JPH04202137A - Production of iridoid glucoside - Google Patents
Production of iridoid glucosideInfo
- Publication number
- JPH04202137A JPH04202137A JP2331519A JP33151990A JPH04202137A JP H04202137 A JPH04202137 A JP H04202137A JP 2331519 A JP2331519 A JP 2331519A JP 33151990 A JP33151990 A JP 33151990A JP H04202137 A JPH04202137 A JP H04202137A
- Authority
- JP
- Japan
- Prior art keywords
- medium
- culturing
- iridoid
- culture
- plant
- 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
- 238000004519 manufacturing process Methods 0.000 title claims description 24
- 229930182491 iridoid glucoside Natural products 0.000 title abstract 5
- 241000196324 Embryophyta Species 0.000 claims abstract description 49
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims abstract description 21
- 229910052799 carbon Inorganic materials 0.000 claims abstract description 21
- 238000012258 culturing Methods 0.000 claims abstract 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 abstract description 15
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims abstract description 15
- 239000008103 glucose Substances 0.000 claims abstract description 15
- 229930006000 Sucrose Natural products 0.000 claims abstract description 12
- 210000000056 organ Anatomy 0.000 claims abstract description 12
- 239000005720 sucrose Substances 0.000 claims abstract description 12
- 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 abstract description 11
- 229930182489 iridoid glycoside Natural products 0.000 claims description 38
- 150000008145 iridoid glycosides Chemical class 0.000 claims description 28
- 239000005648 plant growth regulator Substances 0.000 claims description 17
- ZUKLFFYDSALIQW-MSUKCBDUSA-N Iridoid glycoside Chemical compound [H][C@]12CC[C@H](C(O)=O)[C@@]1([H])[C@H](OC1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O)OC=C2 ZUKLFFYDSALIQW-MSUKCBDUSA-N 0.000 claims description 10
- 235000009411 Rheum rhabarbarum Nutrition 0.000 claims 1
- 241000219063 Rheum rhabarbarum [NCBITaxon:3621] Species 0.000 claims 1
- 239000002609 medium Substances 0.000 abstract description 41
- QIIDATRCGITYRZ-UHFFFAOYSA-N Catalpol Natural products OCC1OC(OC2OC=CC3C(O)C(=C(CO)C23)O)C(O)C(O)C1O QIIDATRCGITYRZ-UHFFFAOYSA-N 0.000 abstract description 7
- LHDWRKICQLTVDL-PZYDOOQISA-N catalpol Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1O[C@H]1[C@@H]2[C@@]3(CO)O[C@H]3[C@@H](O)[C@@H]2C=CO1 LHDWRKICQLTVDL-PZYDOOQISA-N 0.000 abstract description 7
- UXSACQOOWZMGSE-UHFFFAOYSA-N catalposide Natural products OC1C(O)C(O)C(CO)OC1OC1C2C3(CO)OC3C(OC(=O)C=3C=CC(O)=CC=3)C2C=CO1 UXSACQOOWZMGSE-UHFFFAOYSA-N 0.000 abstract description 7
- LHDWRKICQLTVDL-UHFFFAOYSA-N methyl iridoid glycoside Natural products OC1C(O)C(O)C(CO)OC1OC1C2C3(CO)OC3C(O)C2C=CO1 LHDWRKICQLTVDL-UHFFFAOYSA-N 0.000 abstract description 7
- 239000006870 ms-medium Substances 0.000 abstract description 5
- 239000000126 substance Substances 0.000 abstract description 5
- 238000000034 method Methods 0.000 abstract description 4
- 241000405414 Rehmannia Species 0.000 abstract description 2
- 241000405911 Rehmannia glutinosa Species 0.000 abstract description 2
- 230000008635 plant growth Effects 0.000 abstract 1
- 230000001105 regulatory effect Effects 0.000 abstract 1
- SEOVTRFCIGRIMH-UHFFFAOYSA-N indole-3-acetic acid Chemical compound C1=CC=C2C(CC(=O)O)=CNC2=C1 SEOVTRFCIGRIMH-UHFFFAOYSA-N 0.000 description 16
- 239000003617 indole-3-acetic acid Substances 0.000 description 8
- 210000004027 cell Anatomy 0.000 description 7
- 229930000044 secondary metabolite Natural products 0.000 description 7
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 6
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 6
- 150000001413 amino acids Chemical class 0.000 description 5
- 241000220479 Acacia Species 0.000 description 4
- 235000010643 Leucaena leucocephala Nutrition 0.000 description 4
- 210000004748 cultured cell Anatomy 0.000 description 4
- 239000011782 vitamin Substances 0.000 description 4
- 229930003231 vitamin Natural products 0.000 description 4
- 229940088594 vitamin Drugs 0.000 description 4
- 235000013343 vitamin Nutrition 0.000 description 4
- 244000184734 Pyrus japonica Species 0.000 description 3
- 229930195376 Rehmannioside Natural products 0.000 description 3
- 239000004480 active ingredient Substances 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 125000004394 rehmannioside group Chemical group 0.000 description 3
- 210000001519 tissue Anatomy 0.000 description 3
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Chemical group OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 description 2
- GFFGJBXGBJISGV-UHFFFAOYSA-N Adenine Chemical compound NC1=NC=NC2=C1N=CN2 GFFGJBXGBJISGV-UHFFFAOYSA-N 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 2
- WQZGKKKJIJFFOK-QTVWNMPRSA-N D-mannopyranose Chemical group OC[C@H]1OC(O)[C@@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-QTVWNMPRSA-N 0.000 description 2
- 229930091371 Fructose Chemical group 0.000 description 2
- 239000005715 Fructose Chemical group 0.000 description 2
- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical group OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 description 2
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Chemical group O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 description 2
- 241000208422 Rhododendron Species 0.000 description 2
- 239000005708 Sodium hypochlorite Substances 0.000 description 2
- 241000863480 Vinca Species 0.000 description 2
- 239000008272 agar Substances 0.000 description 2
