl-D-glucoside, palatinoce, D-raffinose, No assimilability to salicin, stachyose, D-tagatose, D-xylose, acetic acid, 2'-deoxy adenosine, adenosine-5'-monophosphate, tyrosine-5'-monophosphate, D-fructose-6-phosphate The
Talaromyces flavus PSF-234の形態的、培地上の特徴、生理学的特徴等を以下に示す。
胞子の形態について、分生子形成型はフィアロ型で、亜球形の4μm程度の分生子を形成する。
The morphological, culture characteristics, physiological characteristics and the like of Talaromyces flavus PSF-234 are shown below.
With regard to the morphology of the spore, the conidia forming type is a fiallo type, and forms a subspherical conidia of about 4 μm.
培地に形成されたコロニー表面の特徴について、PDA培地、PSA培地上で初めは白色、その後緑色となり、ビロード状の3〜5cmのコロニーを形成する。 With regard to the characteristics of the colony surface formed in the medium, PDA medium, initially white on PSA medium and then green, form a velvety 3-5 cm colony.
培地に形成されたコロニー裏面の特徴について、灰緑色で、20℃〜30℃でオレンジがかったピンク色の色素を生成した。 The characteristics of the back of the colony formed in the culture medium were grayish green and produced an orange-pink pigment at 20 ° C to 30 ° C.
生育温度について、PDA培地、PSA培地を用いて1週間培養し、最適生育温度は28℃〜33℃であった。 The growth temperature was cultured for 1 week using PDA medium and PSA medium, and the optimum growth temperature was 28 ° C to 33 ° C.
微生物分類・検定同定システムによる資化性調査では、BIOLOG FF MICROPLATEを使用して調査した結果は、以下の通りである。
glucose-1-phosphate、D-saccharic acid、tween 80、salicin、sebacic acid、dextrin、D-glucuronic acid、fumaric acid、L-phenylalanine、N-acetyl-D-glucosamine、erythritol、glycerol、D-sorbitol、L-proline、glycogen、γ-hydroxy-butyric acid、succinic acid 、mono-methyl ester、adonitol、amygdalin、2-keto-D-gluconic acid、alaninamide、gentiobiose、L-alanyl-glycine、D-arabitol、D-gluconic acid、L-asparagine、quinic acidに対して資化性があった。
The results of investigation using BIOLOG FF MICROPLATE in the assimilability research by the microorganism classification and test identification system are as follows.
glucose-1-phosphate, D-saccharic acid,
Talaromyces pinophilus PSF-332の形態的、培地上の特徴、生理学的特徴等を以下に示す。
胞子の形態について、分生子形成型はフィアロ型で、亜球形の3μm程度の分生子を形成する。
The morphological, culture characteristics, physiological characteristics and the like of Talaromyces pinophilus PSF-332 are shown below.
With regard to the morphology of the spore, the conidia forming type is a fiallo type, which forms subspherical conidia of about 3 μm.
培地に形成されたコロニー表面の特徴について、PDA培地、PSA培地上で初めは白色、その後黄緑色または緑色となり、羊毛状の3〜5cmのコロニーを形成する。 With respect to the characteristics of the colony surface formed in the medium, PDA medium, initially white on PSA medium, then yellowish green or green, and form fleecy 3-5 cm colonies.
培地に形成されたコロニー裏面の特徴について、クリーム色で、25℃付近ではPDA培地で紫色、PSA培地でピンク色の色素を生成した。 About the characteristic of the back of the colony formed in the culture medium, a cream color was produced in purple at around 25 ° C. in PDA medium and in pink medium at pink medium.
生育温度について、PDA培地、PSA培地を用いて1週間培養し、最適生育温度は30℃〜35℃であった。 About growth temperature, it culture | cultivated for 1 week using a PDA culture medium and a PSA culture medium, and the optimal growth temperature was 30 degreeC-35 degreeC.
