JPH0568535A - Method for separating and selecting symbiotic microorganism useful for controlling disease injury of dicotyledon - Google Patents
Method for separating and selecting symbiotic microorganism useful for controlling disease injury of dicotyledonInfo
- Publication number
- JPH0568535A JPH0568535A JP3259915A JP25991591A JPH0568535A JP H0568535 A JPH0568535 A JP H0568535A JP 3259915 A JP3259915 A JP 3259915A JP 25991591 A JP25991591 A JP 25991591A JP H0568535 A JPH0568535 A JP H0568535A
- Authority
- JP
- Japan
- Prior art keywords
- microorganisms
- seedling
- hypocotyl
- dicotyledon
- cut
- 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.)
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Links
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
Description
【０００１】[0001]
【産業上の利用分野】本発明は、双子葉植物の生育段階
における病害の防除に用いる共生微生物の分離選抜方法
に関する。より詳しくは、微生物が胚軸からトラップさ
れる現象を、共生微生物の植物組織内定着の手段として
他の植物にも応用し、双子葉植物の生育段階における病
害の防除を図る技術は、本発明者らの開発技術である
が、本発明はその開発技術に用いる共生微生物の分離選
抜の方法に関する。BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method for separating and selecting symbiotic microorganisms used for controlling diseases at the growth stage of dicotyledonous plants. More specifically, the technique of applying the phenomenon in which a microorganism is trapped from the hypocotyl to other plants as a means of colonization of a symbiotic microorganism in plant tissue to control a disease at the growth stage of a dicotyledonous plant is disclosed in the present invention. The present invention relates to a method of separating and selecting symbiotic microorganisms used in the development technique.
【０００２】[0002]
【従来技術とその問題点】病原菌に抗菌活性のある微生
物を利用した生物学的防除手段が世界各国で種々試みら
れている。たしかに、病原菌に何らかの活性がある有用
微生物は土壌中や植物から容易に分離することができ
る。しかしこの有用微生物をそのまま病土に投入した
り、植物に散布した場合、殆ど病害防除の効果が発現し
ない。その理由は、有用微生物が土壌中や植物で安定し
ないことによる。このため有用微生物を担体（キャリア
−）に吸着させて土壌中で安定させる方法、病原菌の改
良による植物で安定化する有用微生物の作出などが試み
られている。しかしこれらの方法は、微生物の種特異性
を無視した微生物の働きが充分発揮されない方法であ
り、また栽培環境や生態系を無視した方法であり、防除
効果については、これまで一般的に実施されて来た土壌
消毒、抵抗性品種の育成、接木栽培等の方法に及ばない
現状にある。このように生物学的防除に用いる有用微生
物は、これまで無作為的に分離されてきたといってよ
く、新らしい分離選抜技術の開発が俟たれている現状に
ある。2. Description of the Related Art Various biological control means using microorganisms having antibacterial activity against pathogenic bacteria have been tried in various countries around the world. Certainly, useful microorganisms having some activity against pathogenic bacteria can be easily separated from soil or plants. However, when this useful microorganism is directly added to disease soil or sprayed on plants, the effect of controlling disease is hardly exhibited. The reason is that useful microorganisms are not stable in soil or plants. Therefore, attempts have been made to adsorb useful microorganisms to a carrier to stabilize them in soil, and to produce useful microorganisms that are stabilized in plants by improving pathogenic bacteria. However, these methods are methods in which the action of microorganisms that ignores the species specificity of the microorganisms is not fully exerted, and methods that ignore the cultivation environment and ecosystem, and the control effect has been generally practiced so far. The current situation is inferior to the existing methods such as soil disinfection, breeding of resistant varieties, and graft cultivation. As described above, it can be said that useful microorganisms used for biological control have been randomly separated so far, and under the present circumstances, the development of a new separation / selection technique is under way.
