JPH104811A - Proliferation of large amount of plant of genus hemerocallis - Google Patents
Proliferation of large amount of plant of genus hemerocallisInfo
- Publication number
- JPH104811A JPH104811A JP8180019A JP18001996A JPH104811A JP H104811 A JPH104811 A JP H104811A JP 8180019 A JP8180019 A JP 8180019A JP 18001996 A JP18001996 A JP 18001996A JP H104811 A JPH104811 A JP H104811A
- Authority
- JP
- Japan
- Prior art keywords
- plant
- medium
- hemerocallis
- culturing
- plants
- 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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- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
Description
【0001】[0001]
【従来の技術】ヘメロカリス属はユリ科の1属で、日本
を含む東アジア各地に分布している。1800年代から
今日にかけて、主にアメリカ合衆国で観賞用に品種改良
が行われてきた。ヘメロカリス属植物は耐寒性、耐乾燥
性に優れ、痩せ地での植栽が可能であり、園芸用植物な
らびに造園用植物として多く用いられている。2. Description of the Related Art The genus Hemerocallis is a genus of the lily family and is distributed throughout East Asia including Japan. From the 1800's to today, breeding has been carried out mainly for ornamental use in the United States. Hemerocallis plants are excellent in cold resistance and drought resistance, can be planted on a thin land, and are widely used as horticultural plants and landscaping plants.
【0002】従来、植物体の生長点や組織の切片を培地
上で無菌的に培養することにより、体細胞胚、不定芽、
カルスなどを誘導し、生育させることにより当該植物を
増殖させる植物の組織培養による増殖方法が、種々の植
物を対象に研究され、これまでに種々の技術が開発され
ている。しかしながら、組織培養の条件は植物種個々に
ついて詳細に検討しなければならず、全ての植物種で容
易に実施できるものではない。ヘメロカリス属植物の増
殖は実生または株分けで行っているのが実状である。[0002] Conventionally, aseptic culture of a growing point of a plant or a section of a tissue on a medium has resulted in somatic embryos, adventitious buds,
Propagation methods by tissue culture of plants in which callus and the like are induced and grown by growing the plants are studied for various plants, and various techniques have been developed so far. However, the conditions for tissue culture must be examined in detail for each plant species and cannot be easily carried out for all plant species. In fact, the growth of Hemerocallis plants is carried out by seedlings or strains.
【0003】[0003]
【発明が解決しようとする課題】本発明の目的は、花卉
植物として有用なヘメロカリス属植物を遺伝的安定性を
保ちつつ短期間に安定して大量増殖する方法を提供する
ものである。SUMMARY OF THE INVENTION It is an object of the present invention to provide a method for stably mass-producing Hemerocallis plants useful as flowering plants in a short period of time while maintaining genetic stability.
【課題を解決するための手段】本発明の構成は、以下の
(1)〜(3)の技術的手段からなる。 (1)無菌的に分離したヘメロカリス属植物の組織を不
定芽形成培地で培養して不定芽を形成させ小植物体を獲
得する工程と、前記獲得した小植物体を多芽体誘導培地
で培養して多芽体を誘導する工程と、前記誘導した多芽
体を分割して植物体再生培地で培養して植物体を再生す
る工程とを含むヘメロカリス属植物の大量増殖方法。 (2)ヘメロカリス属植物の組織が花茎であることを特
徴とする(1)に記載のヘメロカリス属植物の大量増殖
方法。 (3)多芽体誘導培地および植物体再生培地が植物生長
調節物質を含有しない培地であることを特徴とする
(1)又は(2)に記載のヘメロカリス属植物の大量増
殖方法。The constitution of the present invention comprises the following technical means (1) to (3). (1) A step of culturing aseptically isolated Hemerocallis plant tissue in an adventitious bud formation medium to form adventitious buds and obtaining plantlets, and culturing the obtained plantlets in a polyplast induction medium A method for mass-producing Hemerocallis plants, comprising: a step of inducing polyblasts by regenerating the plant; and a step of regenerating the plants by dividing the induced polyblasts and culturing the plants in a plant regeneration medium. (2) The method for large-scale propagation of Hemerocallis plants according to (1), wherein the tissue of Hemerocallis plants is a flower stem. (3) The method for mass-producing Hemerocallis plants according to (1) or (2), wherein the multibud induction medium and the plant regeneration medium are mediums containing no plant growth regulator.
