WO2018186307A1 - ミツアリア属に属する菌株及び該菌株を用いた微生物農薬 - Google Patents
ミツアリア属に属する菌株及び該菌株を用いた微生物農薬 Download PDFInfo
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- WO2018186307A1 WO2018186307A1 PCT/JP2018/013857 JP2018013857W WO2018186307A1 WO 2018186307 A1 WO2018186307 A1 WO 2018186307A1 JP 2018013857 W JP2018013857 W JP 2018013857W WO 2018186307 A1 WO2018186307 A1 WO 2018186307A1
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01C—PLANTING; SOWING; FERTILISING
- A01C1/00—Apparatus, or methods of use thereof, for testing or treating seed, roots, or the like, prior to sowing or planting
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G7/00—Botany in general
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H6/00—Angiosperms, i.e. flowering plants, characterised by their botanic taxonomy
- A01H6/82—Solanaceae, e.g. pepper, tobacco, potato, tomato or eggplant
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/20—Bacteria; Substances produced thereby or obtained therefrom
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
Definitions
- the present invention relates to a bacterial strain belonging to the genus Mitsuaria, a bacterial plant disease control agent, a plant growth regulator, and the like using the bacterial strain.
- Plant diseases caused by pathogenic bacteria
- measures such as cultivating control and the use of disinfectants are being addressed.
- some bacterial plant diseases are difficult to control even using such cultivated control or disinfectant.
- vegetable bacterial wilt is a soil infectious disease caused by Ralstonia solanacerum, a bacterium that inhabits the soil, and tomato, eggplant, pepper, potato, etc. are infected with this bacterial wilt.
- this fungus is distributed around the world mainly in tropical, subtropical, and temperate regions, and more than 200 kinds of crops are infected and damaged mainly in the above solanaceous plants. It is one of serious and important problems.
- bacterial wilt control methods have been devised so far, for example, the use of chemical pesticides, soil disinfection such as solar disinfection, reductive disinfection, breeding of resistant rootstock, etc. Even if bacteria on the surface of the soil are sterilized by soil disinfection, bacterial wilt bacteria can survive for a long time in the deep part of the soil (about 50 cm to 1 m), so there should be no soil disinfectant etc. reaching such depth Therefore, it is difficult to completely eliminate this bacterium from the soil. In addition, resistance varieties are not completely resistant and may not be effective depending on environmental conditions. Furthermore, a bacterial blight resistance inducer (Patent Document 1) has been proposed, but this also does not provide a sufficient effect.
- a method using a biopesticide has been proposed as a control method for suppressing the pathogenesis of a plant body by an antagonistic action under the condition that a pathogen is present in the soil.
- control by microbial pesticides using strains of Pseudomonas genus, Ralstonia genus, Acinetobacter genus, Pythium genus has been studied (for example, Acinetobacter GEBT349).
- Non-patent literature 1) using these strains, and even these biological pesticides do not have sufficient control effects.
- scab disease is a bacterial plant disease that infects tomatoes, ume, kiwi fruit, citrus fruits, and the like, which eventually kills the plant body.
- This is also a soil infectious disease caused by bacteria such as Clavibacter michiganensis, and it is said that the pathogenic bacteria can survive in soil for more than 3 years. It can be said that the disease is extremely difficult to control.
- the present invention provides a plant non-pathogenic strain that can be used safely as a biopesticide and exhibits a stable bacterial plant disease control effect in actual crop production sites, and a bacterial plant disease control using the strain It was made for the purpose of providing agents.
- the embodiment of the present invention is as follows.
- a strain belonging to the genus Mitsuaria which has a bacterial plant disease control action.
- the strain described in (1) which is a strain belonging to Mitsaria chitosanitabida.
- a bacterial plant disease control agent comprising, as an active ingredient, a live cell of the strain described in any one of (1) to (3) or a culture containing the live cell .
- the agent according to (4) which is a vegetable bacterial wilt control agent and / or a scab control agent (for example, a vegetable scab control agent).
- a method for controlling bacterial plant diseases comprising: (8) The method according to (7), which comprises controlling vegetable bacterial wilt and / or scab (for example, vegetable scab).
- a plant growth regulator comprising a viable cell of the strain according to any one of (1) to (3) or a culture containing the viable cell as an active ingredient.
- the present invention has a remarkable effect (practical and stable effect) for bacterial plant disease control, which has conventionally been difficult to control, such as bacterial wilt disease and scab, and at an actual crop production site.
