JPH02163017A - Tissue culture of plant of genus iris - Google Patents
Tissue culture of plant of genus irisInfo
- Publication number
- JPH02163017A JPH02163017A JP63312793A JP31279388A JPH02163017A JP H02163017 A JPH02163017 A JP H02163017A JP 63312793 A JP63312793 A JP 63312793A JP 31279388 A JP31279388 A JP 31279388A JP H02163017 A JPH02163017 A JP H02163017A
- Authority
- JP
- Japan
- Prior art keywords
- plant
- callus
- medium containing
- iris
- tissue
- 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
- 206010020649 Hyperkeratosis Diseases 0.000 claims abstract description 41
- 229930192334 Auxin Natural products 0.000 claims abstract description 20
- 239000002363 auxin Substances 0.000 claims abstract description 20
- UQHKFADEQIVWID-UHFFFAOYSA-N cytokinin Natural products C1=NC=2C(NCC=C(CO)C)=NC=NC=2N1C1CC(O)C(CO)O1 UQHKFADEQIVWID-UHFFFAOYSA-N 0.000 claims abstract description 14
- 239000004062 cytokinin Substances 0.000 claims abstract description 14
- 239000007787 solid Substances 0.000 claims abstract description 12
- 239000003375 plant hormone Substances 0.000 claims abstract description 7
- 230000008929 regeneration Effects 0.000 claims abstract description 6
- 238000011069 regeneration method Methods 0.000 claims abstract description 6
- 238000000034 method Methods 0.000 claims description 26
- 229930191978 Gibberellin Natural products 0.000 claims description 8
- 239000003448 gibberellin Substances 0.000 claims description 8
- IXORZMNAPKEEDV-OBDJNFEBSA-N gibberellin A3 Chemical class C([C@@]1(O)C(=C)C[C@@]2(C1)[C@H]1C(O)=O)C[C@H]2[C@]2(C=C[C@@H]3O)[C@H]1[C@]3(C)C(=O)O2 IXORZMNAPKEEDV-OBDJNFEBSA-N 0.000 claims description 7
- 230000001939 inductive effect Effects 0.000 claims description 4
- 230000001172 regenerating effect Effects 0.000 claims description 3
- 241000196324 Embryophyta Species 0.000 abstract description 52
- SEOVTRFCIGRIMH-UHFFFAOYSA-N indole-3-acetic acid Chemical compound C1=CC=C2C(CC(=O)O)=CNC2=C1 SEOVTRFCIGRIMH-UHFFFAOYSA-N 0.000 abstract description 8
- 241001579180 Matthiola longipetala Species 0.000 abstract 2
- 229940037201 oris Drugs 0.000 abstract 1
- IXORZMNAPKEEDV-UHFFFAOYSA-N gibberellic acid GA3 Natural products OC(=O)C1C2(C3)CC(=C)C3(O)CCC2C2(C=CC3O)C1C3(C)C(=O)O2 IXORZMNAPKEEDV-UHFFFAOYSA-N 0.000 description 5
- 238000004161 plant tissue culture Methods 0.000 description 4
- 230000015572 biosynthetic process Effects 0.000 description 3
- 230000006698 induction Effects 0.000 description 3
- 230000000644 propagated effect Effects 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- HYVABZIGRDEKCD-UHFFFAOYSA-N N(6)-dimethylallyladenine Chemical compound CC(C)=CCNC1=NC=NC2=C1N=CN2 HYVABZIGRDEKCD-UHFFFAOYSA-N 0.000 description 2
- 239000005708 Sodium hypochlorite Substances 0.000 description 2
- WZRRJZYYGOOHRC-UQJCXHNCSA-N gibberellin A8 Chemical compound C([C@@]1(O)C(=C)C[C@@]2(C1)[C@H]1C(O)=O)C[C@H]2[C@@]2(OC3=O)[C@H]1[C@@]3(C)[C@@H](O)[C@@H](O)C2 WZRRJZYYGOOHRC-UQJCXHNCSA-N 0.000 description 2
- SUKJFIGYRHOWBL-UHFFFAOYSA-N sodium hypochlorite Chemical compound [Na+].Cl[O-] SUKJFIGYRHOWBL-UHFFFAOYSA-N 0.000 description 2
- 239000002689 soil Substances 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- PRPINYUDVPFIRX-UHFFFAOYSA-N 1-naphthaleneacetic acid Chemical compound C1=CC=C2C(CC(=O)O)=CC=CC2=C1 PRPINYUDVPFIRX-UHFFFAOYSA-N 0.000 description 1
