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KR100259782B1 - Preparing method of root of ginseng from plant tissue culture - Google Patents

Preparing method of root of ginseng from plant tissue culture Download PDF

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KR100259782B1
KR100259782B1 KR1019970047810A KR19970047810A KR100259782B1 KR 100259782 B1 KR100259782 B1 KR 100259782B1 KR 1019970047810 A KR1019970047810 A KR 1019970047810A KR 19970047810 A KR19970047810 A KR 19970047810A KR 100259782 B1 KR100259782 B1 KR 100259782B1
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ginseng
culture
ginseng root
root
growth hormone
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KR19990025936A (en
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오훈일
이우진
김정혜
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    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
    • A01H4/005—Methods for micropropagation; Vegetative plant propagation using cell or tissue culture techniques
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
    • A01H4/002—Culture media for tissue culture
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    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
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    • C12N5/0025—Culture media for plant cell or plant tissue culture
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    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/04—Plant cells or tissues
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2501/00—Active agents used in cell culture processes, e.g. differentation
    • C12N2501/30—Hormones

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Abstract

PURPOSE: A method for producing Insam(ginseng) root by using plant tissue culture is provided, therefore it is possible to produce high content saponin and Insam(ginseng) extract regardless of weather, soil, or damage from disease and harmful insects. CONSTITUTION: The method for producing Insam(ginseng) root by using plant tissue culture comprises the steps of: (i) inoculating and cultivating germinated seed or the intercept of an undried Insam(ginseng) in an MS solid medium that contains growth hormone mixed of auxin and cytokinin at 25deg.C for some 4 weeks to induce callus; (ii) cultivating the induced callus in an MS solid medium for inducing Insam(ginseng) root which contains growth hormone mixed of auxin 2-4mg/L and cytokinin 0.5-1mg/L; (iii) cultivating the induced Insam(ginseng) root in an SH liquid medium which contains growth hormone mixed of auxin 2-4mg/L and cytokinin 0.5-1mg/L; and (iv) cultivating in an SH liquid medium which contains only auxin 2-4mg/L.

Description

식물조직배양을 이용한 인삼근의 생산방법Production Method of Ginseng Root Using Plant Tissue Culture

본 발명은 식물조직배양법에 의한 고려인삼(Panax ginseng C.A. Meyer) 근(root)의 생산방법, 보다 상세하게는 고려인삼에 들어있는 유효성분인 사포닌 및 기타 유효성분을 식물근 배양법에 의해 생산하는 방법에 관한 것이다.The present invention is a method of producing root ginseng (Panax ginseng CA Meyer) root by the plant tissue culture method, more specifically a method for producing saponin and other active ingredients in the active ginseng in Korea ginseng by plant root culture method It is about.