【0004】[0004]
【発明の実施の形態】本発明に用いられるヘメロカリス
属植物は植物分類学上、ユリ科に属する植物で日本に生
育するものはもちろん、世界に生育する自然交配種およ
び人工交配種を含む。日本に自生する野生種を例示すれ
ば、ノカンゾウ、ハマカンゾウなどを挙げることができ
る。アメリカ合衆国においては、品種改良が盛んに行わ
れ、優れた園芸品種が多く作出されているが、これらも
本発明に用いることができる。欧米で一般にデイリリー
と呼ばれているものもヘメロカリス属植物に属する。BEST MODE FOR CARRYING OUT THE INVENTION The plant of the genus Hemerocallis used in the present invention includes plants belonging to the lily family in terms of plant taxonomy and growing in Japan, as well as natural hybrids and artificial hybrids growing in the world. Examples of wild species that grow naturally in Japan include Norikanzo and Hamakanzo. In the United States, varieties are being actively improved, and many excellent horticultural varieties are produced. These can also be used in the present invention. What is generally called a daily lily in Europe and the United States also belongs to Hemerocallis plants.
【0005】これらヘメロカリス属植物の植物体の切
片、例えば、茎頂、花茎、茎、葉、根などは、必要に応
じてエタノール、次亜塩素酸ナトリウム、塩化ベンザル
コニウムなどの殺菌剤を用いて殺菌して用いられる。あ
るいは、消毒種子を無菌播種して発芽させた無菌幼苗な
ども利用することができる。特に、花茎は取扱いが容易
で殺菌作業上からも好適である。[0005] Sections of these plants of the genus Hemerocallis, for example, shoot apices, flower stalks, stems, leaves, roots, and the like, may be treated with a fungicide such as ethanol, sodium hypochlorite, or benzalkonium chloride, if necessary. It is used after sterilization. Alternatively, aseptic seedlings and the like germinated by aseptically sowing disinfected seeds can also be used. In particular, inflorescences are easy to handle and suitable for sterilization.
【0006】本発明で用いる組織培養用培地には、不定
芽形成培地、多芽体誘導培地、植物体再生培地がある。
これらの具体例として、通常の植物の組織培養に一般に
用いられるムラシゲ−スクーグ(Murasige−S
koog)培地(以下、「MS培地」という。)、ガン
ボーグ−B5(Gamborg−B5)培地などを挙げ
ることができる。また、通常は肥料として用いられてい
るハイポネックス(ハイポネックス・ジャパン社製商品
名)の水溶液なども用いることができる。これらの基本
培地に必要に応じて適当量の炭素源、オーキシン、サイ
トカイニンなどの植物生長調節物質、ココナットウォー
ターなどの植物由来有機物などを添加して用いることが
できる。ここで、多芽体誘導培地と植物体再生培地に
は、植物体に変異が起こる確率を小さくするためにも植
物生長調節物質を加えないことが好ましい。[0006] The tissue culture medium used in the present invention includes an adventitious bud formation medium, a multiple shoots induction medium, and a plant regeneration medium.
As specific examples of these, Murashige-Skoog (Murasige-Skoog) generally used for tissue culture of ordinary plants is used.
koog) medium (hereinafter referred to as “MS medium”), Gamborg-B5 medium, and the like. An aqueous solution of Hyponex (trade name, manufactured by Hyponex Japan), which is usually used as a fertilizer, can also be used. If necessary, an appropriate amount of a carbon source, a plant growth regulator such as auxin, cytokinin, or a plant-derived organic substance such as coconut water can be added to these basic media. Here, it is preferable not to add a plant growth regulator to the multi-bud-induction medium and the plant regeneration medium in order to reduce the probability of occurrence of mutation in the plant.