- WQZGKKKJIJFFOK-PHYPRBDBSA-N alpha-D-galactose Chemical group OC[C@H]1O[C@H](O)[C@H](O)[C@@H](O)[C@H]1O WQZGKKKJIJFFOK-PHYPRBDBSA-N 0.000 description 2
- GUBGYTABKSRVRQ-QUYVBRFLSA-N beta-maltose Chemical group OC[C@H]1O[C@H](O[C@H]2[C@H](O)[C@@H](O)[C@H](O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@@H]1O GUBGYTABKSRVRQ-QUYVBRFLSA-N 0.000 description 2
- 230000001851 biosynthetic effect Effects 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 239000003599 detergent Substances 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 230000004069 differentiation Effects 0.000 description 2
- 230000001882 diuretic effect Effects 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 229930182830 galactose Chemical group 0.000 description 2
- 229930182470 glycoside Natural products 0.000 description 2
- -1 ionone glycosides Chemical class 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 238000004161 plant tissue culture Methods 0.000 description 2
- SUKJFIGYRHOWBL-UHFFFAOYSA-N sodium hypochlorite Chemical compound [Na+].Cl[O-] SUKJFIGYRHOWBL-UHFFFAOYSA-N 0.000 description 2
- 239000008223 sterile water Substances 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 238000002137 ultrasound extraction Methods 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- VELYAQRXBJLJAK-XKKWFBPMSA-N (2s,3r,4s,5s,6r)-2-[[(1s,4ar,5r,7s,7as)-5,7-dihydroxy-7-methyl-4a,5,6,7a-tetrahydro-1h-cyclopenta[c]pyran-1-yl]oxy]-6-(hydroxymethyl)oxane-3,4,5-triol Chemical compound O([C@@H]1OC=C[C@H]2[C@H](O)C[C@@]([C@@H]12)(O)C)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O VELYAQRXBJLJAK-XKKWFBPMSA-N 0.000 description 1
- 229930024421 Adenine Natural products 0.000 description 1
- 229930192334 Auxin Natural products 0.000 description 1
- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 description 1
- 240000001879 Digitalis lutea Species 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- 206010020649 Hyperkeratosis Diseases 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
- 240000004371 Panax ginseng Species 0.000 description 1
- 235000002789 Panax ginseng Nutrition 0.000 description 1
- 208000004880 Polyuria Diseases 0.000 description 1
- 241000206609 Porphyra Species 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 229960000643 adenine Drugs 0.000 description 1
- 208000007502 anemia Diseases 0.000 description 1
- 230000003178 anti-diabetic effect Effects 0.000 description 1
- RJWJHRPNHPHBRN-FKVJWERZSA-N aucubin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1O[C@H]1[C@@H]2C(CO)=C[C@@H](O)[C@@H]2C=CO1 RJWJHRPNHPHBRN-FKVJWERZSA-N 0.000 description 1
- UTDFQMAXCUGNJR-UHFFFAOYSA-N aucubin Natural products OCC1OC(Oc2ccoc2C3C(O)CCC3O)C(O)C(O)C1O UTDFQMAXCUGNJR-UHFFFAOYSA-N 0.000 description 1
- 239000002363 auxin Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 229910052801 chlorine Inorganic materials 0.000 description 1
- 239000000460 chlorine Substances 0.000 description 1
- 230000015271 coagulation Effects 0.000 description 1
- 238000005345 coagulation Methods 0.000 description 1
- 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 1
- 239000004062 cytokinin Substances 0.000 description 1
- 239000002934 diuretic Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 235000008434 ginseng Nutrition 0.000 description 1
- 150000002338 glycosides Chemical class 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 238000003306 harvesting Methods 0.000 description 1
- 241000411851 herbal medicine Species 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 238000005286 illumination Methods 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 229930002839 ionone Natural products 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 239000009569 juzentaihoto Substances 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
- 239000007788 liquid Substances 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- VELYAQRXBJLJAK-UHFFFAOYSA-N myoporoside Natural products C12C(C)(O)CC(O)C2C=COC1OC1OC(CO)C(O)C(O)C1O VELYAQRXBJLJAK-UHFFFAOYSA-N 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 230000001543 purgative effect Effects 0.000 description 1
- 230000001932 seasonal effect Effects 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 125000000185 sucrose group Chemical group 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 239000013076 target substance Substances 0.000 description 1
- 238000012090 tissue culture technique Methods 0.000 description 1
- 230000009385 viral infection Effects 0.000 description 1
Landscapes
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
- Medicines Containing Plant Substances (AREA)
- Compounds Of Unknown Constitution (AREA)
Abstract
Description
【発明の詳細な説明】
（産業上の利用分野）
本発明は、ジオウの組織培養によりイリドイド配糖体を
生産する方法、およびイリドイド配糖体高生産株を作出
する方法に関する。DETAILED DESCRIPTION OF THE INVENTION (Industrial Application Field) The present invention relates to a method for producing iridoid glycosides by tissue culture of Rhizoma spp., and a method for producing a strain that produces high iridoid glycosides.
（従来の技術）
アカヤジオウやカイケイジオウ等のジオウはゴマツバ科
に属する多年草で、根は生薬“地黄゛として用いられて
いる。地黄は血熱などによる出血や湿疹等の治療を目的
に八味地黄丸、当帰六黄湯などの方剤中に配剤される。(Prior art) Rhizophrenia such as Acacia japonicus and Porphyra japonica are perennial plants belonging to the family Rubicaceae, and their roots are used as a herbal medicine called ``Rhioh''. It is used in formulas such as Maru and Tokirokuoto.
また、最も重要な補血薬の１つとして貧血、衰弱等の治
療、改善を目的に十全大補湯、四物湯などに配剤される
。Also, as one of the most important blood supplement drugs, it is added to juzentaihoto, shimotsuto, etc. for the purpose of treating and improving anemia, weakness, etc.