고려인삼(Panax ginseng C.A. Meyer)은 반음지성 숙근초로서 오가피과(Araliaceae) 인삼속에 속하는 다년생 식물로 인삼의 약리적 효능은 건강한 사람에게는 항상성을 유지해 주며 병약자에게는 항상성을 정상화시켜 준다. 이에 대해 과학적으로 입증되고 있는 약리효과를 보면 간기능 보호작용, 항당뇨기능, 심장강화 및 혈압조절, 항암효과, 뇌기능 강화, 면역기능 강화 등이 있다. 인삼 사포닌은 인삼속 식물에만 함유된 특이한 모형의 담마레인(dammarane)계 트리터펜(triterpene) 배당체로 ginsenoside라고 한다. 인삼사포닌은 사포제닌(sapogenin) 인 프로토파낙사디올(protopanaxadiol)과 프로토파낙사트리올(protopanaxatriol)의 알콜성 -OH기에 글루코스(glucose), 람노스(rhamnose), 자일로스(xylose), 아라비노스(arabinose) 등과 같은 당류가 에테르 결합된 것으로 고려인삼에서는 총 26종의 구조가 밝혀졌다. 사포닌은 인삼 뿐만 아니라, 다른 여러 식물에도 함유되어 있는데, 인삼의 경우 주요 약효성분으로 작용을 하지만 식물에 따라 유해성분으로 작용하는 것도 있다. 인삼 엑기스(extract) 및 사포닌은 다년생 식물인 인삼을 재배하여 수확, 추출하여 제품화하고 있는데 기존의 밭에서 재배한 인삼으로부터 엑기스 및 사포닌을 추출하는 방법은 다음과 같은 단점이 있다.Korean ginseng (Panax ginseng C.A. Meyer) is a perennial plant belonging to the genus Araliaceae ginseng. Scientifically proven pharmacological effects include liver function protection, antidiabetic function, heart strengthening and blood pressure control, anticancer effect, brain function enhancement, and immune function enhancement. Ginseng saponin is a unique type of dammarane triterpene glycoside that is contained only in the genus Ginseng. It is called ginsenoside. Ginseng saponins are the alcoholic -OH groups of saponogenin, protopanaxadiol and protopanaxatriol, including glucose, rhamnose, xylose, and arabinose. Sugars such as arabinose and the like were ether-linked, and a total of 26 structures were identified in Korean ginseng. Saponin is contained not only in ginseng, but also in many other plants. Ginseng acts as a major medicinal ingredient, but some plants act as harmful components. Ginseng extract (extract) and saponins are grown, harvested, extracted and commercialized perennial plant ginseng, the method of extracting the extract and saponins from the ginseng grown in the existing field has the following disadvantages.

첫째, 인삼의 생육기간이 4~6년으로 길며 6년간 재배해야 100~150g(fresh wt)의 수삼을 수확할 수 있다.First, the growth period of ginseng is 4 ~ 6 years long and it needs to be grown for 6 years to harvest 100 ~ 150g (fresh wt) ginseng.

둘째, 연작이 불가능하여 재배면적이 줄어들고 있으며 해가림의 특수한 시설 조건 하에서만 재배가 가능하다.Second, because of the impossibility of planting, the cultivation area is decreasing and it is possible to cultivate only under special facility conditions of the sun.

셋째, 재배조건이 까다롭고 기후, 토양, 병충해 등 환경의 지배를 많이 받아 공급이 안정되지 못하다.Third, supply conditions are not stable due to demanding cultivation conditions and a lot of environmental control such as climate, soil and pests.

넷째, 인삼은 이식법을 이용하여 다년간 재배해야 하므로 병충해 예방을 위하여 농약을 사용하기 때문에 농약잔류의 가능성이 높다.Fourth, because ginseng must be grown for many years using a transplantation method, there is a high possibility of pesticide residues because pesticides are used to prevent pests.

위와 같이 밭에서 재배한 인삼에서 엑기스 및 사포닌을 생산하는데 발생하는 문제를 해결하기 위하여 최근 세포배양에 의해 생산된 인삼 캘러스를 재배인삼 대용으로 사용하기 위한 연구가 꾸준히 진행되어 왔다.In order to solve the problems caused by the production of extract and saponin from the ginseng grown in the field as described above, research has been steadily underway to use the ginseng callus produced by cell culture as a substitute for cultivated ginseng.

