WO2015033400A1 - コリモナス属細菌の培養方法及び保存方法 - Google Patents
コリモナス属細菌の培養方法及び保存方法 Download PDFInfo
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- 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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- 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
- A01N63/22—Bacillus
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- 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
- C12N1/04—Preserving or maintaining viable microorganisms
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- the present invention relates to a method for culturing microorganisms, and more particularly, to a method for cultivating a bacterium belonging to the genus Colimonas, which is suitable for long-term storage.
- Bacteria belonging to the genus Collimonas are known as microorganisms that suppress the growth of plant pathogenic bacteria, and various studies have been conducted. If the action of suppressing the growth of plant pathogenic bacteria by the genus Collimonas is clarified, the bacteria can be expected to be used as a microbial pesticide.
- microorganisms contained in the microbial pesticide are desired to have high storage stability in order to exert the action provided by each microorganism.
- the inventors have conducted intensive research in order to obtain a microorganism storage method that can be stored for a long period of time in addition to the conventionally known microorganism storage methods described above.
- an object of the present invention is to provide a novel culture method and storage method suitable for preserving Collimonas bacteria for a long period of time.
- the invention of claim 1 A method for culturing a bacterium belonging to the genus Corimonas characterized by culturing the bacterium belonging to the genus Corimonas in a sealed state in a medium containing rice bran medium and / or okara medium.
- the invention of claim 2 2.
- the invention of claim 3 A method for culturing a bacterium belonging to the genus Corimonas, comprising culturing the bacterium belonging to the genus Corimonas in a medium containing green tuff.
- the invention of claim 4 4.
- the invention of claim 5 5.
- FIG. 2 is a diagram showing the growth state of the D-25 strain after 4 days of culturing from the state shown in FIG.
- FIG. 7 is a diagram showing the growth state of the D-25 strain after inoculating and cultivating the D-25 strain in a medium containing Towada stone, and after vacuum drying treatment, and (a) D- Inoculated 100 ⁇ L of 25 strains of bacterial solution, (b) Inoculated 100 ⁇ L of D-25 strain with 0.7 g of Towada stone powder, and added 200 ⁇ L of sterilized water, (c) 0 100 gL of D-25 strain was added to 5 g of Towada stone powder, and 400 ⁇ L of sterilized water was added.
- Samples were prepared in which the ratio of the weight of Towada stone to the amount of water including the pre-culture solution and sterilized water of the D-25 strain was 7: 3, 5: 5, and each D-25 strain It is the graph which calculated the number of bacteria. This shows the state of D-25 strain inoculated on Towada stone powder. From left, 100 ⁇ L of D-25 strain was inoculated on 0.7 g of Towada stone powder, and 200 ⁇ L of TSB and glycerin 3% were infused.
- the microorganism used in this embodiment was a microorganism belonging to Collimonas sp.
- Examples of microorganisms belonging to Collimonas sp. include D-25 strain, Cal2 strain, and Cal31 strain. Among these, the mycological properties of the D-25 strain are shown in Tables 1 to 3.
- Towada stone as an additive used in this embodiment is a green tuff mined in Hinai-cho, Odate City, Akita Prefecture, the official name is ⁇ quartz andesitic pumice tuff '', and it is called ⁇ Green Tough '' Are known.
- Towada stone is a composite ore containing quartz, feldspar, chlorite, etc. as a mineral composition, and has a characteristic that it contains a lot of minerals.
- Towada stone is porous and has functions such as adsorption and release of substances, and is also used as an agricultural fertilizer.
- Table 4 shows the main composition of Towada stone.
- D-25 strain An attempt was made to preserve D-25 by adding glycerin and Towada stone in rice bran / okara medium of Collimonas sp. D-25 (hereinafter referred to as “D-25 strain”) and R2A medium.
- the medium in the pre-culture was R2A medium
- the medium in the main culture was rice bran / okara medium
- the R2A medium, and glycerin and Towada stone were added to the medium in the main culture.
- the composition of the R2A medium is as follows.
- the D-25 strain was inoculated with 2 toothpick into 2 mL of R2A medium, pre-cultured at 24 ° C. and 200 rpm for 2 days, and the viable count was calculated.
