JPS63248321A - Mass production of medicinal carrot by tissue culture - Google Patents
Mass production of medicinal carrot by tissue cultureInfo
- Publication number
- JPS63248321A JPS63248321A JP62083641A JP8364187A JPS63248321A JP S63248321 A JPS63248321 A JP S63248321A JP 62083641 A JP62083641 A JP 62083641A JP 8364187 A JP8364187 A JP 8364187A JP S63248321 A JPS63248321 A JP S63248321A
- Authority
- JP
- Japan
- Prior art keywords
- medium
- ginseng
- acid
- plant growth
- tissue culture
- 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.)
- Granted
Links
- 238000004519 manufacturing process Methods 0.000 title claims description 14
- 244000000626 Daucus carota Species 0.000 title claims description 5
- 235000002767 Daucus carota Nutrition 0.000 title claims description 5
- 240000004371 Panax ginseng Species 0.000 claims description 86
- 235000008434 ginseng Nutrition 0.000 claims description 75
- 235000003140 Panax quinquefolius Nutrition 0.000 claims description 47
- 206010020649 Hyperkeratosis Diseases 0.000 claims description 46
- 235000005035 Panax pseudoginseng ssp. pseudoginseng Nutrition 0.000 claims description 44
- 239000005648 plant growth regulator Substances 0.000 claims description 37
- 230000000392 somatic effect Effects 0.000 claims description 37
- SEOVTRFCIGRIMH-UHFFFAOYSA-N indole-3-acetic acid Chemical compound C1=CC=C2C(CC(=O)O)=CNC2=C1 SEOVTRFCIGRIMH-UHFFFAOYSA-N 0.000 claims description 32
- 235000002789 Panax ginseng Nutrition 0.000 claims description 31
- 241000196324 Embryophyta Species 0.000 claims description 26
- 210000002257 embryonic structure Anatomy 0.000 claims description 23
- 238000000034 method Methods 0.000 claims description 21
- 239000007787 solid Substances 0.000 claims description 20
- NWBJYWHLCVSVIJ-UHFFFAOYSA-N N-benzyladenine Chemical compound N=1C=NC=2NC=NC=2C=1NCC1=CC=CC=C1 NWBJYWHLCVSVIJ-UHFFFAOYSA-N 0.000 claims description 19
- PRPINYUDVPFIRX-UHFFFAOYSA-N 1-naphthaleneacetic acid Chemical compound C1=CC=C2C(CC(=O)O)=CC=CC2=C1 PRPINYUDVPFIRX-UHFFFAOYSA-N 0.000 claims description 17
- 230000004069 differentiation Effects 0.000 claims description 17
- JTEDVYBZBROSJT-UHFFFAOYSA-N indole-3-butyric acid Chemical compound C1=CC=C2C(CCCC(=O)O)=CNC2=C1 JTEDVYBZBROSJT-UHFFFAOYSA-N 0.000 claims description 17
- 230000006698 induction Effects 0.000 claims description 17
- 239000003617 indole-3-acetic acid Substances 0.000 claims description 16
- 210000001161 mammalian embryo Anatomy 0.000 claims description 15
- IXORZMNAPKEEDV-OBDJNFEBSA-N gibberellin A3 Chemical compound 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 12
- 239000005980 Gibberellic acid Substances 0.000 claims description 11
- 238000012258 culturing Methods 0.000 claims description 11
- 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 claims description 11
- 229910017053 inorganic salt Inorganic materials 0.000 claims description 8
- 230000015572 biosynthetic process Effects 0.000 claims description 6
- 235000002791 Panax Nutrition 0.000 claims description 4
- 241000208343 Panax Species 0.000 claims description 4
- 239000000203 mixture Substances 0.000 claims description 4
- 230000001172 regenerating effect Effects 0.000 claims description 4
- 241000511582 Actinomyces meyeri Species 0.000 claims description 3
- 240000007190 Daucus pusillus Species 0.000 claims description 3
- 235000002196 Daucus pusillus Nutrition 0.000 claims description 3
- 241000180649 Panax notoginseng Species 0.000 claims description 2
- 235000003143 Panax notoginseng Nutrition 0.000 claims description 2
- 230000001939 inductive effect Effects 0.000 claims description 2
- 150000001875 compounds Chemical class 0.000 claims 3
- OVSKIKFHRZPJSS-DOMIDYPGSA-N 2-(2,4-dichlorophenoxy)acetic acid Chemical compound OC(=O)[14CH2]OC1=CC=C(Cl)C=C1Cl OVSKIKFHRZPJSS-DOMIDYPGSA-N 0.000 claims 2
- 241001530105 Anax Species 0.000 claims 1
- 241000784863 Stomatium Species 0.000 claims 1
- 239000010907 stover Substances 0.000 claims 1
- 239000002609 medium Substances 0.000 description 81
- 239000006870 ms-medium Substances 0.000 description 11
- FAIXYKHYOGVFKA-UHFFFAOYSA-N Kinetin Natural products N=1C=NC=2N=CNC=2C=1N(C)C1=CC=CO1 FAIXYKHYOGVFKA-UHFFFAOYSA-N 0.000 description 8
- QANMHLXAZMSUEX-UHFFFAOYSA-N kinetin Chemical compound N=1C=NC=2N=CNC=2C=1NCC1=CC=CO1 QANMHLXAZMSUEX-UHFFFAOYSA-N 0.000 description 8
- 229960001669 kinetin Drugs 0.000 description 8
- 239000007788 liquid Substances 0.000 description 4
- 240000005373 Panax quinquefolius Species 0.000 description 3
- 239000004480 active ingredient Substances 0.000 description 3
- 210000004748 cultured cell Anatomy 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 239000001963 growth medium Substances 0.000 description 3
- 229930000044 secondary metabolite Natural products 0.000 description 3
- 239000002689 soil Substances 0.000 description 3
- UZKQTCBAMSWPJD-UQCOIBPSSA-N trans-Zeatin Natural products OCC(/C)=C\CNC1=NC=NC2=C1N=CN2 UZKQTCBAMSWPJD-UQCOIBPSSA-N 0.000 description 3
- UZKQTCBAMSWPJD-FARCUNLSSA-N trans-zeatin Chemical compound OCC(/C)=C/CNC1=NC=NC2=C1N=CN2 UZKQTCBAMSWPJD-FARCUNLSSA-N 0.000 description 3
- 229940023877 zeatin Drugs 0.000 description 3
- 229920001817 Agar Polymers 0.000 description 2
- 240000002769 Morchella esculenta Species 0.000 description 2
- 235000002779 Morchella esculenta Nutrition 0.000 description 2
- 239000008272 agar Substances 0.000 description 2
- 210000004027 cell Anatomy 0.000 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 description 2
- 239000004062 cytokinin Substances 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- -1 etc. are suitable Substances 0.000 description 2
- 239000012467 final product Substances 0.000 description 2
- 230000008635 plant growth Effects 0.000 description 2
- 239000003375 plant hormone Substances 0.000 description 2
- 230000035755 proliferation Effects 0.000 description 2
- 230000001902 propagating effect Effects 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 238000009331 sowing Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- COVMVPHACFXMAX-OYNOKLRGSA-N (-)-morellic acid Chemical compound C1=CC(C)(C)OC2=C1C(O)=C1C(=O)C3=C[C@@H](C(=O)[C@]4(C\C=C(\C)C(O)=O)OC5(C)C)C[C@@H]5[C@]34OC1=C2CC=C(C)C COVMVPHACFXMAX-OYNOKLRGSA-N 0.000 description 1
- 239000005631 2,4-Dichlorophenoxyacetic acid Substances 0.000 description 1
- 229930192334 Auxin Natural products 0.000 description 1
- 244000025254 Cannabis sativa Species 0.000 description 1
- XNIZIBPYBUCVEU-UHFFFAOYSA-N Morellic acid Natural products CC1Oc2c(CC=C(C)C)c3OC45C6CC(C=C4C(=O)c3c(O)c2C=C1)C(=O)C5(CC=C(C)/C(=O)O)OC6(C)C XNIZIBPYBUCVEU-UHFFFAOYSA-N 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 239000002363 auxin Substances 0.000 description 1
- 210000003323 beak Anatomy 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000012790 confirmation Methods 0.000 description 1
- 239000007799 cork Substances 0.000 description 1
- 238000012364 cultivation method Methods 0.000 description 1
- 238000012136 culture method Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 229940107131 ginseng root Drugs 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 238000003306 harvesting Methods 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- COVMVPHACFXMAX-ZVKSWBPMSA-N isomorellic acid Natural products O=C(O)/C(=C\C[C@]12C(=O)[C@H]3C=C4C(=O)c5c(O)c6c(c(C/C=C(\C)/C)c5O[C@]14[C@@H](C(C)(C)O2)C3)OC(C)(C)C=C6)/C COVMVPHACFXMAX-ZVKSWBPMSA-N 0.000 description 1
- HRRDCWDFRIJIQZ-UHFFFAOYSA-N naphthalene-1,8-dicarboxylic acid Chemical compound C1=CC(C(O)=O)=C2C(C(=O)O)=CC=CC2=C1 HRRDCWDFRIJIQZ-UHFFFAOYSA-N 0.000 description 1
- 238000011017 operating method Methods 0.000 description 1
- 230000002062 proliferating effect Effects 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 229930182490 saponin Natural products 0.000 description 1
- 150000007949 saponins Chemical class 0.000 description 1
- 235000017709 saponins Nutrition 0.000 description 1
- 230000009105 vegetative growth Effects 0.000 description 1
Abstract
(57)【要約】本公報は電子出願前の出願データであるた
め要約のデータは記録されません。(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.