- OVSKIKFHRZPJSS-UHFFFAOYSA-N 2,4-D Chemical compound OC(=O)COC1=CC=C(Cl)C=C1Cl OVSKIKFHRZPJSS-UHFFFAOYSA-N 0.000 description 1
- 239000005972 6-Benzyladenine Substances 0.000 description 1
- GOSWTRUMMSCNCW-HNNGNKQASA-N 9-ribosyl-trans-zeatin Chemical compound C1=NC=2C(NC\C=C(CO)/C)=NC=NC=2N1[C@@H]1O[C@H](CO)[C@@H](O)[C@H]1O GOSWTRUMMSCNCW-HNNGNKQASA-N 0.000 description 1
- OJDCBRZJXYBPFZ-UHFFFAOYSA-N Gibberellin A2 Natural products C1CC(O)C(C)(C2C3C(O)=O)C(=O)OC21C1CCC2CC13CC2(O)C OJDCBRZJXYBPFZ-UHFFFAOYSA-N 0.000 description 1
- 235000015164 Iris germanica var. florentina Nutrition 0.000 description 1
- FAIXYKHYOGVFKA-UHFFFAOYSA-N Kinetin Natural products N=1C=NC=2N=CNC=2C=1N(C)C1=CC=CO1 FAIXYKHYOGVFKA-UHFFFAOYSA-N 0.000 description 1
- NWBJYWHLCVSVIJ-UHFFFAOYSA-N N-benzyladenine Chemical compound N=1C=NC=2NC=NC=2C=1NCC1=CC=CC=C1 NWBJYWHLCVSVIJ-UHFFFAOYSA-N 0.000 description 1
- GRYLNZFGIOXLOG-UHFFFAOYSA-N Nitric acid Chemical compound O[N+]([O-])=O GRYLNZFGIOXLOG-UHFFFAOYSA-N 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 125000003118 aryl group Chemical group 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000012136 culture method Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 239000003205 fragrance Substances 0.000 description 1
- WZRRJZYYGOOHRC-UHFFFAOYSA-N gibberellic acid 8 Natural products OC(=O)C1C2(C3)CC(=C)C3(O)CCC2C2(OC3=O)C1C3(C)C(O)C(O)C2 WZRRJZYYGOOHRC-UHFFFAOYSA-N 0.000 description 1
- OJDCBRZJXYBPFZ-UIEKCWFXSA-N gibberellin A2 Chemical compound C1C[C@H](O)[C@](C)([C@H]2[C@@H]3C(O)=O)C(=O)O[C@]21[C@@H]1CC[C@@H]2C[C@@]13C[C@]2(O)C OJDCBRZJXYBPFZ-UIEKCWFXSA-N 0.000 description 1
- 239000003617 indole-3-acetic acid Substances 0.000 description 1
- -1 indole-3acetic acid -Butyric acid Chemical compound 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 244000023249 iris florentino Species 0.000 description 1
- QANMHLXAZMSUEX-UHFFFAOYSA-N kinetin Chemical compound N=1C=NC=2N=CNC=2C=1NCC1=CC=CO1 QANMHLXAZMSUEX-UHFFFAOYSA-N 0.000 description 1
- 229960001669 kinetin Drugs 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 229910017604 nitric acid Inorganic materials 0.000 description 1
- 239000010662 orris oil Substances 0.000 description 1
- 230000001902 propagating effect Effects 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000012090 tissue culture technique Methods 0.000 description 1
- UZKQTCBAMSWPJD-UQCOIBPSSA-N trans-Zeatin Natural products OCC(/C)=C\CNC1=NC=NC2=C1N=CN2 UZKQTCBAMSWPJD-UQCOIBPSSA-N 0.000 description 1
- UZKQTCBAMSWPJD-FARCUNLSSA-N trans-zeatin Chemical compound OCC(/C)=C/CNC1=NC=NC2=C1N=CN2 UZKQTCBAMSWPJD-FARCUNLSSA-N 0.000 description 1
- 239000000341 volatile oil Substances 0.000 description 1
- 229940023877 zeatin Drugs 0.000 description 1
Abstract
Description
【発明の詳細な説明】
［産業上の利用分野］
本発明は、イリス属植物の組織培養方法に関し、特には
イリス属植物において、そのカルス様組織を誘導する方
法、カルス様組織を継代培養する方法、及びカルス様組
織から植物体へ再生する方法に関する。[Detailed Description of the Invention] [Industrial Application Field] The present invention relates to a method for culturing tissues of plants of the genus Iris, and in particular, a method for inducing callus-like tissue in plants of the genus Iris, and a method for subculturing callus-like tissues. The present invention relates to a method for regenerating a callus-like tissue into a plant body.
