JP2591477B2 - How to Induce Kenaf Callus - Google Patents
How to Induce Kenaf CallusInfo
- Publication number
- JP2591477B2 JP2591477B2 JP6115218A JP11521894A JP2591477B2 JP 2591477 B2 JP2591477 B2 JP 2591477B2 JP 6115218 A JP6115218 A JP 6115218A JP 11521894 A JP11521894 A JP 11521894A JP 2591477 B2 JP2591477 B2 JP 2591477B2
- Authority
- JP
- Japan
- Prior art keywords
- callus
- kenaf
- medium
- seeds
- solid 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.)
- Expired - Fee Related
Links
- 206010020649 Hyperkeratosis Diseases 0.000 title claims description 29
- 240000000797 Hibiscus cannabinus Species 0.000 title claims description 23
- 238000000034 method Methods 0.000 claims description 13
- 230000006698 induction Effects 0.000 claims description 10
- 230000001939 inductive effect Effects 0.000 claims description 8
- 239000007787 solid Substances 0.000 claims description 8
- 239000003375 plant hormone Substances 0.000 claims description 5
- 230000001954 sterilising effect Effects 0.000 claims description 5
- 229930192334 Auxin Natural products 0.000 claims description 4
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 4
- 239000002363 auxin Substances 0.000 claims description 4
- 229910052799 carbon Inorganic materials 0.000 claims description 4
- 229930006000 Sucrose Natural products 0.000 claims description 2
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 2
- 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 description 2
- 239000004062 cytokinin Substances 0.000 claims description 2
- 239000005720 sucrose Substances 0.000 claims description 2
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims 3
- -1 2,4-dichlorophenoxy Chemical group 0.000 claims 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 claims 1
- 229910052791 calcium Inorganic materials 0.000 claims 1
- 239000011575 calcium Substances 0.000 claims 1
- 239000002609 medium Substances 0.000 description 18
- 241000196324 Embryophyta Species 0.000 description 5
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 238000004659 sterilization and disinfection Methods 0.000 description 3
- 239000005631 2,4-Dichlorophenoxyacetic acid Substances 0.000 description 2
- HXKWSTRRCHTUEC-UHFFFAOYSA-N 2,4-Dichlorophenoxyaceticacid Chemical compound OC(=O)C(Cl)OC1=CC=C(Cl)C=C1 HXKWSTRRCHTUEC-UHFFFAOYSA-N 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 2
- 241000218033 Hibiscus Species 0.000 description 2
- 235000005206 Hibiscus Nutrition 0.000 description 2
- 235000015928 Hibiscus cannabinus Nutrition 0.000 description 2
- 235000007185 Hibiscus lunariifolius Nutrition 0.000 description 2
- 239000008272 agar Substances 0.000 description 2
- 238000007796 conventional method Methods 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 230000012010 growth Effects 0.000 description 2
- 239000006870 ms-medium Substances 0.000 description 2
- 241000502561 Acacia irrorata Species 0.000 description 1
- 241000218236 Cannabis Species 0.000 description 1
- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 description 1
- 244000061176 Nicotiana tabacum Species 0.000 description 1
- 235000002637 Nicotiana tabacum Nutrition 0.000 description 1
- 240000007594 Oryza sativa Species 0.000 description 1
- 235000007164 Oryza sativa Nutrition 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 230000007910 cell fusion Effects 0.000 description 1
- 239000000460 chlorine Substances 0.000 description 1
- 229910052801 chlorine Inorganic materials 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 230000035784 germination Effects 0.000 description 1
- QWPPOHNGKGFGJK-UHFFFAOYSA-N hypochlorous acid Chemical compound ClO QWPPOHNGKGFGJK-UHFFFAOYSA-N 0.000 description 1
- SEOVTRFCIGRIMH-UHFFFAOYSA-N indole-3-acetic acid Chemical compound C1=CC=C2C(CC(=O)O)=CNC2=C1 SEOVTRFCIGRIMH-UHFFFAOYSA-N 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 230000008635 plant growth Effects 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 235000009566 rice Nutrition 0.000 description 1
- 238000011282 treatment Methods 0.000 description 1
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Description
【０００１】[0001]
【産業上の利用分野】本発明はケナフ（ハイビスカス
カナビナス、Ｈｉｂｉｓｃｕｓ ｃａｎｎａｂｉｎｕ
ｓ）のカルスの誘導方法に関するものであり、さらに詳
しくは、ケナフの種子を、特定のカルス誘導用固体培地
を用いて、特定の培養条件により培養することにより、
ケナフのカルスを安定かつ高効率で短期間で誘導する方
法に関するものである。The present invention relates to a kenaf (hibiscus)
Cannabis, Hibiscus cannabinu
s) The method for inducing callus, more specifically, by culturing kenaf seeds under a specific culture condition using a specific callus-inducing solid medium,
The present invention relates to a method for stably and efficiently inducing kenaf callus in a short time.