그러나, 식물세포배양은 종종 필요성분의 함량이 낮고 생산성의 불안전성 등 몇가지 문제들에 접하게 되는데, 이러한 문제점들을 극복하기 위하여 최근에는 Agrobacterium rhizogenes 감염에 의한 모상근(hairy root) 배양에 대해 관심을 기울이고 있다. 모상근을 이용한 현재까지의 연구에서는 모상근 유도시 토양세균인 Agrobacterium rhizogenes를 감염시켜 형질전환을 일으키도록 하여 모상근을 유도하였는데(일본특허공보 제 소63-254982, 일본특허공보 제 평3-285690호, 일본특허 공보 제 평4-341194호), 형질전환에 의해 모상근을 유도시킬 경우 형질전환을 위해 감염시킨 균을 제거하기 위해서 반코마이신(vancomycin), 카베니실린(carbenicillin), 테트라싸이클린(tetracycline) 등과 같은 항생제를 사용해야 하며, 균을 제거하는데 많은 시간이 필요하고, 균을 죽인 뒤에 다시 배지를 바꾸어주어야 하는 등의 번거로운 과정을 거쳐야 하는 단점이 있다. 또한 외래 유전자인 Ri 플라스미드(Ri plasmid)가 인삼염색체에 무작위로 혼입되므로 이로 인한 인삼유전자의 발현(gene espression)이 달라져 인삼엑기스의 경우 안전성에 대한 검토가 있어야 할 것으로 사료된다.However, plant cell culture often encounters several problems, such as low content of necessary ingredients and instability of productivity. Recently, attention has been paid to hairy root culture by Agrobacterium rhizogenes infection to overcome these problems. To date, studies using hairy roots have induced hairy roots by inducing transformation by infecting soil bacteria, Agrobacterium rhizogenes, to induce hairy roots (Japanese Patent Publication No. 63-254982, Japanese Patent Application Laid-Open No. 3-285690, Japan Patent Publication No. Hei 4-341194), in the case of induction of hairy root by transformation, antibiotics such as vancomycin, carbenicillin, tetracycline, etc. It has to be used, it takes a lot of time to remove the bacteria, and has to go through a cumbersome process, such as changing the medium after killing the bacteria. In addition, the foreign gene Ri plasmid (Ri plasmid) is randomly mixed into the ginseng chromosome, so the gene expression of the ginseng gene is changed, so the safety of the ginseng extract should be considered.

본 발명자들은 여러가지 방법으로 연구를 거듭한 결과, 지금까지 국내외 학계에서 성공사례가 없었던 균의 감염없이 유도된 인삼근 배양에 의해서 인삼 엑기스 및 사포닌을 생산하는 방법을 개발하여 본 발명을 완성하였다.As a result of repeated studies in various methods, the present inventors have developed a method of producing ginseng extract and saponin by cultivating ginseng root without infection of bacteria which have not been successful in domestic and foreign academia.

따라서, 본 발명의 목적은 밭에서 재배한 인삼으로부터 인삼 엑기스 및 사포닌을 추출하는 종래의 기술이 지니고 있던 문제점들을 해결하고, 다량의 인삼 엑시스 및 사포닌을 단기간에 얻을 수 있는 식물조직배양 방법에 의한 고려인삼근의 생산방법을 제공함에 있다.Accordingly, an object of the present invention is to solve the problems of the conventional techniques of extracting ginseng extract and saponin from the ginseng grown in the field, and by the method of plant tissue culture that can obtain a large amount of ginseng extract and saponin in a short time To provide a production method of triceps.

제1도는 인삼 사포닌(saponin)의 구조를 나타낸 것이며,Figure 1 shows the structure of ginseng saponin (saponin),

제2도는 본 발명에 의해 배양한 인삼근내의 사포닌 성분을 HPLC로 나타낸 것이다.Figure 2 shows the saponin components in the ginseng root cultured by the present invention by HPLC.

본 발명의 인삼근 생산방법은 i) 발아된 씨앗 또는 수삼의 절편을 오옥신과 사이토키닌이 혼합된 식물성장호르몬을 함유한 MS 고체배지에 접종하여 캘러스를 유도하는 공정과, ii) 전기 단계에 의해 유도된 캘러스를 오옥신의 함량이 사이토키닌의 함량에 비하여 상대적으로 과량으로 혼합된 식물성장 호르몬을 함유한 인삼근 유도용 MS 고체배지에서 배양하여 인삼근을 유도하는 공정과, iii) 전기 단계에 의해 유도된 인삼근을 오옥신의 함량이 사이토키닌의 함량에 비하여 상대적으로 과량으로 혼합된 식물성장 호르몬을 함유한 인삼근 배양용 SH 액체배지에서 배양하여 인삼근을 증식배양하는 공정과, iv) 전기 단계에 의해 배양된 인삼근을 식물성장 호르몬으로는 오옥신만을 함유하고 있는 SH 액체배지에서 배양함으로써 사포닌 및 기타 인삼성분 생성을 유도하는 사포닌 유도공정을 포함한다.The ginseng root production method of the present invention comprises the steps of i) inoculating germinated seeds or slices of ginseng into MS solid medium containing plant growth hormone mixed with oxine and cytokinin to induce callus; Inducing ginseng root by culturing the callus induced by incubation in MS solid medium for inducing ginseng root containing plant growth hormone mixed with a relatively large amount of oxine compared to the cytokinin content, and iii) the first step Ginseng root-induced growth by culturing in ginseng root cultured SH liquid medium containing plant growth hormone mixed with excessive amount of oxine compared to cytokinin; and iv Saponin and other ginseng components are produced by cultivating ginseng root cultured in the previous step in SH liquid medium containing only oxine as plant growth hormone. Saponin induction process to induce.