微生物分類・検定同定システムによる資化性調査では、BIOLOG FF MICROPLATE使用) を使用して調査した結果は、以下の通りである。
glucose-1-phosphate、D-saccharic acid、tween 80、dextrin、D-glucuronic acid、D-melezitose、glycerol、L-proline、2-keto-D-gluconic acid、α-keto-glutaric acid、D-gluconic acid、quinic acid、L-glutamic acidに対して資化性があった。
The results of investigation using BIOLOG FF MICROPLATE) are as follows in the assimilability research by the microorganism classification / test identification system.
glucose-1-phosphate, D-saccharic acid,
本発明の微生物は、Streptomyces panaciradicis PSA-107株（受託番号：NITE P-02407）、Bacillus megaterium PSB-139株（受託番号：NITE P-02408）、Bacillus velezensis NA5-3株（受託番号：NITE P-02482）、Bacillus aryabhattai NA6株（受託番号：NITE P-02483）、Talaromyces flavus PSF-234株（受託番号：NITE P-02426）、Talaromyces pinophilus PSF-332株（受託番号：NITE P-02427）として独立行政法人製品評価技術基盤機構特許微生物寄託センター（〒292-0818千葉県木更津市かずさ鎌足2-5-8 122号室）に寄託されている。 The microorganism of the present invention is Streptomyces panacradicis PSA-107 (Accession No .: NITE P-02407), Bacillus megaterium PSB-139 (Accession No .: NITE P-02408), Bacillus velezensis NA5-3 (Accession No .: NITE P) -02482), Bacillus aryabhattai NA6 strain (Accession No .: NITE P-02483), Talaromyces flavus PSF-234 strain (Accession No .: NITE P-02426), Talaromyces pinophilus PSF-332 strain (Accession No .: NITE P-02427) It has been deposited at the Patent Organism Depositary Center of the National Institute of Technology and Evaluation, National Institute of Technology Evaluation (room 2-5-8 122, Kazusa Tsuji, Kisarazu City, Chiba Prefecture 292-0818).
さらに、本発明の微生物には、ジャガイモそうか病防除作用を有する、上記PSA-107株、PSB-139株、NA5-3株、NA6株、PSF-234株、及びPSF-332株の派生株もまた包含される。このような派生株は、親株を長期保存する間に自然変異が生じた株、親株の培養物を紫外線、重イオンビーム等の放射線や、化学変異原で処理することによって遺伝子型が変化した株、等を含む。化学変異原の例は、N-エチル-N-ニトロソウレア、メタンスルホン酸エチル、ニトロソグアニジン等を含む。 Furthermore, the microorganism of the present invention has a potato scab controlling activity, and the aforementioned PSA-107, PSB-139, NA5-3, NA6, PS6, PSF-234 and derivatives of PSF-332. Is also included. Such a derivative strain is a strain in which a spontaneous mutation occurs during long-term storage of the parent strain, or a strain whose genotype has been changed by treating the culture of the parent strain with radiation such as ultraviolet light or heavy ion beam or a chemical mutagen , Etc. Examples of chemical mutagens include N-ethyl-N-nitrosourea, ethyl methanesulfonate, nitrosoguanidine and the like.
本発明の菌株の培養条件は特に限定しない。例えば、PSA-107菌の培養条件（液体培養）の例は、以下のとおりである。
でんぷん寒天培地（Soluble Starch 10g、Yeast Extract 1g、Sucrose 1g、KCl 0.1g、KH2PO4 0.1g、MgSO4・7H2O 0.1g、NaNO3 0.1g、蒸留水 1000mL）上で培養したコロニー片を用いて、TSB培地、30℃、6日間振とうの培養条件で培養する。
The culture conditions of the strain of the present invention are not particularly limited. For example, examples of culture conditions (liquid culture) of PSA-107 bacteria are as follows.
Colony pieces cultured on starch agar (Soluble Starch 10 g, Yeast Extract 1 g, Sucrose 1 g, KCl 0.1 g, KH 2 PO 4 0.1 g, MgSO 4 · 7 H 2 O 0.1 g, NaNO 3 0.1 g, distilled water 1000 mL) The culture is carried out under the culture conditions of shaking at 30 ° C. for 6 days using TSB medium.
PSB-139菌の培養条件（液体培養）の例は、以下のとおりである。
R2A寒天培地（Soluble Starch 0.5g、 Protease Peptone No.3 0.5g、Yeast Extract 0.5g、Sucrose 0.5g、Casamino acid 0.5g、KH2PO4 0.3g、MgSO4・7H2O 0.05g、Sodium pyruvate 0.3g、Ager 15g、蒸留水 1000mL）上で培養したコロニー片を用いて、R2A液体培地で、30℃、6日間振とうの培養条件で培養する。
Examples of culture conditions (liquid culture) of PSB-139 bacteria are as follows.
R2A agar medium (Soluble Starch 0.5 g, Protease Peptone No. 3 0.5 g, Yeast Extract 0.5 g, Sucrose 0.5 g, Casamino acid 0.5 g, KH 2 PO 4 0.3 g, MgSO 4 · 7 H 2 O 0.05 g, Sodium pyruvate 0.3 The colony pieces cultured on g, Ager 15 g, distilled water (1000 mL)) are cultured in R2A liquid medium under culture conditions of shaking at 30 ° C. for 6 days.