- Bacterial plant disease control agents that can be used safely can be provided.
- a secondary product it is possible to provide a plant growth regulator and the like.
- strain photography photograph 14 days after inoculation of tomato bacterial wilt in the tomato bacterial wilt control effect confirmation test of Example 1 is shown (drawing substitute photograph).
- the 5 strains on the left are the untreated section (Control), and the 5 strains on the right are the TWR114 strain treated section (TWR114).
- a strain belonging to the genus Mitsaria is used, for example, a strain belonging to Mitsaria chitosanitabida.
- the term “bacterial plant disease” means damage to a useful plant caused by a plant disease in which the pathogen is a bacterium, and damage caused by a plant disease caused by a filamentous fungus or virus is not included.
- This genus Mitalia is a genus whose bacterial plant disease control action was not known at all by conventional knowledge, and no species having phytopathogenicity are known.
- a strain isolated from green onion rhizosphere soil, 1-1 Yanagito, Gifu City, Gifu Prefecture was identified by bacteriological properties and phylogenetic analysis by 16S rRNA gene genome sequencing. It is more preferable to use a TWR114 strain which has been found to be a pathogenic and a new strain belonging to Mitsaria chitosan nitabida, or a mutant strain of the strain that retains the same properties as the strain.
- the API20NE kit Sysmex Biomelieu Co., Ltd. product
- the TWR114 strain was founded on March 17, 2017 in the National Institute of Technology and Evaluation Microbiology Depositary Center ( ⁇ 292-0818, 2-5-8 Kazusa-Kamashita, Kisarazu-shi, Chiba, Japan) After the domestic deposit, the request for transfer to the international deposit was received on January 22, 2018, and the international deposit number is NITE BP-02445.
- a medium that can be used for culturing a strain belonging to the genus Mitsuaria may be any medium as long as the strain can grow.
- a medium containing glucose, peptone, or yeast extract can be used in addition to a common medium.
- a solid medium such as a slant medium containing agar and a plate medium may be used in addition to the liquid medium.
- any medium that can be assimilated by a strain belonging to the genus Mitsuaria can be used.
- Specific examples include various synthetic or natural carbon sources that can be used by strains belonging to the genus Mitsuaria, such as glucose, arabinose, mannose, starch hydrolyzate, and molasses.
- various synthetic or natural products that can be used by the strain including organic nitrogen-containing substances such as peptone, meat extract, yeast extract, soybean meal, and the like can be used.
- mineral salts such as salt and phosphate
- metal salts such as calcium, magnesium and iron
- trace nutrients such as vitamins, amino acids and nucleic acid-related substances
- various additives such as an antifoaming agent can be added as necessary.
- the culture of a strain belonging to the genus Mitsuaria can be performed under aerobic conditions such as a shaking culture method and an aeration culture method.
- the culture conditions are not limited to these, but the temperature is 20 to 40 ° C., preferably 30 to 35 ° C., the pH is 5 to 8, preferably 6 to 7, and the culture period is 1 to 4 days, preferably 2 to 3 days is recommended.
- the strain belonging to the genus Mitsuaria cultivated as described above can be used as an active ingredient such as a bacterial plant disease control agent in the state of a culture containing living cells without separating the cells. Further, from the above culture, viable cells were separated from the culture by a usual method such as membrane separation or centrifugation, and washed as necessary. Mixtures of other components, etc.) can also be used as active ingredients. Furthermore, it can also be used in the form of a dried product obtained by lyophilizing or spray-drying the culture or the separated viable cells, or a diluted product of these liquids or solids. Moreover, it can also be used in the state formulated into various dosage forms together with various additives according to conventional methods for agricultural chemical formulations. Examples of such dosage forms include granules, emulsions, wettable powders and flowable agents.
- the concentration of viable cells contained in the bacterial plant disease control agent and the like according to the present invention is not particularly limited as long as the desired effect is exhibited. However, if the concentration of bacteria is too low, sufficient results cannot be obtained, and conversely, Since the bacteria may be wasted even if the bacteria concentration is increased, for example, when prepared and used as a liquid agent, it can be appropriately adjusted in the range of 1 ⁇ 10 5 to 1 ⁇ 10 10 cfu / ml, Desirably, the range of 1 ⁇ 10 6 to 1 ⁇ 10 9 cfu / ml is suitable.
- “cfu” means a colony forming unit.