상기에서 오옥신의 함량과 사이토키닌의 함량은 각각 2~4mg/ℓ, 0.5~1mg/ℓ이 내인 것이 바람직하나 이들 식물성장호르몬의 첨가량은 당업자가 필요에 따라 선택하여 증감이 가능한 것이므로 본 발명에 있어서는 상기 수치로의 제한을 반드시 요하는 것은 아니다. 다만 오옥신의 함량이 사이토키닌의 함량에 비하여 상대적으로 과량인 요건을 만족시킨다면 식물성장 호르몬의 함량은 실험의 성질에 따라 가변적일 수 있다.In the above, the content of oxine and cytokinin is preferably in the range of 2 to 4 mg / l and 0.5 to 1 mg / l, respectively, but the amount of these plant growth hormones can be selected and increased as needed by those skilled in the art. In this case, the limitation to the numerical value is not necessarily required. However, the content of plant growth hormone may be variable depending on the nature of the experiment if the content of oxin meets the requirement of excessive excess compared to the content of cytokinin.

본 발명에서는 먼저 고려인삼의 씨앗이나 수삼을 살균하고 멸균증류수로 헹군 뒤 식물성장호르몬이 첨가된 발아용 한천평판배지에 심어서 암조건으로 3개월간 배양하여 발아시키거나 근을 유도한다. 발아후, 근성장이 활발한 세포주를 선발하여 오옥신함량이 사이토키닌 함량에 비해 상대적으로 과량인 식물성장호르몬 농도조성을 갖는 배지에 옮겨 계대배양하면서 근생장률이 높은 세포주가 유도된다.In the present invention, first, sterilize the seeds or ginseng of Korean ginseng and rinse with sterile distilled water, and then plant in agar plate medium for germination with plant growth hormone added to incubate for 3 months under cancer conditions to induce germination or muscle. After germination, cell lines with active muscle growth were selected, and the cell lines with high muscle growth rate were induced while passaged to medium having a plant growth hormone concentration composition in which oxine content was relatively excessive compared to cytokinin content.

위와 같이하여 유도된 근을 오옥신 함량이 사이토키닌 함량에 비해 상대적으로 과량인 식물생장호르몬 농도조성을 갖고, 자당, 질소원, 인삼염이 첨가된 근배양용 액체배지에 접종하여 25℃ 배양기에서 암조건으로 근을 증식시켜 이들 근을 수확한다.Roots induced as above were inoculated in a liquid culture medium for cultivation of hormonal growth hormone with an excess amount of oxine compared to cytokinin content, and added to sucrose, nitrogen source, and ginseng salt in a culture medium at 25 ° C. in a dark condition. These muscles are harvested by multiplying them.

이와 같이 증식된 근으로부터 사포닌 형성을 유도하기 위하여 인삼근 배양용 배지조성중 인산칼륨(potassium phosphate)와 항산암모늄(ammonium sulfate)를 사용하여 질소원의 농도를 조정하고, 칼륨염을 사용하여 인산염의 농도를 조정, 에너지원으로 사용되는 자당(sucrose)의 농도를 조정하며, 식물성장 호르몬인 오옥신을 첨가, 배지내 수소이온 농도를 조정하여 제조된 배양배지에서 증식되어 수확된 근을 25℃배양기에서 암조건으로 4주간 배양하여 사포닌을 유도하여 인삼근으로부터 사포닌을 얻는다.Potassium phosphate and ammonium sulfate were used to adjust the concentration of the nitrogen source in the culture of ginseng root culture to induce saponin formation from the proliferated muscles. Adjust the concentration of sucrose used as energy source, adjust the concentration of hydrogen ions in the medium by adding oxine, a plant growth hormone, and adjust the concentration of hydrogen ions in the medium. Incubate for 4 weeks to induce saponin to obtain saponin from ginseng root.

