JPH0584028A - Plant body of strawberry - Google Patents
Plant body of strawberryInfo
- Publication number
- JPH0584028A JPH0584028A JP12688591A JP12688591A JPH0584028A JP H0584028 A JPH0584028 A JP H0584028A JP 12688591 A JP12688591 A JP 12688591A JP 12688591 A JP12688591 A JP 12688591A JP H0584028 A JPH0584028 A JP H0584028A
- Authority
- JP
- Japan
- Prior art keywords
- culture
- strawberry
- medium containing
- cytokinin
- liquid medium
- 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.)
- Pending
Links
Landscapes
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
Description
【０００１】[0001]
【産業上の利用分野】本発明はイチゴの葉芽増殖方法に
よる植物体に関し、とくにイチゴの植物組織の通気培養
による植物体の大量増殖に関するものである。BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a plant by a method for growing leaf buds of strawberries, and more particularly to mass-growing plants by aeration culture of strawberry plant tissue.
【０００２】[0002]
【従来の技術】イチゴのウイルスフリー苗の増殖は一般
にランナーを利用している。しかし、ランナーによる増
殖方法は、ランナーの発生する時期が限られているの
で、短期間に大量の増殖を行うには設備、労力が必要で
ある。そこで、それに代わる方法として組織培養による
増殖方法が研究されている。そのなかで、遺伝的変異の
発生率が少ない茎頂培養による腋芽の大量増殖方法の研
究が高野、赤木により報告されている（「農業および園
芸」第６３巻（１９８８年）第１号（株）養賢堂発行、
ｐ１５９〜１６０）。前記高野らの増殖方法は、ランナ
ーの先端から茎頂を切り取り、ショ糖を含む培地を用い
て照明下に１〜２ケ月培養すると発根し、葉が２〜３枚
展開してくるので、この幼植物体をベンジルアミノプリ
ン（以下、ＢＡと記す。）を含む増殖培地へ移植して、
イチゴ苗を増殖させるというものである。また、川合
は、イチゴの寒天培養、ジャーファーメンター培養につ
いて報告している（ＢＩＯ ＩＮＤＵＳＴＲＹ Ｖｏ
ｌ．３ Ｎｏ．４（１９８６）（株）シーエムシー発行
ｐ７３）。2. Description of the Related Art Generally, runners are used to grow strawberry virus-free seedlings. However, in the breeding method by the runner, since the time when the runner is generated is limited, equipment and labor are required to carry out a large amount of breeding in a short period of time. Therefore, a proliferating method by tissue culture has been studied as an alternative method. Among them, Takano and Akagi reported a study on a mass-proliferation method of axillary buds by shoot apical culture with a low incidence of genetic mutation ("Agriculture and Horticulture" Vol. 63 (1988) No. 1 (strain)). ) Published by Yokendo,
p159-160). Takano et al.'S method of multiplication cuts the shoot apex from the tip of the runner, and roots it when cultivated for 1-2 months under illumination using a medium containing sucrose, so that 2-3 leaves develop, This seedling was transplanted to a growth medium containing benzylaminopurine (hereinafter referred to as BA),
The idea is to grow strawberry seedlings. Kawai has also reported on strawberry agar culture and jar fermenter culture (BIO INDUSTRY Vo.
l. 3 No. 4 (1986) CMC Publishing, p73).
【０００３】さらに、本発明者は先にイチゴの植物組織
から葉芽の増殖をする方法について特許出願をした（特
願平３−６２５８５号）。その方法はイチゴの植物組織
を無菌的に摘出して栄養分を含む培地で培養して幼葉を
展開する前または幼葉を展開し始めた段階の大きさに育
成させ、これをサイトカイニンを含む培地で培養して葉
芽を増殖させるというものである。Further, the present inventor has previously filed a patent application for a method for growing leaf buds from strawberry plant tissue (Japanese Patent Application No. 3-62585). The method is aseptically removing plant tissue of strawberries and culturing them in a medium containing nutrients and growing them to the size of the stage before the development of young leaves or the stage of starting to develop young leaves, and this is a medium containing cytokinin. It is to culture and grow leaf buds.