Description
【発明の詳細な説明】
〈産業上の利用分野〉
この発明は、組織培養により薬用人参（オタ不ニンジン
、チクセツニンジン、アメリカニンジン。[Detailed Description of the Invention] <Industrial Application Field> This invention is directed to the production of medicinal ginseng (Ginseng ginseng, ginseng ginseng, American ginseng) through tissue culture.
三七ニンジンなど）の植物体の一部（たとえば１生長点
１葉、茎、根９種子等）由来のカルスから植物体を再生
して薬用人参を大量生産する方法に関するものである。The present invention relates to a method for mass-producing medicinal ginseng by regenerating a plant from a callus derived from a part of a plant (for example, one leaf, stem, root, and nine seeds from one growing point) of ginseng (Ginseng ginseng, etc.).
〈発明の背景および従来の技術〉 薬用人参（オタネニンジン、チクセツニンジン。<Background of the invention and conventional technology> Medicinal ginseng (panax ginseng, chikusetsu ginseng).
アメリカニンジン、三七ニンジンなど、以下同じ）は、
古来より強壮強精を中心に万能高貴薬として珍重されて
きている。従来の薬用人参組織培養物の培養法として、
特開昭５９−１６９４８６号、特開昭５９−１６９４８
７号、特開昭５９−１６９４８８号等があるが、いずれ
も薬用人参の特定の細胞を組織培養して増殖させる点に
関するものである。American carrots, 37 carrots, etc.) are
Since ancient times, it has been prized as a versatile noble medicine, mainly for tonicity and tonicity. As a conventional cultivation method for medicinal ginseng tissue culture,
JP 59-169486, JP 59-16948
No. 7, JP-A No. 59-169488, etc., all of which relate to tissue culture and proliferation of specific cells of medicinal ginseng.
〈発明が解決しようとする問題点〉
薬用人参の植物体の一部を組織培養して得られる培養細
胞に含まれる構成成分は、栽培した植物体に含まれる構
成成分と相異することが多い。たとえ培養細胞に有効成
分が含まれていたとしても、その含有量は少ない場合が
多い。また、培養細胞に含まれる薬効成分等が仮に抽出
・分離されたとしても、我国の関連法規では薬物として
認められない。また、一般に薬効成分は二次代謝産物に
多いといわれている。しかし、組織培養法は、−次代謝
産物を得るのには優れた手段となり得るが、薬効成分等
の二次代謝産物を得るには不適当であるという問題があ
る。<Problems to be solved by the invention> The components contained in cultured cells obtained by tissue culturing a part of a medicinal ginseng plant are often different from the components contained in the cultivated plant. . Even if cultured cells contain active ingredients, the content is often small. Furthermore, even if the medicinal components contained in cultured cells were extracted and separated, they would not be recognized as drugs under the relevant laws and regulations of Japan. Additionally, it is generally said that many of the medicinal components are found in secondary metabolites. However, although the tissue culture method can be an excellent means for obtaining secondary metabolites, there is a problem in that it is inappropriate for obtaining secondary metabolites such as medicinal ingredients.
ところで、薬用人参は、その種子を蒔いてから収穫まで
５〜６年を要する。薬用人参の種子を蒔く時期も限定さ
れ四季を通じて短期間であり、しかも種子が発芽し生長
させる条件作りおよび栽培が困難であるという問題があ
る。By the way, medicinal ginseng takes five to six years from sowing seeds to harvesting. The period for sowing medicinal ginseng seeds is limited and is short throughout the four seasons, and there is a problem in that it is difficult to create and cultivate conditions for the seeds to germinate and grow.
さらに、栽培薬用人参は固体差が大きく、栽培条件の他
に固体差によってもたとえばサポニン類等の有効成分の
質的および量的変異が大きく、産地による品質の不均一
性が大きいという問題がある。したがって、薬用人参の
有効成分の構成物およびその含量が安定した品質の植物
体を得るのが至難である。Furthermore, cultivated medicinal ginseng has large individual differences, and there are large qualitative and quantitative variations in active ingredients such as saponins due to individual differences in addition to cultivation conditions, and there is a problem that quality is highly heterogeneous depending on the production area. . Therefore, it is extremely difficult to obtain a plant body of medicinal ginseng with a stable composition and content of active ingredients.
この発明は、上記問題点を解決すべく薬用人参の植物体
の一部由来のカルスから植物体を再生することにより、
均一の性質を有するなど品質の安定した優良品種の薬用
人参を四季を問わず、〜年中安定して大量に供給する方
法を確立提供し、もって薬用人参の優良品種の育成と増
殖とを図り資源確保を目的とする。In order to solve the above problems, this invention regenerates a plant from a callus derived from a part of a medicinal ginseng plant.
Establish and provide a method for stably supplying high-quality medicinal ginseng varieties with uniform properties and stable quality throughout the year, regardless of the four seasons, and thereby aim to cultivate and multiply high-quality medicinal ginseng varieties. The purpose is to secure resources.
く問題点を解決するための手段〉
この発明は、上記目的を達成するために、所定の基準培
地を用いて薬用人参の植物体の一部を組織培養して当該
カルスから植物体を再生する方法であって、
（ａ） 前記薬用人参の植物体の一部を植物生長調節
物質の添加した前記基準培地で培養してカルスを誘導す
るカルス誘導工程と、
山） 誘導された前記カルスを一定期間毎に植物化長調
Ｗ＠質の添加した前記基準培地で継代培養して不定胚を
形成させる不定胚形成工程と、最終生産物（再生植物体
）の増大を図るために、必要ならば植物生長調節物質の
無添加または低濃度添加の液体基準培地中で前記不定胚
を増殖させた後、
ｆｃｌ 増殖させた不定胚を前記基準培地または基準
培地の無機塩濃度を減少さ廿た改良培地に植物生長調節
物質をそれぞれ添加した培地で培養して前記不定胚を茎
葉分化さ廿る茎葉分化工程と、（ｄｌ 茎葉分化した
前記茎葉固体を植物生長＃Ｉ１１節物質の添加した基準
培地に移植して発根させる発根工程
とを含ませることとした。Means for Solving the Problems> In order to achieve the above object, the present invention involves tissue culturing a part of a medicinal ginseng plant using a predetermined reference medium and regenerating the plant from the callus. A method comprising: (a) a callus induction step of culturing a part of the medicinal ginseng plant in the standard medium supplemented with a plant growth regulator to induce callus; A somatic embryo formation step in which somatic embryos are formed by subculturing in the standard medium supplemented with vegetative growth W@ quality every period, and if necessary, in order to increase the final product (regenerated plant body). After propagating the somatic embryos in a liquid standard medium with no or low concentration of plant growth regulators added, the fcl-propagated somatic embryos are grown in the standard medium or an improved medium in which the inorganic salt concentration of the standard medium is reduced. and (dl) a shoot differentiation step in which the somatic embryos are differentiated into shoots by culturing them in a medium supplemented with a plant growth regulating substance, and (dl) the shoot solids differentiated into shoots are transplanted to a standard medium supplemented with a plant growth #I11 node substance. It was decided to include a rooting process in which the roots are grown.