- concentration even when using a culture, what is necessary is just to design suitably according to the said living cell density
- the present invention relates to a solanaceous plant (eg, solanaceous vegetables such as eggplant, tomato, green pepper, paprika, potato), a cucurbitaceae plant, whose pathogen is a bacterium belonging to the phytopathogenic Ralstonia solanacerum.
- solanaceous plants eg, solanaceous vegetables such as eggplant, tomato, green pepper, paprika, potato
- a cucurbitaceae plant whose pathogen is a bacterium belonging to the phytopathogenic Ralstonia solanacerum.
- Bacterial diseases such as (cucumber, bitter gourd and other Cucurbitaceae vegetables), Clavibacter michiganensis, Pseudomonas syringae, and Xanthomonas tomato (Xanthomonas tomato) Widespread against various bacterial plant diseases such as scabs such as ume, kiwifruit, citrus (oranges, grapefruits, mandarin oranges, citrus citrus fruits, etc.) A suitable control effect is exhibited. In particular, it is characterized by being very effective against bacterial wilt and eggplant tomato scab of solanaceous vegetables (solanum crops).
- control means avoiding plant diseases and the like by preventing the crops (plants) from being infected with the target bacterial plant diseases.
- the present invention provides seed germination promoting action and root, stem and leaf growth promoting action of eggplant crops and cruciferous crops (eg Chinese cabbage, cabbage, radish, turnip, broccoli, cauliflower, chingensai, komatsuna, mizumina). It can also be used as a plant growth regulator.
- eggplant crops and cruciferous crops eg Chinese cabbage, cabbage, radish, turnip, broccoli, cauliflower, chingensai, komatsuna, mizumina.
- the plant disease control agent or the like according to the present invention can be applied directly as it is, or diluted with water or the like.
- the application method as a drug is not particularly limited, for example, a method of directly attaching to crops or seeds by spraying or dipping, a method of spraying on soil, a method of adding to water or fertilizer added to crops or soil, or a farm equipment.
- the method of processing is mentioned.
- medical agent to the water added to soil, and the method of processing to an agricultural implement are more suitable.
- various bacterial plant diseases are suppressed, It also regulates plant growth.
- the drug application rate of the present invention varies depending on the applied crop, target disease, application method, occurrence tendency, degree of damage, environmental conditions, dosage form to be used, etc. For example, a method of treating 1 ml to 10 L, preferably 2 ml to 3 L, more preferably 2.5 ml to 1 L of a liquid agent per crop can be exemplified.
- the application time varies depending on the target disease, dosage form, etc., but if an effect at the seed stage is desired, before sowing and / or after sowing, if an effect at the growth stage is desired, before planting. And / or it is preferable to carry out treatment appropriately up to about 4 weeks after planting.
- the present invention is a microbial pesticide that is free from the appearance of resistant bacteria, it can be used continuously for several days or continuously.
- inorganic copper compounds such as basic copper sulfate, anhydrous copper sulfate, cupric hydroxide, basic copper chloride, etc.
- organic copper compounds for example, organic copper, copper nonylphenol sulfonate, inorganic sulfur compounds such as sulfur, total sulfide sulfur, organic sulfur compounds such as dineb, manneb, propineb, thiadiazine, thiuram, polycarbamate, anilinopyrimidine compounds such as cyprodinil , Pyrimethanil, mepanipyrim, etc., phenylpyrrole compounds, such as fludioxonil, organochlorine compounds, such as chlorothalonil, captan, triazine, fluazinam, sulfenic acid, fusalide, etc., bicarbonates such as sodium bicarbonate, potassium bicarbonate, etc.