【0007】オーキシンとしては、2,4−ジクロロフ
ェノキシ酢酸(2,4−D)、α−ナフタレン酢酸(N
AA)、インドール酢酸(IAA)などを使用すること
ができる。またサイトカイニンとしては、6−フルフリ
ルアミノプリン、6−ベンジルアミノプリン(BA
P)、ゼアチンなどを使用することができる。Auxins include 2,4-dichlorophenoxyacetic acid (2,4-D) and α-naphthaleneacetic acid (N
AA), indoleacetic acid (IAA) and the like can be used. Examples of cytokinins include 6-furfurylaminopurine and 6-benzylaminopurine (BA
P), zeatin and the like can be used.
【0008】固体培地で培養を行う場合は、植物培養用
基材として、寒天、ジェランガムなどを0.2〜2.0
重量%程度添加して用いる。When culturing on a solid medium, agar, gellan gum or the like is used as a substrate for plant cultivation in an amount of 0.2 to 2.0.
It is used by adding about weight%.
【0009】本発明で用いる培養容器としては、その形
状に特に制限はなく、光を透過するガラス、ポリカーボ
ネート製などの容器であればよい。また培養容器は無菌
状態、湿度を保つため蓋や栓などで密閉されていること
が望ましい。[0009] The culture vessel used in the present invention is not particularly limited in its shape, and may be any vessel made of glass, polycarbonate or the like that transmits light. Further, it is desirable that the culture vessel is sealed with a lid, a stopper, or the like in order to maintain a sterile condition and humidity.
【0010】再生植物体を得るまでの培養温度は、20
〜30℃、好ましくは23〜25℃である。また、光条
件としては、蛍光灯などを連続的または断続的に照射す
ることが望ましい。液体培地で培養を行う場合は、ロー
タリーシェーカーなどを用いて振盪培養してもよい。The cultivation temperature until a regenerated plant is obtained is 20
-30 ° C, preferably 23-25 ° C. As for the light condition, it is desirable to irradiate a fluorescent lamp or the like continuously or intermittently. When culturing in a liquid medium, shaking culturing may be performed using a rotary shaker or the like.
【0011】[0011]
(実施例1)本発明の一実施例を添付した図面を参照し
て以下に説明する。図1は、本発明の処理工程概略図で
ある。図中Aが花茎組織を採取する工程、Bが不定芽を
形成させ小植物体を獲得する工程、Cが多芽体を誘導す
る工程、Dが植物体を再生する工程、Eが苗を馴化する
工程を示している。なお、全工程に要する日数は約6ヶ
月であった。(Embodiment 1) An embodiment of the present invention will be described below with reference to the accompanying drawings. FIG. 1 is a schematic diagram of the processing steps of the present invention. In the figure, A is a step of collecting flower stem tissue, B is a step of forming adventitious buds to obtain small plantlets, C is a step of inducing polyblasts, D is a step of regenerating plants, and E is a habit of seedlings. FIG. The number of days required for the entire process was about 6 months.
【0012】花茎組織を採取する工程Aにおいて、ヘメ
ロカリスの野生種であるノカンゾウ(Hemerocallis thun
bergii var. logituba Miq.)、およびハマカンゾウ(Hem
erocallis aurantiaca var. littorea Makino)から花蕾
が緑色の未熟な花茎1を採取し、花蕾部を切除した後、
5cm程度の長さに切りそろえ、有効塩素濃度1%の次
亜塩素酸ナトリウムで表面を滅菌後、2〜3mmの輪切
りにして花茎切片2を調製した。In step A of collecting flower stem tissue, Hemerocallis thun , a wild species of Hemerocallis, is used.
bergii var.logituba Miq.), and licorice ( Hem )
erocallis aurantiaca var. littorea Makino) from the flower buds are harvested green immature flower stalks 1, after you cut the florets part,
The pieces were cut into lengths of about 5 cm, sterilized on the surface with sodium hypochlorite having an effective chlorine concentration of 1%, and cut into 2 to 3 mm pieces to prepare flower stem sections 2.