イリス属植物は、その根茎部に高級天然精油の一種であ
るオリス油を蓄積する、重要な香料植物である。Plants of the genus Iris are important aromatic plants that accumulate orris oil, a type of high-grade natural essential oil, in their rhizomes.
従来、イリス属植物は畑地で栽培され、その繁殖は地下
茎による栄養繁殖に頼っていた。しかし、畑地栽培によ
るイリス属植物の繁殖方法は、天候、土質等の自然環境
の影響を受けやすく、その繁殖は不安定であった。又、
ウィルスにおかされることも避けられず、健全な植物体
の生育が困難であった。Traditionally, plants of the genus Iris were cultivated in fields, and their reproduction relied on vegetative propagation via underground rhizomes. However, the method of propagating plants of the genus Iris through upland cultivation is susceptible to the influence of the natural environment such as weather and soil quality, and the propagation has been unstable. or,
Infection with viruses was unavoidable, making it difficult to grow healthy plants.
近年、組織培養技術によるイリス属植物の繁殖も報告さ
れている（Ｈａｒｔ ５ｃｉｅｎｃｅ 、 １０巻第５
号、１９７５年）。しかし、その和文でカルス化に成功
したのはイリス属植物の花部を用いた場合だけであり、
その他の部位（例えば根茎部、幼葉部）ではカルス化が
起きなかった。又、花部切片から得られたカルスを継代
培養した場合に、そのカルスから植物体への再生を安定
して実施できる継代培養の期間についても不明である。In recent years, propagation of Iris plants using tissue culture techniques has also been reported (Hart 5science, Vol. 10, No. 5).
No., 1975). However, in the Japanese version, callus formation was only successful when using the flower parts of plants of the genus Iris.
Callus formation did not occur in other parts (for example, rhizomes and young leaves). In addition, when callus obtained from flower sections is subcultured, it is also unclear how long the subculture can stably regenerate the callus into a plant.
以上のように、前記和文に記載の技術は、切片を採取す
ることのできる時期が制限されている点、及び植物体へ
の再生可能な継代培養期間が不明である点で、安定的な
大量繁殖方法としては不適切であった。As mentioned above, the technique described in the above Japanese paper has the disadvantages that the time when sections can be collected is limited and the period of subculture during which plants can be regenerated is unknown. It was inappropriate as a mass breeding method.
〔発明が解決しようとする課題］
従って、自然環境の影響を受けることなく、ウィルスフ
リーのイリス属植物の安定した大量繁殖を実現するため
に、カルス誘導用切片の採取時間に制限が−なく、しか
も継代培養を長期間行ってもカルスから植物体への再生
を安定に実施することのできるイリス属植物の組織培養
方法の開発が望まれでいた。[Problems to be Solved by the Invention] Therefore, in order to achieve stable mass propagation of virus-free plants of the genus Iris without being affected by the natural environment, there is no limit to the time for collecting sections for callus induction. Moreover, it has been desired to develop a tissue culture method for plants of the genus Iris that can stably regenerate callus into plants even after long-term subculturing.
本発明者は、驚くべきことに、カルス誘導用切片として
イリス属植物の葉のつけ根部分を用い、そしてこれをオ
ーキシン類含有培地に置床するとカルスが誘導され、し
かもこのカルスをオーキシン類含有培地で継代培養する
と植物体への再生能を長期間維持させることができ、更
に、そのカルスを特定の再生培地に移植することによっ
て植物体へ再生することができることを見出した。Surprisingly, the present inventors have found that when the base of a leaf of a plant belonging to the genus Iris is used as a section for callus induction, and when this is placed on an auxin-containing medium, callus is induced. It has been found that by subculturing, the ability to regenerate a plant can be maintained for a long period of time, and furthermore, by transplanting the callus to a specific regeneration medium, it can be regenerated into a plant.