- Organophosphorus compounds such as EDDP Fosetyl, tolcrofosmethyl, IBP, etc., benzimidazole compounds such as carbendazim, thiophanate methyl, thiabendazole, benomyl, fuberidazole, etc., dicarboximide compounds such as iprodione, procimidone, vinclozoline, etc., azole compounds such as fenbuconazole, cimeconazole , Diclobutrazole, triticonazole, ipconazole, fluconazole, microbutanyl, penconazole, viteltanol, bromconazole, oxpoconazole, cyproconazole, difenoconazole, diniconazole, epoxiconazole, fenbuconazole, fluquinconazole, flusilazole , Flutriahole, hexaconazole, imibenconazole, metcon
- mepronil, dichromedin, flutolanil, pencyclon, furametopyl, tifluzamide and the like acylalanine compounds such as oxadixyl, Metalaxyl, methoxy acrylate compounds such as azoxystrobin, cresoxime methyl, metminostrobin, trifloxystrobin, picoxystrobin, pyraclostrobin, orysastrobin, quinoxaline compounds such as quinomethionate, hydroxyanilide compounds such as Fenhexamide, etc., cyanoacetamide compounds, such as simoxanil, cyanoimidazole compounds, such as cyazofamide, etc.Famoxadone, spiroxamine, triazoxide, pyrazophos, fluorimide, dimethomorph, iprovalicarb, fenamidone, ethaboxam, cyflufenamide, dithianone, car
- a living microbial cell of a genus Mitsuaria genus for example, Mituaria chitosan nitabida TWR114, or a culture containing the microbial cell
- bacterial properties such as vegetable bacterial wilt and scab
- Tomato bacterial wilt control effect confirmation test 1 Tomatoes (variety: Ponterosa) were grown up to 4 compound leaf stages in a 9 cm diameter plastic pot (150 g of seedling culture in the bottom layer, 20 g of river sand in the middle layer, 150 g of seedling culture soil in the upper layer) filled with seedling culture soil. And after mixing 30 ml of Mitsaria chitosan nitavida TWR114 strain suspension with 70 ml of sterilized water, it treated with the bottom surface water supply to the vinyl pot, and stored for 3 days in a 30 degreeC greenhouse. Thereafter, 100 ml of the tomato bacterial wilt suspension was inoculated by the same bottom surface water supply treatment.
- the tomato bacterial wilt suspension treated with the bottom surface water supply was also carried out.
- the TWR114 strain suspension was NB medium (meat extract 0.5%, peptone 1.5%, sodium chloride 0.5%, potassium monohydrogen phosphate 0.5%, pH 7.0) at 27 ° C.
- Centrifugal collection (5000 ⁇ g, 15 minutes) was performed after shaking culture at 120 rpm for 48 hours, and this was suspended twice in sterile water.
- the absorbance at 600 nm (OD600) was 1.0 (3 ⁇ 3 ⁇ ). 10 9 cfu / ml).
- the suspension of tomato bacterial wilt fungus was centrifuged for 24 hours in CPG medium and then centrifuged and diluted with MgCl 2 (10 mM), and the 600 nm absorbance (OD600) was 1.0 (about 1 ⁇ 10 9). cfu / ml).
- Tomato bacterial wilt control effect confirmation test 2 Tomato (variety: Ponterosa) was grown in a pot (9 cm ⁇ 9 cm) filled with horticultural soil until the 4th compound leaf stage. Then, 30 ml of the TWR114 strain suspension was irrigated and stored in a greenhouse at 30 ° C. for 4 days. Thereafter, 50 ml of the tomato bacterial wilt suspension was inoculated by the same irrigation treatment. As a comparison, there are also those obtained by irrigating only the suspension of tomato bacterial wilt (non-treated group) and those obtained by spraying 10 ml of the suspension of Acinetobacter GEBT349 described in Non-Patent Document 1 (control group). Carried out.
- the TWR114 strain suspension and the GEBT349 strain suspension are the NB medium (meat extract 0.5%, peptone 1.5%, sodium chloride 0.5%, potassium monohydrogen phosphate 0.5 %, PH 7.0) at 27 ° C., 120 rpm, 48 hours of shaking culture, and then collected by centrifugation (5000 ⁇ g, 15 minutes) and suspended in sterilized water twice.
- the 600 nm absorbance (OD600) was adjusted to 1.0 (3 ⁇ 10 9 cfu / ml).
- the tomato bacterial wilt suspension was used after diluting 100 times with distilled water, which was collected by centrifugation after shaking culture in YP medium for 24 hours.
- the degree of occurrence of tomato bacterial wilt was indexed according to the following criteria 10 days after inoculation with tomato bacterial wilt, and the disease severity and control value were calculated.
- the test results are shown in Table 2 below.
- the untreated group has a disease severity of 58.3, the control group has a disease severity of 41.7 and a control value of 28.6, whereas the TWR114 strain treated group has a very low disease severity (8.3) and A high control value (85.7) was shown, and it was shown that the strain exhibited a very high control effect against tomato bacterial wilt even when compared with the control group.