이하, 실시예를 통하여 본 발명을 보다 상세하게 설명한다. 그러나, 본 발명이 이들 실시예에 국한되는 것은 아니다.Hereinafter, the present invention will be described in more detail with reference to Examples. However, the present invention is not limited to these examples.

[실시예 1] 살균 및 캘러스 유도공정Example 1 Sterilization and Callus Induction Process

고려 인삼의 씨앗을 개갑한 것이나, 수삼의 절편을 70% 에탄올 용액과 4% NaOCl 용액으로 살균한 뒤 멸균증류수로 세척한 다음 하기의 표 1과 같은 조성의 평판배지에 식물생장호르몬으로서 1-키네틴(kinetin)과 1-나프탈렌 아세틱 액시드(NAA), 6-벤질아미노퓨린(BAP)과 인돌아세틱액시드(IAA), 그리고 6-벤질아미노퓨린과 1-나프탈렌아세틱액시드 조합으로 첨가된 캘러스 유도용 한천배지에 심어 25℃의 배양기에서 암조건으로 3개월간 배양시켜 캘러스를 유도하였다.The seeds of Korean ginseng were re-opened, and sections of fresh ginseng were sterilized with 70% ethanol solution and 4% NaOCl solution, washed with sterile distilled water, and then 1-kinetine as a plant growth hormone in a plate medium of the composition shown in Table 1 below. callus added with (kinetin), 1-naphthalene acetic acid (NAA), 6-benzylaminopurine (BAP) and indoleacetic acid (IAA), and 6-benzylaminopurine with 1-naphthaleneacetic acid combination Planted in agar medium for induction was incubated for 3 months in a dark condition in an incubator at 25 ℃ to induce callus.

[실시예 2] 인삼근 유도공정Example 2 Ginseng Muscle Induction Process

실시예 1에서 유도된 캘러스를 하기의 표 2와 같은 조성의 배지에 식물성장호르몬으로서 키네틴과 1-나프탈렌아세틱액시드, 6-벤질아미노퓨린과 인돌아세틱액시드, 그리고 6-벤질아미노퓨린과 1-나프탈렌아세틱액시드 조합으로 첨가된 한천배지에 심어 25℃의 배양기에서 암조건으로 3개월간 배양시켜 근을 유도시켰다.Callus derived from Example 1 was kinetin and 1-naphthaleneacetic acid, 6-benzylaminopurine and indoleacetic acid, and 6-benzylaminopurine as plant growth hormone in a medium having a composition as shown in Table 2 below. -Induced muscle by incubating in agar medium at 25 ℃ incubator for 3 months in agar medium added with naphthalene acetic acid combination.

[실시예 3] 인삼근의 증식배양공정Example 3 Growth Process of Ginseng Root

실시예 2에서 유도된 생장이 활발한 근을 표 2의 배지에 식물생장호르몬으로서 6-벤질아미노퓨린과 1-나프탈렌아세틱액시드가 첨가된 액체배지에 옮겨 25℃ 원심 회전 진탕배양기에서 암조건, 60rpm 조건으로 배양하여 4주간격으로 계대배양하였는데, 산택된 세포주의 배증시간(doubling time)은 약 10일이었다.The active growth induced in Example 2 was transferred to a liquid medium to which 6-benzylaminopurine and 1-naphthaleneacetic acid were added as a plant growth hormone in the medium of Table 2, in a 25 ° C. centrifugal rotation incubator, at 60 rpm. Conditions were subcultured at 4 week intervals, and the doubling time of the selected cell lines was about 10 days.