【０００２】[0002]
【従来の技術】植物のカルスの誘導は、細胞融合、外来
遺伝子の導入など、植物に対する種々の生物化学的取扱
いにおいて必要である。カルスの誘導は、タバコやイネ
をはじめとして種々の植物により行われ、カルスの誘導
方法について特開平４−５３４９５号公報、特開平２−
１２８６３１号公報において開示されている。2. Description of the Related Art Induction of plant callus is necessary in various biochemical treatments on plants, such as cell fusion and introduction of foreign genes. The callus is induced by various plants including tobacco and rice, and the callus induction method is disclosed in JP-A-4-53495 and JP-A-2-53.
No. 1,861,631.
【０００３】[0003]
【発明が解決しようとする課題】特開平４−５３４９５
号公報にはハイビスカスのカルスの誘導方法について示
されているが、植物種が異なるとカルスの誘導条件が全
く異なるため、従来のこの方法ではケナフのカルスを高
効率で短期間で誘導することはできなかった。Problems to be Solved by the Invention
The publication discloses a method for inducing the callus of hibiscus, but the callus induction conditions are completely different for different plant species.Therefore, with this conventional method, it is possible to induce kenaf callus with high efficiency in a short period of time. could not.
【０００４】また、特開平２−１２８６３１号公報には
ケナフのカルスの誘導方法について示されているが、ケ
ナフの種子を寒天培地上で発芽させた後に、その苗条の
茎の一片を植物ホルモンを含有する別の培地に置床して
カルスを誘導するため、操作が煩雑で、誘導期間に４０
日と長期間を要するという問題点があった。Japanese Patent Application Laid-Open No. 2-128631 discloses a method for inducing kenaf callus. After germinating kenaf seeds on an agar medium, a piece of the stem of the shoot is planted with a plant hormone. Since the callus is induced by placing the medium on another medium containing the medium, the operation is complicated, and 40
There is a problem that it takes days and a long time.
【０００５】本発明の目的は、上記問題点を解決し、ケ
ナフのカルスを安定かつ高効率で、容易に短期間で誘導
する方法を提供することである。An object of the present invention is to solve the above problems and to provide a method for guiding kenaf callus stably, with high efficiency and easily in a short period of time.
【０００６】[0006]
【課題を解決するための手段】本発明の第１の発明は、
ケナフの種子に滅菌処理を施した後、その種子を、炭素
源と、オーキシン類とサイトカニン類とからなる植物ホ
ルモンを含有し、ｐＨ調整された合成固体培地に置床
し、無菌的に培養するケナフのカルスの誘導方法であっ
て、前記合成固体培地が、炭素源としてショ糖３重量％
と、オーキシン類として２，４−ジクロロフェノキシ酢
酸３００μｇ／ｌとサイトカイニン類としてカイチネン
３００μｇ／ｌとからなる植物ホルモンを含有し、ｐＨ
７に調整された合成固体培地であり、２５℃の温度で１
１時間日長の培養条件で無菌的に培養することを特徴と
するケナフのカルスの誘導方法である。Means for Solving the Problems A first invention of the present invention is:
After sterilizing kenaf seeds, the seeds are placed on a pH-adjusted synthetic solid medium containing a carbon source and a plant hormone consisting of auxins and cytocanins, and the culture is aseptically cultured. A method for inducing kenaf callus, wherein the synthetic solid medium contains 3% by weight of sucrose as a carbon source.