【０００３】[0003]
【発明が解決しようとする課題】本発明者らは、これま
での研究成果として、植物の健全組織内は一部の植物を
除き無菌状態にあること、その例外植物としてシクラメ
ン、サツマイモ、トウモロコシ、コンニャクなどがある
こと、しかしこれら有菌植物も種子そのものは無菌状態
にあり、発芽後、子葉が展開するとその胚軸から微生物
をトラップして有菌状態になることを知見している。そ
こでこの、微生物が胚軸からトラップされる現象を、共
生微生物の植物組織内定着の手段として他の植物にも応
用し、双子葉植物の生育段階における病害の防除を図る
技術を本発明者らは開発し、別途特許出願中であるが、
本発明はその開発技術に用いる共生微生物の分離選抜の
方法を提供するものである。DISCLOSURE OF INVENTION Problems to be Solved by the Invention As a result of the research so far, the present inventors have found that the healthy tissue of a plant is in a sterile state except for some plants, and its exceptional plants include cyclamen, sweet potato, corn, It has been found that konjak, etc. exist, but the seeds themselves of these fungal plants are in a sterile state, and after germination, the cotyledon expands to trap the microorganisms from their hypocotyls and become germinated. Therefore, the present inventors have developed a technique for controlling the disease at the growth stage of dicotyledonous plants by applying the phenomenon that the microorganisms are trapped from the hypocotyl to other plants as a means of colonizing the symbiotic microorganisms in plant tissues. Has been developed and a patent is pending,
The present invention provides a method for separating and selecting a symbiotic microorganism used in the development technique.
【０００４】[0004]
（着想）双子葉植物の種子発芽は最初に胚軸が伸長し根
を伸長させる。次に、子葉を展開させる。この胚軸伸長
と子葉展開に要する栄養分は、種子に蓄えられた栄養分
が利用される（種子栄養分に従属する）。子葉展開後か
ら本葉１〜３枚に成長すると双子葉植物苗は根から養分
を吸収したり、葉で同化するなど独立栄養に移行する。
この子葉展開後から本葉２〜３枚展開までの、独立栄養
に移行する期間は、植物の種としての本来の特性がまだ
備わっていない時期にある。このため双子葉植物苗を、
その胚軸で切断して微生物を接種すると組織内にトラッ
プされると考えられる。しかし、種としての特異性は存
在する。このためトラップされた微生物は、その植物に
選ばれた微生物になるものと考えられる。そこで、この
非特異性と、特異性の両性質を、組織内定着能力のある
有用微生物の分離選抜に利用したところ、胚軸接種した
挿し木苗から、各種双子葉植物の組織内に定着できる有
用な微生物（共生微生物）の分離選抜方法が開発され
た。(Concept) In seed germination of dicotyledons, the hypocotyl first extends and the roots extend. Next, the cotyledons are expanded. The nutrients required for this hypocotyl elongation and cotyledon expansion utilize the nutrients stored in the seeds (subordinate to the seed nutrients). After the cotyledon expansion, when growing to 1 to 3 true leaves, the dicotyledonous plant seedlings absorb nutrients from the roots and are assimilated by the leaves, thereby shifting to autotrophic.
The period from the development of cotyledons to the development of 2 to 3 true leaves is a period in which autotrophic shift occurs at a time when the original characteristics as a seed of a plant are not yet provided. Therefore, dicot seedlings
It is considered that when cut by the hypocotyl and inoculated with a microorganism, it is trapped in the tissue. However, there are species specificities. Therefore, the trapped microorganisms are considered to be the microorganisms selected by the plant. Therefore, by utilizing both these non-specificity and specificity in the separation and selection of useful microorganisms capable of colonizing the tissue, it is possible to establish them in the tissues of various dicotyledonous plants from the hypocotyl-inoculated cutting seedlings. A method for separating and selecting various microorganisms (symbiotic microorganisms) has been developed.