地黄の成分の化学的研究としては、１９７１年に北用ら
が日本産カイケイジオウの新鮮根から主イリドイド配糖
体としてカタルポールの単離を報告した〔北用勲、西村
正、薬誌９１巻第５９３頁（１９７１））。以来、日本
産カイケイジオウおよびアカヤジオウの新鮮根の含有成
分についての検討が行われている。これまでに糖類、ア
ミノ酸類、イリドイド配糖体、ヨノン配糖体などが知ら
れており、中でもイリドイド配糖体は含量が比較的高く
、地黄の品質評価の指標物質とされている。As for the chemical research on the components of Rhizoma, in 1971, Kitayo et al. reported the isolation of catalpol as the main iridoid glycoside from the fresh roots of Kaikeijiou from Japan [Isao Kitayo, Tadashi Nishimura, Yakushu 91 Vol. 593 (1971)). Since then, studies have been carried out on the components contained in the fresh roots of Japanese-produced Acacia japonicus and Aca-yajiou. So far, saccharides, amino acids, iridoid glycosides, ionone glycosides, etc. have been known, and among them, iridoid glycosides have a relatively high content and are used as indicator substances for evaluating the quality of rhizoma.
地黄のイリドイド配糖体としては、カタルボールが主で
あり、他にレーマンニオシド（ｒｅｈｍａｎｎｉｏｓｉ
ｄｅ ） Ａ、 Ｂ、 ＣおよびＤ、レオヌリド（Ｉｅ
ｏｎｕｌｉｄｅ）、オークビン（ａｕｃｕｂｉｎ ）等
がある。特にカタルボールは利尿作用や緩和な瀉下作用
が認められ、地黄の有効成分として最近注目されている
。The main iridoid glycosides in rhizome are catarrhbole, and other iridoid glycosides include rehmannioside.
de) A, B, C and D, leonuride (Ie
onulide, aucbin, etc. In particular, catarrhbor has been recognized to have diuretic and mild purgative effects, and has recently attracted attention as an active ingredient in rhizome.
このように生薬の材料や有効成分の抽出材料として有用
なジオウは、わが国では奈良、長野でわずかに栽培され
ているだけで、そのほとんどを中国や韓国から輸入して
おり、またジオウの栽培は天候に左右される等の欠点が
あるため、安定に供給することが望まれている。In this way, geophyte, which is useful as a material for crude drugs and for extracting active ingredients, is only cultivated in small quantities in Nara and Nagano in Japan, and most of it is imported from China and South Korea. Since it has drawbacks such as being affected by the weather, a stable supply is desired.
また、アカヤジオウはウィルス罹患率が高く、ウィルス
罹患株はウィルスに感染していない株番土比べてカタル
ポール含量が少ない、あるいはカタルボールを全く産出
しないことが知られている〔西岡五夫、生薬学雑誌、４
２ （１）、１〜１１頁（１９８Ｂ））。In addition, it is known that the virus infection rate is high in Acacia japonicus, and that virus-affected strains contain less catarrhpol or do not produce catarrhpol at all than strains that are not infected with the virus [Goo Nishioka, Pharmaceutical journal, 4
2 (1), pp. 1-11 (198B)).
従って、高品質の材料を天候の影響や地域的、季節的な
制約を受けることなく、高効率で安定に供給するため、
組織培養を利用した大量増殖を行うことが考えられるが
、植物体の増殖だけでなく、・ 植物体が産生ずる有用
物質の採取をも目的とする場合、次のような問題点があ
る。Therefore, in order to supply high-quality materials stably and efficiently without being affected by weather or regional or seasonal constraints, we
It is conceivable to perform mass propagation using tissue culture, but if the purpose is not only to propagate the plants, but also to collect useful substances produced by the plants, there are the following problems.
組織培養技術を用いて培養細胞を作出し、その植物の２
次代謝産物を培養細胞中に産生させる方法が、医薬品を
中心とした分野で検討されているが、十分な有用物質生
産能を有する培養細胞は、数例を除きほとんどの薬用植
物では得られていない。これは、下記の点に起因すると
考えられる。Cultured cells are created using tissue culture technology, and 2
Methods for producing secondary metabolites in cultured cells are being studied in the pharmaceutical field, but cultured cells with sufficient ability to produce useful substances have not been obtained for most medicinal plants, with the exception of a few cases. do not have. This is considered to be due to the following points.
・一般に２次代謝物質の生合成系は複雑であり、植物成
長調節物質の添加や他の培養条件により生合成系のある
分野が阻害を受けると、目的物質の産生能は著しく低下
してしまうこと。・Generally, the biosynthetic system of secondary metabolites is complex, and if a certain area of the biosynthetic system is inhibited by the addition of plant growth regulators or other culture conditions, the ability to produce the target substance will be significantly reduced. thing.
・多くの植物の２次代謝物質の産生能は、器官分化との
関連が強く、一般には特定の器官にのみ認められること
が多いのに対し、培養細胞は植物成長調節物質によって
強制的に脱分化させた細胞であり、２次代謝活性のよう
な一種の生理的分化の性質を失っている可能体が強いこ
と。・The ability to produce secondary metabolites in many plants is strongly related to organ differentiation, and is generally found only in specific organs, whereas cultured cells are forced to desorb by plant growth regulators. They are differentiated cells, and there is a strong possibility that they have lost a type of physiological differentiation property such as secondary metabolic activity.
そのため、植物の器官培養により親植物の２次代謝産物
の生産能を維持した例が、オタネニンジン、ジギタリス
、ニチニチソウではあるが、ジオウについての検討は全
くなされていない。Therefore, Panax ginseng, digitalis, and periwinkle are examples of maintaining the ability of the parent plant to produce secondary metabolites through plant organ culture, but no studies have been conducted on periwinkle.