【0013】不定芽を形成させ小植物体を獲得する工程
Bにおいて、該切片2をα−ナフタレン酢酸(NAA)
0.1mg/l、6−ベンジルアミノプリン(BAP)
0.1mg/lを含む1/2濃度のMS固体培地(不定
芽形成培地3)へ着床し、25℃、連続照明下(36μ
mol/m2/s)で不定芽4を形成させた。形成され
た不定芽の数を表1に示す。In the step B of forming adventitious buds and obtaining plantlets, the section 2 is treated with α-naphthaleneacetic acid (NAA).
0.1 mg / l, 6-benzylaminopurine (BAP)
Implanted in a 濃度 concentration MS solid medium (adventitious bud formation medium 3) containing 0.1 mg / l, and under continuous lighting (36 μm) at 25 ° C.
mol / m 2 / s) to form adventitious buds 4. Table 1 shows the number of adventitious shoots formed.
【0014】[0014]
【表1】 [Table 1]
【0015】ノカンゾウとハマカンゾウの不定芽形成効
率(不定芽を形成した切片数/置床切片数×100)は
各々19.0%と9.1%であった。成功率に種間差が
生じたが、これはそれぞれの種の遺伝子型の違いによる
ものであると考えられた。図2は、ノカンゾウの不定芽
を示す写真である。得られたノカンゾウの不定芽は、発
根して完全な小植物体となるまで培養を継続した。The adventitious bud formation efficiencies (the number of sections that formed adventitious buds / the number of planted sections × 100) of Norikanzo and Hamakanzo were 19.0% and 9.1%, respectively. There was an interspecies difference in the success rate, which was thought to be due to the genotype difference of each species. FIG. 2 is a photograph showing adventitious buds of Norizo. The obtained adventitious shoots of Noctuids were continued to culture until rooting and complete plantlets were obtained.
【0016】つづく多芽体を誘導する工程Cにおいて、
上記得られたノカンゾウの小植物体を、表2に示す各多
芽体誘導培地5へ移植し、25℃、連続照明下(36μ
mol/m2/s)でロータリーシェーカー(90〜1
00rpm)を用いて振盪培養を行い、多芽体6を誘導
した。In the subsequent step C for inducing polyblasts,
The obtained plantlets of Noctuids elegans were transplanted into each of the multiple shoots induction mediums 5 shown in Table 2, and were continuously illuminated at 25 ° C. (36 μm).
mol / m 2 / s) with a rotary shaker (90 to 1).
(00 rpm) to induce polyblasts 6.
【0017】[0017]
【表2】 [Table 2]
【0018】多芽体誘導培地5の最適条件は、ショ糖濃
度10g/l、植物生長調節物質無添加、ココナットウ
ォーター無添加であり、該培地で2回の継代を行い、3
ヶ月培養した時点での多芽体誘導効率(多芽体を構成す
る芽条の合計数/培養植物体数×100)は275.0
%であった。図3は、ノカンゾウの多芽体を示す写真で
ある。The optimal conditions for the multi-blast medium 5 are sucrose concentration of 10 g / l, no addition of plant growth regulator and no coconut water.
The multibud induction efficiency at the time of culturing for months (total number of shoots constituting the multibud / number of cultured plants × 100) is 275.0.
%Met. FIG. 3 is a photograph showing polyblasts of Norizo.