- Potatoes (variety: Digima) were grown up to 4 compound leaves in a 9 cm diameter plastic pot filled with horticultural soil. Then, 30 ml of the TWR114 strain suspension was irrigated into a vinyl pot and stored in a greenhouse at 30 ° C. for 4 days. Thereafter, 50 ml of potato bacterial wilt suspension was inoculated by the same irrigation treatment. As a comparison, an irrigated treatment of only the potato bacterial wilt fungus suspension (non-treated group) and a product of 10 ml of Acinetobacter GEBT349 strain suspension (control group) were also carried out.
- the TWR114 strain suspension and the GEBT349 strain suspension are the NB medium (meat extract 0.5%, peptone 1.5%, sodium chloride 0.5%, potassium monohydrogen phosphate 0.5 %, PH 7.0) at 30 ° C., 120 rpm, 24 hours of shaking culture, and then collected by centrifugation (5000 ⁇ g, 10 minutes) and suspended in sterilized water twice.
- the 600 nm absorbance (OD600) was adjusted to 1.0 (3 ⁇ 10 9 cfu / ml).
- the potato bacterial wilt suspension was used after diluting 100-fold with distilled water after shaking culture in a YP medium for 24 hours.
- the degree of potato bacterial wilt disease was indexed according to the following criteria 14 days after the potato bacterial wilt inoculation, and the disease severity and control value were calculated.
- the test results are shown in Table 3 below.
- the untreated group has a disease severity of 50 and the control group has a higher disease severity (75.0) than the untreated group, whereas the TWR114 strain treated group has a disease severity of 16.7 and a control value of 66.7. There was also a very good control effect against potato wilt.
- Tomato tomato wilt disease control effect confirmation test
- Tomato tomato wilt disease control effect confirmation test
- Tomato tomato wilt disease control effect confirmation test
- the degree of disease of each compound leaf was indexed according to the following criteria 28 days after inoculation with tomato scab disease fungus, and the disease severity and control value were calculated from the following formulas.
- TWR114 strain suspension was obtained by centrifuging (5000 ⁇ g, 15 minutes) after culturing with shaking in NB medium at 27 ° C., 120 rpm for 6 days, and suspending this in sterile water twice. Used, and 600 nm absorbance (OD600) was adjusted to 1.0.
- TWR114 strain suspension was prepared in the same manner as in Example 1.
- test results are shown in Table 7 below. While the tap water added group has a germination rate of 63.3%, the TWR114 strain treated group has an improved germination rate (73.3%) compared to the untreated group, and the strain promotes cabbage seed germination. It was shown that the effect is also exerted.