NA5-3菌の培養条件の例は、以下のとおりである。
R2A寒天培地上で培養したコロニー片を用いて、R2A液体培地で30℃、6日間振とう培養を行う。
Examples of culture conditions of NA5-3 bacteria are as follows.
Using colony pieces cultured on R2A agar medium, shake culture is carried out at 30 ° C. for 6 days in R2A liquid medium.
NA6菌の培養条件の例は、以下のとおりである。
R2A寒天培地上で培養したコロニー片を用いて、R2A液体培地で30℃、6日間振とう培養を行う。
Examples of culture conditions of the NA6 bacteria are as follows.
Using colony pieces cultured on R2A agar medium, shake culture is carried out at 30 ° C. for 6 days in R2A liquid medium.
PSF-234菌の培養条件の例は、以下のとおりである。
ポテトデキストロース寒天培地(Potato Extract 100ml、Sucrose20g、Ager15g、蒸留水 1000mL)上で培養したコロニー片を用いて、R2A液体培地で30℃、6日間振とう培養を行う。
Examples of culture conditions of PSF-234 bacteria are as follows.
Using a colony piece cultured on potato dextrose agar medium (
PSF-332菌の培養条件の例は、以下のとおりである。
ポテトデキストロース寒天培地上で培養したコロニー片を用いて、R2A液体培地で30℃、6日間振とう培養を行う。
Examples of culture conditions of PSF-332 bacteria are as follows.
The colony pieces cultured on potato dextrose agar medium are subjected to shaking culture at 30 ° C. for 6 days in R2A liquid medium.
２．ジャガイモそうか病防除資材及び防除方法
本発明の微生物は、寒天培地又は液体培地を用いて、培養・増殖される。増殖された微生物は、白金耳等により採取、遠心分離等の操作により集菌して採取、あるいは培養液の状態として、ジャガイモそうか病防除資材の製造に用いることができる。
2. Potato scab control material and control method The microorganism of the present invention is cultured and grown using an agar medium or a liquid medium. The grown microorganisms can be collected by operations such as platinum ear, collected by centrifugation and the like, and collected or used as a culture solution for producing potato scum controlling material.
ジャガイモそうか病防除資材には、Streptomyces panaciradicis PSA-107株（受託番号：NITE P-02407）、Bacillus megaterium PSB-139株（受託番号：NITE P-02408）、Bacillus velezensis NA5-3株（受託番号：NITE P-02482）、Bacillus aryabhattai NA6株（受託番号：NITE P-02483）、Talaromyces flavus PSF-234株（受託番号：NITE P-02426）、及びTalaromyces pinophilus PSF-332株（受託番号：NITE P-02427）、並びにそれらの上記派生株からなる群から選択される少なくとも１種の菌株を含有させることができる。 Materials for controlling potato scab include Streptomyces panacradicis PSA-107 (Accession No .: NITE P-02407), Bacillus megaterium PSB-139 (Accession No .: NITE P-02408), and Bacillus velezensis NA5-3 (Accession No.) : NITE P-02482), Bacillus aryabhattai NA 6 strain (Accession number: NITE P-02483), Talaromyces flavus PSF-234 strain (Accession number: NITE P-02426), and Talaromyces pinophilus PSF-332 strain (Accession number: NITE P) And at least one strain selected from the group consisting of the above derivatives thereof.
ジャガイモそうか病防除資材の製造において、培養・増殖して得られた本発明の微生物は、PSA-107株、PSB-139株、NA5-3株、NA6株、PSF-234株、及びPSF-332、並びにこれらの派生株のいずれかを単独で用いることもできるし、又はこれらの株を2種以上組み合わせて用いることもできる。 The microorganism of the present invention obtained by culturing and growing in the production of a material for controlling potato scab is PSA-107 strain, PSB-139 strain, NA5-3 strain, NA6 strain, PSF-234 strain, and PSF- 332, and any of these derivatives may be used alone, or two or more of these strains may be used in combination.