【０００４】[0004]
【発明が解決しようとする課題】前記高野らのイチゴの
腋芽増殖法における課題として、次のようなことが挙げ
られている。すなわち、培養期間が長いこと、増殖した
腋芽の大きさが均一でないこと、基部が硬くて分割しに
くく、機械による自動化も困難であること等である。ま
た、川合の報告によると前記ジャーファーメンター培養
により得られた腋芽の茎葉長はばらつきが大きく、特に
平均より小さい個体の割合が多いことが指摘されてい
る。このようにイチゴの葉芽の増殖方法についての増殖
期間、得られる植物体の大きさが小さいとかばらばらで
ある等の問題点があった。Problems to be solved by the above-mentioned Takano et al.'S strawberry axillary bud growth method include the following. That is, the culture period is long, the size of the axillary buds grown is not uniform, the base is hard and difficult to divide, and automation by a machine is also difficult. According to Kawai's report, the foliage length of the axillary buds obtained by the jar fermenter culture varies widely, and in particular, the proportion of individuals smaller than the average is high. As described above, there are problems with the method for growing strawberry leaf buds, such as the growth period, the size of the obtained plant being small and the plants being scattered.
【０００５】そこで、本発明者の発明である特願平３−
６２５８５号の発明は短い培養期間で、均一の葉芽を持
ち、操作性の改善されたイチゴの葉芽の増殖方法として
提案したものである。上記特願平３−６２５８５号の発
明では根の誘導については検討をしていないが、この葉
芽を常法どおり栄養分を含む固体培地（ショ糖３０ｇ／
リットル、ゲルライト２ｇ／リットルを添加し、無機塩
濃度を１／２としたＭＳ改変培地）に置床して培養する
ことにより根の誘導を行った場合、培養２１日間後の発
根率は６４％であった。また、根を持たない葉芽をその
まま土壌に移植した場合は全く活着しなかった。Therefore, Japanese Patent Application No. 3-
The invention of No. 62585 is proposed as a method for growing strawberry leaf buds having uniform leaf buds and improved operability in a short culture period. In the invention of Japanese Patent Application No. 3-62585 mentioned above, the induction of roots is not examined, but the leaf buds are solid medium containing nutrients (sucrose 30 g /
When the roots were induced by culturing by adding 1 liter of gellite and 2 g / liter of gellite and culturing by placing the medium on an MS modified medium with an inorganic salt concentration of 1/2, the rooting rate after 21 days of culture was 64%. Met. In addition, when rootless leaf buds were directly transplanted to soil, they did not survive at all.
【０００６】本発明は上記従来技術の欠点を解決して、
簡単な操作で、発根率を向上させた増殖方法によるイチ
ゴの植物体を提供することを目的とする。The present invention solves the above-mentioned drawbacks of the prior art,
It is an object of the present invention to provide a strawberry plant body by a growth method with an improved rooting rate by a simple operation.
【０００７】[0007]
【課題を解決するための手段】本発明の上記目的は次の
構成で達成される。すなわち、イチゴの植物組織を無菌
的に摘出して栄養分を含む培地で培養して後、これをサ
イトカイニンを含む液体培地で振とう培養または通気撹
拌培養して葉芽を増殖させ、この葉芽をサイトカイニン
を含まない液体培地で通気撹拌培養することにより根を
誘導して得られるイチゴの植物体である。The above objects of the present invention can be achieved by the following constitutions. That is, after aseptically removing strawberry plant tissue and culturing it in a medium containing nutrients, it is shake-cultured or aeration-agitation culture in a liquid medium containing cytokinin to grow leaf buds, and the leaf buds are cultivated with cytokinin. It is a strawberry plant obtained by inducing roots by culturing with aeration and stirring in a liquid medium not containing it.
【０００８】サイトカイニンを含む液体培地で振とう培
養または通気撹拌培養する植物組織は幼葉を展開する前
または幼葉を展開しはじめた段階、例えば０．５〜５ｍ
ｍ、好ましくは１〜２ｍｍの大きさに育成した植物組織
を用いることが望ましい。The plant tissue to be shake-cultured or aerated and agitated in a liquid medium containing cytokinin is before the development of young leaves or at the stage when the young leaves have begun to be developed, for example, 0.5 to 5 m.