この発明で使用できる前記基準培地としては、ムラシゲ
アンドスクーグ（Ｍｕｒａｓｈｉｇｅ ＆Ｓｋｏｏｇ氏
培地（以下ｒＭＳＭＳ培地いう）、リンスマイヤーアン
ドスクーグ（Ｌｉｎｓｍａｉｅｒ ＆Ｓｋｏｏｇ）氏培
地（以下「ＬＳ培地」という）、ホワイト（Ｗｈｉｔｅ
）氏培地（以下「Ｗ培地」という）。Examples of the reference medium that can be used in the present invention include Murashige &Skoog's medium (hereinafter referred to as rMSMS medium), Linsmaier &Skoog's medium (hereinafter referred to as ``LS medium''), and White (hereinafter referred to as ``LS medium''). White
) medium (hereinafter referred to as "W medium").
ガウスレット（Ｇａｕｔｈｅｒｅｔ ）氏培地（以下
「 □Ｇ培地」という）、チュレッケ（Ｔｕｌ
ｅｃｋａ ）氏培地（以下「Ｔ培地」という）、モー
レル（Ｍｏｒｅｌ ）氏培地（以下「Ｍ培地」という）
、ヒルデブラント（Ｈ３ｌｄｅｂｒａｎｄｔ ）氏培地
（以下「Ｈ培地」という）等々が利用できる。なかで
もＭＳ培地、ＬＳ培地およびＷ培地等が好適であり、特
に、ＭＳ培地、ＬＳ培地１等が最適である。Gautheret's medium (hereinafter referred to as "□G medium"), Tulecke's medium (hereinafter referred to as "□G medium"),
ecka)'s medium (hereinafter referred to as "T medium"), Mr. Morel's medium (hereinafter referred to as "M medium")
, Hildebrandt's medium (hereinafter referred to as "H medium"), etc. can be used. Among them, MS medium, LS medium, W medium, etc. are suitable, and MS medium, LS medium 1, etc. are particularly suitable.
また、この発明にかかる組織培養で利用できる植物生長
調節物質として、種々のオーキシン、サイトカイニン等
その他が利用できる。この植物生長調節物質の濃度は、
通常使用される濃度範囲の１０＝ｓｇ／ １〜１０２ｍ
Ｂ７ Ｎで利用できる。Furthermore, various auxins, cytokinins, and others can be used as plant growth regulators that can be used in the tissue culture according to the present invention. The concentration of this plant growth regulator is
Usually used concentration range 10=sg/1-102m
Available in B7N.
以下この発明にかかる組織培養による薬用人参の大量生
産方法を説明する。The method for mass-producing medicinal ginseng by tissue culture according to the present invention will be described below.
この発明にかかる組織培養による薬用人参の大量生産方
法の大要は、薬用人参の植物体の一部（たとえば生長点
１葉、茎、根１種子など）由来のカルスを誘導し２ こ
のカルスを縫代培養して不定胚に形成させ、さらに必要
ならばこの不定胚を増殖させた後、茎葉分化させ、この
茎葉分化した茎葉固体を発根させて最終的には植物体を
再生するものである。The outline of the method for mass production of medicinal ginseng by tissue culture according to the present invention is to induce callus derived from a part of the medicinal ginseng plant (for example, one growing point, one leaf, one stem, one root, one seed, etc.), The somatic embryo is formed through seam culture, and if necessary, the somatic embryo is multiplied, then differentiated into shoots and leaves, and the differentiated shoots and leaves are rooted to finally regenerate the plant body. be.
まず、薬用人参〔オタネニンジン（Ｐａｎａ−ｘ−７ｇ
二ｊｎ８刈ユＩＬＣ，八、門ＥＹＥＩ？ ） 、チクセ
゛ンニンジン（ム促、ｘ−ｊ町刈Ｉ皿ｍ Ｃ，Ａ、門
ＥＹＥＲ＞ 、アメリカニンジン（ＰａｕＬ １頂１
硯ｍｌ舅ユ叩−Ｌ、）、三七ニンジン（Ｐａｎａｘ
胚豆訂Ｍ堕ｌ Ｆ、Ｈ・Ｃ）！ＥＮ） 等〕の植物
体の一部（たとえば、生長点。First, medicinal ginseng [Panax ginseng (Pana-x-7g
2jn8 Kariyu ILC, 8, Gate EYEI? ), Chikusen carrots (Mu prompt, x-j Machikari I plate m C, A, gate EYER>, American carrots (PauL 1 top 1
Inkstone ml 舅ゆうた-L, ), 37 carrots (Panax
Germame revision M fell F, H・C)! A part of a plant (e.g., a growing point).
葉、茎、根１種子など）由来のカルスを誘導する。Induce callus derived from leaves, stems, roots, seeds, etc.).
すなわち、このカルスの誘導には、前記基準培地に添加
する植物生長調節物質としては、２，４−ジクロロフェ
ノキシ酸！（以下ｒ２，４− Ｄ Ｊという）。That is, for the induction of callus, the plant growth regulator to be added to the reference medium is 2,4-dichlorophenoxy acid! (hereinafter referred to as r2,4-D J).
ナフタリン酢酸（以下ｒＮＡＡＪという）、インドール
酪酸（以下ｒｒＢＡＪという）、インドール酢酸（以下
ｒＩ ＡＡＪという）等が利用できる。Naphthalene acetic acid (hereinafter referred to as rNAAJ), indolebutyric acid (hereinafter referred to as rrBAJ), indoleacetic acid (hereinafter referred to as rIAAJ), etc. can be used.
なかでも、２．４−Ｄ、ＮＡＡが特に好適である。また
サイトカイニンとして、力・イネチン、ベンジルアデニ
ン（以下ｒＢＡＪという）、ゼアチン（以下ｒＺＪとい
う）などが利用できる。カイネチンを２．４−Ｄまたは
ＩＡＡと組み合わせて使用するか。Among them, 2.4-D and NAA are particularly preferred. Further, as cytokinin, inetin, benzyladenine (hereinafter referred to as rBAJ), zeatin (hereinafter referred to as rZJ), etc. can be used. Should kinetin be used in combination with 2.4-D or IAA?
またはＢＡｔ−ＮＡＡまたはｒＢＡと組み合わせて使用
するか、２を２．４−ＤまたはＩＡＡまたはＩＢＡまた
はＮＡＡと組み合わせて使用すると、前記薬用人参（オ
タネニンジンはか）のカルス誘導が促進される。Alternatively, when used in combination with BAt-NAA or rBA, or when 2 is used in combination with 2.4-D or IAA or IBA or NAA, callus induction of the medicinal ginseng (Panax ginseng) is promoted.
このカルス誘導工程で使用される前記植物生長調節物質
の濃度は、通常使用される濃度範囲の１０＝ｍｇ／ｊ！
〜１０” ｍｇ／ Ｉｔが適当である。この発明にがか
るカルス誘導工程では、２．４−Ｄおよびカイネチンを
それぞれの濃度範囲を０．１ｍｇ／ｆｆ〜１０ｍｇ／Ｉ
！添加したとき最良の効果を得る。もっとも、カイネチ
ン、ベンジルアデニン（ＢＡ）、ゼアチン（Ｚ）は必須
の植物ホルモンではないが、これらを付加的に添加する
とカルスの誘導が促進される。必須の植物ホルモンとし
ては、２．４−Ｄ、 ＮＡＡ、ＩＡＡ、ＩＢＡのうちの
少なくともいずれか一種が存在すれば薬用人参（オタネ
ニンジンなど）の植物体の一部由来のカルスは誘導され
る。The concentration of the plant growth regulator used in this callus induction step is within the commonly used concentration range of 10=mg/j!
~10'' mg/It is appropriate. In the callus induction process according to the present invention, the concentration range of 2.4-D and kinetin is 0.1 mg/ff~10 mg/It.
! Get the best effect when added. However, although kinetin, benzyladenine (BA), and zeatin (Z) are not essential plant hormones, additional addition of these promotes callus induction. As essential plant hormones, callus derived from a part of a medicinal ginseng (panax ginseng, etc.) plant body can be induced if at least one of 2.4-D, NAA, IAA, and IBA is present.
