JP2846355B2 - Soil Disease Prevention Law - Google Patents
Soil Disease Prevention LawInfo
- Publication number
- JP2846355B2 JP2846355B2 JP1212651A JP21265189A JP2846355B2 JP 2846355 B2 JP2846355 B2 JP 2846355B2 JP 1212651 A JP1212651 A JP 1212651A JP 21265189 A JP21265189 A JP 21265189A JP 2846355 B2 JP2846355 B2 JP 2846355B2
- Authority
- JP
- Japan
- Prior art keywords
- soil
- cells
- disease prevention
- soil disease
- antibiotic
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Description
【発明の詳細な説明】 〔産業上の利用分野〕 本発明は土壌病害防止法に関し、詳しくは、土壌伝染
病の防除を目的とした、抗生物質生産菌による土壌病害
防止法に関するものである。The present invention relates to a method for preventing soil diseases, and more particularly, to a method for preventing soil diseases caused by antibiotic-producing bacteria for the purpose of controlling soil infectious diseases.
現在、農作物の周年栽培や連作栽培による土壌病原菌
の増殖を阻止するために臭化メチルやクロルピクリン等
の化学農薬が使用されている。このため土壌中では、有
益な微生物も死滅してしまい土壌の活力が失なわれ生産
性が低下しつつある。さらにその土壌を再生させるため
に施用される不熟有機肥料により、線虫やダニ等の病害
虫が以前にもまして増大し、そのためにさらに農薬を用
いるという悪循環がくり返されている。At present, chemical pesticides such as methyl bromide and chloropicrin are used to prevent the growth of soil pathogenic bacteria due to year-round cultivation or continuous cultivation of crops. For this reason, beneficial microorganisms are also killed in the soil, the vitality of the soil is lost, and productivity is decreasing. In addition, pests such as nematodes and mites are increasing more than ever before due to immature organic fertilizers applied to regenerate the soil, and the vicious cycle of using more pesticides has been repeated.
近年これら化学農薬に変わるものとして、微生物の拮
抗作用を利用して土壌病害を防止する方法が行なわれる
ようになり、現在土壌改良剤及び拮抗微生物資材として
市販されている。しかしこれらは、流通時、保存時にお
いて安定性に欠け、菌数の減少弱体化により土壌に施用
した際に十分な効果を発揮できないという問題を抱えて
いる。さらに十分量の有用菌で土壌を処理しても、定着
性が悪い為に有効な効果を示さないという問題があっ
た。In recent years, as a substitute for these chemical pesticides, a method for preventing soil diseases by using the antagonistic action of microorganisms has been carried out, and is currently marketed as a soil conditioner and an antagonistic microorganism material. However, they have a problem that they lack stability during distribution and storage and cannot exert a sufficient effect when applied to soil due to a decrease in the number of bacteria and weakening. Further, even if the soil is treated with a sufficient amount of useful bacteria, there is a problem that an effective effect is not exhibited due to poor fixation.
本発明らは、上記課題を解決するために鋭意研究を行
なった結果、ポリヒドロキシブチレート（以下PHBと略
記する）を菌体内に蓄積させた抗生物質生産菌を飢餓状
態にすることにより、土壌施用の際に混合するキャリヤ
ーとの吸着性が向上し、有効に土壌病害を防止すること
ができることを見出し本発明を完成するに至った。The present inventors have conducted intensive studies in order to solve the above-mentioned problems, and as a result, by starving the antibiotic-producing bacteria in which polyhydroxybutyrate (hereinafter abbreviated as PHB) has been accumulated in the cells, soil The present inventors have found that the adsorptivity to a carrier mixed during application is improved, and soil disease can be effectively prevented, and the present invention has been completed.
すなわち本発明は、抗生物質生産菌を栄養源を含む培
地で培養して菌体内にPHBを蓄積させた後、洗菌し、飢
餓状態とし、キャリヤーと混合して土壌に施用すること
を特徴とする土壌病害防止法を提供するものである。That is, the present invention is characterized in that an antibiotic-producing bacterium is cultured in a nutrient-containing medium to accumulate PHB in the cells, washed, starved, mixed with a carrier, and applied to soil. To provide a soil disease prevention law.