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[실시예 4] 인삼 사포닌 유도배양 공정 (2차 대사산물의 유도 공정)Example 4 Ginseng Saponin Induction Culture Process (Induction Process of Secondary Metabolite)

실시예 3에서 증식하여 수확한 인삼근으로부터 사포닌을 유도하기 위하여, 실시예 2에서의 표 2와 같은 조성의 배지 1ℓ에 자당(sucrose), NH4H2PO4, 질소원, 오옥신을 첨가하고, 배지의 수소이온농도를 조절하여 제조한 인삼 사포닌 유도용 액체배지에 실시예 3에서 증식시켜 수확한 근을 1.5%씩 접종하여 25℃ 원심회전 진탕배양기에서 암조건, 60 rpm으로 4주간 배양한 결과, 생체중량(fresh weight) 2.69 g을 얻어 배양초기보다 약 4.5배 성장하였고, 사포닌 함량은 0.27%로 총 사포닌 함량은 7.26mg 이었다.In order to induce saponins from the ginseng roots grown and harvested in Example 3, sucrose, NH 4 H 2 PO 4 , a nitrogen source, and oxine are added to 1 L of a medium having the composition shown in Table 2 in Example 2, Ginseng saponin induction liquid medium prepared by controlling the hydrogen ion concentration of the medium was inoculated in 1.5% each of the roots harvested by growing in Example 3 and cultured for 4 weeks at 60 rpm in a dark condition centrifugal rotation incubator at 25 ℃ , 2.69 g of fresh weight was obtained to grow about 4.5 times than the initial stage of culture. The saponin content was 0.27% and the total saponin content was 7.26 mg.

위의 실시예와 같은 방법으로 인삼의 근을 배양하여 얻은 사포닌을 정량분석한 결과 도 1과 같은 결과를 얻었으며, 본 발명에 의한 사포닌 성분은 5년근 재배 인삼의 사포닌 성분과 일치하는 인삼 사포닌 임을 확인하였다.As a result of quantitative analysis of saponins obtained by culturing the roots of ginseng in the same manner as in the above example, the result was obtained as shown in FIG. 1, and the saponin component according to the present invention was ginseng saponin which is identical to the saponin component of 5-year-old cultivated ginseng. Confirmed.

본 발명의 생산방법에 의하면 균의 감염없이 유도된 인삼근 배양에 의해서 단기간에 기후, 토양, 병충해 등 자연환경의 지배를 받지 않고 고함량의 사포닌 및 인삼엑기스를 안정적으로 생산할 수 있다.According to the production method of the present invention it is possible to stably produce a high content of saponin and ginseng extracts without being controlled by the natural environment such as climate, soil, pests by the ginseng root culture induced without bacteria infection.

Claims (5)