また、本発明の微生物は、当該微生物のみで本発明のジャガイモそうか病防除資材とすることもできるが、他の各種資材、例えばゼオライト、バーミキュライト、イソライト、モンモリロナイト等の無機質鉱物、珪藻土、焼成珪藻土、サンゴ砂、木炭粉末、活性炭等の多孔質物、硫酸アンモニウム、塩化アンモニウム、硝酸アンモニウム、尿素、リン酸アンモニウム、過リン酸石灰、硫酸カリウム、塩化カリウム、硫酸カルシウム、炭酸カルシウム等の化成肥料、魚粕、肉粕、骨粉、カニガラ、フェザーミール、蒸製毛粉、皮粉等の動物質肥料、なたね油かす、ひまし油かす、だいず油かす、米ぬか等の植物質肥料、牛糞堆肥、豚ぷん堆肥、鶏糞堆肥等の堆肥類、火山灰土壌、褐色森林土、灰色低地土、赤色土、黄色土等の土壌のいずれもとも組み合わせて、ジャガイモそうか病防除資材として、及び、肥料、土壌改良資材、育苗培土等として、使用することもできる。また、粒剤、粉剤、錠剤、乳剤、水和剤、微生物固定化剤、等の任意の剤型として使用することもできる。このような剤型の製造において、農業分野で通常使用される添加剤（例えば、担体、希釈剤、界面活性剤、分散剤、展着剤、防かび剤、着色剤、等）を適宜使用しうる。 In addition, the microorganism of the present invention can be used as a potato scab control material of the present invention only by the microorganism, but other various materials such as zeolite, vermiculite, isolite, mineral minerals such as montmorillonite, diatomaceous earth, calcined diatomaceous earth , Coral sand, charcoal powder, porous materials such as activated carbon, ammonium sulfate, ammonium chloride, ammonium nitrate, urea, ammonium phosphate, calcium superphosphate, potassium sulfate, potassium chloride, calcium sulfate, calcium carbonate, etc. Animal manure such as meat meal, bone meal, crab meal, feather meal, steamed hair powder, skin meal etc. Plant fertilizer such as rapeseed meal, rapeseed meal residue, castor oil meal, dain oil cake, rice bran, cow manure compost, pig manure compost, chicken manure compost etc Soil such as compost, volcanic ash soil, brown forest soil, gray lowland soil, red soil and yellow soil Combination, as potato scab control agents, and, fertilizers, soil improving material, as nursery bed soil and the like, may also be used. Moreover, it can also be used as arbitrary dosage forms, such as a granule, a powder, a tablet, an emulsion, a wettable powder, the microorganisms fixing agent, etc. In the production of such formulations, additives (for example, carriers, diluents, surfactants, dispersants, spreaders, fungicides, colorants, etc.) usually used in the agricultural field are appropriately used. sell.
本発明の微生物は、ジャガイモそうか病に対し防除作用（例えば、抑制能）を有するので、本発明の微生物又はジャガイモそうか病防除資材を用いてジャガイモ（すべての品種）の種子、種イモ、葉及び栽培土壌等の少なくとも一つを処理すれば、ジャガイモそうか病の防除効果が発揮される。この処理方法としては、ジャガイモを栽培する際の、土壌への施用、育苗培土への添加、作物種子にバクテリゼーション処理、種イモ浸漬処理、葉面散布、養液栽培における養液への添加、土耕栽培における株元への添加・潅注等をあげることができる。本発明の微生物の処理により、ジャガイモそうか病の防除を行うことはもとより、ジャガイモそうか病汚染圃場の伝染を阻止、あるいは予防対策として使用することができる。施用する微生物（菌）用量は、ジャガイモそうか病を防除することが可能であれば特に制限されないし、また、ジャガイモそうか病による汚染又は被害の状況に応じて適宜、用量及び処理回数を決定しうる。用量は、例えば、104〜109 cfu/g土壌又は104〜109 cfu/ml液剤であるが、この範囲外であってもよい。 Since the microorganism of the present invention has a control action (for example, suppressing ability) against potato scab, seeds of potato (all varieties), seeds, potato, etc., using the microorganism of the present invention or the potato scab control material If at least one of leaves and cultivated soil is treated, the control effect of potato scab is exhibited. This treatment method includes: application to soil, addition to nursery culture, bacteriization treatment to crop seeds, seed potato immersion treatment, foliar application, addition to nutrient solution in hydroponic culture, when growing potatoes Addition and irrigation to the stock origin in soil culture can be mentioned. By treating the microorganisms of the present invention, it is possible not only to control potato scab, but also to prevent the spread of potato scab contaminated fields or to use it as a preventive measure. The amount of microbe (microbe) to be applied is not particularly limited as long as it can control potato scab, and the dosage and the number of treatments are appropriately determined depending on the status of contamination or damage by potato scab. It can. The dose is, for example, 10 4 to 10 9 cfu / g soil or 10 4 to 10 9 cfu / ml solution, but may be outside this range.