- each of the four sample tubes has a glycerin concentration of 0% (sterile distilled water only), 10% (W / W), 30% (W / W), 50 1 mL of% (W / W) aqueous solution was added, sealed with parafilm, and allowed to stand at 24 ° C.
- the cells from the R2A medium and rice bran / okara medium on the sixth day of the culture were obtained, and two sample tubes containing only each of the cells were prepared separately. 200 ⁇ L of sterilized water was added, and 0.5 g and 1.0 g of sterilized Towada stone were added, well vortexed, and the opening was tightly wrapped with parafilm and allowed to stand at 24 ° C.
- the number of viable cells on the 0th day, the 2nd day, the 4th day, and the 6th day of the sample cultured in the R2A medium and rice bran / okara medium was calculated.
- the sample to which the glycerin was added was used to calculate the viable cell count on the 3rd and 6th days.
- the sample to which the Towada stone was added was subjected to viable cell count on the 6th day.
- Bacteria belonging to the genus Collimonas are known as microorganisms that grow in an oligotrophic environment, for example, microorganisms that grow on an R2A medium and are difficult to grow on a medium rich in nutrients. Moreover, regarding the storage, storage at a low temperature of 4 ° C. or less is known.
- the sterilized Towada stone is put in a container that can be sealed, D-25 strain is added, and a small amount of water is added, so that it can be stored in powder form (Table 7). If this method is used, it is possible to prevent the microbial cells from being killed by vacuum drying or freeze drying.
- the medium in the preculture is the same as the R2A medium described in Example 1.
- the D-25 strain was inoculated with 2 toothpick into 2 mL of R2A medium, and precultured at 24 ° C. and 200 rpm for 2 days. After pre-culture, the culture solution was centrifuged at 6000 rpm for 3 minutes, the supernatant was removed, and an equal amount of physiological saline was added to make a bacterial solution for inoculation.
- FIG. 1 shows the results of calculating the viable cell count of the D-25 strain in the three culture solutions.
- Cal2 strain Collimonas sp. Cal2 strain
- Collimonas sp. Collimonas sp. (Collimonas sp.) Cal31 strain
- Cal31 strain Tried to save.
- the medium in the preculture is the same as the R2A medium described in Example 1.
- Both Cal2 and Cal31 strains were found to grow to 10 9 cfu / ml when cultured in rice bran / okara medium for 3 days.
- the D-25 strain also showed a slightly higher number of viable bacteria than the Cal2 and Cal31 strains.
- the strain belonging to Collimonas sp. Can be grown in rice bran or okara medium and the number of viable bacteria can be maintained at a constant number.
- a powder obtained by treating Towada stone in powder form was used as a carrier for fixing the D-25 strain.
- Towada stone powder that had been subjected to dry heat sterilization treatment was prepared, and 5 g of the powder was added to the centrifuge tube containing the culture solution of D-25 strain and stirred in a mothers small reactor. Stirring was performed 3 times in total for 1 minute, changing the upper and lower mounting positions. During the stirring, a large lump of Towada stone was formed, but it was processed by crushing with platinum ears.
- the number of viable bacteria of the D-25 strain to which Towada stone was added was 5.5 ⁇ 10 7 cfu / g, and a microorganism preparation of about 10 7 cfu / g could be prepared.
- the reason why the number of viable bacteria has decreased is that stirring was too intense.
- Collimononas sp. Can secure a large amount of bacteria in a short time, it can be covered by increasing the initial input amount.
- Example 1 it was considered that when Towada stone was added to the culture solution of the D-25 strain, it was possible to preserve the D-25 strain in powder form. Therefore, in this example, the bacteria of the genus Collimonas were used. Towada stone powder suitable for growth and water content were examined.
- the bacterium belonging to the genus Collimonas used in this example was the D-25 strain, and Towada stone treated in a powder form of 1 ⁇ m to 80 ⁇ m was used as a carrier for fixing the D-25 strain.
- Towada stone powder was weighed 0.9 g, 0.7 g, 0.5 g, dispensed into test tubes, and then sterilized with a dry heat sterilizer.
- Table 9 shows the viable count calculation results for the D-25 strain.
- the number of viable cells indicates a value when 3 mL of sterilized water is added.
- the state of D-25 stock at the time of calculation is shown in FIG.
- sample 2 and sample 3 growth of D-25 strain was confirmed.