발아된 씨앗 또는 수삼의 절편을 오옥신과 사이토키닌이 혼합된 식물성장호르몬을 함유한 MS 고체배지에 접종하여 캘러스를 유도하는 공정과, 전기 단계에 의해 유도된 캘러스를 오옥신의 함량이 2~4mg/ℓ와 사이토키닌의 함량이 0.5~1mg/ℓ로 혼합된 식물성장 호르몬을 함유한 인삼근 유도용 MS 고체 배지에서 배양하여 인삼근을 유도하는 공정과, 전기 단계에 의해 유도된 인삼근을 오옥신의 함량이 2~4mg/ℓ와 사이토키닌의 함량이 0.5~1mg/ℓ로 혼합된 식물성장 호르몬을 함유한 인삼근 배양용 SH 액체 배지에서 배양하여 인삼근을 증식배양하는 공정과, 전기 단계에 의해 배양된 인삼근을 식물성장 호르몬으로는 오옥신만을 2~4mg/ℓ 함유하고 있는 SH 액체배지에서 배양함으로써 사포닌 및 기타 인삼성분 생성을 유도하는 사포닌 유도공정을 포함하는 식물조직배양을 이용한 인삼근의 생산방법.Inoculate germinated seeds or fresh ginseng slices into MS solid medium containing plant growth hormone mixed with oxine and cytokinin to induce callus, and callus induced by the previous step contains 2 ~ 4mg of oxine. Induced ginseng root by culturing in MS solid medium for ginseng root inducing plant growth hormone mixed with / l and cytokinin content of 0.5 ~ 1mg / l, and ginseng root induced by electric step The process of proliferating and cultivating ginseng root by culturing in SH liquid medium for cultivating ginseng root containing plant growth hormone mixed with 2 ~ 4mg / l of oxine and 0.5 ~ 1mg / l of cytokine Plant tank including saponin induction process to induce the production of saponins and other ginseng components by culturing the ginseng root cultured by the step in SH liquid medium containing only 2 ~ 4mg / L of oxine as plant growth hormone Method of Production of the ginseng root culture. 제1항에 있어서, 캘러스 유도공정은 암조건에서 25℃로 4주간 배양하는 것을 특징으로 하는 식물조직배양을 이용한 인삼근의 생산방법.The method of claim 1, wherein the callus induction process is cultured for 4 weeks at 25 ° C. under dark conditions. 제1항에 있어서, 상기 증식배양공정은 암조건에서 60rpm의 원심배양으로 4주간 배양하는 것을 특징으로 하는 식물조직배양을 이용한 인삼근의 생산방법.The method of claim 1, wherein the proliferation culture process is a culture of ginseng root using plant tissue culture, characterized in that culture for 4 weeks in a centrifugal culture of 60rpm under dark conditions. 제1항에 있어서, 사포닌 유도공정은 인삼근 배양용 액체배지에 증식배양되어 수확된 근세포를 암조건에서 60rpm으로 원심배양하여 4주간 배양하는 것을 특징으로 하는 식물조직배양을 이용한 인삼근의 생산방법.The method for producing ginseng root using plant tissue culture according to claim 1, wherein the saponin induction process is cultured for 4 weeks by centrifugation of myocytes grown and cultured in liquid medium for ginseng root culture at 60 rpm under dark conditions. . 제1항에 있어서, 증식된 근으로부터 사포닌 형성을 유도하기 위하여 인삼근 배양용 배지조성 중 질소원의 농도를 조성하기 위하여는 인산칼륨과 황산암모늄을 첨가하고, 인삼염의 농도를 조정하기 위하여는 칼륨염을 첨가하며, 에너지원으로는 자당을 첨가하여 주는 것을 특징으로 하는 식물조직배양법을 이용한 인삼근의 생산방법.The method according to claim 1, wherein potassium phosphate and ammonium sulfate are added to form a nitrogen source in the composition of culture medium for ginseng root culture to induce saponin formation from the proliferated muscle, and potassium salt for adjusting the concentration of ginseng salt. The method of producing ginseng root using the plant tissue culture method, characterized in that by adding sucrose as an energy source.
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KR100353636B1 (en) * 2001-01-19 2002-09-28 백기엽 The method for mass production and proliferation of adventitious roots by plant tissue culture in ginseng
KR101377545B1 (en) 2012-09-13 2014-03-25 한방바이오 주식회사 Medium for improving growth of cultured root of ginseng

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KR20010057525A (en) * 1999-12-23 2001-07-04 한상욱 Tissue Culture Method of Wild Ginseng Using Liquid Culture System

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KR890016182A (en) * 1988-04-28 1989-11-28 황영규 Production method of saponin by transformed ginseng cells
KR930000004A (en) * 1991-06-15 1993-01-15 황백 Massive growth of ginseng roots (Chairy roots)

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KR890016182A (en) * 1988-04-28 1989-11-28 황영규 Production method of saponin by transformed ginseng cells
KR930000004A (en) * 1991-06-15 1993-01-15 황백 Massive growth of ginseng roots (Chairy roots)

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Publication number Priority date Publication date Assignee Title
KR100353636B1 (en) * 2001-01-19 2002-09-28 백기엽 The method for mass production and proliferation of adventitious roots by plant tissue culture in ginseng
KR101377545B1 (en) 2012-09-13 2014-03-25 한방바이오 주식회사 Medium for improving growth of cultured root of ginseng

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