- the present invention provides a plant non-pathogenic strain that can be used safely as a biopesticide and that exhibits a stable bacterial plant disease control effect in an actual crop production site, and a bacterial plant disease using the strain It aims at providing a control agent etc.
- Bacterial plant disease control agents such as agents and scab control agents can be provided.
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Abstract
Description
(1)細菌性植物病害防除作用を有するミツアリア属に属する菌株。
(2)ミツアリア・キトサンニタビダ(Mitsuaria chitosanitabida)に属する菌株である、(1)に記載の菌株。
(3)ミツアリア・キトサンニタビダ TWR114菌株(NITE BP-02445)。
(4)(1)~(3)のいずれか1つに記載の菌株の生菌体又は該生菌体を含む培養物を有効成分として含有することを特徴とする、細菌性植物病害防除剤。
(5)野菜類青枯病防除剤及び/又はかいよう病防除剤(例えば、野菜類かいよう病防除剤)である、(4)に記載の剤。
(6)ナス科植物の防除剤である、(4)又は(5)に記載の剤。
(7)(1)~(3)のいずれか1つに記載の菌株の生菌体又は該生菌体を含む培養物を、植物体及び/又は土壌(特に根圏土壌)に接触させるステップを含んでなる、細菌性植物病害防除方法。
(8)野菜類青枯病及び/又はかいよう病(例えば、野菜類かいよう病)を防除することを特徴とする、(7)に記載の方法。
(9)ナス科植物の病害を防除することを特徴とする、(7)又は(8)に記載の方法。
(10)(1)~(3)のいずれか1つに記載の菌株の生菌体又は該生菌体を含む培養物を有効成分として含有することを特徴とする、植物成長調節剤。
(11)野菜類種子発芽促進剤及び/又は野菜類成長促進剤である、(10)に記載の剤。
(12)(1)~(3)のいずれか1つに記載の菌株の生菌体又は該生菌体を含む培養物を、植物体及び/又は土壌(特に根圏土壌)に接触させるステップを含んでなる、植物成長調節方法。
(13)野菜類種子発芽促進及び/又は野菜類成長促進をすることを特徴とする、(12)に記載の方法。
形態:桿菌
大きさ:幅0.9~1.9μm、長さ0.9~3.0μm
運動性:+
(B)培養的性質
コロニーの色:ベージュ色~薄いピンク色
ブイヨン寒天平板培養:ベージュ色~薄いピンク色のコロニーを形成し、表面は光沢がある。
(C)生理学的性質
グラム染色性:-
硝酸塩の還元:+
インドール生成(トリプトファン):-
グルコース発酵:-
アルギニンジヒドロラーゼ:-
ウレアーゼ:-
加水分解(β-グルコシダーゼ):-
加水分解(プロテアーゼ):+
最適生育pH:中性域
最適生育温度:30~35℃
同化(グルコース):+
同化(アラビノース):+
同化(マンノース):+
同化(マンニトール):-
同化(N-アセチル-グルコサミン):+
同化(マルトース):+
同化(グルコン酸カリウム):+
同化(カプリン酸):-
同化(アジピン酸):-
同化(リンゴ酸塩):+
同化(クエン酸三ナトリウム):-
同化(酢酸フェニル):-
なお、本発明において「防除」とは、農作物(植物)が対象とする細菌性植物病害菌に感染すること等を防止することにより、当該植物病害等を回避することを意味する。
トマト(品種:ポンテローザ)を、育苗培土を充填した直径9cmのビニールポット(底層に育苗培土150g、中層に川砂20g、上層に育苗培土150g)で4複葉期まで生育させた。そして、ミツアリア・キトサンニタビダ TWR114菌株懸濁液30mlを70mlの滅菌水と混合した後、ビニールポットに底面給水によって処理し、30℃の温室で3日間保管した。その後、トマト青枯病菌懸濁液100mlを同じ底面給水処理により接種した。比較として、トマト青枯病菌懸濁液のみを底面給水処理したもの(無処理区)も実施した。なお、TWR114菌株懸濁液は、NB培地(肉エキス0.5%、ペプトン1.5%、塩化ナトリウム0.5%、リン酸一水素カリウム0.5%、pH7.0)で27℃、120rpm、48時間振盪培養後に遠心集菌(5000×g、15分)し、これを滅菌水で懸濁した操作を2回したものを使用し、600nm吸光度(OD600)が1.0(3×109cfu/ml)となるように調製した。また、トマト青枯病菌懸濁液は、CPG培地で24時間振とう培養後に遠心集菌したものをMgCl2(10mM)で希釈し、600nm吸光度(OD600)が1.0(約1×109cfu/ml)となるように調製して使用した。
トマト(品種:ポンテローザ)を、園芸培土を充填したポット(9cm×9cm)で4複葉期まで生育させた。そして、TWR114菌株懸濁液30mlを灌注処理し、30℃の温室で4日間保管した。その後、トマト青枯病菌懸濁液50mlを同じ灌注処理により接種した。比較として、トマト青枯病菌懸濁液のみを灌注処理したもの(無処理区)及び非特許文献1に記載のアシネトバクター(Acinetobacter)GEBT349菌株懸濁液10mlを茎葉散布処理したもの(対照区)も実施した。なお、TWR114菌株懸濁液及びGEBT349菌株懸濁液は、当該菌株をそれぞれNB培地(肉エキス0.5%、ペプトン1.5%、塩化ナトリウム0.5%、リン酸一水素カリウム0.5%、pH7.0)で27℃、120rpm、48時間振盪培養後に遠心集菌(5000×g、15分)し、これを滅菌水で懸濁した操作を2回したものを使用し、いずれも600nm吸光度(OD600)が1.0(3×109cfu/ml)となるように調製した。また、トマト青枯病菌懸濁液は、YP培地で24時間振とう培養後に遠心集菌したものを蒸留水で100倍に希釈して使用した。
