KR100710857B1 - Mass production method of high-quality cultured root of wild ginseng - Google Patents

Mass production method of high-quality cultured root of wild ginseng Download PDF

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KR100710857B1
KR100710857B1 KR1020040081476A KR20040081476A KR100710857B1 KR 100710857 B1 KR100710857 B1 KR 100710857B1 KR 1020040081476 A KR1020040081476 A KR 1020040081476A KR 20040081476 A KR20040081476 A KR 20040081476A KR 100710857 B1 KR100710857 B1 KR 100710857B1
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진희옥
김용기
양영국
장석광
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주식회사 진산삼바이오
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Abstract

본 발명은 산삼시료 및 배양 중에 철저한 오염방지와 신선한 O2 및 영양분의 적기 보급을 통하여 인삼사포닌 함량이 높고 품질도 우수한 양질의 산삼배양근을 대량으로 생산할 수 있는 방법을 제공하기 위한 것으로서, 엄선된 산삼시료의 무균화와 MS배지를 이용한 캘러스의 유도, 상기 캘러스를 IBA가 함유된 4/5배지를 이용하여 산삼배양근을 배양함에 있어서, 산삼시료의 무균화는 멸균수에 2% 차아염소산소다가 첨가된 용액에 침지한 후 70% 에탄올이 투입된 진공감압세척기에서의 살균으로 진행하고, 캘로스의 유도는 초음파로 진동시켜 진행하며, 배양 중간에 O2를 공급하며, IBA가 함유된 배지에 인삼조사포닌 함량증가촉진제를 첨가하고, 배양 5∼6주의 pH를 5.8로 조절함과 동시에 4/5 MS 배지를 투입하여 배양하는 고품질 산삼배양근 대량 생산방법을 특징으로 한다.The present invention is to provide a method for mass production of high quality ginseng saponin and high quality ginseng culture roots through thorough pollution prevention and timely diffusion of fresh O 2 and nutrients during wild ginseng samples and cultures. Sterilization of wild ginseng samples by adding 2% sodium hypochlorite to sterile water in sterilization of samples and induction of callus using MS medium, and cultivating wild ginseng cultured roots using 4/5 medium containing IBA. After immersing in the prepared solution and proceeding to sterilization in a vacuum pressure washer with 70% ethanol, the induction of callus is vibrated by ultrasonic vibration, supplying O 2 in the middle of the culture, ginseng tank in the medium containing IBA A method of mass production of high-quality wild ginseng cultured roots was added by adding a saponin increase accelerator, adjusting the pH of the culture for 5 to 6 weeks to 5.8, and adding 4/5 MS medium. It features.

Description

고품질 산삼배양근(부정근)의 대량 생산방법{Mass production method of high-quality cultured root of wild ginseng}Mass production method of high-quality cultured root of wild ginseng}

본 발명은 산삼배양근(부정근)을 배양할 때 철저한 오염방지로 품질을 높이고, 생산량 극대화도 달성할 수 있는 고품질 산삼배양근의 대량 생산방법에 관한 것이다.The present invention relates to a mass production method of high-quality wild ginseng cultured root which can improve the quality by thorough pollution prevention and achieve maximum production when cultivating wild ginseng cultured (negative muscle).

지금까지는 특정 기계와 기구를 이용하여 산삼배양근(부정근)을 배양해 왔다. 실제로 산삼배양근을 대량으로 배양했을 때 상품가치가 있는 산삼배양근의 최종 수확량은 이론치와는 달리 매우 낮다. 낙후된 배양기술과 온도, 습도, 계절별 환경변화 등 생장환경변화에 유효 적절히 대처하지 못한 것도 원인이지만, 보다 근본적인 문제는 미생물 등에 의한 산삼시료 및 배양 중의 미생물 감염과 성장에 필요한 영양분의 부족, pH 조절 실패 등이다.Up to now, wild ginseng cultured muscles have been cultivated using specific machinery and equipment. In fact, when ginseng cultured in large quantities, the final yield of commodity-priced wild ginseng was very low. It is caused by the lack of effective coping with growth conditions such as cultivation technology and temperature, humidity, and seasonal environmental changes, but more fundamental problems are the lack of nutrients necessary for microbial infection and growth in microorganisms and growth and pH control by microorganisms. Failure.

따라서 본 발명의 목적은 산삼시료 및 배양 중에 철저한 오염방지와 신선한 O2 및 영양분의 적기 보급을 통하여 인삼사포닌 함량이 높고 품질도 우수한 양질의 산삼배양근을 대량으로 생산할 수 있는 방법을 제공하는 것이다. Therefore, an object of the present invention is to provide a method for producing a large amount of high quality ginseng saponin and high quality ginseng cultured roots through a thorough prevention of wild ginseng samples and cultivation of fresh O 2 and nutrients in a timely manner.