- viable bacteria were confirmed up to 10 4 cfu / mL.
- Example 4 D-25 strain was inoculated into Towada stone powder, and when sterilized water was added, the growth of D-25 strain was confirmed. Therefore, in this example, it was suitable for the growth of D-25 strain. An experiment was conducted to confirm the weight ratio of Towada stone and water content.
- Example Method 1 The D-25 strain was cultured for 1 week in a medium containing Towada stone, and then vacuum-dried for 3 days (once reduced pressure treatment, once for about 30 seconds) to remove moisture. After drying, 3 mL of sterile water was added and suspended, and the number of viable bacteria in the suspension was calculated.
- Example Method 2 Samples were prepared by adjusting the ratio of the weight of Towada stone to the amount of water containing the pre-culture solution and sterilized water of D-25 strains to 7: 3, 5: 5, and each D-25 strain The number of bacteria was calculated.
- Sample 1 Inoculated 100 ⁇ L of D-25 strain preculture into 0.7 g of Towada stone powder, and added 200 ⁇ L of TSB and 3% glycerin (weight of Towada stone and D-25 strain preculture The ratio of the water content to the water content is 7: 3)
- Sample 2 100 g of the preculture of D-25 strain was inoculated to 0.6 g of Towada stone powder, and 300 ⁇ L of TSB and 3% of glycerin were added (the weight of Towada stone and the preculture of D-25 strain The ratio of the water content to the water content is 6: 4)
- Sample 3 100 g of the preculture of D-25 strain was inoculated to 0.5 g of Towada stone powder, and 400 ⁇ L of TSB and 3% of glycerin were added (the weight of Towada stone and the preculture of D-25 strain The ratio of the water content to the water content is 5
- Table 10 shows the results of viable count calculation.
- D-25 strain was inoculated into Towada stone powder, and when sterilized water was added, the growth of D-25 strain was confirmed.
- Towada stone powder D-25 strain were used.
- an experiment was conducted to observe the inhibitory effect on the phytopathogenic fungi of the genus Collimonas.
- Sample 1 100 ⁇ L of D-25 strain preculture solution added to 0.7 g of Towada stone powder, and 200 ⁇ L of sterilized water added
- Sample 2 100 ⁇ L of D-25 strain preculture solution, sterilized water
- Sample 3 100 g of the preculture solution of D-25 strain was inoculated into 0.7 g of Towada stone powder that was not sterilized, and 300 ⁇ L of sterilized water was added
- Sample 4 0.7 g 100 ⁇ L of the preculture solution of D-25 strain was added to Towada stone powder that had been sterilized, and 300 ⁇ L of sterilized water was added.
- Colimonas strain D-25 was precultured in 1/10 TSB medium for 4 days at 24 ° C. with shaking.
- the viable cell count after the preculture was 2.6 ⁇ 10 7 cfu / mL.
- each sample was prepared, it was left to stand at room temperature for 2 days, and 3 mL of sterilized water was added to each sample and stirred using a vortex. For D-25 of the stirred culture, the viable cell count was calculated using 1/10 TSA medium.
- the culture solution according to each sample was inoculated into 10 ⁇ LTSA and 1/10 TSA medium, and at the same time, Rhizoctonia agar sections were inoculated. Cultivation was performed at room temperature, and it was observed whether it was suppressed against D-25 strain Rhizoctonia.
- the bacteria belonging to the genus Collimonas can be stored for a long period of time, so that the bacteria can be used as a microbial pesticide.
- green tuff can be used as a carrier, it can be used as a coating material for plant seeds.