【0019】つづく植物体を再生する工程Dにおいて、
ノカンゾウの多芽体6を多芽体誘導培地5から表3に示
す各植物体再生培地7へ分割移植した。1〜2ヶ月の間
に芽条が伸長し、容易に発根して再生植物体8を得た。In the following step D for regenerating the plant,
The multi shoots 6 of the licorice were divided and transplanted from the multi shoot induction medium 5 to each plant regeneration medium 7 shown in Table 3. The buds elongated during 1-2 months, easily rooted, and the regenerated plant 8 was obtained.
【0020】[0020]
【表3】 [Table 3]
【0021】植物体再生培地7の最適条件は、基本培地
として0.3%(w/v)ハイポネックス水溶液培地を
使用し、ショ糖濃度10g/l、寒天濃度14g/lで
あった。ここで得られた、再生植物体8を、該最適条件
の培地に植え継いで培養を継続したところ、1ヶ月で高
さ10cmまで成長した。図4は多芽体を植物体再生培
地に移植することによって伸長したノカンゾウの芽条を
示す写真である。The optimum conditions for the plant regeneration medium 7 were as follows: a 0.3% (w / v) Hyponex aqueous medium was used as a basic medium, and the sucrose concentration was 10 g / l and the agar concentration was 14 g / l. When the regenerated plant body 8 obtained here was subcultured in a medium under the optimum conditions and continued to be cultured, it grew to a height of 10 cm in one month. FIG. 4 is a photograph showing shoots of licorice elongated by transplanting the multiple shoots into a plant regeneration medium.
【0022】つづく苗を馴化する工程Eにおいて、再生
植物体8をバーミキュライトの苗床に移植し、外環境へ
の馴化を行った。図5は再生したノカンゾウ植物体の馴
化を示す写真である。In the following step E for acclimating the seedlings, the regenerated plant 8 was transplanted to a vermiculite seedbed to acclimate to the external environment. FIG. 5 is a photograph showing the acclimation of the regenerated licorice plant.
【0023】[0023]
【発明の効果】本発明により、これまで実生、株分けな
どによっていたヘメロカリス属植物の増殖が、短期間
で、大量に、かつ遺伝的に均一に行えるようになる。Industrial Applicability According to the present invention, the growth of Hemerocallis plants, which have been conventionally performed by seedlings or strains, can be performed in a short period, in a large amount, and genetically uniformly.
【図1】本発明の一実施例である処理工程概略図であ
る。FIG. 1 is a schematic view of a processing step according to an embodiment of the present invention.
【図2】ノカンゾウの不定芽を示す写真である。FIG. 2 is a photograph showing adventitious buds of Norizo.
【図3】ノカンゾウの多芽体を示す写真である。FIG. 3 is a photograph showing multiple shoots of licorice.
【図4】多芽体を植物体再生培地に移植することによっ
て伸長したノカンゾウの芽条を示す写真である。FIG. 4 is a photograph showing shoots of licorice elongated by transplanting a multi-bud body into a plant regeneration medium.
【図5】再生したノカンゾウ植物体の馴化を示す写真で
ある。FIG. 5 is a photograph showing habituation of a regenerated licorice plant.
1 花茎 2 花茎切片 3 不定芽形成培地 4 不定芽 5 多芽体誘導培地 6 多芽体 7 植物体再生培地 8 再生植物体 9 苗 REFERENCE SIGNS LIST 1 flower stem 2 flower stem section 3 adventitious bud formation medium 4 adventitious bud 5 multi-bud induction medium 6 multi-bud 7 plant regeneration medium 8 regenerated plant 9 seedling
Claims (3)
組織を不定芽形成培地で培養して不定芽を形成させ小植
物体を獲得する工程と、前記獲得した小植物体を多芽体
誘導培地で培養して多芽体を誘導する工程と、前記誘導
した多芽体を分割して植物体再生培地で培養して植物体
を再生する工程とを含むヘメロカリス属植物の大量増殖
方法。1. A step of culturing a tissue of a genus Hemerocallis isolated aseptically in an adventitious bud formation medium to form adventitious buds to obtain plantlets, and transforming the obtained plantlets into a polyblast induction medium. A method for mass-producing Hemerocallis plants, comprising the steps of: culturing in S. cerevisiae to induce polyblasts; and dividing the induced polyblasts and culturing in a plant regeneration medium to regenerate plants.