0:発病なし
1:一部の小葉が萎凋
2:半数未満の複葉が萎凋
3:半数以上の複葉が萎凋
4:枯死
<発病度及び防除価>
発病度=Σ(発病指数×該当株数)/(調査株数×4)×100
防除価=100-(処理区の発病度/無処理区の発病度)×100
バレイショ(品種:デジマ)を、園芸培土を充填した直径9cmのビニールポットで4複葉期まで生育させた。そして、TWR114菌株懸濁液30mlをビニールポットに灌注処理し、30℃の温室で4日間保管した。その後、バレイショ青枯病菌懸濁液50mlを同じ灌注処理により接種した。比較として、バレイショ青枯病菌懸濁液のみを灌注処理したもの(無処理区)及びアシネトバクター GEBT349菌株懸濁液10mlを茎葉散布処理したもの(対照区)も実施した。なお、TWR114菌株懸濁液及びGEBT349菌株懸濁液は、当該菌株をそれぞれNB培地(肉エキス0.5%、ペプトン1.5%、塩化ナトリウム0.5%、リン酸一水素カリウム0.5%、pH7.0)で30℃、120rpm、24時間振盪培養後に遠心集菌(5000×g、10分)し、これを滅菌水で懸濁した操作を2回したものを使用し、いずれも600nm吸光度(OD600)が1.0(3×109cfu/ml)となるように調製した。また、バレイショ青枯病菌懸濁液は、YP培地で24時間振とう培養したものを蒸留水で100倍に希釈して使用した。
0:発病なし
1:一部の小葉が萎凋
2:半数未満の複葉が萎凋
3:半数以上の複葉が萎凋
4:枯死
<発病度及び防除価>
発病度=Σ(発病指数×該当株数)/(調査株数×4)×100
防除価=100-(処理区の発病度/無処理区の発病度)×100
トマト(品種:桃太郎エイト)を、実施例1と同様に育苗培土を充填した直径9cmのビニールポットで4複葉期まで生育させた。そして、トマトかいよう病菌懸濁液をハサミに霧吹きで1回噴霧後、TWR114菌株懸濁液をハサミの両面に各1回噴霧した。その後ハサミでトマト第1本葉を葉柄の付け根から切除し、28℃のガラス温室で1ヵ月栽培した。比較として、トマトかいよう病菌懸濁液のみをハサミに噴霧して切除処理したもの(無処理区)も実施した。なお、TWR114菌株懸濁液は、実施例1と同様の方法で調製し、また、トマトかいよう病菌懸濁液は、PSB培地で4日間振とう培養後に遠心集菌したものを滅菌水に懸濁し、約6×108cfu/mlに調製して使用した。
0:無病徴
1:複葉の一部に壊死・萎凋症状あり
2:複葉の大部分が壊死・萎凋
<発病度及び防除価>
発病度(%)={Σ(発病指数×複葉数)/全複葉数×2}×100
防除価={1-(処理区の発病度/無処理区の発病度)}×100
直径60mmのシャーレ2枚にそれぞれ濾紙を敷き、各60粒のトマト種子(品種:ポンテローザ)を濾紙上に播種した。更に、TWR114菌株懸濁液又は水道水を2ml添加し、夜間20℃、日中25℃の恒温器内に静置した。なお、TWR114菌株懸濁液は、NB培地で27℃、120rpm、6日間振盪培養後に遠心集菌(5000×g、15分)し、これを滅菌水で懸濁した操作を2回したものを使用し、600nm吸光度(OD600)が1.0となるように調製した。
直径9cmのビニールポットに滅菌土壌(育苗培土:川砂:バーミキュライト=1:1:1)を充填し、ここでトマト(品種:ポンテローザ)を4複葉期まで生育させた。そして、TWR114菌株懸濁液30mlを滅菌水70mlと混合した後、ビニールポットに底面給水により処理し、30℃の温室で28日間保管した。その間、適宜底面給水を行った。比較として、底面給水処理のみしたもの(無処理区)も実施した。なお、TWR114菌株懸濁液は、実施例1と同様の方法により調製した。
直径60mmのシャーレ2枚にそれぞれ濾紙を敷き、水道水を1ml添加した後、各30粒のキャベツ種子(品種:四季獲)を濾紙上に播種した。更に、TWR114菌株懸濁液、又は水道水を1ml添加し、夜間20℃、日中25℃の恒温器内に静置した。なお、TWR114菌株懸濁液は、NB培地で27℃、120rpm、6日間振盪培養後に遠心集菌(5000×g、15分)し、これを滅菌水で懸濁した操作を2回したものを使用し、600nm吸光度(OD600)が0.32となるように調製した。
(1)ミツアリア・キトサンニタビダ(Mitsuaria chitosanitabida)TWR114菌株(NITE BP-02445)。
Claims (13)
- 細菌性植物病害防除作用を有するミツアリア(Mitsuaria)属に属する菌株。
- ミツアリア・キトサンニタビダ(Mitsuaria chitosanitabida)に属する菌株である、請求項1に記載の菌株。
- ミツアリア・キトサンニタビダ(Mitsuaria chitosanitabida)TWR114菌株(NITE BP-02445)。
- 請求項1~3のいずれか1項に記載の菌株の生菌体又は該生菌体を含む培養物を有効成分として含有することを特徴とする、細菌性植物病害防除剤。
- 野菜類青枯病防除剤及び/又はかいよう病防除剤である、請求項4に記載の剤。
- ナス科植物の防除剤である、請求項4又は5に記載の剤。
- 請求項1~3のいずれか1項に記載の菌株の生菌体又は該生菌体を含む培養物を、植物体及び/又は土壌に接触させるステップを含んでなる、細菌性植物病害防除方法。
- 野菜類青枯病及び/又はかいよう病を防除することを特徴とする、請求項7に記載の方法。
- ナス科植物の病害を防除することを特徴とする、請求項7又は8に記載の方法。
- 請求項1~3のいずれか1項に記載の菌株の生菌体又は該生菌体を含む培養物を有効成分として含有することを特徴とする、植物成長調節剤。
- 野菜類種子発芽促進剤及び/又は野菜類成長促進剤である、請求項10に記載の剤。
- 請求項1~3のいずれか1項に記載の菌株の生菌体又は該生菌体を含む培養物を、植物体及び/又は土壌に接触させるステップを含んでなる、植物成長調節方法。
- 野菜類種子発芽促進及び/又は野菜類成長促進をすることを特徴とする、請求項12に記載の方法。