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Abstract
Description
米ぬか培地及び/又はおから培地を含む培地にてコリモナス属細菌を密封状態で培養することを特徴とするコリモナス属細菌の培養方法である。
前記米ぬか培地及び/又はおから培地を含む培地に、添加剤として緑色凝灰岩を添加することを特徴とする請求項1記載のコリモナス属細菌の培養方法である。
緑色凝灰岩を含む培地にてコリモナス属細菌を培養することを特徴とするコリモナス属細菌の培養方法である。
請求項3において、前記緑色凝灰岩の重量と培養液の水分量との比率を7:3に調整してコリモナス属細菌を培養することを特徴とするコリモナス属細菌の培養方法である。
請求項1乃至4何れか一項において、培養後のコリモナス属細菌を常温下で保存することを特徴とするコリモナス属細菌の保存方法である。
表1~表3に記載の菌額的性質を有するD-25株をコリモナス(Collimonas)属分類群に帰属するものと推定した。この菌株は、平成23年6月9日付けで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(日本国千葉県木更津市かずさ鎌足2-5-8)に受託番号NITE P-1104として寄託されている。
ペプトン:0.5g
酵母エキス:0.5g
カザミノ酸:0.5g
グルコース:0.5g
可溶性デンプン:0.5g
ピルビン酸Na :0.5g
K2HPO4:0.3g
MgSO4:0.05g
R2A培地2mLにD-25株を爪楊枝で植菌し、2日間、24℃、200 rpmで前培養をし、生菌数算定を行った。
R2A培地で前培養を行ったD-25株について、培養2日目の生菌数は、5.67×109cfu/mlであった。表5~表7それぞれに、前記R2A培地、米ぬか・おから培地で本培養をしたサンプル、前記グリセリンを加えたサンプル、前記十和田石を加えたサンプルにおけるD-25株の生菌数算定結果を示す。
(考察)
米ぬか・おから培地とR2A培地でD-25株を培養したものを比較すると、全体的に米ぬか・おから培地における増殖が良く、生菌数も長期間維持できている(表5)。更に、常温下で、8ヶ月経過後も108cfu/mlを維持している。これは、米ぬか・おから培地において、遠心分離をした際に培地成分の持込(沈殿として米ぬか・おからの一部が沈殿する)が多いため、生菌数を長く維持できたと考えられる。
R2A培地2mLにD-25株を爪楊枝で植菌し、2日間、24℃、200 rpmで前培養を行った。前培養後、培養液を6000 rpmで3分間遠心分離し、上清を取り除き、等量の生理食塩水を加え、これを植菌用の菌液とした。
おから・米ぬか培地では6日目に1010cfu/mlまで増殖し(図1(a))、おから培地、米ぬか培地ではそれぞれ109cfu/mlまで増殖した(図1(b)、(c))。
R2A培地、4℃で保存していたD-25株、Cal2株、Cal31株それぞれを新しいR2A液体培地に植菌し、24℃で2日間前培養を行った。
R2A培地で前培養を行ったCal2株、Cal31株、D-25株について、前培養後の生菌数はそれぞれ以下の通りであった。
D-25株 : 4.3×109 cfu/mL
Cal2株:1.8×109 cfu/mL
Cal31株 : 5.3×109 cfu/mL
表8に、Cal2株、Cal31株、D-25株の生菌数算定結果を示す。
Cal2株、Cal31株について、どちらも米ぬか・おから培地で3日間培養すると109cfu/mlまで生育することが分かった。D-25株についてもCal2株、Cal31株に比べてやや高い生菌数を示した。
<D-25株を使用した微生物製剤の作製>
実施例3において米ぬか・おから培地で培養したD-25株の培養液を用いて、微生物製剤作製のための実験を行った。
実施例3で作製した米ぬか・おから培地によるD-25株の培養液を2mL 遠心チューブ(15mL容)に分注した。生菌数は1.5×1010 cfu/2mLであった。
十和田石を添加したD-25株の生菌数は5.5×107cfu/gとなり、107cfu/g程の微生物製剤を作製することができた。生菌数が低下してしまった要因としては、攪拌が激しすぎたことが挙げられる。しかしコリモナス スピーシーズ(Collimonas sp.)は菌量を短時間で多量に確保できるので、初期の投入量を増やすことでカバーできると考えられる。
十和田石の粉末を0.9g、 0.7g、 0.5g 秤量し、試験管に分注後、乾熱滅菌器にて滅菌処理を行った。
サンプル1については、D-25株の生育が見られなかった。図3(a)中白く見えるものは、十和田石の粉末だと思われる。
十和田石を含む培地でD-25株を一週間培養後、真空乾燥を3日間行い(減圧処理は1回、30秒程度)水分を除去した。乾燥後、滅菌水を3mL加え懸濁し、懸濁液中の生菌数を算定した。
0.9gの十和田石粉末にD-25株の前培養液を100μL植菌したもの
0.7gの十和田石粉末にD-25株の前培養液を100μL植菌し、減菌水200μL加えたもの
0.5gの十和田石粉末にD-25株の前培養液を100μL植菌し、減菌水400μL加えたもの
生菌数算定の結果、どのサンプルでも、図4図示のようにD-25株の生育を確認することはできなかったので、死滅したと考えられる。
十和田石の重量とD-25株の前培養液及び減菌水を含む水分の水分量との比を7:3、5:5に調整したサンプルを作製し、それぞれのD-25株の生菌数を算定した。
図5図示のように、一週間以上の生存が認められ、106cfu/mL程度の生菌数が保持されていることが確認された。