상기의 과제는, 엄선된 산삼시료의 무균화와 MS배지를 이용한 캘러스의 유도, 상기 캘러스를 IBA가 함유된 4/5배지를 이용하여 산삼배양근을 배양함에 있어서, 산삼시료의 무균화는 멸균수에 2% 차아염소산소다가 첨가된 용액에 침지한 후 70% 에탄올이 투입된 진공감압세척기에서의 살균으로 진행하고, 캘로스의 유도는 초음파로 진동시켜 진행하며, 배양 중간에 O2를 공급하며, IBA가 함유된 배지에 인삼조사포닌 함량증가촉진제를 첨가하고, 배양 5∼6주의 pH를 5.8로 조절함과 동시에 4/5 MS 배지를 투입하여 배양하는 고품질 산삼배양근 대량 생산방법으로 달성할 수 있다.
The above problem is the sterilization of wild ginseng samples in the sterilization of wild ginseng samples in sterilization of selected wild ginseng samples and induction of callus using MS medium, and in cultivating wild ginseng culture roots using 4/5 medium containing IBA. After immersing in a solution to which 2% sodium hypochlorite was added to the solution, 70% ethanol was added to sterilization in a vacuum pressure washer, induction of callus was vibrated by ultrasonic vibration, and O 2 was supplied in the middle of the culture. It can be achieved by mass production method of high quality wild ginseng cultured root cultured by adding ginseng irradiated ponin increasing agent to IBA-containing medium, adjusting pH of cultivated 5-6 weeks to 5.8 and adding 4/5 MS medium. .

배양할 산삼의 시료는 강원도 오대산에서 채취한 자연산 야생 산삼으로서 산삼 감정 인증된 것으로 한다. 생산단계는 산삼의 소독, 캘러스(미원시분화세포괴)유도, 액체배양, 산삼배양근(부정근)의 수확 등으로 이뤄진다.The sample of wild ginseng to be cultured is wild wild ginseng collected from Odaesan, Gangwon-do. The production stage consists of disinfection of wild ginseng, induction of callus, and culture of liquid and harvesting wild ginseng root.

산삼시료 소독방법은 산삼의 표면과 뿌리사이의 이물질을 흐르는 수돗물에 깨끗이 세척하고, 멸균수로 3회 이상 세척한다.The method of sterilizing wild ginseng sample is to wash the foreign substances between the surface and the root of wild ginseng in tap water and clean it more than 3 times with sterile water.

산삼을 비이커에 넣고 멸균수와 2%의 차아염소산소다(NaOCl)를 넣어 교반기에서 약 10분간 교반한 후, 멸균수로 3회 이상 세척하는 방법과, 재세척된 산삼 시 료를 클린벤치 내에서 진공감압세척기에 넣고 70% 에탄올을 첨가하고 진공 감압하여 기포를 발생시키면서 뿌리 사이에 존재하는 미생물 및 기타 오염물질을 완전히 제거하고 약 15분간 지속시키는 방법으로 한다.Put the wild ginseng into the beaker, add sterilized water and 2% sodium hypochlorite (NaOCl) and stir in the stirrer for about 10 minutes, wash it three times or more with sterilized water, and clean the rewashed wild ginseng sample in the clean bench. Place it in a vacuum pressure washer and add 70% ethanol and decompress in vacuum to create bubbles while completely removing microorganisms and other contaminants between roots and lasting for about 15 minutes.

산삼 시료를 빼내어 멸균수로 3회 이상 세척한다.Remove the wild ginseng sample and wash it three times or more with sterile water.

상기한 소독방법에다 2% 차아염소산소다, 또는 70% 에틸렌알콜과 진공감압세척방법을 적용한 산삼 시료의 오염도 측정결과는 75%와 35%로 큰 차이가 있음을 확인하였다.Contamination of wild ginseng samples using 2% sodium hypochlorite, or 70% ethylene alcohol and vacuum vacuum cleaning method was 75% and 35%.

<표1> 소독방법에 따른 오염도 비교<Table 1> Comparison of Pollution Degrees According to Disinfection Methods

소독액       Disinfectant 소독방법Disinfection Method 오염도(%)Pollution degree (%) 차아염소산      Hypochlorous acid 2% 차아염소산소다용액에 5∼10분 침지 Soak 5-10 minutes in 2% sodium hypochlorite solution 75%   75% 차아염소산,70% 에탄올Hypochlorous acid, 70% ethanol 2% 차아염소산소다용액으로 5∼10분 침지 후 70% 에탄올이 첨가된 진공감압기에서 2∼5분 살균 5 to 10 minutes immersion with 2% sodium hypochlorite solution and sterilization for 2 to 5 minutes in a vacuum reducer with 70% ethanol 35%   35%