本発明に用いられる抗生物質生産菌としてはプソイド
モナス（Pseudomonas）属又はバチルス（Bacillus）属
の菌が挙げられる。Pseudomonas属としてはPseudomonas
（以下P.と略す）cepacia、P.gladioli、P.mallei、P.p
seudomallei、P.picketti、P.eolanacearum、P.voran
s、P.steroni、P.delafieldii、P.facilis、P.saccharo
phila、P.flava、P.pseudoflava、P.palleronii、P.car
yophylli、P.diminuta、P.vesicularis等が挙げられる
が、好ましくはP.cepacia、P.gladioliである。またBac
illus属では、Bacillus（以下B.と略す）cereus、B.ant
hracis、B.azotoformans、B.mycoides、B.thuringiensi
s等が挙げられ、好ましくはB.cereus、B.mycoidesであ
る。Examples of the antibiotic-producing bacteria used in the present invention include bacteria belonging to the genus Pseudomonas or the genus Bacillus. Pseudomonas as a genus of Pseudomonas
(Hereinafter abbreviated as P.) cepacia, P. gladioli, P. mallei, Pp
seudomallei, P.picketti, P.eolanacearum, P.voran
s, P.steroni, P.delafieldii, P.facilis, P.saccharo
phila, P.flava, P.pseudoflava, P.palleronii, P.car
Yophylli, P. diminuta, P. vesicularis and the like are preferable, and P. cepacia and P. gladioli are preferable. Also Bac
In the genus illus, Bacillus (hereinafter abbreviated as B.) cereus, B.ant
hracis, B. azotoformans, B. mycoides, B. thuringiensi
s and the like, and preferably B. cereus and B. mycoides.
本発明における飢餓状態は、菌体を非栄養性の物質と
菌体濃度が10８〜14）個/g程度になる様に混合すること
により形成される。尚、非栄養性の物質としては、水あ
るいは炭素源や窒素源を含んでいない水溶液、軽量多孔
質無機担体、ゼオライト、ケイソウ土、パーライト、ベ
ントナイト、大谷石、アンスラ石、石灰石、バーミキュ
ライト等の単品または組み合わせが挙げられるが、栄養
源を含んでいない物質であれば上記に制限されない。し
かしながらこれらの中で特に好ましいのは水である。The starved state in the present invention is formed by mixing the cells with a non-nutritive substance so that the concentration of the cells is about 10 8 to 14 ) cells / g. Non-nutritive substances include water or aqueous solutions containing no carbon or nitrogen sources, lightweight porous inorganic carriers, zeolites, diatomaceous earth, perlite, bentonite, Otani stone, anthracite, limestone, vermiculite, etc. Alternatively, a combination may be mentioned, but the substance is not limited to the above as long as the substance does not contain a nutrient source. However, among these, water is particularly preferred.
本発明に用いられるキャリヤーとしては、稲ワラ、モ
ミガラ、ヒル石、貝化石、ピートモス、乾燥畜糞、米ぬ
か、石膏、骨粉、草木灰、無機担体等が挙げられるが、
その中でも稲ワラやモミガラがより好ましい。Examples of the carrier used in the present invention include rice straw, rice hulls, hill stones, shell fossils, peat moss, dried animal dung, rice bran, gypsum, bone powder, plant ash, inorganic carriers, and the like.
Among them, rice straw and firgrass are more preferable.