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CN201880036862.6A CN111094544B (zh) | 2017-04-04 | 2018-03-30 | 属于松江菌属的菌株和使用了该菌株的微生物农药 |
BR112019020762A BR112019020762A2 (pt) | 2017-04-04 | 2018-03-30 | cepa pertencente ao gênero mitsuaria, agente de controle de doenças bacterianas em plantas, método de controle de doenças bacterianas de plantas, regulador de crescimento de plantas, e, método para regular o crescimento de plantas. |
US16/500,681 US20200359635A1 (en) | 2017-04-04 | 2018-03-30 | Strain belonging to genus mitsuaria and microbial pesticide using said strain |
KR1020197029640A KR102448442B1 (ko) | 2017-04-04 | 2018-03-30 | 미츠아리아속에 속하는 균주 및 그 균주를 사용한 미생물 농약 |
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WO2023288294A1 (en) | 2021-07-16 | 2023-01-19 | Novozymes A/S | Compositions and methods for improving the rainfastness of proteins on plant surfaces |
FR3127675A1 (fr) * | 2021-10-04 | 2023-04-07 | Biointrant | composition comprenant une souche de Mitsuaria noduli |
WO2023225459A2 (en) | 2022-05-14 | 2023-11-23 | Novozymes A/S | Compositions and methods for preventing, treating, supressing and/or eliminating phytopathogenic infestations and infections |
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JP2012211124A (ja) | 2011-03-24 | 2012-11-01 | National Agriculture & Food Research Organization | 青枯病抵抗性誘導剤及び青枯病防除方法 |
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SOMEYA, N. ET AL.: "Diversity of culturable chitinolytic bacteria from rhizospheres of agronomic plants in japan", MICROBES ENVIRONMENTS, vol. 26, no. 1, 2011, pages 7 - 14, XP055559357 * |
Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2023288294A1 (en) | 2021-07-16 | 2023-01-19 | Novozymes A/S | Compositions and methods for improving the rainfastness of proteins on plant surfaces |
FR3127675A1 (fr) * | 2021-10-04 | 2023-04-07 | Biointrant | composition comprenant une souche de Mitsuaria noduli |
WO2023057375A1 (fr) * | 2021-10-04 | 2023-04-13 | Biointrant | Composition comprenant une souche de mitsuaria noduli |
WO2023225459A2 (en) | 2022-05-14 | 2023-11-23 | Novozymes A/S | Compositions and methods for preventing, treating, supressing and/or eliminating phytopathogenic infestations and infections |
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KR102448442B1 (ko) | 2022-09-27 |
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US20200359635A1 (en) | 2020-11-19 |
JP7296588B2 (ja) | 2023-06-23 |
BR112019020762A2 (pt) | 2020-04-28 |
KR20190136009A (ko) | 2019-12-09 |
JPWO2018186307A1 (ja) | 2020-05-14 |
CN111094544A (zh) | 2020-05-01 |
CN111094544B (zh) | 2023-10-24 |
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