カルス誘導の基本的な操作手順は、薬用人参（オタネニ
ンジン、アメリカニンジン、チクセツニンジン１三七ニ
ンジン等）の植物体の一部（たと記植物生長調節物質（
２，４−Ｄ、ＮＡＡ、ＩＢＡまたはＩＡＡ（以上の４種
類は必須の植物生長調節物質）と、必要に応じてカイネ
チン、ベンジルアデニンまたはゼアチンなど〕を添加し
た基準寒天培地に置床し、１５〜３０℃、明所または暗
所で数週間（通常３週間〜６週間程度）培養してカルス
を誘導することができる。The basic operating procedure for callus induction is to extract a part of the plant body of medicinal ginseng (panax ginseng, American ginseng, ginseng ginseng, ginseng 137 ginseng, etc.).
2,4-D, NAA, IBA, or IAA (the above four types are essential plant growth regulators), and kinetin, benzyladenine, zeatin, etc. as necessary] were placed on a standard agar medium, and the plate was placed on a standard agar medium supplemented with 15~ Callus can be induced by culturing at 30° C. in the light or in the dark for several weeks (usually about 3 to 6 weeks).
つぎに、前記誘導カルスから不定胚を形成させる。すな
わち、薬用人参（オタネニンジンなど））の植物体の一
部から誘導したカルスを、前記基準培地に必須の植物生
長調節物質として２．４−Ｄ。Next, somatic embryos are formed from the induced callus. That is, callus derived from a part of a medicinal ginseng (panax ginseng, etc.) plant body is used as an essential plant growth regulator in the reference medium using 2.4-D.
ＩＢＡ、ＩＡＡ、ＮＡＡの４種類のうちの少なくとも一
つを添加（濃度範囲＝１０＝ｍｇ／ｌ〜１０ｍｇ ／ｌ
程度）した固体培地に、約１５〜２５℃、明所下で一定
期間ごと（約４週間毎〜約８週間毎）に継代培養し、一
定期間（約１年間、４８週間〜５４週間）継代培養を繰
り返して前記誘導カルスを不定胚に形成させる。その後
、最終生産物である再生植物体の増大を図るために、必
要に応じてこの不定胚は、植物生長調節物質無添加また
は低濃度添加（約１Ｏ−２ａ＋ｇ／ｊ！〜１０−’ ｔ
ａｇ／ Ａ’程度）の液体基準培地に移植して、１５〜
２５℃、明所下、約２０日間振盪（回転または往復）培
養してこの不定胚を増殖させる。この場合、誘導カルス
を前記液体培地で培養した方が固体培地で培養するより
も、増殖を促進することができる。Addition of at least one of the four types of IBA, IAA, and NAA (concentration range = 10 = mg/l to 10 mg/l
The cells were subcultured on a solid medium at approximately 15 to 25°C under the light at regular intervals (approximately every 4 weeks to approximately 8 weeks) for a specified period of time (approximately 1 year, 48 weeks to 54 weeks). Subculturing is repeated to form the induced callus into somatic embryos. Thereafter, in order to increase the number of regenerated plants that are the final product, the somatic embryos may be treated without the addition of plant growth regulators or with the addition of a low concentration of plant growth regulators (approximately 1O-2a+g/j!~10-'t
ag/A') into a liquid standard medium for 15~
The somatic embryos are grown by culturing with shaking (rotation or reciprocation) at 25° C. in the light for about 20 days. In this case, culturing the induced callus in the liquid medium can promote proliferation more than culturing it in the solid medium.
つぎに、この不定胚を茎葉分化さセる。すなわち、基準
培地または基準培地の無機塩の含量を減少（１／１未満
１／２０以上）させた改良培地に植物生長調節物質をそ
れぞれ添加した固体培地（基準培地または改良培地）に
この増殖させた不定胚を移植し、一定の条件下（約１５
〜２５℃、明所下。Next, this somatic embryo is differentiated into stems and leaves. That is, this growth is performed on a solid medium (standard medium or improved medium) in which a plant growth regulator is added to a standard medium or an improved medium in which the content of inorganic salts is reduced (less than 1/1 to 1/20 or more). The somatic embryos were transplanted under certain conditions (approximately 15
~25°C, under light.
３週間〜６週間程度）で培養して前記不定胚を茎葉分化
させる。なお、この茎葉分化工程で使用できる植物生長
調節物質としては、ジベレリン酸（以下「ＧＡ」という
）、とベンジルアデニン（ＢＡ）との組合せ、ＮＡＡと
ＢＡとの組合せ、ＩＢＡとＢＡとの組合せ、またはＩＡ
ＡとＢＡとの組合せの以上４［Ｊｌの組合せ群から選択
されるいずれかの組合せが利用できる。特に、前記ＧＡ
とＢＡとの組合せは、薬用人参（オタネニンジンなど）
の増殖不定胚を茎葉分化させるのに好適である。The somatic embryos are cultured for about 3 to 6 weeks) to differentiate into stems and leaves. In addition, plant growth regulators that can be used in this shoot and leaf differentiation step include a combination of gibberellic acid (hereinafter referred to as "GA") and benzyladenine (BA), a combination of NAA and BA, a combination of IBA and BA, or IA
Any combination selected from the above 4 [Jl combinations of combinations of A and BA can be used. In particular, the GA
The combination of and BA is medicinal ginseng (panax ginseng, etc.)
It is suitable for causing stem and leaf differentiation of proliferating somatic embryos.
また、この茎葉分化工程で使用する植物生長調節物質の
添加量は、それぞれの添加１ｌ１０−２ｎｉ／ １〜１
０ｍｇ／ｌ程度が好適できる。In addition, the amount of the plant growth regulator used in this foliage differentiation step is 1 liter 10-2 ni/1 to 1 for each addition.
Approximately 0 mg/l is suitable.
つぎに、前記茎葉分化した固体を発根させる。Next, the stem-and-leaf differentiated solid is rooted.
すなわち、前記茎葉分化した固体を、植物生長調節物質
を添加した基準固体培地に移植して約１５〜２０℃、明
所下で一定期間（約４週間〜１０週間程度）培養する。That is, the stem-and-leaf differentiated solid is transplanted to a reference solid medium supplemented with a plant growth regulator and cultured at about 15 to 20° C. in the light for a certain period of time (about 4 to 10 weeks).
この茎葉分化した固体の発根工程中に使用される植物生
長調節物質としては、インドール酪酸（ＩＢＡ）、ナフ
タリン酢酸（ＮＡＡ）。Plant growth regulators used during the rooting process of this foliage-differentiated solid include indolebutyric acid (IBA) and naphthaleneacetic acid (NAA).
インドール酢酸（ＩＡＡ）またはジベレリン酸（ＧＡ）
から選択されるいずれかの植物生長調節物質が利用でき
る。特に、前記ＩＢＡおよびＮＡＡは好適である。また
、前記植物生長調節物質の基準培地への添加量は、通常
に用いられる範囲すなわち１０’ ｍｇ／ ｌ〜１０嘴
ｇ／ｌが好適である。Indole acetic acid (IAA) or gibberellic acid (GA)
Any plant growth regulator selected from: In particular, the above-mentioned IBA and NAA are suitable. Further, the amount of the plant growth regulator added to the standard medium is preferably within the commonly used range, that is, 10' mg/l to 10 beak g/l.
〈実施例〉
（１）この発明にかかる薬用人参の大量培養方法におい
て、まずムラシゲアンドスクーグ（Ｍｕｒａｓｈｉｇｅ
＆ Ｓｋｏｏｇ ／ＭＳ）氏培地を基準培地として
使用した場合の実施例について述べる。<Example> (1) In the method for mass culturing medicinal ginseng according to the present invention, first, Murashige and Skoog
&Skoog/MS)'s medium was used as the standard medium.
（ａ） カルス誘導の実施例
薬用人参（オタネニンジン、 Ｐａｎａｘ幻ｌ狙邸−
（：、Ａ、ＭＥＹＥＲ）の根からカルスを誘導する場合
の実施例はつぎのとおりである。(a) Example of callus induction Medicinal ginseng (Panax ginseng, Panax ginseng)
An example of inducing callus from the roots of (:, A, MEYER) is as follows.