従って、第一の本発明方法は、イリス属植物の葉のつけ
根部分の切片をオーキシン類含有固体培地上に置床する
ことを特徴とする、イリス属植物のカルス様組織の誘導
方法に関する。Therefore, the first method of the present invention relates to a method for inducing a callus-like tissue of a plant of the genus Iris, which comprises placing a section of the base of a leaf of a plant of the genus Iris on a solid medium containing auxins.
第二の本発明方法は、イリス属植物のカルス様組織をオ
ーキシン類含有培地で継代培養することを特徴とする、
イリス属植物のカルス様組織の継代培養方法に関する。The second method of the present invention is characterized in that callus-like tissue of a plant of the genus Iris is subcultured in a medium containing auxins.
This invention relates to a method for subculturing callus-like tissues of plants of the genus Iris.
第三の本発明方法は、植物ホルモンを含まない培地、サ
イトカイニン類含有培地、ジベレリン類含有培地、サイ
トカイニン類とオーキシン類とを含有する培地、及びジ
ベレリン類とオーキシン類とを含有する培地から成る群
から選んだ植物体再生培地に、イリス属植物のカルス様
組織を移植することを特徴とする、イリス属植物におけ
るカルス様組織から植物体への再生方法に関する。The third method of the present invention provides a group consisting of a medium containing no plant hormones, a medium containing cytokinins, a medium containing gibberellins, a medium containing cytokinins and auxins, and a medium containing gibberellins and auxins. The present invention relates to a method for regenerating a callus-like tissue from a plant of the genus Iris to a plant, the method comprising transplanting the callus-like tissue of a plant of the genus Iris to a plant regeneration medium selected from .
以下、本発明をより詳細に説明する。The present invention will be explained in more detail below.
第一の本発明方法においては、香料植物切片として、イ
リス属植物の葉のつけ根部分を用いる。In the first method of the present invention, the base portion of a leaf of a plant belonging to the genus Iris is used as a fragrance plant section.
これを通常の方法で滅菌した後、オーキシン類及び場合
によりサイトカイニン類を添加した植物組織培養用固形
培地上に置床してカルス様組織を誘導する。植物組織培
養用固形培地としては、ムラシゲとスクーグ（Ｍｕｒａ
ｓｈｉｇｅ ＆ Ｓｋｏｏｇ）培地を始め、リンスマイ
ヤーとスクーグ（Ｌｉｎｓｍａｉｅｒ ＆ Ｓｋｏｏｇ
Ｈ以下ＬＳと略す）、ホワイト（Ｗｈｉｔｅ）、ガンボ
ルグ（Ｇａｎｂｏｒｇ）、−１−ｙチ（Ｎ ｉ ｔｓｃ
ｈ）、ヘラ−（Ｈｅｌｌｅｒ）、シェンクとヒルデブラ
ンドＣ５ｃｈｅｎｋ ＆ １ｌｉｌｄｅｂｒａｎｔ）、
−’−７チとエッチ（Ｎｉｔｓｃｈ ＆ Ｎ１ｔｓｃｈ
）、コーレンバッハとシュミット（Ｋｏｈｌｅｎｂａｃ
ｈ ＆ Ｓｃｈｍｉｄｔ）などのいずれの培地を用いて
もよい。カルス８Ｗｒ Ｎのためには光を照射する必要
はない。After sterilizing this by a conventional method, it is placed on a solid medium for plant tissue culture supplemented with auxins and optionally cytokinins to induce callus-like tissue. Solid media for plant tissue culture include Murashige and Skoog (Mura
Shige & Skoog medium, Linsmaier & Skoog (Linsmaier & Skoog)
H (hereinafter abbreviated as LS), White, Ganborg, -1-y Chi (N i tsc
h), Heller (C5chenk & 1lildebrant),
-'-7 Chi and Ecchi (Nitsch & N1tsch
), Kohlenbach and Schmidt (Kohlenbach
Any medium such as H & Schmidt may be used. There is no need for light irradiation for callus 8WrN.