It is desirable to use a plant tissue grown to a size of m, preferably 1 to 2 mm.
【０００９】本発明に用いる植物組織は茎頂組織がウイ
ルスフリーであり、好ましい。その茎頂組織を無菌的に
摘出して培養し、０．５〜５ｍｍ好ましくは１〜２ｍｍ
の大きさに育成し、これをサイトカイニンを含む液体培
地で振とう培養または通気撹拌培養して葉芽を増殖さ
せ、この葉芽をサイトカイニンを含まない液体培地で通
気撹拌培養して根を誘導することによって植物体を再生
させることができる。この液体培地で根を誘導するた
め、ジャーファメンタを利用することができる。The plant tissue used in the present invention is preferable because the shoot apex tissue is virus-free. The shoot apical tissue is aseptically removed and cultured, 0.5 to 5 mm, preferably 1 to 2 mm
By culturing the leaf buds in a liquid medium containing cytokinin by shaking culture or aeration stirring culture to grow the leaf buds, and aerating the leaf buds in a liquid medium containing no cytokinin by aeration stirring to induce roots. The plant can be regenerated. Jarfamentors can be used to induce roots in this liquid medium.
【００１０】ここで、サイトカイニンとして、例えばベ
ンジルアミノプリン（以下、ＢＡという。）を用いる場
合はＢＡを０．０１〜１０ｍｇ／リットル、好ましくは
０．０１〜５ｍｇ／リットルを含む液体培地で振とう培
養または通気撹拌培養させる。このＢＡの含有割合が下
限値未満だと葉原基が増殖しないため、植物体一個体の
みしか得られない。また前記ＢＡ添加量が上限値を超え
ると葉原基の形成が阻害されるため、葉芽が得られな
い。また、本発明に用いるサイトカイニンはＢＡまたは
ＢＡと同様な効果を示すカイネチンまたはホルクロロフ
ェニュロン液剤（４ＰＵ）を用いることができる。Here, for example, when benzylaminopurine (hereinafter referred to as BA) is used as cytokinin, BA is shaken in a liquid medium containing 0.01 to 10 mg / liter, preferably 0.01 to 5 mg / liter. Incubate or culture with aeration and stirring. If the content of BA is less than the lower limit value, the leaf primordia do not grow, and only one plant can be obtained. Further, if the amount of BA added exceeds the upper limit, the formation of leaf primordia is inhibited, so that leaf buds cannot be obtained. As the cytokinin used in the present invention, BA or kinetin or forchlorophenuron liquid (4PU) having the same effect as BA can be used.
【００１１】上記本発明に用いる培地は特に制限はな
く、一般的植物培養用の培地が用いられる。The medium used in the present invention is not particularly limited, and a general plant culture medium is used.
【００１２】[0012]
【作用】本発明の第一段の茎頂組織の培養は固体培地で
行い、ある程度育成したら液体培地で育成させることが
望ましい。その理由は、はじめから液体培地中で振とう
培養または通気撹拌培養すると、植物組織が培地になじ
んでないため、液体培地の表面に浮いてしまい、振とう
培養または通気撹拌培養中に培養容器の壁面に付着して
枯死するので、それを避けるためである。また、液体培
地を用いると固体培地に比較して、植物組織の成長が早
くなる。サイトカイニンを含む液体培地で振とう培養し
て葉芽を増殖させ、この葉芽をサイトカイニンを含まな
い液体培地で通気撹拌培養して発根率を約７７％に向上
させることができる。It is desirable that the first-stage shoot apical tissue of the present invention is cultured in a solid medium, and after being grown to some extent, it is grown in a liquid medium. The reason is that when shaking culture or aeration-agitation culture is performed in the liquid medium from the beginning, the plant tissue does not adapt to the culture medium and floats on the surface of the liquid medium, which causes the wall surface of the culture vessel during shaking culture or aeration-agitation culture. This is because it adheres to and die, so to avoid it. In addition, when a liquid medium is used, the plant tissue grows faster than a solid medium. The leaf buds can be propagated by shaking culture in a liquid medium containing cytokinin, and the leaf buds can be aerated and stirred in a liquid medium containing no cytokinin to improve the rooting rate to about 77%.