薬用人参（オタネニンジン、 Ｐａｎａｘ員μ咀邦−
Ｃ，Ａ、ＭＥＹＥＲ）の根ヲ無菌的にコルクポーラ−で
打抜き、これを約２〜３Ｉ１１−の厚さにメスで切断し
て円盤状の外植片を調製する。Medicinal ginseng (Panax ginseng)
The root of C.A. MEYER) is aseptically punched out with a cork polar and cut into a thickness of about 2 to 3 I11 with a scalpel to prepare a disc-shaped explant.
一方、植物生長調節物質として２．４−Ｄおよびカイネ
チン（Ｋｉｎｅｔｉｎ ）の濃度が、 ０．０ 。On the other hand, the concentration of 2.4-D and kinetin as plant growth regulators was 0.0.
０．１ 、１．０ 、１０ 剛ｇ／ｌ各４種類のもの
を組み合わせて添加した合計１６種類のＭＳ固定培地を
調製する。そして、この固定培地上に前記薬用人参の円
盤状の外植片を置床しく第１図）、２５℃、明所下で６
週間培養を行いオタネニンジンの根由来のカルスを誘導
する。（第２図）このカルス誘導での成績は、第１表に
示すとおりである。第１表から明らかなように２．４−
Ｄおよびカイネチンがそれぞれ１．０ ｍｇ／ｌの組
合せの添加区のカルス形成が最適である。A total of 16 types of MS fixed media are prepared by adding combinations of four types each at 0.1, 1.0, and 10 g/l. Then, place the medicinal ginseng disc-shaped explants on this fixed medium (Fig. 1), and store them at 25°C in the light for 6 hours.
Culture is performed for weeks to induce callus derived from Panax ginseng roots. (Figure 2) The results of this callus induction are shown in Table 1. As is clear from Table 1, 2.4-
Callus formation in the area where D and kinetin were added in combination at 1.0 mg/l each was optimal.
第１表ニオタネニンジンの根のカルス誘導成績（基準培
地：ＭＳ培地）
植物生長調節物質としてＩＡＡ、ＮＡＡまたはＩＢＡと
カイネチンとのそれぞれの組合せを用いた場合の前記カ
ルス誘導成績は、はぼおなし傾向を示す。この場合のカ
ルス誘導成績は、第２表に示すとおりである。Table 1 Results of callus induction in roots of Panax ginseng (standard medium: MS medium) The results of callus induction when using the respective combinations of IAA, NAA, or IBA and kinetin as plant growth regulators were not significant. Show trends. The callus induction results in this case are as shown in Table 2.
第２表ニオタネニンジンの根のカルス誘導成績（基準培
地二ＭＳ培地）
（各濃度単位：ｍｇ／１）
（ｂｌ 前記誘導カルスからの不定胚形成の実施例。Table 2 Results of callus induction of roots of Panax ginseng (standard medium and MS medium) (Each concentration unit: mg/1) (bl Example of somatic embryo formation from the above-mentioned induced callus.
前記（ａ）で誘導したカルスを植物生長調節物質として
２．４−Ｄを１．０ｍｇ／ ｌ 、カイネチンを０．０
ｍｇ／ｐ、添加したＭＳ培地（固体培地）に移植し、２
０℃、明所下で６週間毎に約１年間（約４８週間〜５４
週間）継代培養を繰り返す。誘導当初は淡白色の軟らか
いカルスは、この約１年間の継代培養により不定胚の黄
色の部分が次第に認められるようになる。（第３図）
このようにして得られる不定胚は、植物生長調節物質無
添加または低濃度添加（約１０−２１１ｇ／ｌ〜１０’
ｍｇ／ １ ）のＭＳ培地（液体培地）に移植し、２
５℃、明所下で回転振盪培養（１１０−１１５ｒｐｔａ
）または往復振盪培養（１１０〜１３０往復／分）の
条件で約２０日間培養して、不定胚を増殖させる。The callus induced in (a) above was treated with 2.4-D at 1.0 mg/l and kinetin at 0.0 as plant growth regulators.
mg/p, transferred to MS medium (solid medium) supplemented with 2
0℃, under light, every 6 weeks for about 1 year (about 48 weeks to 54 weeks)
Weeks) Repeat subculture. At the beginning of induction, the callus is pale white and soft, but after about one year of subculturing, the yellow part of the somatic embryo gradually becomes visible. (Figure 3) The somatic embryos obtained in this way are produced without the addition of plant growth regulators or with the addition of low concentrations of plant growth regulators (approximately 10-211 g/l to 10'
mg/1) into MS medium (liquid medium), and
Rotary shaking culture (110-115 rpta) in the light at 5°C.
) or reciprocating shaking culture (110 to 130 reciprocations/min) for about 20 days to proliferate somatic embryos.
第２表は、植物生長調節物質に２．４−Ｄ、 ＨＡＡ
、ＩＢＡ、ＮＡＡを用いた場合の不定胚の形成成績を対
比して示す。Table 2 shows plant growth regulators such as 2.4-D and HAA.
The results of somatic embryo formation when using , IBA, and NAA are shown in comparison.
〔本頁以下余白〕 （各濃度単位：ｍｇ／ｊｌり （Ｃ） 前記不定胚の茎葉分化の実施例。[Margins below this page] (Each concentration unit: mg/jl (C) Example of stem and leaf differentiation of the somatic embryo.
前記（ｂ）で得た不定胚を、ＭＳ培地またはＭＳ培地の
含有無機塩の濃度を１／１未満１／２０以上の濃度に減
少させたＭＳ改良培地に植物生長調節物質としてジベレ
リン酸（ＧＡ）をＯ，Ｏｎ＋ｇ／ １． １．０ｍｇ／
１．５．０ ｍｇ／ Ｌ １０．０ｍｇ／ ７！およ
びベンジルアデニン（ＢＡ）を０゜Ｑｍｇ ／ ｆ！
。The somatic embryos obtained in (b) above were placed in an MS medium or an MS improved medium in which the concentration of inorganic salts contained in the MS medium was reduced to less than 1/1 and 1/20 or more, and gibberellic acid (GA) was added as a plant growth regulator. ) to O, On+g/1. 1.0mg/
1.5.0 mg/L 10.0 mg/7! and benzyladenine (BA) at 0°Qmg/f!
.
０．１ｍｇ／ １２ 、１．０ ｍｇ／ Ｉ！、 ｌｏ
、ｏｎ＋ｇ／ Ｉ！をそれぞれ組み合わせて添加した合
計１６種類の各固体培地に移植して、２０℃、明所下で
３０日間培養し、前記（ｂ）で得た不定胚を茎葉分化さ
せる。（第４図）
第４表は、植物生長調節物質の各試験濃度に対する不定
胚の茎葉分化成績を示す。0.1 mg/12, 1.0 mg/I! , lo
,on+g/I! The somatic embryos obtained in (b) above are transplanted onto a total of 16 types of solid media containing a combination of the following and cultured in the light at 20° C. for 30 days to differentiate into stems and leaves. (Figure 4) Table 4 shows the results of stem and leaf differentiation of somatic embryos for each tested concentration of the plant growth regulator.
（各濃度単位：ｒａｇ／ｊ）
第４表の結果より、ＧＡを１．０ｍｇ／ １〜５．０ｍ
ｇ／７！、ＢＡを０．１ ｍｇ／ １〜１．０ ｍｇ／
Ａそれぞれ添加したものが好適であることがわかる。(Each concentration unit: rag/j) From the results in Table 4, GA at 1.0 mg/1 to 5.0 m
g/7! , BA at 0.1 mg/1-1.0 mg/
It can be seen that it is preferable to add each of A.
なお、植物生長調節物質として、ＮＡＡ、ＩＡＡまたは
ＩＢＡの３種類とＢＡとのそれぞれの組合せを用いた場
合、前記不定胚の茎葉分化の成績はいずれもほぼ似た１
項向を示す。第５表は、この場合の不定胚の茎葉分化成
績を示している。In addition, when each combination of three types of NAA, IAA, or IBA and BA was used as a plant growth regulator, the results of stem and leaf differentiation of the somatic embryos were almost similar.
Indicates direction. Table 5 shows the results of stem and leaf differentiation of somatic embryos in this case.
（各濃度単位：ｍｇ／ｌ）
第６表は、培地の無機塩の含有濃度を基準培地（ＭＳ培
地）の無機塩濃度組成の１／１〜１／３０にしたときの
それぞれの組成に対する不定胚の茎葉分化成績を示す。(Each concentration unit: mg/l) Table 6 shows the indefinite values for each composition when the concentration of inorganic salt in the medium is set to 1/1 to 1/30 of the inorganic salt concentration composition of the standard medium (MS medium). The results of stem and leaf differentiation of embryos are shown.