温度は、一般には１５°Ｃ〜３０°Ｃの範囲、好ましく
は２０°Ｃ〜３０°Ｃ１更に好ましくは２５°Ｃ〜２７
°Ｃである。又、オーキシン類としては、２゜４−ジク
ロロフェノキシ酢酸（以下、２．４−Ｄと略す）、α−
ナフタレン酢酸、インドール−３酢酸、インドール−３
−酪酸（以下、ＩＢＡと略す）、２．４．５−）ジクロ
ロフェノキシ酢酸などのいずれを添加しても良い。サイ
トカイニン類としてはゼアチン、６−ベンジルアデニン
、カイネチン（以下、ＫＴと略す）、リボシルゼアチン
、イソペンテニルアデニンなどのいずれを添加しても良
い。オーキシン類としては２．４−Ｄが好ましく、サイ
トカイニン類としてはＫＴが好ましい。添加するオーキ
シン類の濃度は一般には０、１ ｔａｒ／ｒｔｆｌ 〜
１０ ｕｇ／ｍｌの範囲であり、好ましくは０．５μｇ
／ ｍｌ〜２μｇ ／ ｍｌである。オーキシン類の
濃度が０．１μｇ ／ ｍ１未満だと、カルスが誘導さ
れる確率は極端に低下する。オーキシン類の濃度が１０
Ｉ！ｇ／ｄを超えてもカルスの誘導率が低下する。The temperature is generally in the range 15°C to 30°C, preferably 20°C to 30°C, more preferably 25°C to 27°C.
It is °C. In addition, as auxins, 2゜4-dichlorophenoxyacetic acid (hereinafter abbreviated as 2.4-D), α-
naphthaleneacetic acid, indole-3acetic acid, indole-3acetic acid
-Butyric acid (hereinafter abbreviated as IBA), 2.4.5-)dichlorophenoxyacetic acid, etc. may be added. As the cytokinins, any of zeatin, 6-benzyladenine, kinetin (hereinafter abbreviated as KT), ribosylzeatin, isopentenyladenine, etc. may be added. As the auxin, 2.4-D is preferred, and as the cytokinin, KT is preferred. The concentration of auxins to be added is generally 0.1 tar/rtfl ~
in the range of 10 ug/ml, preferably 0.5 μg
/ml to 2 μg/ml. When the concentration of auxins is less than 0.1 μg/ml, the probability of inducing callus is extremely reduced. The concentration of auxins is 10
I! Even if it exceeds g/d, the callus induction rate decreases.
サイトカイニン類を添加するとカルス化が促進される。Addition of cytokinins promotes callus formation.
サイトカイニン類の濃度は一般には０．０１ｐｇ／ ｍ
１〜１鱈／ ｍｌの範囲であり好ましくは０．０１屑／
ｍ２〜０．５硝／ ｍｌである。The concentration of cytokinins is generally 0.01 pg/m
The range is 1 to 1 cod/ml, preferably 0.01 pieces/ml.
m2 to 0.5 nitric acid/ml.
第二の本発明方法では、イリス属植物のカルス、好まし
くは前記の第一の本発明方法で誘導されたカルス様組織
を、カルス様組織を誘導させた条件と同じ条件で月に１
回程度の間隔で継代培養する。In the second method of the present invention, the callus of a plant of the genus Iris, preferably the callus-like tissue induced by the first method of the present invention, is grown once a month under the same conditions as the conditions under which the callus-like tissue was induced.
Subculture at intervals of approximately 100 times.
下記の実施例で示すとおり、本発明方法によって継代培
養されているカルス様組織は、１８ケ月後であっても植
物体への再生能を維持している。従って必要に応じて、
カルス様組織から植物体への再生を行うことができる。As shown in the Examples below, the callus-like tissue subcultured by the method of the present invention maintains the ability to regenerate into plants even after 18 months. Therefore, if necessary,
It is possible to regenerate callus-like tissue into a plant body.