本発明のジャガイモそうか病の防除方法としては、上記のように本発明の微生物（Streptomyces panaciradicis PSA-107株（受託番号：NITE P-02407）、Bacillus megaterium PSB-139株（受託番号：NITE P-02408）、Bacillus velezensis NA5-3株（受託番号：NITE P-02482）、Bacillus aryabhattai NA6株（受託番号：NITE P-02483）、Talaromyces flavus PSF-234株（受託番号：NITE P-02426）、及びTalaromyces pinophilus PSF-332株（受託番号：NITE P-02427）、並びにそれらの上記派生株からなる群から選択される少なくとも１種の菌株）又はジャガイモそうか病防除資材単独で処理することもできるが、各種土壌消毒と組み合わせて処理すると、ジャガイモそうか病の防除効果はさらに高まる。土壌消毒法としては、クロールピクリン処理、太陽熱消毒、還元消毒があげられる。クロールピクリン処理の場合はガス抜き後に、あるいは太陽熱消毒や還元消毒の場合は消毒後に、本発明の微生物処理を行うことができ、処理の方法としては種子、種イモ、葉及び栽培土壌等の少なくとも一つに上記微生物の処理方法と同様にして行うことができる。 As a control method of potato scab of the present invention, the microorganism of the present invention (Streptomyces panacradicis PSA-107 strain (Accession number: NITE P-02407), Bacillus megaterium PSB-139 strain (Accession number: NITE P) as described above -02408), Bacillus velezensis strain NA5-3 (Accession number: NITE P-02482), Bacillus aryabhattai NA 6 strain (Accession number: NITE P-02483), Talaromyces flavus PSF-234 strain (Accession number: NITE P-02426), And / or Talaromyces pinophilus PSF-332 (Accession No .: NITE P-02427), and at least one strain selected from the group consisting of the above derivatives thereof) or potato scab control materials alone can also be used. However, when treated in combination with various soil disinfections, the control effect of potato scab is further enhanced. Soil disinfection methods include chloropicrin treatment, solar heat disinfection and reduction disinfection. The microorganism treatment of the present invention can be carried out after degassing in the case of chloropicrin treatment, or after disinfection in the case of solar heat disinfection or reduction disinfection, and the method of treatment is at least seeds, seeds, leaves, cultivated soil, etc. One can be carried out in the same manner as the method for treating the above-mentioned microorganism.
以下に本発明の実施形態を示すが、本発明は、これら実施例に限定されるものではない。 Hereinafter, embodiments of the present invention will be described, but the present invention is not limited to these examples.
［実施例１］
＜ジャガイモそうか病に対するポット試験1＞
1. 汚染土の作成
Streptomyces. scabiei S58株（鹿児島県農業開発総合センターより分譲）をTSB培地で振とう培養（30℃、3日間）したものをバーミキュライト・オートミール培地に接種・混合して25℃で4週間培養し、培養物を作成した。この培養物を2mmの篩を通した土壌に化成肥料（N-P-K：12-12-12kg/10a）を混合してStreptomyces.scabiei S58株が104cfu/g 土壌のそうか病汚染土壌を作成した。
Example 1
<Pot test 1 for potato scab>
1. Create contaminated soil
A shaking culture (30 ° C., 3 days) of Streptomyces. Scabiei S58 strain (distributed from Kagoshima Prefectural Agricultural Development Center) in TSB medium is inoculated into and mixed with vermiculite oatmeal medium and cultured at 25 ° C. for 4 weeks, Cultures were made. Streptomyces. Scabiei S58 strain made a scab-contaminated soil of 10 4 cfu / g soil by mixing this culture with soil passing through a 2 mm sieve and converting fertilizer (NPK: 12-12-12 kg / 10 a). .
２．有用菌の処理
Streptomyces panaciradicis PSA-107については、培養物（液体培地で30℃・6日間振とうした培養液を鉱物質固体培地に接種し25℃・4日間培養）をポットあたり30g施用し種イモを植え付けた。
2. Treatment of useful bacteria
About Streptomyces panacradicis PSA-107, 30 g per pot was applied and the seed potato was planted by applying a culture (a culture broth shaken at 30 ° C. for 6 days with liquid medium was inoculated to a mineral solid medium and cultured at 25 ° C. for 4 days) .