ただし、植物生長調節物質の添加量は、ＧＡが１．０ｍ
ｇ／ ｌ 、 Ｂ Ａが０．１ｍｇ／ｊ！のときの成績
結果を示す。However, the amount of plant growth regulator added is 1.0 m
g/l, BA is 0.1mg/j! Shows the results of the results.
第６表：不定胚の茎葉分化成績
なお、この茎葉分化した固体の一部には、茎葉分化と同
時に発根するものが認められるが、茎葉分化にとどまる
ものが多い。Table 6: Results of stem and leaf differentiation of somatic embryos Although some of these individuals that undergo shoot and leaf differentiation are observed to develop roots at the same time as shoot and leaf differentiation, many remain in shoot and leaf differentiation.
（ｄ） 茎葉分化固体の発根の実施例。(d) Example of rooting of a stem-and-leaf differentiated solid.
前記（ｅ）で茎葉分化した固体を、ＭＳ培地に植物生長
調節物質としてインドール酪酸（ＩＢＡ）、ナフタリン
酸（ＮＡＡ）、 インドール酢酸（ＩＡＡ）、 ジベレ
リン酸（ＯＡ）をそれぞれ添加した固体培地に移植し、
２０ｔ、明所下で８迎間培養して、前記茎葉分化した固
体の発根させる。（第５図）その後、発根した前記植物
体を必要に応じて土壌に移植する。（第６図）第７表は
、前記ＩＢＡ添加培地、ＮＡＡ添加培地、ｒＡＡ添加培
地に対する茎葉分化した固体の発根成績を示す。The solids differentiated into leaves in step (e) above are transplanted to a solid medium in which indolebutyric acid (IBA), naphthalic acid (NAA), indoleacetic acid (IAA), and gibberellic acid (OA) are added as plant growth regulators to MS medium. death,
The plants were cultured at 20 tons in the light for 8 hours to root the stem-and-leaf-differentiated solids. (FIG. 5) Thereafter, the rooted plants are transplanted to soil as necessary. (FIG. 6) Table 7 shows the rooting results of the shoot-and-leaf differentiated solids in the IBA-added medium, NAA-added medium, and rAA-added medium.
（２） この発明に利用できる基準培地について、前
記（１）の（ａ）カルスの誘導から（ｄ）茎葉分化した
固体の発根にいたる薬用人参の大量生産方法の各工程の
試験結果は、第８表に示すとおりである。(2) Regarding the reference medium that can be used in this invention, the test results of each step of the method for mass production of medicinal ginseng from (a) induction of callus to (d) rooting of foliage-differentiated solids in (1) above are as follows: As shown in Table 8.
なお、試験にかかる基準培地は、前記ＭＳ培地のほか、
リンスマイヤーアンドスクーグ培地（Ｌｉｎｓｍａｉｅ
ｒ ＆ Ｓｋｏｏｇ氏培地−ＬＳ培地）。In addition to the above-mentioned MS medium, the reference medium used for the test is
Linsmaie and Skoog medium
r &Skoog's medium-LS medium).
ホワイ！・培地（Ｗｈｉｔｅ氏培地・Ｗ培地）、ガウス
レット培地（Ｇａｕｔｈｅｒｅｔ氏培地−Ｇ培地）。why! - Medium (White's medium/W medium), Gautheret medium (Gautheret's medium-G medium).
チュレッケ培地（Ｔｕｌｅｃｋｅ氏培地・Ｔ培地）。Tuecke's medium (Tulecke's medium/T medium).
モーレル培地（Ｍｏｒｅ１氏培地・Ｍ培地）、ヒルデブ
ラント培地（Ｈｉｌｄｅｂｒａｎｄｔ氏培地・Ｈ培地）
について試験する。Morel medium (More1 medium/M medium), Hildebrandt medium (Hildebrandt medium/H medium)
Test for.
また、茎葉分化に用いた基準培地の無機塩の濃度は、各
基準培地の１／２の濃度のものを利用する。Further, the concentration of inorganic salt in the standard medium used for stem and leaf differentiation is 1/2 that of each standard medium.
第８表：基準培地の試験結果 〔本頁以下余白〕 〈発明の効果〉 薬用人参（オタネニンジン、チクセツニンジン。Table 8: Test results of reference medium [Margins below this page] <Effect of the invention> Medicinal ginseng (panax ginseng, chikusetsu ginseng).
アメリカニンジン、三七ニンジンなど）の植物体の一部
由来カルスから植物体を再生したので、四季に関係なく
品質の安定した優良品種の薬用人参を大量に得ることが
でき、しかも１つの植物体から大量の植物体が再生でき
るから、再生された植物体に固体差が殆どなく品質の安
定した薬用人参を大量に得ることができ、薬用人参の優
良品種の育成と増殖による資源確保が図れる等々、発明
目的を達成する顕著な効果を奏する。By regenerating the plant from callus derived from a part of the plant (American ginseng, Sanchi ginseng, etc.), it is possible to obtain a large amount of medicinal ginseng of excellent variety with stable quality regardless of the four seasons, and moreover, from one plant. Since a large amount of plants can be regenerated from the plant, a large amount of medicinal ginseng with stable quality can be obtained with almost no individual differences between the regenerated plants, and resources can be secured by cultivating and propagating superior medicinal ginseng varieties. , has a remarkable effect of achieving the purpose of the invention.
第１図はオタネニンジンの根切片を基準（固体）培地に
置床したときの図、第２図はオタネニンジンの根から誘
導したカルス、第３図はオタネニンジンの根カルスから
形成した不定胚、第４図はオタネニンジンの不定胚から
分化した茎葉、第５図はオタネニンジンの分化した茎葉
を発根させた幼植物、第６図はオタネニンジンの根由来
のカルスから得た幼植物を土壌へ移植した植物体。
第１図ニオタネニンジンの根切片を固体培地上に置床し
た図。
Ｘ＾）へ−へ−〜
第２図ニオタネニンジンの根から誘導したカルス。
第３図ニオタネニンジンの根カルスから形成した不定胚
。
Ｊシー＋＋（Ｖへ＾１情−＾い１ｑ１シｎ〜マ心−）４
−−”” −” ＃−′％ Ｉｌ＋ ／ ／ Ｋ ７
”’ ＞ ’＊：ｌ” ｌ ’、、” ’、ｌ、ｒ
ｌ： ’：’、、’、ｌ’、：：’、’：、Ｌ・・−一
一ゝ−−′第４図ニオタネニンジンの不定胚から分化し
た茎叱第５図ニオタネニンジンの分化した茎葉を発根さ
せた珈１九
ψｖ１）１１
第６図ニオタネニンジンの根由来カルスから得た幼植物
を土壌へ移植した再生植物体。
Ｊハノ
、−手続ネ市正書（方力
１１訃０６２年０７月ｂｇ口
１、事件の表示 昭和６２年特許願第８３６４１
号２、発明の名称 組織培養による薬用人参の大
■生産方法３、 ７Ｊｉ正をする者
゛１丁件との（又Ｉ（系 特許出■臥住 所
〒９３６ 富山県滑用市清水町１２番１５８−・１
１代理人
〒５３０ 人反市北区西天満３丁目１１番９号−５、
禄１正命令のロイ’ｔ ＩＩ鼾ｎ ６２年０６月０
３８（発送口： 昭和６２年０６月３０日）６、 ？
ｌｌｉ＋Ｔ：の対架 ■ イリ１１潅を証明する
書面■ 明８旺許中、「図面の簡単な説明の欄、：■
図面
７、補正の内容
■代理権を証明する書面は、昭和６γ問４月２４日付手
続補正書く自発〉の提出とシ窪こ委任状（全員の分：合
計４名分−４通〕を提出致しました。ご確認の４、図面
の簡単な説明
第１図はオタネニンジンの根切片を基′９（固体）培地
に置床したときの生物形態を示す写真、第２図はオタネ
ニンジンの根から誘導したカルスの生物形態を示す写真
、第３図はオタネニンジンの根カルスから形成した不定
胚の生物形態を示す写真、第４図はオタネニンジンの不
定胚から分化した茎葉の生物形態を示す写真、第５図は
オタネニンジンの分化した草葉を発根させた幼ＩＬケ物
の生物形態を示す写真、第６図はオタネニンジンの恨由
来のカルスから得た幼植物を土壌へ移ｔｒｒ したｔＭ
’ｔ”Ｊ体の生物形態を示す写真である。
以上
第１図
第２図
第３図
第４図
Ｉ
；
第５図
雰
第６図Figure 1 shows a root section of Panax ginseng placed on a standard (solid) medium, Figure 2 shows a callus derived from roots of Panax ginseng, Figure 3 shows a somatic embryo formed from a root callus of Panax ginseng, and Figure 4 Fig. 5 shows a seedling obtained by rooting the differentiated stem and leaves of Panax ginseng, and Fig. 6 shows a plant obtained from a callus derived from the roots of Panax ginseng and transplanted into soil. FIG. 1 is a diagram showing a root section of Panax ginseng placed on a solid medium. X^) He-He-~ Figure 2 Callus derived from the roots of Panax ginseng. Figure 3: Somatic embryo formed from root callus of Panax ginseng. J C++ (V to ^1 jo-^i 1q1 shin~mashin-) 4
−−”” −” ＃−′% Il+ / / K 7
"'>'*:l" l ',,"', l, r
l: ':',,',l',::',':,L...-11ゝ--'Fig. Fig. 6 A regenerated plant obtained by transplanting a young plant obtained from root-derived callus of Panax ginseng into soil. J Hano, - Procedural City Official Book (Fangli 11 death July 062 bg mouth 1, Incident indication 1988 Patent Application No. 83641
No. 2, Title of the invention: Production method of medicinal ginseng by tissue culture 3, 7Ji corrective person (also I) (related patent address)
12-158-1 Shimizucho, Nameyo City, Toyama Prefecture 936
1 Agent 3-11-9-5 Nishitenma, Kita-ku, Hitozori City, 530
Roy't II of the Roku 1 Positive Order June 0, 62
38 (Shipping port: June 30, 1986) 6.?