【０００５】（構成）双子葉植物を播種し、発芽、生育
した苗の未分化組織であるその胚軸の中間から切断す
る。その切断苗を無殺菌の土壌に挿し木して、土壌中か
ら前記双子葉植物を好む微生物を組織内にトラップす
る。その挿し木苗を育苗する。発根したその育成苗を採
集して、これを常法により表面殺菌後培養し、培養によ
り生じたコロニ−を釣菌して分離する。この分離した共
生微生物の浮遊液に、胚軸を切断した前記双子葉植物苗
を浸漬接種後、挿し木して育苗する。その育成苗から、
先の共生微生物を再分離する。以上の過程を経る双子葉
植物の病害防除に用いる共生微生物の分離選抜方法であ
る（図１参照）。分離された微生物は、シュ−ドモナス
（Ｐｓｅｕｄｏｍｏｎａｓ），キサントモナス（Ｘａｎ
ｔｈｏｍｏｎａｓ），エルヴィニア（Ｅｒｗｉｎｉ
ａ），バシラス（Ｂａｃｉｌｌｕｓ），放線菌、コリネ
型細菌に属する細菌である。(Constitution) A dicotyledon is sowed and cut from the middle of its hypocotyl, which is an undifferentiated tissue of a germinated and grown seedling. The cut seedlings are planted in unsterilized soil, and the dicotyledonous microorganisms are trapped in the tissue from the soil. Raise the cuttings. The rooted grown seedlings are collected, surface-sterilized by a conventional method, and then cultured, and colonies produced by the culture are picked up and separated. The separated suspension of symbiotic microorganisms is dipped and inoculated with the dicotyledonous plant seedlings from which the hypocotyls have been cut, and then cuttings are taken to raise seedlings. From the raised seedlings,
Reseparate the previous symbiotic microorganisms. This is a method for separating and selecting symbiotic microorganisms used for disease control of dicotyledonous plants that have undergone the above processes (see FIG. 1). The isolated microorganisms are Pseudomonas and Xanthomonas (Xan).
thomonas), Erwinia
a), Bacillus, actinomycetes, coryneform bacteria.
【０００６】[0006]
ナス科植物・・・・・トマト、ナス、ピ−マン ウリ科植物・・・・・ユウガオ、キュウリ、プリンスメ
ロン、マスクメロン、シロウリ、マクワウリ、ニガウ
リ、ヘチマ、スイカ、カボチャ アブラナ科植物・・・ハクサイ、キャベツ、ブロッコリ
−、ダイコン マメ科植物・・・・・ダイズ、インゲン アカザ科植物・・・・ホウレンソウ セリ科植物・・・・・ニンジン キンポウゲ科植物・・デルヒニウム バラ科植物・・・・・イチゴ 上記各植物について、以下の通り本発明を実施した。 （１）播種して子葉が展開してから本葉１〜３枚のステ
−ジになった苗の胚軸を切断する。 （２）その切断苗を無殺菌の土壌に挿し木して、土壌中
から前記双子葉植物を好む微生物を組織内にトラップす
る。 （３）その挿し木苗を育苗する。 （４）発根したその育成苗を採集して、これを７０％エ
タノ−ルで一分間及び塩化第二水銀１０００倍で一分間
表面滅菌する。 （５）次いでこれを１％ペプトン水中で磨砕する。 （６）その磨砕液をブイヨン寒天倍地、ジャガイモ寒天
倍地に画線または塗付する。 （７）２５゜Ｃの定温器で１週間前後培養後、生じたコ
ロニ−を釣菌する。 （８）分離した微生物（共生微生物）の浮遊液に、胚軸
を切断した前記双子葉植物苗を浸漬接種後、挿し木して
育苗する。 （９）その育成苗は生育状況を調べ、再び前記（４）、
（５）、（６）、（７）の作業を繰返して、組織内から
先の共生微生物を再分離する。Solanaceae plant ・ ・ ・ Tomato, eggplant, peumman Cucurbitaceae plant ・ ・ ・ Yugao, cucumber, prince melon, muskmelon, shirouri, makuwauri, bitter gourd, loofah, watermelon, pumpkin Brassicaceae plant ・ ・ ・Chinese cabbage, cabbage, broccoli, Japanese radish legumes ・ ・ ・ Soybeans, green beans azaleas ・ ・ ・ ・ ・ ・ Spinach / Aceraceae ・ ・ ・ Carrots buttercups ・ Delhinium roses ・ ・ ・Strawberries The present invention was carried out as follows for each of the above plants. (1) After seeding and developing cotyledons, cut the hypocotyl of a seedling having 1 to 3 stages of true leaves. (2) The cut seedlings are planted in unsterilized soil, and the dicotyledonous microorganisms are trapped in the tissue from the soil. (3) Raise the cuttings. (4) The rooted grown seedlings are collected and surface-sterilized with 70% ethanol for 1 minute and mercuric chloride 1000 times for 1 minute. (5) Then, this is ground in 1% peptone water. (6) Stroke or apply the ground solution to broth agar and potato agar. (7) After culturing for about one week in a 25 ° C. incubator, the resulting colonies are picked up. (8) The suspension of the separated microorganisms (symbiotic microorganisms) is immersed and inoculated with the dicotyledonous plant seedlings having the hypocotyls cut off, and then cuttings are taken to raise seedlings. (9) The growing seedlings are examined for the growth condition, and the above (4),
The operations of (5), (6), and (7) are repeated to re-separate the above-mentioned symbiotic microorganism from within the tissue.