PHBを菌体内に蓄積させる培養法としては公知の方法
が採用される。即ち肉エキス培地やポテトデキストロー
ス培地などの窒素源の少ない培地で培養される。この培
養後、更に炭素源のみの培地で培養すると更に効果的に
PHBが蓄積される。上記のようにして培養された菌体
は、上記の非栄養性物質として混合した後、数時間〜数
ケ月、好ましくは１日〜７日程度常温にて放置する。そ
の後、菌体濃度が108個/g程度になる様に水で希釈し、
それとキャリヤーとが重量比で1:0.1〜1:20、好ましく
は1:1〜1:5程度になる様に上記キャリヤーと混合する。
その後すぐにあるいは１日〜７日、より好ましくは１日
〜２日程度常温にて放置した後土壌に施用する。A known method is employed as a culture method for accumulating PHB in cells. That is, it is cultured in a medium having a small nitrogen source such as a meat extract medium or a potato dextrose medium. After this cultivation, further culturing in a medium containing only a carbon source is more effective.
PHB accumulates. After the cells cultured as described above are mixed as the above non-nutritive substance, they are allowed to stand at room temperature for several hours to several months, preferably for about one to seven days. After that, dilute with water so that the bacterial cell concentration is about 10 8 / g,
It is mixed with the carrier so that the weight ratio with the carrier is about 1: 0.1 to 1:20, preferably about 1: 1 to 1: 5.
Immediately thereafter, or after leaving for 1 to 7 days, more preferably for 1 to 2 days at room temperature, it is applied to the soil.
以下の実施例により本発明をより具体的に説明する
が、本発明はこれらの実施例に限定されるものではな
い。The present invention will be described more specifically with reference to the following examples, but the present invention is not limited to these examples.
尚、表１に実施例で使用した代表的な微生物を示す
が、これらはすべて抗生物質を生産する公知株である。Table 1 shows typical microorganisms used in Examples, which are all known strains producing antibiotics.
実施例１〜５及び比較例１〜５ 表１に示す各種菌体を肉エキス−ペプトン液体培地
（ディフコ社製）で30℃で16〜48時間振とうあるいは静
置培養した後、遠心分離法によって集菌し、さらに純水
にて５回洗菌した後、最終菌体濃度が1012個/g程度にな
る様に純水を加えて混合し、５日間常温にて放置し飢餓
状態とした（実施例１〜５）。 Examples 1 to 5 and Comparative Examples 1 to 5 The various bacterial cells shown in Table 1 were cultured in a meat extract-peptone liquid medium (manufactured by Difco) at 30 ° C. for 16 to 48 hours or static culture, followed by centrifugation. After the cells were collected and washed 5 times with pure water, pure water was added and mixed so that the final cell concentration was about 10 12 cells / g, and the mixture was left at room temperature for 5 days to be starved. (Examples 1 to 5).
また上記の飢餓状態の代わりに、栄養源として肉エキ
ス−ペプトン溶液を純水の変わりに用いて栄養豊富状態
とした（比較例１〜５）。Further, instead of the above starvation state, a meat extract-peptone solution was used as a nutrient source instead of pure water to make a nutrient-rich state (Comparative Examples 1 to 5).
その後さらに飢餓状態下の菌体には純水、また栄養豊
富状態の菌体には同じ液体培地を用いて1000倍希釈した
後、あらかじめ滅菌しておいた稲ワラを重量比が1:1に
なるように混合し、シャーレ上にて20℃で保存して経時
的に生菌数を測定し、菌体の生存率を求めた。After that, the cells under starvation were diluted with pure water, and the cells in the nutrient-rich state were diluted 1000-fold using the same liquid medium, and the rice straw that had been sterilized in advance was reduced to a weight ratio of 1: 1. After mixing, the cells were stored on a Petri dish at 20 ° C., and the number of viable cells was measured over time to determine the survival rate of the cells.
生菌数測定は、普通寒天培地（日水製薬社製）に試料
を一定の割合で希釈して植菌し、48時間後のコロニー数
より求めた。The viable cell count was determined by diluting the sample at a fixed rate on a normal agar medium (manufactured by Nissui Pharmaceutical Co., Ltd.), inoculating the sample, and determining the number of colonies 48 hours later.
結果を表２に示す。 Table 2 shows the results.