lli+T: counter-shelf ■ Document certifying Ili 11 ■ Mei 8 Wangxu, ``Column for brief explanation of drawings,: ■
Drawing 7, Contents of amendment ■ Documents proving power of attorney: Submission of ``Spontaneous amendment to the procedures dated April 24, 1939'' and Shikuboko power of attorney (for all members: 4 copies for 4 people in total) Confirmation 4. Brief explanation of the drawings Figure 1 is a photograph showing the biological morphology of a Panax ginseng root section placed on a base '9 (solid) medium, and Figure 2 is a photograph derived from the roots of Panax ginseng. Figure 3 is a photograph showing the biological form of a somatic embryo formed from the root callus of Panax ginseng, Figure 4 is a photograph showing the biological form of a stem and leaf differentiated from a somatic embryo of Panax ginseng, and Figure 5 is a photograph showing the biological form of a callus differentiated from a somatic embryo of Panax ginseng. The figure is a photograph showing the biological form of a young IL plant that has taken root from differentiated grass leaves of Panax ginseng.
These are photographs showing the biological morphology of the 't'J body.
Claims (8)
を組織培養して当該カルスから植物体を再生する方法で
あって、 前記薬用人参の植物体の一部を植物生長調節物質の添加
した前記基準培地に培養してカルスを誘導するカルス誘
導工程と、 誘導された前記カルスを一定期間毎に植物生長調節物質
の添加した前記基準培地で継代培養して不定胚を形成さ
せる不定胚形成工程と、増殖させた不定胚を前記基準培
地または基準培地の無機塩濃度を減少させた改良培地に
植物生長調節物質をそれぞれ添加した培地で培養して前
記不定胚を茎葉分化させる茎葉分化工程と、茎葉分化し
た前記茎葉固体を植物生長調節物質の添加した基準培地
に移植して発根させる発根工程と を含むことを特徴とする組織培養による薬用人参の大量
生産方法。(1) A method of tissue culturing a part of a medicinal ginseng plant using a predetermined reference medium and regenerating the plant from the callus, wherein the part of the medicinal ginseng plant is treated with a plant growth regulator. a callus induction step of inducing callus by culturing on the standard medium supplemented with a plant growth regulator, and subculturing the induced callus at regular intervals on the standard medium supplemented with a plant growth regulator to form somatic embryos. a somatic embryo formation step, and a stomatium in which the proliferated somatic embryos are cultured in the standard medium or an improved medium with a reduced inorganic salt concentration of the standard medium to which a plant growth regulator has been added, and the somatic embryos are differentiated into stovers. 1. A method for mass production of medicinal ginseng by tissue culture, comprising a differentiation step and a rooting step of transplanting the differentiated foliage solid to a standard medium containing a plant growth regulator and causing the roots to form.
￥￥ｇｉｎｓｅｎｇ￥Ｃ．Ａ．ＭＥＹＥＲ）またはチク
セツニンジン（￥Ｐａｎａｘ￥ ￥ｊａｐｏｎｉｃｕｍ
￥Ｃ．Ａ．ＭＥＹＥＲ）またはアメリカニンジン（￥Ｐ
ａｎａｘ￥ ￥ｑｕｉｎｑｕｅｆｏｌｉｕｍ￥Ｌ．）または三七ニン
ジン（￥Ｐａｎａｘ￥ ￥ｎｏｔｏｇｉｎｓｅｎｇ￥Ｆ
．Ｈ．ＣＨＥＮ）である特許請求の範囲第１項記載の組
織培養による薬用人参の大量生産方法。(2) The medicinal ginseng is Panax ginseng (￥Panax
￥￥ginseng￥C. A. MEYER) or Panax ginseng (￥Panax￥ ￥japonicum
¥C. A. MEYER) or American carrots (¥P
anax￥￥quinquefolium￥L. ) or 37 carrots (￥Panax￥ ￥notoginseng￥F
．． H. A method for mass production of medicinal ginseng by tissue culture according to claim 1, wherein the method is: CHEN).
リンスマイヤーアンドスクーグ培地、ガウスレット培地
、チュレッケ培地、モーレル培地ヒルデブラント培地の
うちから選択されたいずれかの培地である特許請求の範
囲第１項記載の組織培養による薬用人参の大量生産方法
。(3) The reference medium is Murashige and Skoog medium,
The method for mass production of medicinal ginseng by tissue culture according to claim 1, wherein the medium is any one selected from Linsmeyer and Skoog medium, Gauslett medium, Zurecke medium, Morrell medium, and Hildebrand medium.
長調節物質は、２，４−ジクロロフェノキシ酢酸、ナフ
タリン酢酸、インドール酢酸およびインドール酪酸のう
ちから選択されるいずれかの化合物である特許請求の範
囲第１項記載の組織培養による薬用人参の大量生産方法
。(4) The plant growth regulator added to the reference medium in the callus induction step is any compound selected from 2,4-dichlorophenoxyacetic acid, naphthaleneacetic acid, indoleacetic acid, and indolebutyric acid. A method for mass production of medicinal ginseng by tissue culture according to scope 1.
長調節物質は、２，４−ジクロロフェノキシ酢酸、ナフ
タリン酢酸、インドール酢酸またはインドール酪酸のう
ちから選択されるいずれかの化合物である特許請求の範
囲第１項記載の組織培養による薬用人参の大量生産方法
。(5) A patent claim in which the plant growth regulator added to the reference medium in the somatic embryo formation step is any compound selected from 2,4-dichlorophenoxyacetic acid, naphthaleneacetic acid, indoleacetic acid, or indolebutyric acid. A method for mass production of medicinal ginseng by tissue culture according to item 1.
加する植物生長調節物質は、ジベレリン酸とベンジルア
デニンとの組合せ、ナフタリン酢酸とベンジルアデニン
との組合せ、インドール酢酸とベンジルアデニンとの組
合せ、およびインドール酪酸とベンジルアデニンとの組
合せからなる組合群より選択されるいずれかの組合せで
ある特許請求の範囲第１項記載の組織培養による薬用人
参の大量生産方法。(6) The plant growth regulators added to the standard medium or improved medium in the shoot and leaf differentiation step include a combination of gibberellic acid and benzyladenine, a combination of naphthalene acetic acid and benzyladenine, a combination of indoleacetic acid and benzyladenine, and The method for mass production of medicinal ginseng by tissue culture according to claim 1, wherein the combination is any combination selected from the group consisting of combinations of indolebutyric acid and benzyladenine.