（発明が解決しようとする課題）
前記のように、植物の組織培養や器官培養により２次代
謝産物の産生量を増加させるのに成功した例もあるが、
２次代謝産物の産生については植物毎にその培養条件に
大きく影響を受け、特定の２次代謝産物の産生能を高め
ることは困難であった。(Problems to be Solved by the Invention) As mentioned above, there are examples of success in increasing the production amount of secondary metabolites through tissue culture and organ culture of plants.
The production of secondary metabolites is greatly affected by the culture conditions of each plant, and it has been difficult to increase the production ability of specific secondary metabolites.
本発明の目的は、ジオウにおいて組織培養の手法を用い
てイリドイド配糖体、特にカタルボールの生産能を高め
た植物体を得て、これより高効率でイリドイド配糖体を
得ることにある。An object of the present invention is to obtain a plant with enhanced production ability of iridoid glycosides, particularly catabolbol, using a tissue culture technique, and to obtain iridoid glycosides with higher efficiency.
（課題を解決するための手段）
本発明者は、ジオウの植物体を特定の培養条件下での培
養によってイリドイド配糖体高生産株を作出し、イリド
イド配糖体を効率よく得ることができることを見出し、
本発明を完成させた。(Means for Solving the Problems) The present inventors have discovered that iridoid glycosides can be efficiently obtained by creating a strain that produces high iridoid glycosides by culturing the plants of Rhizoma spp. under specific culture conditions. heading,
The present invention has been completed.
即ち、本発明は、ジオウの細胞、組織あるいは器官から
培養によって誘導、成長させた植物体を、植物成長調節
物質としてナフタレン酢酸を０．５〜４ｍｇ／ｌ、炭素
源としてシュークロースを含む第１の培地で培養後、炭
素源としてグルコースを含む第２の培地で培養し、得ら
れた植物体からイリドイド配糖体を回収することを特徴
とする特許イド配糖体の生産方法を要旨とする。That is, the present invention is directed to culturing and growing a plant body derived from cells, tissues, or organs of Rhododendron japonica using a first plant containing 0.5 to 4 mg/l of naphthaleneacetic acid as a plant growth regulator and sucrose as a carbon source. The gist of this invention is a patented method for producing iridoid glycosides, which comprises culturing in a second medium containing glucose as a carbon source, and recovering iridoid glycosides from the resulting plant. .
上記方法において、第１の培地での培養後、イリドイド
配糖体高生産能の株を選抜してから、第２の培地で培養
を行えば、より効果的である。In the above method, it is more effective if, after culturing in the first medium, a strain with high iridoid glycoside production ability is selected and then cultured in the second medium.
さらに、本発明は、ジオウの細胞、組織あるいは器官か
ら培養によって誘導、成長させた植物体を、植物成長調
節物質としてナフタレン酢酸を０゜５〜４ ｍｇ／ ｌ
、炭素源としてシュークロースを含む第１の培地で培
養後、炭素源としてグルコースを含む第２の培地で培養
することによりイリドイド配糖体高生産株を作出する方
法にも関する。Furthermore, the present invention provides a plant body induced and grown by culture from cells, tissues, or organs of Rhododendron japonica, treated with naphthalene acetic acid as a plant growth regulator at a concentration of 0.5 to 4 mg/l.
The present invention also relates to a method for producing a strain with high iridoid glycoside production by culturing in a first medium containing sucrose as a carbon source and then culturing in a second medium containing glucose as a carbon source.
本発明において使用する植物は、ジオウであり、その栽
培種としては例えば、アカヤジオウ（Ｒｅｈｍａｎｎｉ
ａ ｇｌｕｔｉｎｏｓａ ＬＩＢ、ｖａｒ、ｐｕｒ
ｐｕｒｅａ ＭＡＫＩＮＯ） 、カイケイジオウ（
Ｒｅｈｍａｎｎｉａ ｇｌｕｔｉｎｏｓａ ｈｕｅｉｃ
ｈｉｎｇｅｎｓｉｓ）があるが、その他、イリドイド配
糖体産生能を有する植物であれば特に限定されない。The plant used in the present invention is Rehmanni, and its cultivated species include, for example, Rehmanni
a glutinosa LIB, var, pur
purea MAKINO), Kaikeijiou (
Rehmannia glutinosa hueic
hingensis), but other plants are not particularly limited as long as they have the ability to produce iridoid glycosides.
上記植物は、有効成分としてイリドイド配糖体を含むも
のであり、これは抗糖尿病作用、血液凝固抑制作用、利
尿作用を有する。このイリドイド配糖体は、カタルポー
ルを主成分とし、他にレーマンニオシド（ｒｅｈｍａｎ
ｎｉｏｓｉｄｅ ） Ａ、 Ｂ、 ＣおよびＤルオヌリ
ド（ｌｅｏｎｕｌｉｄｅ ） 、オークビン（ａｕｃｕ
ｂｉｎ ）を含む。The above-mentioned plants contain iridoid glycosides as active ingredients, which have anti-diabetic effects, anti-blood coagulation effects, and diuretic effects. This iridoid glycoside has catalpol as its main component, and also contains rehmannioside (rehman
nioside) A, B, C and D leonulide, aucu
bin).
本発明においては、上記ジオウの細胞、組織あるいは器
官から培養により誘導、成長させた植物体を特定の第１
の培地で培養し、次いで特定の第２の培地で培養するこ
とにより、圃場で栽培された植物体よりもイリドイド配
糖体産生能が大きい植物体を作出でき、この高生産株よ
りイリドイド配糖体、特にカタルポールを高収率で採取
できる。In the present invention, a plant body induced and grown by culture from the cells, tissues, or organs of the above-mentioned Rhizoma spp.
By culturing in this medium and then culturing in a specific second medium, it is possible to create plants that have a greater ability to produce iridoid glycosides than plants grown in the field. body, especially catalpol, can be collected with high yield.