【００１３】[0013]
【実施例】本発明の実施例を説明する。 実施例１ 栽培されているイチゴから摘出した茎頂組織を次のよう
な条件で培養した。EXAMPLES Examples of the present invention will be described. Example 1 A shoot apex tissue extracted from a cultivated strawberry was cultured under the following conditions.
【００１４】まず、イチゴの茎頂組織をＢＡ１ｍｇ／リ
ットル、ショ糖１０ｇ／リットルおよびゲルライト２ｇ
／リットルを含むＭＳ改変培地で１４日間培養する（図
１の（１））。次いで、この茎頂組織をＢＡ１ｍｇ／リ
ットルおよびショ糖１０ｇ／リットルを含むＭＳ改変培
地で２１日間、１８０ｒｐｍで振とう培養する（図１
（２））。ここで、約１５０株分の葉芽が得られるが、
この培養を繰り返すことでその後は４n〜６n倍（ｎ＝継
代回数）と指数関数的に増殖することができる。図２に
示すシャーレは直径９ｃｍのものである。その後、増殖
した葉芽をショ糖３０ｇ／リットルを含み、無機塩濃度
１／２としたＭＳ改変培地を収容したビン中に入れ、フ
ィルタ除菌した空気を送り込みながら通気撹拌培養す
る。２１日間通気撹拌培養して、イチゴの植物体が再生
できた。First, the shoot apex tissue of strawberry was BA 1 mg / liter, sucrose 10 g / liter and gellite 2 g.
Culture is performed for 14 days in the MS modified medium containing 1 / liter ((1) in FIG. 1). Then, this shoot apex tissue is shake-cultured at 180 rpm for 21 days in an MS modified medium containing BA 1 mg / liter and sucrose 10 g / liter (FIG. 1).
(2)). Here, about 150 strains of leaf buds are obtained,
By repeating this culture, it is possible to grow exponentially with 4 n to 6 n times (n = passage number) thereafter. The petri dish shown in FIG. 2 has a diameter of 9 cm. Then, the grown leaf buds are placed in a bottle containing MS-modified medium containing 30 g / l of sucrose and having an inorganic salt concentration of ½, and aerobically agitated culture while feeding filtered air. After culturing with aeration and stirring for 21 days, strawberry plants could be regenerated.
【００１５】実施例２ 実施例１と同様に栽培されているイチゴから摘出した茎
頂組織をＢＡ１ｍｇ／リットル、ショ糖１０ｇ／リット
ルおよびゲルライト２ｇ／リットルを含むＭＳ改変培地
で１４日間培養し、次いで、この茎頂組織をＢＡ１ｍｇ
／リットルおよびショ糖１０ｇ／リットルを含むＭＳ改
変培地で２１日間培養する。このとき振とう培養をせず
にフィルタ除菌した空気を送り込みながら通気撹拌方法
を用いる。その後、増殖した葉芽をショ糖３０ｇ／リッ
トルを含み、無機塩濃度１／２としたＭＳ改変培地を収
容したビン中に入れ、フィルタ除菌した空気を送り込み
ながら通気撹拌培養して、イチゴの植物体が再生でき
た。Example 2 A shoot apex tissue extracted from a strawberry grown in the same manner as in Example 1 was cultivated for 14 days in an MS modified medium containing BA 1 mg / liter, sucrose 10 g / liter and gellite 2 g / liter, and then , 1mg of this shoot apex tissue
Culture for 21 days in MS modified medium containing 1 g / l and 10 g / l sucrose. At this time, the aeration and agitation method is used while feeding the filter-sterilized air without shaking culture. Then, the grown leaf buds were placed in a bottle containing MS-modified medium containing sucrose at 30 g / liter and having an inorganic salt concentration of 1/2, and cultivated with aeration and agitation while feeding filter-sterilized air to produce a strawberry plant. I was able to regenerate my body.