And a plant hormone consisting of 300 μg / l of 2,4-dichlorophenoxyacetic acid as an auxin and 300 μg / l of kaitinene as cytokinins;
7 is a synthetic solid medium adjusted to 7 and 1 at 25 ° C.
It is a method for inducing kenaf callus, which comprises aseptically culturing under a culture condition of one hour photoperiod.
【０００７】[0007]
【０００８】[0008]
【０００９】[0009]
【実施例】次に本発明の一実施例を説明する。ケナフ
（Ｈｉｂｉｓｃｕｓ ｃａｎｎａｂｉｎｕｓ）としては
中国産の青皮三号等を用いてカルスの誘導実験を行っ
た。Next, an embodiment of the present invention will be described. Callus induction experiments were carried out using kenaf (Hibiscus cannabinus), such as Chinese blue skin No.3.
【００１０】ケナフの種子を有効塩素濃度１％の次亜塩
素酸溶液に２０分浸漬することにより、滅菌を行った。
種子は厚い殻に包まれているため、殻内部のカルスの誘
導に必要な細胞を破壊することなく滅菌操作を行え、葉
や茎からカルスを誘導する場合に比べ滅菌操作上有利で
ある。[0010] Sterilization was performed by immersing kenaf seeds in a hypochlorous acid solution having an effective chlorine concentration of 1% for 20 minutes.
Since the seeds are wrapped in thick shells, the sterilization operation can be performed without destroying the cells necessary for the induction of callus inside the shell, which is advantageous in sterilization operation as compared with the case where the callus is induced from leaves or stems.
【００１１】培地としてはＭＳ培地（ムラシゲ・スクー
グ培地）を基本培地とし、これに２，４−ジクロロフェ
ノキシ酢酸３００μｇ／ｌとカイチネン３００μｇ／を
この濃度割合で添加し、水酸化ナトリウム溶液を加える
ことによりｐＨを７に調整した。この培地に寒天１重量
％を添加して常法により加圧蒸気殺菌した培地を、直径
４０ｍｍの植物培養用試験管に２０ｍｌづつ分注した。
培地のｐＨを調整する本発明の方法によれば、培地を固
体化させることができ、そのｐＨは中性付近が好まし
い。なお、従来の方法で使用される培地およびＭＳ培地
はｐＨが５．６〜５．８と酸性側であり、ｐＨ調整はさ
れておらず、このｐＨ領域では培地を固体化することは
できなかった。As a medium, an MS medium (Murasige-Skoog medium) is used as a basic medium, and 2,4-dichlorophenoxyacetic acid (300 μg / l) and kaitinene (300 μg /) are added at this concentration ratio, and a sodium hydroxide solution is added. The pH was adjusted to 7 with. 1% by weight of agar was added to this medium, and the medium was sterilized by pressurized steam in a conventional manner, and 20 ml of the medium was dispensed into test tubes for plant culture having a diameter of 40 mm.
According to the method of the present invention for adjusting the pH of a medium, the medium can be solidified, and its pH is preferably near neutral. The medium and the MS medium used in the conventional method have an acidic pH of 5.6 to 5.8, are not adjusted in pH, and cannot be solidified in this pH range. Was.
【００１２】ついで、上記ケナフの種子を上記試験管内
の合成固体培地上に置床し、２５℃で、照度１２００Ｌ
ｕｘの植物育成用蛍光灯による１１時間日長の照射条件
下で培養すると、２４時間程度の後にケナフの種子が発
芽する。発芽した種子の成長点部分は増殖速度が高いた
め、葉や茎を用いてカルスを誘導する場合と異なり発芽
直後から細胞の変異が始まり、カルスの誘導が行われ
る。本方法により、カルス細胞塊が観察され、カルスの
誘導が確認されるまでに要した期間は３日間であり、１
００％の誘導効率で安定にカルスを誘導することができ
た。この誘導期間は、従来のカルスの誘導に要する時間
と比較してかなり短いものである。図１に、本発明によ
り誘導されたケナフのカルスの写真を示す。Next, the kenaf seeds are placed on the synthetic solid medium in the test tube, and the illuminated light is 1200 L at 25 ° C.