実施例６及び比較例６〜７ Pseudomonas cepaciaを用い、実施例１及び比較例１
に示す方法で菌体を調製し、さらに同様に滅菌していな
い稲ワラに吸着させ、５日後に土壌重量の１％量を土壌
に施用して各種土壌病害の発生株率を下記式により求め
た（実施例６及び比較例６）。 Example 6 and Comparative Examples 6 and 7 Using Pseudomonas cepacia, Example 1 and Comparative Example 1
The bacterial cells are prepared by the method shown in (1), and are similarly adsorbed on unsterilized rice straw. After 5 days, 1% of the soil weight is applied to the soil, and the occurrence rate of various soil diseases is determined by the following formula. (Example 6 and Comparative Example 6).
また菌体を施用しない土壌についても同様に各種土壌
病害の発生株率を求めた（比較例７）。The rate of occurrence of various soil diseases was similarly determined for soil to which no cells were applied (Comparative Example 7).
結果を表３に示す。 Table 3 shows the results.
───────────────────────────────────────────────────── フロントページの続き (56)参考文献 特開 昭59−62509（ＪＰ，Ａ) 特開 昭60−186230（ＪＰ，Ａ) 特開 昭64−16579（ＪＰ，Ａ) 特開 昭63−291578（ＪＰ，Ａ) 特開 昭62−148413（ＪＰ，Ａ) (58)調査した分野(Int.Cl.6，ＤＢ名) A01N 63/02 C12N 1/20 ＣＡ（ＳＴＮ) ＲＥＧＩＳＴＲＹ（ＳＴＮ) ＷＰＩＤＳ（ＳＴＮ)──────────────────────────────────────────────────続 き Continuation of the front page (56) References JP-A-59-62509 (JP, A) JP-A-60-186230 (JP, A) JP-A-64-16579 (JP, A) JP-A-63-163 291578 (JP, A) JP-A-62-148413 (JP, A) (58) Fields investigated (Int. Cl. 6 , DB name) A01N 63/02 C12N 1/20 CA (STN) REGISTRY (STN) WPIDS (STN)
Claims (2)
して菌体内にポリヒドロキシブチレートを蓄積させた
後、洗菌し、飢餓状態とし、キャリヤーと混合して土壌
に施用することを特徴とする土壌病害防止法。1. An antibiotic-producing bacterium is cultured in a nutrient-containing medium to accumulate polyhydroxybutyrate in the cells, washed, starved, mixed with a carrier and applied to soil. Soil disease prevention law characterized by the following.
domonas）属又はバチルス（Bacillus）属の菌である請
求項１記載の土壌病害防止法。2. The antibiotic-producing bacterium is Pseudomonas (Pseu).
The soil disease prevention method according to claim 1, wherein the soil disease is a bacterium belonging to the genus domonas or the genus Bacillus.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP1212651A JP2846355B2 (en) | 1989-08-18 | 1989-08-18 | Soil Disease Prevention Law |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP1212651A JP2846355B2 (en) | 1989-08-18 | 1989-08-18 | Soil Disease Prevention Law |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPH0377803A JPH0377803A (en) | 1991-04-03 |
| JP2846355B2 true JP2846355B2 (en) | 1999-01-13 |
Family
ID=16626157
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP1212651A Expired - Fee Related JP2846355B2 (en) | 1989-08-18 | 1989-08-18 | Soil Disease Prevention Law |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JP2846355B2 (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0734738B2 (en) * | 1991-10-02 | 1995-04-19 | 有機質肥料生物活性利用技術研究組合 | Plant pathogen-suppressing microorganism and method of using the same |
| CN105601449A (en) * | 2015-12-30 | 2016-05-25 | 顺达康（苏州）农业科技发展有限公司 | Special continuous cropping resisting complexing agent for cucumbers |
-
1989
- 1989-08-18 JP JP1212651A patent/JP2846355B2/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| JPH0377803A (en) | 1991-04-03 |
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