高生産株の作出のために使用する植物体としては、ジオ
ウの外植片、例えば葉、茎、根等の器官の切片または細
胞から培養により誘導した植物体を使用するため、イリ
ドイド配糖体産生能を維持しているとともに、一定の遺
伝形質をもち、ウィルスフリーの植物体を安定に大量に
供給できるという利点も有する。The plants used for the production of high-producing strains are those derived by culture from explants of P. aeruginosa, such as sections of organs such as leaves, stems, and roots, or cells, so the iridoid glycosides are used. It has the advantage of maintaining productivity, having certain genetic traits, and being able to stably supply a large amount of virus-free plants.
外植片からの植物体の誘導について以下に説明する。外
植片としては、これまで使用していた茎、根の他に、葉
を使用することができる。外植片は、例えば洗剤で洗浄
後、流水ですすぎ、ついで次亜塩素酸ナトリウム溶液に
浸漬し、滅菌水ですすぐ等の操作により洗浄、滅菌を行
ったものを使用する。次いで、この外植片を適宜培地上
に置床し、培養を行うことにより植物体を誘導し、さら
に成長させる。The induction of plants from explants will be explained below. As explants, leaves can be used in addition to the stems and roots that have been used so far. The explant used is one that has been washed and sterilized by, for example, washing with a detergent, rinsing with running water, then immersing in a sodium hypochlorite solution, and rinsing with sterile water. Next, this explant is placed on a suitable medium and cultured to induce a plant body and further grow.
この培養で使用する培地は、通常植物の組織培養に用い
られている培地であればよく、無機成分および炭素源を
含み、必要に応じ植物成長調節物質、ビタミン類、アミ
ノ酸類を添加した培地である。具体的にはムラシゲ・ス
クーグの培地（ＭＳ培地）、リンスマイヤー・スクーグ
の培地、ホワイトの培地、ニッチ・ニッチの培地等の、
無機成分からなる基本培地にシュークロース等の炭素源
、植物成長調節物質、ビタミン類、アミノ酸類を添加し
た培地が例示される。植物成長調節物質にはナフタレン
酢酸（ＮＡＡ） 、インドール酢酸（ＩＡＡ）等のオー
キシン類、ヘンシルアデニン（ＢＡ）、カイネチン等の
サイトカイニン類があるが、ここで植物体の誘導、成長
に用いる培地にはＩＡＡおよびＢＡを添加したものが好
ましい。特に、ＩＡＡおよびＢＡをそれぞれ１ｍｇ／ｌ
程度添加した培地が望ましい。植物体の誘導、成長に用
いる培地としては、寒天やゼラチンを０．５〜１％添加
した固形培地が好ましい。The medium used for this culture may be any medium normally used for plant tissue culture, and may contain inorganic components and carbon sources, and may be supplemented with plant growth regulators, vitamins, and amino acids as necessary. be. Specifically, Murashige-Skoog medium (MS medium), Linsmeyer-Skoog medium, White's medium, Niche Niche medium, etc.
An example is a medium in which a carbon source such as sucrose, plant growth regulators, vitamins, and amino acids are added to a basic medium consisting of inorganic components. Plant growth regulators include auxins such as naphthalene acetic acid (NAA) and indole acetic acid (IAA), and cytokinins such as hensyl adenine (BA) and kinetin. It is preferable to add IAA and BA. In particular, IAA and BA at 1 mg/l each
It is preferable to use a medium supplemented with some amount. The medium used for inducing and growing plants is preferably a solid medium to which 0.5 to 1% of agar or gelatin is added.
培養条件は特に限定されないが、通常、温度１０〜４０
’Ｃ１照明下で行う。Culture conditions are not particularly limited, but usually the temperature is 10-40°C.
'Do it under C1 lighting.
上記培地および培養条件下で培養後、２週間〜２箇月、
通常１箇月程度で植物体が誘導される。2 weeks to 2 months after culturing in the above medium and culture conditions,
Plants are usually induced in about one month.
さらに培養を続け、１〜３箇月、通常２箇月程度で植物
体が約２〜３ｃｍに成長する。The culture is continued, and the plant grows to about 2 to 3 cm in 1 to 3 months, usually about 2 months.
このようにして得られた植物体から、次の方法でイリド
イド配糖体高生産株を作出する。From the plants thus obtained, a strain with high iridoid glycoside production is produced by the following method.
まず、植物成長調節物質としてＮＡＡｏ、５〜４ｍｇ／
］、炭素源としてシュークロースを含有する第１の培地
において植物体の培養を行う。First, as a plant growth regulator, NAAo, 5 to 4 mg/
], the plant is cultured in a first medium containing sucrose as a carbon source.
第１の培地の具体例としては前述のＭＳ培地等の基本培
地に、植物成長調節物質としてＮＡＡを０．５〜４ ｍ
ｇ／ ｌ 、炭素源としてシェークロースを。A specific example of the first medium is a basic medium such as the above-mentioned MS medium, and 0.5 to 4 m of NAA as a plant growth regulator.
g/l, shakerose as carbon source.
１〜１０％、好ましくは約３％添加したものである。It is added in an amount of 1 to 10%, preferably about 3%.
必要に応じ、ビタミン類、アミノ酸類を添加してもよい
。Vitamins and amino acids may be added as necessary.
培養条件は特に限定されないが、通常、温度１゜翳１（
Ｌｌ＄
〜４０°Ｃ，量孝２５°Ｃで、３〜６週間程度培養する
。Culture conditions are not particularly limited, but are usually at a temperature of 1°
Culture at 40°C and 25°C for about 3 to 6 weeks.
後述の実施例で示すように、培地中に植物成長調節物質
としてＮＡＡを０．５〜４ｍｇ／ｌ含有する場合に、他
の植物成長調節物質添加の場合に比べ、集中のイリドイ
ド配糖体含有量が高い。As shown in the Examples below, when the medium contains 0.5 to 4 mg/l of NAA as a plant growth regulator, compared to the case where other plant growth regulators are added, the content of iridoid glycosides is more concentrated. The quantity is high.