【０００７】以上によって分離選抜された共生微生物
は、シュ−ドモナス（Ｐｓｅｕｄｏｍｏｎａｓ），キサ
ントモナス（Ｘａｎｔｈｏｍｏｎａｓ），エルヴィニア
（Ｅｒｗｉｎｉａ），バシラス（Ｂａｃｉｌｌｕｓ），
放線菌，コリネ型細菌に属する細菌であり、そのトラッ
プ状況および組織内定着性について調べたところ、表１
の結果が得られた。本発明方法により分離選抜される植
物は、双子葉植物に限られるが、広く双子葉植物全般に
及び、表１に掲げた植物名に限られるものでない。The symbiotic microorganisms separated and selected as described above are Pseudomonas, Xanthomonas, Erwinia, Bacillus,
It is a bacterium belonging to actinomycetes and coryneform bacteria, and its trap state and tissue colonization properties were examined.
The result was obtained. The plants to be separated and selected by the method of the present invention are limited to dicotyledons, but are not limited to the dicotyledons in general and the plant names listed in Table 1.
【０００８】[0008]
【表１】 [Table 1]
【０００９】[0009]
【発明の効果】本発明は以上のようで、双子葉植物の胚
軸切断挿し木苗から、各種双子葉植物の組織内に定着で
きる有用な微生物（共生微生物）の分離選抜を行うもの
であるから、微生物の種特異性、胚軸特性を利用して微
生物の働きを充分発揮させられ、また栽培環境や生態系
を維持した有効な方法である。本発明によって分離選抜
された共生微生物の細菌属は前記のとおり類別され、そ
れらは特許微生物寄託センタ−に、微工研菌寄第１１６
２５号〜第１１６３０号として寄託しているところであ
る。これら菌株を、胚軸接種により植物組織内に定着さ
せる双子葉植物の病害防除技術に用いたところ、植物の
生育が著しく促進され、また土壌病害に優れた抵抗性を
示した。Industrial Applicability As described above, the present invention separates and selects useful microorganisms (symbiotic microorganisms) that can settle in the tissues of various dicotyledonous plants from hypocotyl-cut cuttings of dicotyledonous plants. It is an effective method that utilizes the species-specificity and hypocotyl characteristics of microorganisms to fully exert the functions of microorganisms and maintains the cultivation environment and ecosystem. The genus of symbiotic microorganisms separated and selected according to the present invention are categorized as described above.
No. 25 to No. 11630 have been deposited. When these strains were used for a disease control technique for dicotyledonous plants that colonize in plant tissues by inoculating hypocotyls, the growth of plants was significantly promoted, and excellent resistance to soil diseases was exhibited.
【図１】本発明方法の過程を略図的に表わした概念図FIG. 1 is a conceptual diagram schematically showing a process of the method of the present invention.
Claims (1)
胚軸の中間から切断し、その切断苗を無殺菌の土壌に挿
し木して、土壌中から前記双子葉植物を好む微生物を組
織内にトラップし、その発根育成苗を採集して、これを
常法により表面殺菌後培養し、培養により生じたコロニ
−を釣菌して分離し、この分離した共生微生物の浮遊液
に、胚軸を切断した前記双子葉植物苗を浸漬接種後、挿
し木して育苗し、その育成苗から先の共生微生物を再分
離することを特徴とする双子葉植物の病害防除に用いる
共生微生物の分離選抜方法。1. A dicotyledonous plant seedling is cut from the middle of its hypocotyl, which is an undifferentiated tissue, and the cut seedling is planted in unsterilized soil to form a microorganism that prefers the dicotyledonous plant from the soil. Trap inside, collect the rooting grown seedlings, culture this after surface sterilization by a conventional method, isolate colonies produced by culturing by colonization, in the suspension of the separated symbiotic microorganisms, Isolation of the dicotyledonous plant seedlings after cutting the hypocotyl by immersion inoculation, cutting and raising seedlings, and reseparating the above-mentioned symbiotic microorganisms from the seedlings for separation of symbiotic microorganisms used for disease control of dicotyledonous plants Selection method.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP3259915A JPH088858B2 (en) | 1991-09-10 | 1991-09-10 | Separation and selection method of symbiotic microorganisms used for disease control of dicotyledonous plants |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP3259915A JPH088858B2 (en) | 1991-09-10 | 1991-09-10 | Separation and selection method of symbiotic microorganisms used for disease control of dicotyledonous plants |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPH0568535A true JPH0568535A (en) | 1993-03-23 |
| JPH088858B2 JPH088858B2 (en) | 1996-01-31 |