When cultivated under the irradiation conditions of 11 hours of photoperiod with a fluorescent light for plant growth of ux, kenaf seeds germinate after about 24 hours. Since the growth point of the germinated seeds has a high growth rate, the cell mutation starts immediately after germination, and the callus is induced, unlike the case where the callus is induced using leaves or stems. According to this method, the time required for callus cell clusters to be observed and callus induction to be confirmed was 3 days.
Callus could be induced stably with an induction efficiency of 00%. This induction period is considerably shorter than the time required for the conventional callus induction. FIG. 1 shows a photograph of kenaf callus induced according to the present invention.
【００１３】[0013]
【発明の効果】本発明の方法によれば、ケナフのカルス
を安定かつ高効率で、容易に短期間で誘導することがで
きる。According to the method of the present invention, the callus of kenaf can be induced stably, with high efficiency and easily in a short period of time.
【図１】本発明により誘導されたケナフのカルスの写
真。FIG. 1 is a photograph of kenaf callus induced according to the present invention.
なし None
Claims (1)
種子を、炭素源と、オーキシン類とサイトカニン類とか
らなる植物ホルモンを含有し、ｐＨ調整された合成固体
培地に置床し、無菌的に培養するケナフのカルスの誘導
方法であって、 前記合成固体培地が、前記炭素源としてショ糖３重量％
と、前記オーキシン類として２，４−ジクロロフェノキ
シ酢酸３００μｇ／ｌと前記サイトカイニン類としてカ
イチネン３００μｇ／ｌとからなる植物ホルモンを含有
し、ｐＨが中性付近に調整された合成固体培地であり、
２５℃の温度で１１時間日長の培養条件で無菌的に培養
する ことを特徴とするケナフのカルスの誘導方法。1. After sterilizing kenaf seeds, the seeds are placed on a synthetic solid medium having a pH adjusted and containing a carbon source and a plant hormone consisting of auxins and cytocanins, Induction of kenaf callus cultured aseptically
The method, wherein the synthetic solid medium comprises 3% by weight of sucrose as the carbon source.
And 2,4-dichlorophenoxy as the auxins
300 μg / l of acetic acid and calcium as the cytokinins
Contains plant hormone consisting of 300 µg / l of itinene
And a synthetic solid medium whose pH has been adjusted to near neutrality,
Aseptically cultured at a temperature of 25 ° C under a culture condition of 11 hours photoperiod
A method for inducing kenaf callus, comprising:
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP6115218A JP2591477B2 (en) | 1994-05-27 | 1994-05-27 | How to Induce Kenaf Callus |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP6115218A JP2591477B2 (en) | 1994-05-27 | 1994-05-27 | How to Induce Kenaf Callus |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPH07313149A JPH07313149A (en) | 1995-12-05 |
| JP2591477B2 true JP2591477B2 (en) | 1997-03-19 |
Family
ID=14657292
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP6115218A Expired - Fee Related JP2591477B2 (en) | 1994-05-27 | 1994-05-27 | How to Induce Kenaf Callus |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JP2591477B2 (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6927192B2 (en) * | 2002-02-15 | 2005-08-09 | Regents Of The University Of Minnesota | Process to improve the quality of grains and seeds |
-
1994
- 1994-05-27 JP JP6115218A patent/JP2591477B2/en not_active Expired - Fee Related
Non-Patent Citations (1)
| Title |
|---|
| ＰＬＡＮＴ ＣＥＬＬ ＲＥＰＯＲＴＳ，ＶＯＬ．１１，ＮＯ．１０（１９９２）Ｐ．５３２−５３４ |
Also Published As
| Publication number | Publication date |
|---|---|
| JPH07313149A (en) | 1995-12-05 |
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