Bacillus megaterium PSB-139については、培養液（液体培地で30℃・10日間振とう）中の菌数を血球計数盤で計数後、1.8×104cfu/g土壌となるように接種し、種イモを植え付けた。また、汚染土に植え付けた区を無処理1区、健全土に植え付けた区を無処理2区とした。いずれの試験区とも6連でパイプハウス内の圃場に埋設して約3ヶ月間栽培（2015年9/16 資材浸漬区種イモ処理、9/18菌株選抜区、資材浸漬区処理・植付け、9/24資材施用区処理・植付け、12/21調査）を行った。 For Bacillus megaterium PSB-139, count the number of bacteria in the culture solution (liquid medium and shake for 10 days at 30 ° C) with a hemocytometer, and inoculate so that it will be 1.8 x 10 4 cfu / g soil. I planted a potato. In addition, the area planted in contaminated soil was defined as untreated area 1 and the area planted in healthy soil was classified as untreated area 2. Buried in the field in the pipe house in 6 test stations in all series and cultivate for about 3 months (2015.9 / 16 Material Immersion Zone Seed Imo Treatment, 9/18 Strain Selection Zone, Material Immersion Zone Treatment / Planting 24 material application zone treatment and planting, 12/21 survey) was conducted.
３．調査
1cm以上に肥大した塊茎を全て収穫し、収量（重量）を調査した。そのうち、重さ10g以上の塊茎については発病程度を6段階で評価し、発病度（指数）を算出した。
3. Research
All tubers enlarged to 1 cm or more were harvested, and the yield (weight) was examined. Among them, for tubers having a weight of 10 g or more, the degree of onset was evaluated in 6 levels, and the onset (index) was calculated.
発病程度は、発病なしを0、発病面積が3％以下を「1」、4〜13%を「2」、14〜25%を「3」、26〜50%を「4」、51〜75%を「5」、76%以上を「6」とし、発病度（Σ（発病程度別塊茎数×発病程度）／（調査塊茎数×6）×100）を算出した。また、発病している塊茎の割合（%）を発病塊茎率とした。 The incidence is 0: no onset, 3% or less of the affected area is “1”, 4 to 13% is “2”, 14 to 25% is “3”, 26 to 50% is “4”, 51 to 75 The incidence (5 (number of tubers according to the degree of onset of disease × degree of onset of disease) / (number of surveyed tubers × 6) × 100) was calculated, with% as 5 and 76% or more as 6. In addition, the proportion (%) of tubers that develop disease was defined as the diseased tuber rate.
４．結果
発病度は無処理１区(病原菌接種区)を100とした場合、PSA-107区が6.4、PSB-139区が39.7、発病塊茎率はPSA-107区が7.5、PSB-139区が44.4となり、いずれも発病を抑制した（図１）。
4. Results When the untreated diseased zone 1 (pathogenic bacteria inoculated zone) is assumed to be 100, the PSA-107 zone is 6.4, the PSB-139 zone is 39.7, the diseased tuber rate is 7.5, and the PSB-139 zone is 44.4. And all suppressed the onset (Fig. 1).
［実施例２］
＜ジャガイモそうか病に対するポット試験2＞
１．培養
Bacillus velezensis NA5-3については培養液（液体培地で30℃・５日間振とう）中の菌数を血球計数盤で計数後、１×105cfu/g土壌となるように接種し、種イモを植え付けた。
Example 2
<Pot test 2 for potato scab>
1. culture
For Bacillus velezensis NA5-3, count the number of bacteria in the culture broth (liquid medium and shake for 5 days at 30 ° C) with a hemocytometer, and inoculate so that it will be 1 x 10 5 cfu / g soil. Planted
Bacillus aryabhattai NA6については培養液（液体培地で30℃・５日間振とう）中の菌数を血球計数盤で計数後、１×105cfu/g土壌となるように接種し、種イモを植え付けた。 For Bacillus aryabhattai NA6, count the number of bacteria in the culture broth (liquid medium and shake for 5 days at 30 ° C) with a hemocytometer, inoculate 1 x 10 5 cfu / g soil, and plant seed potato The
Talaromyces flavus PSF-234については培養液（液体培地で30℃・10日間振とう）中の菌数を血球計数盤で計数後、1×105cfu/g土壌となるように接種し、種イモを植え付けた。 For Talaromyces flavus PSF-234, count the number of bacteria in the culture solution (liquid medium and shake for 10 days at 30 ° C) with a hemocytometer, and inoculate so that it will be 1 x 10 5 cfu / g soil. Planted
Talaromyces pinophilus PSF-332については、培養液（液体培地で30℃・10日間振とう）中の菌数を血球計数盤で計数後、1×105cfu/g土壌となるように接種し、種イモを植え付けた。 For Talaromyces pinophilus PSF-332, count the number of bacteria in the culture solution (liquid medium with shaking at 30 ° C for 10 days) with a hemocytometer, and inoculate so that it will be 1 x 10 5 cfu / g soil. I planted a potato.