ここで、葉中のイリドイド配糖体の含有量が特に高いイ
リドイド配糖体高生産株を選抜して次の培養に供すると
、本発明の目的により適した高生産株が最終的に得られ
る。Here, if a high iridoid glycoside-producing strain with a particularly high iridoid glycoside content in its leaves is selected and subjected to the next culture, a high-producing strain more suitable for the purpose of the present invention will finally be obtained.
第２の培地は、炭素源としてグルコースを含有すること
を特徴とする特にグルコース３〜７％の濃度で含有する
場合にイリドイド配糖体の生産が良好である。具体的に
は、前述と同様のＭＳ培地等の基本培地に炭素源として
グルコースを添加、し、必要に応じ植物成長調節物質、
ビタミン類、アミノ酸類等を添加したものである。植物
成長調節物質としてＮＡＡを第１の培地と同様０．５〜
４ｍｇ／ｌ添加するとより好ましい。The second medium is characterized by containing glucose as a carbon source. Particularly when glucose is contained at a concentration of 3 to 7%, production of iridoid glycosides is good. Specifically, glucose is added as a carbon source to a basic medium such as the MS medium as described above, and if necessary, plant growth regulators,
It contains added vitamins, amino acids, etc. As a plant growth regulator, NAA is added at 0.5 to 0.5% as in the first medium.
It is more preferable to add 4 mg/l.
培養は前記第１の培地による培養と同様に行い、培養期
間は通常６〜８週間程度である。The culture is carried out in the same manner as the culture using the first medium, and the culture period is usually about 6 to 8 weeks.
後の実施例において示すように、炭素源としてグルコー
スを用いた場合、グルコース濃度３〜７％で葉中のカタ
ルボール含有量は約３０〜５０Ｂ／ １であり、他の炭
素源を用いて培養した場合、および圃場で栽培したもの
の相中の含有量に比べ著しく高含有量である。最初から
グルコースをふくむ培地で培養すると、成育が悪い。As shown in the later examples, when glucose is used as the carbon source, the catarrhal content in the leaves is about 30-50 B/1 at a glucose concentration of 3-7%, and when cultured with other carbon sources. The content is significantly higher than that in the phase when grown in the field and when grown in the field. If cultured in a medium containing glucose from the beginning, growth will be poor.
植物体からのイリドイド配糖体の採取は、葉、根、茎等
より次のような抽出方法により行う。例えば、刻んだ試
料にアセトニトリル：水（２：８）混液を加え、３０分
程度超音波抽出を行う。ジエチルエーテルで数回洗浄後
、遠心分離により得た上澄みをフィルターで濾過してイ
リドイド配糖体含有液を得る。Iridoid glycosides are collected from plants by the following extraction method from leaves, roots, stems, etc. For example, a mixture of acetonitrile and water (2:8) is added to a chopped sample, and ultrasonic extraction is performed for about 30 minutes. After washing several times with diethyl ether, the supernatant obtained by centrifugation is filtered to obtain an iridoid glycoside-containing solution.
実施例
植生体生誘盪
アカヤジオウ（Ｒｅｈｍａｎｎｉａ ｇｌｕｔｉｎｏｓ
ａ ＬＩＢ、ｖａｒ。Examples Rehmannia glutinos
a LIB, var.
ｐｕｒｐｕｒｅａ ＭＡＫＩＮＯ）の葉を中性洗剤で洗
浄後、流水でよくすすぎ、次いで有効態塩素１％の次亜
塩素酸ナトリウム溶液に２０分間浸漬し、クリーンベン
チ内で滅菌水により３回洗浄した。この葉を１ｃｍ角に
カミソリで切断し、ＭＳ基本培地にシュークロース３％
、Ｉ Ａ Ａ １ ｍｇ／ Ｉ、ＢＡ１ｍｇ／ｌおよび
寒天０．８培養添加した培地に置床した。Purpurea MAKINO) leaves were washed with a neutral detergent, rinsed thoroughly with running water, then immersed in a sodium hypochlorite solution containing 1% effective chlorine for 20 minutes, and washed three times with sterile water in a clean bench. Cut this leaf into 1 cm squares with a razor and add 3% sucrose to MS basic medium.
, IAA 1 mg/I, BA 1 mg/l, and agar 0.8.
培養は２５°Ｃで白色蛍光灯の連続照明（約２５００
ＬＵＸ）下で行った。培養約１箇月後、カルスより植物
体が誘導され、さらに２箇月培養を続けると約３ｃｍの
植物体に成長した。Culture was performed at 25°C under continuous illumination of white fluorescent lamps (approximately 2,500
LUX). After about 1 month of culture, a plant was induced from the callus, and when the culture was continued for an additional 2 months, the plant grew to about 3 cm.
−Ｊ１の１立ｌによるｌ立１
上記で得られた植物体を、ＭＳ基本培地に植物成長調節
物質としてＮＡＡを０．５〜５ ｍｇ／ｌ 、炭素源と
してシュークロースを３％添加した培地に移植し培養を
■週間行った。比較のためにＮＡＡに代えてＩＡＡを０
．５〜５’ｍｇ／ｌ添加した培地においても培養を行っ
た。- 1 liter of J1 (1 liter) The above-obtained plants were grown in an MS basic medium containing 0.5 to 5 mg/l of NAA as a plant growth regulator and 3% sucrose as a carbon source. The cells were transplanted and cultured for several weeks. For comparison, IAA is 0 instead of NAA.
．． Culture was also carried out in a medium supplemented with 5-5'mg/l.