ことを特徴とする請求項1に記載のヘメロカリス属植物
の大量増殖方法。2. The method according to claim 1, wherein the tissue of the Hemerocallis plant is a flower stem.
植物生長調節物質を含有しない培地であることを特徴と
する請求項1又は請求項2に記載のヘメロカリス属植物
の大量増殖方法。3. The method according to claim 1, wherein the multibud induction medium and the plant regeneration medium are mediums containing no plant growth regulator.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP8180019A JPH104811A (en) | 1996-06-19 | 1996-06-19 | Proliferation of large amount of plant of genus hemerocallis |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP8180019A JPH104811A (en) | 1996-06-19 | 1996-06-19 | Proliferation of large amount of plant of genus hemerocallis |
Publications (1)
Publication Number | Publication Date |
---|---|
JPH104811A true JPH104811A (en) | 1998-01-13 |
Family
ID=16076046
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP8180019A Pending JPH104811A (en) | 1996-06-19 | 1996-06-19 | Proliferation of large amount of plant of genus hemerocallis |
Country Status (1)
Country | Link |
---|---|
JP (1) | JPH104811A (en) |
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CN101897297A (en) * | 2010-07-16 | 2010-12-01 | 浙江清华长三角研究院生物技术与医药研究所 | Two-step seedling quick propagation method for hemerocallis tissue culture by using tender pedicel as explant |
CN102668986A (en) * | 2012-05-30 | 2012-09-19 | 唐山师范学院 | Direct rooting method for tissue culture cluster seedlings of hemerocallis fulva |
CN103430850A (en) * | 2013-09-09 | 2013-12-11 | 中邦园林股份有限公司 | Tissue culture method for polyploid hemerocallis middendorfii and culture medium |
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-
1996
- 1996-06-19 JP JP8180019A patent/JPH104811A/en active Pending
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CN102668986A (en) * | 2012-05-30 | 2012-09-19 | 唐山师范学院 | Direct rooting method for tissue culture cluster seedlings of hemerocallis fulva |
CN103430850A (en) * | 2013-09-09 | 2013-12-11 | 中邦园林股份有限公司 | Tissue culture method for polyploid hemerocallis middendorfii and culture medium |
CN103461135A (en) * | 2013-09-18 | 2013-12-25 | 宁波市农业科学研究院 | Method for propagating hemerocallis hybridus |
CN103609444A (en) * | 2013-11-17 | 2014-03-05 | 浙江大学 | Tissue culture method for hemerocallis sempervirens araki |
CN103704134A (en) * | 2013-12-12 | 2014-04-09 | 北京林业大学 | Method for inducing day lily red rum callus tissues by taking filaments as explant |
CN105706926A (en) * | 2016-02-04 | 2016-06-29 | 河南农业大学 | Day lily rooting culture method |
CN109717075A (en) * | 2017-10-27 | 2019-05-07 | 上海市农业科学院 | A method of hemerocailis middendorffi regeneration plant is obtained by evoking adventive bud |
CN113170732A (en) * | 2021-05-31 | 2021-07-27 | 上海应用技术大学 | Culture medium and method for obtaining day lily callus through inducing mature embryo |
CN113170733A (en) * | 2021-05-31 | 2021-07-27 | 上海应用技术大学 | Culture medium and method for vitrifying day lily callus and unglassing adventitious buds |
CN113491239A (en) * | 2021-08-27 | 2021-10-12 | 黑龙江卉研农业发展有限公司 | Polyploid hemerocallis middendorfii tissue culture and culture medium |
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