２．栽培試験
（１） 汚染土の作成
バーミキュライト・オートミール培地をシナノパックに2Lずつ充填し、オートクレーブ滅菌後、Streptomyces scabiei S58株培養液（TSB培地、30℃、4日間振とう培養）を30mL接種し、室温（25℃）で4週間培養した。5mmの篩を通した土壌に培養物と化成肥料（N-P-K：12-12-12kg/10a）を混合してStreptomyces scabiei S58株が104cfu/g 土壌のそうか病汚染土壌を作成した。
2. Cultivation test (1) Preparation of contaminated soil 2 L each of vermiculite oatmeal medium is filled in Shinano pack, and after autoclave sterilization, 30 mL of Streptomyces scabiei S58 strain culture solution (TSB medium, 30 ° C., shaking culture for 4 days) is inoculated The cells were cultured at room temperature (25 ° C.) for 4 weeks. The culture and chemical fertilizer (NPK: 12-12-12 kg / 10 a) were mixed with the 5 mm sieved soil to prepare a scab-contaminated soil of 10 4 cfu / g soil of Streptomyces scabiei S58 strain.
（２）栽培試験
Bacillus velezensis NA5-3、 Bacillus aryabhattai NA6、Talaromyces flavus PSF-234、又はTalaromyces pinophilus PSF-332をそうか病汚染土2.5kgと混合後、径16cmの不織布ポットに充填し、種イモを植付けた。また、汚染土に植え付けた区を無処理1区、健全土に植え付けた区を無処理2区とした。いずれの試験区とも6連でパイプハウス内の圃場に埋設して栽培し、収量及びそうか病発病程度を調査した（2016年9月12日 資材処理・植付け、12月5日 調査）。地上部病害防除等の管理は慣行に従った。
(2) Cultivation test
After mixing Bacillus velezensis NA5-3, Bacillus aryabhattai NA6, Talaromyces flavus PSF-234, or Talaromyces pinophilus PSF-332 with 2.5 kg of scab-contaminated soil, it was filled in a non-woven cloth pot with a diameter of 16 cm and planted with potato seeds. In addition, the area planted in contaminated soil was defined as untreated area 1 and the area planted in healthy soil was classified as untreated area 2. Both test areas were buried and cultivated in a field in a pipe house at 6 stations, and the yield and the degree of scab disease incidence were investigated (September 12, 2016 Material processing and planting, December 5 survey). Management of epilepsy disease control etc. followed the practice.
３．発病調査
発病調査は重さ10g以上の塊茎を対象に行った。
発病程度は、発病なしを「0」、面積が3％以下を「1」、面積が4〜13%を「2」、面積が14〜25%を「3」、面積が26%以上を「4」とし、発病度（Σ（発病程度別塊茎数×発病程度）／（調査塊茎数×4）×100）を算出した。また、発病している塊茎の割合（%）を発病塊茎率とした。
3. Pathogenesis investigations were conducted on tubers weighing more than 10 g.
As for the degree of onset, there is no onset of disease "0", area 3% or less "1", area 4-13% "2", area 14-25% "3", area 26% or more " The incidence was calculated as follows: Σ (Σ (number of tubers according to the degree of onset of disease × degree of onset of disease) / (number of surveyed tubers × 4) × 100). In addition, the proportion (%) of tubers that develop disease was defined as the diseased tuber rate.
４．結果
無処理１区(病原菌接種区)を100とした場合、発病度はPSF-234区46.9、PSF-332区が44.5、NA5-3区が40.9、NA6区が36.0であり、発病塊茎率はPSF-234区が69.6、PSF-332区が67.9、NA5-3区が64.3、NA6区が64.3であり、いずれも発病を抑制した（図２）。
4. Result The incidence is PSF-234 46.9, PSF-332 44.5, NA 5-3 40.9, NA 6 6 36.0, and the diseased tuber rate is 100. PSF-234 was 69.6, PSF-332 was 67.9, NA 5-3 was 64.3, and NA 6 was 64.3, both of which suppressed the onset (FIG. 2).