培養８週間後、得られた植物体の葉よりカタルボールを
抽出した。抽出は次のように行った。即ち、刻んだ試料
１ｇにアセトニトリル：水（２：８）混液を加え、３０
分間超音波抽出を行った後、ジエチルエーテルで３回洗
浄し、３５’００ｒｐｍで３分間遠心分離を行った。得
られた上澄み２ｍｌを０．４５ｍのフィルターで濾過し
てイリドイド配糖体含有溶液を得た。After 8 weeks of culture, catarrhboles were extracted from the leaves of the resulting plants. Extraction was performed as follows. That is, a mixture of acetonitrile and water (2:8) was added to 1 g of a chopped sample, and 30
After performing ultrasonic extraction for one minute, the mixture was washed three times with diethyl ether and centrifuged at 35'00 rpm for three minutes. 2 ml of the obtained supernatant was filtered through a 0.45 m filter to obtain an iridoid glycoside-containing solution.
得られた植物体の葉中のカタルボール含量を高速液体マ
イクログラフィーを用いて調べた結果を第１図に示す。The catarrhal content in the leaves of the obtained plants was investigated using high-performance liquid micrography, and the results are shown in FIG.
同図より明らかなように、ＮＡＡ濃度が０．５〜４ ｍ
ｇ／ ｌの場合、葉中のカタルボール含有量は１〜５ｍ
ｇ／ｌと良好であり、ＩＡＡを用いた場合は約半分の含
量であった。また、ＮＡＡ、ＩＡＡともに濃度が５ｍｇ
／ｌを超えるとカタルボールの生産はみられない。As is clear from the figure, the NAA concentration is 0.5 to 4 m
g/l, the catarrhal content in the leaves is 1-5 m
g/l, which was good, and when IAA was used, the content was about half. In addition, the concentration of both NAA and IAA is 5 mg.
/l, no production of catarrhal bole is observed.
ここで、葉中のカタルボール含有量が高い、カタルボー
ル高生産株を高速液体クロマトグラフィーで含有量を測
定し、選抜した。Here, a high catarrhal producing strain with a high catarrhal content in its leaves was selected by measuring the content using high performance liquid chromatography.
ｌ立 −１立
前記で得たカタルボール高生産株を、ＭＳ培地に植物成
長調節物質としてＮＡＡ １〜２ｍｇ／ｌ、炭素源とし
てグルコースを添加した培地に移植して８週間培養した
。比較のために炭素源をシュークロース、ガラクトース
、フラクトース、マンノース、マルトースに代えた培地
にも移植して培養した。The highly productive Catarbor strain obtained above was transplanted to an MS medium supplemented with 1 to 2 mg/l of NAA as a plant growth regulator and glucose as a carbon source and cultured for 8 weeks. For comparison, the cells were also transplanted and cultured in a medium in which the carbon source was replaced with sucrose, galactose, fructose, mannose, or maltose.
得られた植物体の葉中のカタルポール含有量を調べた結
果、ガラクトース、フラクト−ス、マンノース、マルト
ースを使用した場合はかなり低い。As a result of examining the catalpol content in the leaves of the obtained plants, it was found that it was quite low when galactose, fructose, mannose, and maltose were used.
本発明のグルコース添加の場合と、植物体の組織培養に
普通使用するシュークロースを添加した場合のカタルボ
ール含有量について第２図に示す。Figure 2 shows the catarrhbol content in the case of adding glucose according to the present invention and in the case of adding sucrose, which is commonly used in plant tissue culture.
同図より明らかなように、グルコースを添加した培地で
は、シュークロース添加の場合および圃場栽培の場合に
比較し、全般的にカタルポール含有量が増加し、特にグ
ルコース濃度３〜７％で効果が大きいことが判る。即ち
、シュークロース添加培地の場合に比べ８〜１０倍の生
産量であり、圃場栽培のアカヤジオウの相中のカタルポ
ール含有量の約４（＠の値であるというように、非常に
高い含有量である。As is clear from the figure, in the medium supplemented with glucose, the content of catalpol increases overall compared to the case of sucrose addition and the case of field cultivation, and the effect is particularly high at glucose concentrations of 3 to 7%. It turns out it's big. In other words, the production amount is 8 to 10 times higher than in the case of sucrose-added medium, and the content of catalpol in the phase of field-grown Acacia spp. is about 4 (value of @), which is very high. It is.
（発明の効果）
以上詳述したように、本発明によれば、圃場の栽培に比
ベイリドイド配糖体高生産性の植物体を作出でき、この
イリドイド配糖体高生産株よりイリドイド配糖体を抽出
して採取すれば、通常の栽培に比べ極めて短期間でしか
も高収率でイリドイド配糖体を大量に得ることができ、
非常に効率がよい。(Effects of the Invention) As described in detail above, according to the present invention, it is possible to produce plants with high productivity of beiridoid glycosides in field cultivation, and to extract iridoid glycosides from this high production strain of iridoid glycosides. By harvesting iridoid glycosides, it is possible to obtain large amounts of iridoid glycosides in an extremely short period of time and at a high yield compared to conventional cultivation.
Very efficient.
第１図は、第１の培地で培養後カタルボール含有量を測
定した結果を示す図であり、
第２図は、第２の培地で培養後カタルボール含有量を測
定した結果を示す図である。Figure 1 is a diagram showing the results of measuring the catarrhal content after culturing in the first medium, and Figure 2 is a diagram showing the results of measuring the catarrhal content after culturing in the second medium. be.
Claims (5)
誘導、成長させた植物体を、植物成長調節物質としてナ
フタレン酢酸を０．５〜４ｍｇ／ｌ、炭素源としてシュ
ークロースを含む第１の培地で培養後、炭素源としてグ
ルコースを含む第２の培地で培養し、得られた植物体か
らイリドイド配糖体を回収することを特徴とするイリド
イド配糖体の生産方法。(1) Plants induced and grown by culture from cells, tissues, or organs of Rhizoma spp. are grown in a first medium containing 0.5 to 4 mg/l of naphthalene acetic acid as a plant growth regulator and sucrose as a carbon source. A method for producing iridoid glycosides, which comprises, after culturing, culturing in a second medium containing glucose as a carbon source, and recovering iridoid glycosides from the resulting plant.