第三の本発明方法では、イリス属植物のカルス（例えば
、前記の第一の本発明方法又は第二の本発明方法によっ
て得られたカルス様組織）から植物体を再生することが
できる。植物体への再生は、カルス様Ｍｉ職を植物ホル
モン無添加の通常の固形培地（植物組織培養用固形培地
として前記したもの）に移植して実施することもできる
が、好ましくは植物ホルモンとしてサイトカイニン類０
．１〜１０μｇ／１ｒｄｌ好ましくは０．１〜１．０鱈
／ｄを含む培地、又はジベレリン１０．１〜５０μｇ
／ ｍｌ好ましくは０．１〜１０μｇ ／ ｍｌを含む
培地、或いはサイトカイニンｉｏ、１〜１０μｇ／ ｒ
ｒｄｌとそのサイトカイニン類の１ ／１０量のオーキ
シン類とを含む培地、又はジベレリンｌ１ｊ（０，１〜
５０ｔｎｒ／ｍｌとそのジベレリン類の１／１０量〜１
／１００量のオーキシン類とを含む通常の固形培地（
植物組織培養用固形培地として前記したもの）に移植す
ると、根及び葉を同時に発生した植物体を再生する。ジ
ベレリン類としては、例えばジベレリンＡ８、ジベレリ
ンＡ２、ジベレリンＡ３（以下、ＧＡ３と略す）などの
いずれを添加してもよい。再生した植物体は植物ホルモ
ン無添加の培地で培養を行い、馴化させた後、畑地に移
植する。In the third method of the present invention, a plant body can be regenerated from the callus of a plant of the genus Iris (for example, the callus-like tissue obtained by the above-mentioned first method of the present invention or second method of the present invention). Regeneration into a plant can also be carried out by transplanting the callus-like Mi to a normal solid medium (described above as a solid medium for plant tissue culture) without addition of plant hormones, but it is preferable to use cytokinin as the plant hormone. Class 0
．． Medium containing 1 to 10 μg/1rdl, preferably 0.1 to 1.0 cod/d, or 10.1 to 50 μg of gibberellin
/ml preferably containing 0.1-10 μg/ml, or cytokinin io, 1-10 μg/r
A medium containing rdl and auxin in an amount of 1/10 of its cytokinin, or gibberellin l1j (0.1~
50 tnr/ml and 1/10 amount of gibberellin to 1
/100 amount of auxins (normal solid medium containing
When transplanted to a solid medium for plant tissue culture (as described above), a plant body that has simultaneously developed roots and leaves is regenerated. As the gibberellin, for example, any of gibberellin A8, gibberellin A2, gibberellin A3 (hereinafter abbreviated as GA3), etc. may be added. The regenerated plants are cultured in a medium without plant hormones, and after acclimation, they are transplanted to a field.
次に実施例によって本発明を具体的に説明する。 Next, the present invention will be specifically explained with reference to Examples.
ただし、本発明は以下の態様に限られるものではない。However, the present invention is not limited to the following embodiments.
遺Ｊ引性上
イリス・バリダ（穐朋上四〇、）の根茎の葉のつけ根部
分を切りとり、水洗後に１ ｃｍ立方に細分し、７０％
エタノールに数分間浸漬した。水洗後、５％次亜塩素酸
ナトリウム液に２０分間浸漬し、水洗後更に１〜２ ｍ
ｍの厚さに細分した。再度１％次亜塩素酸ナトリウム液
に１０分間浸漬した後に、更に２ｍ四方に細分し、２
、４−Ｄ １．０μｇ／Ｉｄ、及びＫ Ｔ ０．１ ｔ
ｎｒ／ ｍｌを含むＬＳ固形培地上に置床した。２６°
Ｃにて暗所で培養したところ約１ケ月後にカルス様組織
が出現した。得られた培養物を同条件で約１ケ月に１回
の間隔で植え継ぐことにより、継代培養を行った。この
培養物の継代培養を６ケ月間続けた後に、表１に示した
各種植物ホルモンを含むＬＳ培地に移植したところ、約
１〜２ケ月後に根及び葉を発生し植物体を再生した。そ
の再生頻度を表１に示す。ＫＴｌ、Ｏｎ／ｍｌ及びＩＢ
Ａｏ、１μｇ ／ ｍｌを含む培地で植物体が最もよく
再生した。再生した植物体を畑地に移植したところ、健
全な植物体として順調に生育を続けた。Cut off the base of the leaves of the rhizome of Iris barida (Akiho 140), wash it with water, then divide it into 1 cm cubes, and remove 70% of the leaves.