上述した実験方法1において、D-25株の培養後、真空乾燥処理を行うとコリモナスが死滅するため、滅菌水の代わりにTSB及びグリセリン3%を添加し培養を試みた。なお、D-25株の前培養液の生菌数は2.6×107cfu/mLであった。
サンプル1:0.7gの十和田石粉末にD-25株の前培養液を100μL植菌し、TSB及びグリセリン3%を200μL添加したもの
(十和田石の重量とD-25株の前培養液を含む水分の水分量との比が7:3)
サンプル2:0.6gの十和田石粉末にD-25株の前培養液を100μL植菌し、TSB及びグリセリン3%を300μL添加したもの
(十和田石の重量とD-25株の前培養液を含む水分の水分量との比が6:4)
サンプル3:0.5gの十和田石粉末にD-25株の前培養液を100μL植菌し、TSB及びグリセリン3%を400μL添加したもの
(十和田石の重量とD-25株の前培養液を含む水分の水分量との比が5:5)
図5図示されているように、0.5gの十和田石粉末にD-25株の前培養液を100μL植菌し、減菌水400μL加えたものでは、108cfu/mL程の生菌数が得られているため、本実施例におけるサンプル1ではこれと同等の結果が得られている。
実施例4及び5において、十和田石粉末にD-25株を植菌し、減菌水を加えるとD-25株の生育が確認できたので、参考例として、十和田石粉末、D-25株の条件を変えてD-25株の生育状態を確認すると共に、コリモナス(Collimonas)属細菌の植物病原菌に対する抑制効果を観察する実験を行った。
サンプル1:0.7gの十和田石粉末にD-25株の前培養液を100μL植菌し、減菌水を200μL添加したもの
サンプル2:D-25株の前培養液100μLに、減菌水を900μL添加したもの
サンプル3:0.7gの減菌処理していない十和田石粉末にD-25株の前培養液を100μL植菌し、減菌水を300μL添加したもの
サンプル4:0.7gの減菌処理した十和田石粉末にD-25株の前培養液を100μL植菌し、減菌水を300μL添加したもの
コリモナスD-25株を1/10TSB培地で4日間、24℃で振とう前培養を行った。前培養後の生菌数は2.6×107cfu/mLであった。
サンプル2でも増殖が認められることから、十和田石からの栄養分の溶出よりも、前培養の1/10TSBの持ちこみの栄養分による増殖と考える方が適当であると思われる。
十和田石でD-25を培養し植物病原菌に対する抑制効果を確認すると、D-25のみに比べ阻止円の形成がよりはっきりとしていた。
Claims (5)
- 米ぬか培地及び/又はおから培地を含む培地にてコリモナス属細菌を密封状態で培養する
ことを特徴とするコリモナス属細菌の培養方法。 - 前記米ぬか培地及び/又はおから培地を含む培地に、添加剤として緑色凝灰岩を添加する
ことを特徴とする請求項1記載のコリモナス属細菌の培養方法。 - 緑色凝灰岩を含む培地にてコリモナス属細菌を培養する
ことを特徴とするコリモナス属細菌の培養方法。 - 請求項3において、前記緑色凝灰岩の重量と培養液の水分量との比率を7:3に調整してコリモナス属細菌を培養する
ことを特徴とするコリモナス属細菌の培養方法。 - 請求項1乃至4何れか一項において、培養後のコリモナス属細菌を常温下で保存する
ことを特徴とするコリモナス属細菌の保存方法。
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PCT/JP2013/073781 WO2015033400A1 (ja) | 2013-09-04 | 2013-09-04 | コリモナス属細菌の培養方法及び保存方法 |
EP13893103.5A EP3042949B1 (en) | 2013-09-04 | 2013-09-04 | Cultivation method for bacteria belonging to genus collimonas and storage method |
DK13893103.5T DK3042949T3 (en) | 2013-09-04 | 2013-09-04 | PROCEDURE FOR CULTIVATING BACTERIES BELONGING TO COLLIMONAS GENES AND PROCEDURES FOR STORAGE |
JP2015535200A JP6336453B2 (ja) | 2013-09-04 | 2013-09-04 | コリモナス属細菌の培養方法及び保存方法 |
US14/916,255 US20160194601A1 (en) | 2013-09-04 | 2013-09-04 | Culturing method for bacteria belonging to genus collimonas and storage method |
US16/213,639 US10487306B2 (en) | 2013-09-04 | 2018-12-07 | Culturing method for bacteria belonging to genus Collimonas and storage method |
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US14/916,255 A-371-Of-International US20160194601A1 (en) | 2013-09-04 | 2013-09-04 | Culturing method for bacteria belonging to genus collimonas and storage method |