本発明は、ジャガイモの主要な病害の一つであるジャガイモそうか病の防除に有効な方法を提供するものであるため、農業上有用である。 The present invention is useful in agriculture because it provides an effective method for controlling potato scab, which is one of the major diseases of potato.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113322192A (en) * | 2021-07-15 | 2021-08-31 | 云南省农业科学院药用植物研究所 | Polygonatum sibiricum endophytic fungi escherichia coli inhibiting agent and preparation method and application thereof |
| CN114025612A (en) * | 2019-06-27 | 2022-02-08 | 出光兴产株式会社 | Plant disease control agent and plant disease control method |
| CN116179446A (en) * | 2023-03-22 | 2023-05-30 | 天津坤禾生物科技集团股份有限公司 | Microbial agent for preventing and treating citrus root rot yellowing disease, and preparation method and application thereof |
Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0248509A (en) * | 1988-08-09 | 1990-02-19 | Nippon Kayaku Co Ltd | Method for controlling plant disease injury by kf-44 strain belonging to genus bacillus |
| JP2002138005A (en) * | 2000-10-27 | 2002-05-14 | Idemitsu Kosan Co Ltd | Material for controlling soil pest and method for controlling the same |
| JP2004137239A (en) * | 2002-10-21 | 2004-05-13 | Bio Oriented Technol Res Advancement Inst | Agent and method for controlling soil blight |
| JP2005060287A (en) * | 2003-08-11 | 2005-03-10 | Takasaki Kasei Kk | Scab-controlling material for potato or the like and controlling method |
| JP2005295924A (en) * | 2004-04-14 | 2005-10-27 | Idemitsu Kosan Co Ltd | Actinomycetes for controlling plant disease damage and controller for plant disease damage using the same |
| JP2009136216A (en) * | 2007-12-06 | 2009-06-25 | National Institute Of Advanced Industrial & Technology | Microbial preparation for controlling common scab |
| JP2011229443A (en) * | 2010-04-27 | 2011-11-17 | Nagasaki Prefecture | Method for controlling potato-cyst nematode and potato scab with fusarium oxysporum |
| JP2014224102A (en) * | 2013-04-19 | 2014-12-04 | 独立行政法人農業・食品産業技術総合研究機構 | Potato common scab inhibitor |
| WO2015114552A1 (en) * | 2014-01-29 | 2015-08-06 | University Of Pretoria | Plant growth promoting rhizobacterial strains and their uses |
-
2017
- 2017-08-28 JP JP2017163553A patent/JP6997561B2/en active Active
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0248509A (en) * | 1988-08-09 | 1990-02-19 | Nippon Kayaku Co Ltd | Method for controlling plant disease injury by kf-44 strain belonging to genus bacillus |
| JP2002138005A (en) * | 2000-10-27 | 2002-05-14 | Idemitsu Kosan Co Ltd | Material for controlling soil pest and method for controlling the same |
| JP2004137239A (en) * | 2002-10-21 | 2004-05-13 | Bio Oriented Technol Res Advancement Inst | Agent and method for controlling soil blight |
| JP2005060287A (en) * | 2003-08-11 | 2005-03-10 | Takasaki Kasei Kk | Scab-controlling material for potato or the like and controlling method |
| JP2005295924A (en) * | 2004-04-14 | 2005-10-27 | Idemitsu Kosan Co Ltd | Actinomycetes for controlling plant disease damage and controller for plant disease damage using the same |
| JP2009136216A (en) * | 2007-12-06 | 2009-06-25 | National Institute Of Advanced Industrial & Technology | Microbial preparation for controlling common scab |
| JP2011229443A (en) * | 2010-04-27 | 2011-11-17 | Nagasaki Prefecture | Method for controlling potato-cyst nematode and potato scab with fusarium oxysporum |
| JP2014224102A (en) * | 2013-04-19 | 2014-12-04 | 独立行政法人農業・食品産業技術総合研究機構 | Potato common scab inhibitor |
| WO2015114552A1 (en) * | 2014-01-29 | 2015-08-06 | University Of Pretoria | Plant growth promoting rhizobacterial strains and their uses |
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| CN114025612A (en) * | 2019-06-27 | 2022-02-08 | 出光兴产株式会社 | Plant disease control agent and plant disease control method |
| CN114025612B (en) * | 2019-06-27 | 2023-04-07 | 日本史迪士生物科学株式会社 | Plant disease control agent and plant disease control method |
| CN113322192A (en) * | 2021-07-15 | 2021-08-31 | 云南省农业科学院药用植物研究所 | Polygonatum sibiricum endophytic fungi escherichia coli inhibiting agent and preparation method and application thereof |
| CN116179446A (en) * | 2023-03-22 | 2023-05-30 | 天津坤禾生物科技集团股份有限公司 | Microbial agent for preventing and treating citrus root rot yellowing disease, and preparation method and application thereof |
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