Soaked in ethanol for several minutes. After washing with water, immerse in 5% sodium hypochlorite solution for 20 minutes, and after washing with water, soak for 1 to 2 m.
It was subdivided into a thickness of m. After immersing it again in 1% sodium hypochlorite solution for 10 minutes, it was further subdivided into 2 m squares.
, 4-D 1.0 μg/Id, and K T 0.1 t
The cells were placed on LS solid medium containing nr/ml. 26°
When cultured in the dark at C, callus-like tissue appeared after about one month. Subculture was performed by subculturing the obtained culture under the same conditions at intervals of about once a month. After subculturing this culture for 6 months, it was transplanted to an LS medium containing the various plant hormones shown in Table 1. After about 1 to 2 months, roots and leaves were generated and the plants were regenerated. Table 1 shows the playback frequency. KTl, On/ml and IB
Plants regenerated best in a medium containing 1 μg/ml of Ao. When the regenerated plant was transplanted to a field, it continued to grow smoothly as a healthy plant.
実Ｊｌｄ達え
イリス・バリダ（Ｉｒｉｓ 卯月ｊ鎖）及びイリス・フ
ローレンチナ（通ｆｌｏｒｅｎｔｉｎａ）の根茎の葉の
つけ根部分より実施例１に示した方法により、カルス様
組織を誘導した。この培養物を継代１８ケ月後にＫＴｌ
、Ｏｎ／Ｉｎ１及びＩ Ｂ Ａ ０．１ ｐｇ／１ｔｄ
ｌを含むＬＳ培地に置床したところ、約１〜２ケ月後に
根及び葉を発生し植物体を再生した。再生した植物体は
畑地に移植したところ、健全な植物体として順調に生育
を続けた。A callus-like tissue was induced from the base of the leaves of the rhizomes of Iris barida and Iris florentina by the method shown in Example 1. After passage of this culture for 18 months, KTl
, On/In1 and I B A 0.1 pg/1td
When the plants were placed on an LS medium containing 1 to 10% of the total amount of LS, roots and leaves were generated after about 1 to 2 months, and the plants were regenerated. When the regenerated plant was transplanted to a field, it continued to grow smoothly as a healthy plant.
本発明方法によれば、天候、土質等の自然環境の影響を
うけることなく、安定的にイリス属植物を増殖すること
ができ、しかも無菌的に増殖させるため、ウィルスフリ
ーの健全な植物を増殖させることができる。According to the method of the present invention, plants of the genus Iris can be stably propagated without being affected by the natural environment such as weather and soil quality, and since they are propagated aseptically, virus-free and healthy plants can be propagated. can be done.
Claims (1)
類含有固体培地上に置床することを特徴とする、イリス
属植物のカルス様組織の誘導方法。 ２、イリス属植物のカルス様組織をオーキシン類含有固
体培地で継代培養することを特徴とする、イリス属植物
のカルス様組織の継代培養方法。 ３、植物ホルモンを含まない培地、サイトカイニン類含
有培地、ジベレリン類含有培地、サイトカイニン類とオ
ーキシン類とを含有する培地、及びジベレリン類とオー
キシン類とを含有する培地から成る群から選んだ植物体
再生培地に、イリス属植物のカルス様組織を移植するこ
とを特徴とする、イリス属植物におけるカルス様組織か
ら植物体への再生方法。[Scope of Claims] 1. A method for inducing callus-like tissue of a plant of the genus Iris, which comprises placing a section of the base of a leaf of a plant of the genus Iris on a solid medium containing auxins. 2. A method for subculturing a callus-like tissue of a plant of the genus Iris, which comprises subculturing the callus-like tissue of a plant of the genus Iris on a solid medium containing auxins. 3. Plant regeneration selected from the group consisting of a medium containing no plant hormones, a medium containing cytokinins, a medium containing gibberellins, a medium containing cytokinins and auxins, and a medium containing gibberellins and auxins. A method for regenerating a callus-like tissue from a plant of the genus Iris to a plant body, the method comprising transplanting the callus-like tissue of a plant of the genus Iris into a medium.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP63312793A JPH02163017A (en) | 1988-12-13 | 1988-12-13 | Tissue culture of plant of genus iris |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP63312793A JPH02163017A (en) | 1988-12-13 | 1988-12-13 | Tissue culture of plant of genus iris |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPH02163017A true JPH02163017A (en) | 1990-06-22 |