US16/213,639 Division US10487306B2 (en) | 2013-09-04 | 2018-12-07 | Culturing method for bacteria belonging to genus Collimonas and storage method |
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WO2015033400A1 true WO2015033400A1 (ja) | 2015-03-12 |
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US (2) | US20160194601A1 (ja) |
EP (1) | EP3042949B1 (ja) |
JP (1) | JP6336453B2 (ja) |
DK (1) | DK3042949T3 (ja) |
WO (1) | WO2015033400A1 (ja) |
Cited By (1)
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JP5909695B1 (ja) * | 2015-06-24 | 2016-04-27 | 学校法人東京農業大学 | 植物の細菌性病害に対する微生物防除剤および種子コーティング剤並びに該種子コーティング剤をコートした種子 |
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JPH10243781A (ja) | 1997-03-03 | 1998-09-14 | Shimizu Corp | 微生物の長期保存方法 |
JP2000229802A (ja) * | 1999-02-10 | 2000-08-22 | Hiroshi Kawai | 植物土壌病害予防用微生物資材およびその施用方法 |
WO2013038542A1 (ja) * | 2011-09-15 | 2013-03-21 | 一般社団法人 新環境技術評議会 | コリモナス(Collimonas)属細菌を用いた植物病原菌の増殖抑制方法 |
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KR100430761B1 (ko) * | 2000-10-02 | 2004-05-10 | (주)에스비바이오테크 | 고밀도 길항 미생물 기재의 제조방법 |
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JP6210560B2 (ja) * | 2012-10-10 | 2017-10-11 | 十和田グリーンタフ・アグロサイエンス株式会社 | トリコデルマ(Trichoderma)属菌を用いた植物病原菌の増殖抑制方法 |
-
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- 2013-09-04 JP JP2015535200A patent/JP6336453B2/ja active Active
- 2013-09-04 EP EP13893103.5A patent/EP3042949B1/en not_active Not-in-force
- 2013-09-04 DK DK13893103.5T patent/DK3042949T3/en active
- 2013-09-04 US US14/916,255 patent/US20160194601A1/en not_active Abandoned
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Cited By (2)
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JP5909695B1 (ja) * | 2015-06-24 | 2016-04-27 | 学校法人東京農業大学 | 植物の細菌性病害に対する微生物防除剤および種子コーティング剤並びに該種子コーティング剤をコートした種子 |
JP2017007985A (ja) * | 2015-06-24 | 2017-01-12 | 学校法人東京農業大学 | 植物の細菌性病害に対する微生物防除剤および種子コーティング剤並びに該種子コーティング剤をコートした種子 |
Also Published As
Publication number | Publication date |
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US10487306B2 (en) | 2019-11-26 |
EP3042949A1 (en) | 2016-07-13 |
US20160194601A1 (en) | 2016-07-07 |
JP6336453B2 (ja) | 2018-06-06 |
US20190177682A1 (en) | 2019-06-13 |
DK3042949T3 (en) | 2019-01-21 |
EP3042949A4 (en) | 2017-05-17 |
EP3042949B1 (en) | 2018-11-14 |
JPWO2015033400A1 (ja) | 2017-03-02 |
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