Family
ID=17340699
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP3259915A Expired - Lifetime JPH088858B2 (en) | 1991-09-10 | 1991-09-10 | Separation and selection method of symbiotic microorganisms used for disease control of dicotyledonous plants |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH088858B2 (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5935571A (en) * | 1996-05-20 | 1999-08-10 | Taki Chemical Co., Ltd. | Plug mixture for raising seedlings and method for producing it, and method for raising disease tolerant seedlings |
| EP1449427A4 (en) * | 2001-10-22 | 2010-09-01 | Taki Chemical | Rooted cutting and method of inoculating rooted cutting with microbial strain |
| JP2015530093A (en) * | 2012-09-19 | 2015-10-15 | バイオディスカバリー・ニュージーランド・リミテッド | Methods for screening microorganisms that confer beneficial properties on plants |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS55118390A (en) * | 1979-02-14 | 1980-09-11 | Mosse B | Production of mycorrhiza bacterial cell |
| JPH0276572A (en) * | 1988-09-14 | 1990-03-15 | Snow Brand Milk Prod Co Ltd | Medium for regenerating biffidobacterium protoplast and regeneration thereof using same medium |
-
1991
- 1991-09-10 JP JP3259915A patent/JPH088858B2/en not_active Expired - Lifetime
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS55118390A (en) * | 1979-02-14 | 1980-09-11 | Mosse B | Production of mycorrhiza bacterial cell |
| JPH0276572A (en) * | 1988-09-14 | 1990-03-15 | Snow Brand Milk Prod Co Ltd | Medium for regenerating biffidobacterium protoplast and regeneration thereof using same medium |
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5935571A (en) * | 1996-05-20 | 1999-08-10 | Taki Chemical Co., Ltd. | Plug mixture for raising seedlings and method for producing it, and method for raising disease tolerant seedlings |
| EP1449427A4 (en) * | 2001-10-22 | 2010-09-01 | Taki Chemical | Rooted cutting and method of inoculating rooted cutting with microbial strain |
| JP2015530093A (en) * | 2012-09-19 | 2015-10-15 | バイオディスカバリー・ニュージーランド・リミテッド | Methods for screening microorganisms that confer beneficial properties on plants |
| JP2018121649A (en) * | 2012-09-19 | 2018-08-09 | バイオディスカバリー・ニュージーランド・リミテッド | Method for screening microorganism imparting beneficial properties to plant |
| US10526599B2 (en) | 2012-09-19 | 2020-01-07 | Biodiscovery New Zealand Limited | Accelerated directed evolution of microbial consortia for the development of desirable plant phenotypic traits |
| JP2020156489A (en) * | 2012-09-19 | 2020-10-01 | バイオディスカバリー・ニュージーランド・リミテッド | Methods of screening for microorganisms that impart beneficial properties to plants |
| US10900029B2 (en) | 2012-09-19 | 2021-01-26 | Bioconsortia New Zealand Limited | Methods of screening for microorganisms that impart beneficial properties to plants |
| JP2022000048A (en) * | 2012-09-19 | 2022-01-04 | バイオディスカバリー・ニュージーランド・リミテッド | Screening method of microorganism in which useful properties are imparted to plant |
| US11466266B2 (en) | 2012-09-19 | 2022-10-11 | Bioconsortia, Inc. | Accelerated directed evolution of microbial consortia for the development of desirable plant phenotypic traits |
| JP2023089056A (en) * | 2012-09-19 | 2023-06-27 | バイオディスカバリー・ニュージーランド・リミテッド | Methods of screening for microorganisms that impart beneficial properties to plants |
| US11866698B2 (en) | 2012-09-19 | 2024-01-09 | Bioconsortia, Inc. | Accelerated directed evolution of microbial consortia for the development of desirable plant phenotypic traits |
Also Published As
| Publication number | Publication date |
|---|---|
| JPH088858B2 (en) | 1996-01-31 |
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