Family
ID=18033474
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP63312793A Pending JPH02163017A (en) | 1988-12-13 | 1988-12-13 | Tissue culture of plant of genus iris |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH02163017A (en) |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH09124499A (en) * | 1995-09-07 | 1997-05-13 | L'oreal Sa | Extract of sweet flag family and composition containing it |
| ES2304295A1 (en) * | 2006-07-31 | 2008-10-01 | Uiniversidade De Santiago De Compostela | Method for the micropropagation of iris boissieri and its applications. (Machine-translation by Google Translate, not legally binding) |
| CN102893872A (en) * | 2012-11-03 | 2013-01-30 | 云南省农业科学院花卉研究所 | Tissue culture method for domesticated seedlings of iris pallida |
| CN103734020A (en) * | 2014-01-26 | 2014-04-23 | 上海上房园艺有限公司 | Tissue culture method for German iris tectorum |
| CN104996302A (en) * | 2015-08-20 | 2015-10-28 | 浙江大学 | Method for remarkably improving callus inductivity of iris pseudacorus L. |
| CN105052743A (en) * | 2015-08-20 | 2015-11-18 | 浙江大学 | Method for effectively preserving yellow flag embryonic callus |
| CN108496800A (en) * | 2018-04-10 | 2018-09-07 | 黑龙江省科学院自然与生态研究所 | Chinese small iris high frequency regenerating system method is established based on callus induction |
| CN109452170A (en) * | 2018-11-07 | 2019-03-12 | 东北林业大学 | A kind of method of sword-like iris seed root evoked callus culture |
| CN111567401A (en) * | 2020-04-20 | 2020-08-25 | 上海植物园 | Tissue culture rapid propagation method of butterfly flower' Mengdian |
-
1988
- 1988-12-13 JP JP63312793A patent/JPH02163017A/en active Pending
Non-Patent Citations (1)
| Title |
|---|
| GERHARAD REUTHER BERICHTE DER DEUTSCHEN BOTANISCHEN GESELLSCHAFT=1977 * |
Cited By (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH09124499A (en) * | 1995-09-07 | 1997-05-13 | L'oreal Sa | Extract of sweet flag family and composition containing it |
| US6471997B1 (en) * | 1995-09-07 | 2002-10-29 | Societe L'oreal S.A. | Iridaceae extract and compositions containing it |
| ES2304295A1 (en) * | 2006-07-31 | 2008-10-01 | Uiniversidade De Santiago De Compostela | Method for the micropropagation of iris boissieri and its applications. (Machine-translation by Google Translate, not legally binding) |
| CN102893872A (en) * | 2012-11-03 | 2013-01-30 | 云南省农业科学院花卉研究所 | Tissue culture method for domesticated seedlings of iris pallida |
| CN103734020A (en) * | 2014-01-26 | 2014-04-23 | 上海上房园艺有限公司 | Tissue culture method for German iris tectorum |
| CN104996302A (en) * | 2015-08-20 | 2015-10-28 | 浙江大学 | Method for remarkably improving callus inductivity of iris pseudacorus L. |
| CN105052743A (en) * | 2015-08-20 | 2015-11-18 | 浙江大学 | Method for effectively preserving yellow flag embryonic callus |
| CN108496800A (en) * | 2018-04-10 | 2018-09-07 | 黑龙江省科学院自然与生态研究所 | Chinese small iris high frequency regenerating system method is established based on callus induction |
| CN108496800B (en) * | 2018-04-10 | 2021-07-16 | 黑龙江省科学院自然与生态研究所 | Method for establishing iris high-frequency regeneration system based on callus induction |
| CN109452170A (en) * | 2018-11-07 | 2019-03-12 | 东北林业大学 | A kind of method of sword-like iris seed root evoked callus culture |
| CN109452170B (en) * | 2018-11-07 | 2021-10-15 | 东北林业大学 | Method for callus culture induced by cordyceps sobolifera roots |
| CN111567401A (en) * | 2020-04-20 | 2020-08-25 | 上海植物园 | Tissue culture rapid propagation method of butterfly flower' Mengdian |
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