【００１６】実施例１の方法では葉芽の増殖培養過程で
振とう培養を行うため、例えば３００ｍｌの三角フラス
コに１００ｍｌまでしか培地を入れることができないの
で、培養容器の１／３のスペースしか利用できない。し
かし、本実施例の通気撹拌法を用いると、培養スペース
を有効利用した大量増殖が可能になる。具体的には、３
００ｍｌ三角フラスコ中の１００ｍｌ培地を用いる培養
方法に比較して、本実施例の方法では１４００ｍｌ培地
を２０００ｍｌの培養びんを用いて培養できるので、同
じスペースで２倍以上の組織を培養できる。なお、様々
なセンサや機械をつけたジャーファメンタにも本実施例
は応用できる。また、ジャーファンタで自動的に培地の
みを入れ換えることによって自動化ができる。In the method of Example 1, since shaking culture is carried out in the process of growing and growing leaf buds, for example, since a medium of up to 100 ml can be put in a 300 ml Erlenmeyer flask, only 1/3 of the space of the culture container can be used. .. However, when the aeration and stirring method of this example is used, it is possible to mass-proliferate by effectively utilizing the culture space. Specifically, 3
Compared to the culture method using a 100 ml medium in a 00 ml Erlenmeyer flask, in the method of the present example, 1400 ml medium can be cultured using a 2000 ml culture bottle, so that twice or more tissues can be cultured in the same space. It should be noted that this embodiment can be applied to a jar-famentor equipped with various sensors and machines. In addition, it can be automated by automatically replacing only the medium with a jar phanta.
【００１７】[0017]
【発明の効果】本発明により葉芽を液体培地で通気撹拌
培養することで、培養２１日間後の発根率が約７７％に
向上する。また、液体培地で根を誘導するため、ジャー
ファメンタを利用することができ、葉芽を１コずつ固体
培地に移植する手間が省ける。さらに、液体培地中で葉
芽から根を誘導した植物体は直接土壌に移植しても活着
し、２〜５ｃｍとそろっているので均質な苗を得ること
ができる。EFFECTS OF THE INVENTION By cultivating leaf buds in a liquid medium with aeration and stirring according to the present invention, the rooting rate after 21 days of culturing is improved to about 77%. Further, since the roots are induced in the liquid medium, jarfamenta can be used, and the labor of transplanting leaf buds one by one to the solid medium can be omitted. Furthermore, even if the plant body in which roots are induced from leaf buds in the liquid medium is directly transplanted to the soil, it will survive and have a size of 2 to 5 cm, so that a uniform seedling can be obtained.
【図１】本発明の実施例の葉芽増殖方法を概念的に示す
図である。FIG. 1 is a diagram conceptually showing a leaf bud proliferation method according to an embodiment of the present invention.
【図２】本発明の実施例の葉芽増殖方法で増殖したイチ
ゴの植物体に関する生物の形態を示す写真である。FIG. 2 is a photograph showing the morphology of organisms related to strawberry plants grown by the leaf bud growth method of the example of the present invention.
Claims (1)
養分を含む培地で培養して後、これをサイトカイニンを
含む液体培地で振とう培養または通気撹拌培養して、葉
芽を増殖させ、この葉芽をサイトカイニンを含まない液
体培地で通気撹拌培養することにより根を誘導して得ら
れるイチゴの植物体。1. A strawberry plant tissue is aseptically isolated and cultured in a nutrient-containing medium, and then this is shake-cultured or aeration-agitation-cultured in a liquid medium containing cytokinin to grow leaf buds. A strawberry plant body obtained by inducing roots by culturing leaf buds in a liquid medium containing no cytokinin with aeration and stirring.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP12688591A JPH0584028A (en) | 1991-04-30 | 1991-04-30 | Plant body of strawberry |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP12688591A JPH0584028A (en) | 1991-04-30 | 1991-04-30 | Plant body of strawberry |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPH0584028A true JPH0584028A (en) | 1993-04-06 |
Family
ID=14946260
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP12688591A Pending JPH0584028A (en) | 1991-04-30 | 1991-04-30 | Plant body of strawberry |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH0584028A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104686323A (en) * | 2014-12-31 | 2015-06-10 | 四川省农业科学院园艺研究所 | Method for cultivating strawberry seedlings by secondary detoxification method |
-
1991
- 1991-04-30 JP JP12688591A patent/JPH0584028A/en active Pending
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104686323A (en) * | 2014-12-31 | 2015-06-10 | 四川省农业科学院园艺研究所 | Method for cultivating strawberry seedlings by secondary detoxification method |
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