株を選抜してその後の培養に使用する請求項１記載の生
産方法。(2) The production method according to claim 1, wherein after culturing in the first medium, a strain with high iridoid glycoside production is selected and used for subsequent culturing.
植物体を用いる請求項１または２記載の生産方法。(3) The production method according to claim 1 or 2, which uses a plant that is induced and grown by organ culture from the leaves of Rhubarb.
求項１ないし３のいずれかの項記載の生産方法。(4) The production method according to any one of claims 1 to 3, wherein the iridoid glycoside is recovered from leaves of the plant.
誘導、成長させた植物体を、植物成長調節物質としてナ
フタレン酢酸０．５〜４ｍｇ／ｌを、炭素源としてシュ
ークロースを含む第１の培地で培養後イリドイド配糖体
高生産株を選抜し、次いでこれを、炭素源としてグルコ
ースを含む第２の培地で培養することによりイリドイド
配糖体高生産株を得ることを特徴とする、イリドイド配
糖体高生産株の作出方法。(5) Plants induced and grown by culture from cells, tissues, or organs of Rhizoma spp. are grown in a first medium containing 0.5 to 4 mg/l of naphthalene acetic acid as a plant growth regulator and sucrose as a carbon source. High production of iridoid glycosides, characterized in that a strain with high production of iridoid glycosides is obtained after culturing, and then this is cultured in a second medium containing glucose as a carbon source. How to create stocks.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2331519A JPH04202137A (en) | 1990-11-29 | 1990-11-29 | Production of iridoid glucoside |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2331519A JPH04202137A (en) | 1990-11-29 | 1990-11-29 | Production of iridoid glucoside |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPH04202137A true JPH04202137A (en) | 1992-07-22 |
Family
ID=18244558
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2331519A Pending JPH04202137A (en) | 1990-11-29 | 1990-11-29 | Production of iridoid glucoside |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH04202137A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102657090A (en) * | 2012-05-17 | 2012-09-12 | 福建农林大学 | Rapid propagation method of rehmannia by tissue culture |
| JP2014505070A (en) * | 2011-01-31 | 2014-02-27 | セダーマ | Novel use of extracts from globularia plants and methods for obtaining said extracts by in vitro plant culture |
| CN108034681A (en) * | 2017-12-08 | 2018-05-15 | 新乡医学院 | Utilize the method for the stem of Radix Codonopsis lanceolatae forming layer stem cell production Catalpol for the culture that suspends |
-
1990
- 1990-11-29 JP JP2331519A patent/JPH04202137A/en active Pending
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2014505070A (en) * | 2011-01-31 | 2014-02-27 | セダーマ | Novel use of extracts from globularia plants and methods for obtaining said extracts by in vitro plant culture |
| JP2017039749A (en) * | 2011-01-31 | 2017-02-23 | セダーマ | Novel use of extract of plant origin of globularia and method for obtaining extract by in vitro plant culture |
| CN102657090A (en) * | 2012-05-17 | 2012-09-12 | 福建农林大学 | Rapid propagation method of rehmannia by tissue culture |
| CN108034681A (en) * | 2017-12-08 | 2018-05-15 | 新乡医学院 | Utilize the method for the stem of Radix Codonopsis lanceolatae forming layer stem cell production Catalpol for the culture that suspends |
| CN108034681B (en) * | 2017-12-08 | 2021-08-10 | 新乡医学院 | Method for producing catalpol by using suspension culture rehmannia stem cambium stem cells |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN113025552B (en) | Isolated culture method of ginseng stem cells | |
| EP0525914A1 (en) | Synthetic seed | |
| KR100506625B1 (en) | Mass production method of cultured mountain ginseng by tissue culture | |
| JP2023525203A (en) | Standardized plant extracts from biomass of in vitro cultures, methods of preparation and uses thereof | |
| JPH04202137A (en) | Production of iridoid glucoside | |
| CN112956415B (en) | Liquid culture medium for culturing ginseng adventitious roots and culture method | |
| CN113647324A (en) | Liquid culture medium for culturing ginseng adventitious roots and culture method | |
| JP2545359B2 (en) | Plant culture cells | |
| JP3302853B2 (en) | Culture method of medicinal carrot callus | |
| JP2937435B2 (en) | Tissue culture method of woody plant | |
| RU2762431C1 (en) | Method for isolation of biologically active substances of antimicrobial action from cell cultures of spiny eleuterococcus eleutherococcus senticosus rupr. maxim. | |
| JP2873023B2 (en) | Method for producing podophyllotoxin compounds | |
| JPS61192284A (en) | Production of dried product of plant tissue culture substance of gynostemma pentaphyllum makino | |
| JP2631845B2 (en) | Culture production method of root-like tissue of Panax plants | |
| JPH0335790A (en) | Production of flavonoid glycoside having isoprenoid side chain | |
| JPH0420596B2 (en) | ||
| JPH02191292A (en) | Production of acteoside | |
| JPS61124391A (en) | Production of arbutin | |
| JPH03279333A (en) | Method for preparing acanthopanax senticosus tissue culture product containing eleutherosides and other physiologically active substance | |
| KR0140787B1 (en) | Preparation process of sanguinarine and its derivatives by culturing cells from macleaya species | |
| JPH02249495A (en) | Production of glycyrrhizin | |
| JPS6227797B2 (en) | ||
| Mihalko | Establishing Tissue Culture of King of Bitters (Andrographis paniculata) and Comparison of Callus Versus Plant Neoandrographolide Production | |
| JPH04273888A (en) | New antioxidative glycoside, its production and use | |
| SU967484A1 (en) | Method of producing alpha-fucosospecific lectine |