せた改良培地は、当該培地の無機塩濃度が前記基準培地
の無機塩の含有組成の１／１未満１／２０以上である特
許請求の範囲第１項記載の組織培養による薬用人参の大
量生産方法。(7) A patent claim in which the improved medium with reduced inorganic salt concentration used in the shoot and leaf differentiation step has an inorganic salt concentration of less than 1/1 and 1/20 or more of the inorganic salt content composition of the reference medium. A method for mass production of medicinal ginseng by tissue culture according to item 1.
生長調節物質は、ナフタレン酢酸、インドール酪酸、イ
ンドール酢酸およびジベレリン酸のうちから選択される
いずれかの化合物である特許請求の範囲第１項記載の組
織培養による薬用人参の大量生産方法。(8) The plant growth regulator added to the reference medium used in the rooting step is any compound selected from naphthaleneacetic acid, indolebutyric acid, indoleacetic acid, and gibberellic acid. A method for mass production of medicinal ginseng by tissue culture as described in .
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP62083641A JPS63248321A (en) | 1987-04-03 | 1987-04-03 | Mass production of medicinal carrot by tissue culture |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP62083641A JPS63248321A (en) | 1987-04-03 | 1987-04-03 | Mass production of medicinal carrot by tissue culture |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS63248321A true JPS63248321A (en) | 1988-10-14 |
| JPH0430809B2 JPH0430809B2 (en) | 1992-05-22 |
Family
ID=13808075
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP62083641A Granted JPS63248321A (en) | 1987-04-03 | 1987-04-03 | Mass production of medicinal carrot by tissue culture |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS63248321A (en) |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS6416582A (en) * | 1987-07-11 | 1989-01-20 | Wakunaga Pharma Co Ltd | Culture and production of root-like tissue of plant of panax genus |
| CN102067849A (en) * | 2011-02-11 | 2011-05-25 | 吉林农业大学 | Method for promoting growth of ginseng root by using mixture of choline chloride, benzyl aminopurine and indolebutyric acid |
| CN104126394A (en) * | 2014-07-25 | 2014-11-05 | 普定县银丰农业科技发展有限公司 | Method for planting pseudo-ginseng |
| WO2017020769A1 (en) * | 2015-08-05 | 2017-02-09 | 广西壮族自治区药用植物园 | Method for increasing content of notoginsenoside r1 in panax notoginseng |
| CN107409690A (en) * | 2017-06-20 | 2017-12-01 | 合肥卓畅农业科技有限公司 | A kind of high-yield planting method of pseudo-ginseng |
| CN108029491A (en) * | 2018-02-12 | 2018-05-15 | 丽江滇西本草药业有限公司 | A kind of High aititude notoginseng planting method |
| CN109964771A (en) * | 2019-03-21 | 2019-07-05 | 文山高田三七中药饮片有限公司 | A kind of implantation methods of Radix Notoginseng |
| CN109964770A (en) * | 2019-03-21 | 2019-07-05 | 文山高田三七中药饮片有限公司 | A kind of implantation methods improving notoginseng obstacle |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS62151117A (en) * | 1985-08-29 | 1987-07-06 | 武田薬品工業株式会社 | Method for creating regeneration plant of otane carrot |
| JPS63133922A (en) * | 1986-11-27 | 1988-06-06 | 日本鉱業株式会社 | Tissue culture of ginseng |
-
1987
- 1987-04-03 JP JP62083641A patent/JPS63248321A/en active Granted
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS62151117A (en) * | 1985-08-29 | 1987-07-06 | 武田薬品工業株式会社 | Method for creating regeneration plant of otane carrot |
| JPS63133922A (en) * | 1986-11-27 | 1988-06-06 | 日本鉱業株式会社 | Tissue culture of ginseng |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS6416582A (en) * | 1987-07-11 | 1989-01-20 | Wakunaga Pharma Co Ltd | Culture and production of root-like tissue of plant of panax genus |
| CN102067849A (en) * | 2011-02-11 | 2011-05-25 | 吉林农业大学 | Method for promoting growth of ginseng root by using mixture of choline chloride, benzyl aminopurine and indolebutyric acid |
| CN104126394A (en) * | 2014-07-25 | 2014-11-05 | 普定县银丰农业科技发展有限公司 | Method for planting pseudo-ginseng |
| WO2017020769A1 (en) * | 2015-08-05 | 2017-02-09 | 广西壮族自治区药用植物园 | Method for increasing content of notoginsenoside r1 in panax notoginseng |
| CN107409690A (en) * | 2017-06-20 | 2017-12-01 | 合肥卓畅农业科技有限公司 | A kind of high-yield planting method of pseudo-ginseng |
| CN108029491A (en) * | 2018-02-12 | 2018-05-15 | 丽江滇西本草药业有限公司 | A kind of High aititude notoginseng planting method |
| CN109964771A (en) * | 2019-03-21 | 2019-07-05 | 文山高田三七中药饮片有限公司 | A kind of implantation methods of Radix Notoginseng |
| CN109964770A (en) * | 2019-03-21 | 2019-07-05 | 文山高田三七中药饮片有限公司 | A kind of implantation methods improving notoginseng obstacle |
Also Published As
| Publication number | Publication date |
|---|---|
| JPH0430809B2 (en) | 1992-05-22 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Agrawal et al. | Effective protocol for in vitro shoot production through nodal explants of Simmondsia chinensis | |
| Faisal et al. | In vitro rapid regeneration of plantlets from nodal explants of Mucuna pruriens–a valuable medicinal plant | |
| Barra-Jiménez et al. | Cloning mature holm oak trees by somatic embryogenesis | |
| Zhou et al. | High efficiency plant production of North American ginseng via somatic embryogenesis from cotyledon explants | |
| Shen et al. | Effects of genotype, explant source, and plant growth regulators on indirect shoot organogenesis in Dieffenbachia cultivars | |
| Samosir et al. | Improving acclimatization through the photoautotrophic culture of coconut (Cocos nucifera) seedlings: an in vitro system for the efficient exchange of germplasm | |
| Yang et al. | Callus induction and high-efficiency plant regeneration via somatic embryogenesis in Papaver nudicaule L., an ornamental medicinal plant | |
| JPS63248321A (en) | Mass production of medicinal carrot by tissue culture | |
| Benmahioul et al. | In vitro embryo germination and proliferation of pistachio (Pistacia vera L.) | |
| Sankar-Thomas et al. | Plant regeneration via somatic embryogenesis of Camptotheca acuminata in temporary immersion system (TIS) | |
| JAYAKUMAR et al. | Influence of additives on enhanced in vitro shoot multiplication of Orthosiphon aristatus (Blume) Miq. | |
| KR102602790B1 (en) | Method for mass propagation of Cnidium officinale Makino through tissue culture | |
| Ranjan Deb et al. | Callus mediated indirect somatic embryogenesis and plant regeneration of Saurauia punduana Wallich (Actinidiaceae) from in vitro cotyledonary leaves | |
| Xu et al. | Efficient in vitro plant regeneration of Pinellia ternata (Thunb) Breit | |
| US10028505B2 (en) | Micropropagation of alexandrian laurel (Danae racemosa L) | |
| Deb et al. | Rapid multiplication and induction of early in vitro flowering in Dendrobium primulinum Lindl | |
| CA1270647A (en) | Process for regenerating soybeans | |
| Gopi et al. | In vitro plant regeneration through somatic embryogenesis in medicinally important leaf explants of Coleus forskohlii Briq | |
| Ohki et al. | The effects of inorganic salts and growth regulators on in vitro shoot proliferation and leaf chlorophyll content of Delphinium cardinale | |
| Khanna et al. | Regeneration via organogenesis in callus cultures of Argyrolobium roseum | |
| Koilpillai et al. | In vitro propagation of Graptophyllum pictum L.(Acanthaceae)-a medicinal plant. | |
| HISAJIMA et al. | Microplant propagation of rice plant in vitro | |
| Jha et al. | In vitro culture of Cuminum cyminum regeneration of flowering shoots from calli of hypocotyl and leaf explants | |
| Xiang-Hui et al. | An efficient system for the production of the medicinally important plant: Asparagus cochinchinensis (Lour.) Merr. | |
| JPH08224051A (en) | Larg scale growth of seedling of medical carrot |