산삼 시료의 배양비지 조제는 산삼 시료의 조직배양에 보편적으로 사용되는 MS배지의 4/5MS배지를 만들고 설탕 3%와 한천 0.8%를 첨가하였으며, 식물생장호르몬으로는 오옥신류중에서 1BA 1ppm 및/또는 코코넛밀크를 추가 첨가하고 멸균처리 후 클린벤치에서 멸균된 페트리 디쉬(Petri-dish)에 20ml씩 분주하여 사용하였다.Preparation of culture ginseng of wild ginseng sample was made of 4/5 MS medium of MS medium which is commonly used for tissue culture of wild ginseng sample, and 3% of sugar and 0.8% of agar were added. As plant growth hormone, 1ppm of 1BA and / or Coconut milk was further added and sterilized, and then, 20 ml of petri dish was sterilized in a clean bench.

산삼 시료는 클린벤치 내에서 멸균된 유리접시에 넣고 멸균된 메스로 뿌리의 껍질을 약간 벗겨낸 후 2∼3㎜ 크기로 절단하여 배양배지가 있는 페트리 디쉬에 4∼5개씩 치상하여 파라필름(parafilm)으로 밀봉한 후 실온 23℃의 인큐베이터에서 암(暗)배양한다.The wild ginseng sample is placed in a sterilized glass dish in a clean bench, and the roots are peeled off slightly with a sterilized scalpel, cut into 2 to 3 mm in size, and then cut into 4 to 5 pieces in a petri dish containing culture medium. ) And then incubated in an incubator at room temperature of 23 ° C.

산삼 시료의 페트리 디쉬의 1차 확인 및 재소독은 3∼5일 경과 후 인큐베이 터에서 배양된 페트리 디쉬를 꺼내어 육안으로 세균 및 곰팡이 등에 의한 오염상태 등을 확인한다.For the first verification and re-sterilization of petri dishes of wild ginseng samples, after 3-5 days, petri dishes cultured in an incubator are taken out and visually checked for contamination by bacteria and mold.

오염된 배양조직 산삼 시료를 꺼내어 2% 차아염소산소다용액에서 약 10분간 침지하여 소독한 후 멸균수로 3회 이상 세척하고 새로운 배지에다 재차 치상하여 인큐베이터 내에서 배양하는 방법으로 반복 확인하면서 배양오염을 줄이고, 캘러스를 유도과정 중에는 배양된 페트리 디쉬를 꺼내어 초음파를 3일 간격으로 15∼20분 간 68KHz 이상 3∼4주간 진동하면서 캘러스를 유도한다.Take out the contaminated culture ginseng sample and soak it in 2% sodium hypochlorite solution for about 10 minutes, disinfect it, wash it three times or more with sterile water and inoculate it again in a fresh medium and repeat the culture in an incubator. During the callus induction process, the cultured petri dishes are taken out and the callus is induced while the ultrasound vibrates for 3 to 4 weeks at 68 KHz for 15 to 20 minutes at 3 days intervals.

캘러스(미원시분화세포괴)가 유도된 산삼 시료를 인큐베이터에서 꺼내어 양호한 캘러스를 선발하고, 선발된 캘러스를 클린벤치 내에서 멸균된 메스로 0.1㎝ 미만으로 잘게 자른 후 4/5MS배지에 접종한다. 또는 액체 현탁액 배지에 접종하여 2∼3주 후 접종된 4/5MS배지와 액체 현탁액 배지에서 산삼배양근이 유도된 것을 선별하여 메스로 0.2㎝ 길이로 자른 후 멸균된 4/5MS배지에 있는 액체 현탁액 배지 2ℓ용 유리용기에 접종한다.The wild ginseng samples derived from callus (U.S. differentiated cell mass) were taken out of the incubator to select good callus, and the selected callus was chopped to less than 0.1 cm with a sterile scalpel in a cleanbench and then inoculated into 4 / 5MS medium. Alternatively, inoculated in liquid suspension medium 2 to 3 weeks later, the inoculated 4/5 MS medium and the ginseng cultured root derived from the liquid suspension medium were cut into 0.2 cm lengths with a scalpel, and then the liquid suspension medium in sterilized 4/5 MS medium. Inoculate 2 liter glass containers.

2주일 경과 후, 산삼배양근이 유도된 것을 페트리 디쉬에서 꺼내어 클린벤치에서 우수한 배양근을 멸균된 메스로 흰색, 노란색 부분을 0.5㎝ 이하의 길이로 잘라내 4/5MS 액체 배지에 넣어서 배양한다.After 2 weeks, the induction of wild ginseng culture roots was removed from the petri dish, and the excellent culture roots were removed from the cleanbench with a sterile scalpel and cut to a length of 0.5 cm or less and incubated in 4 / 5MS liquid medium.

배양은 자르는 방법과 찢어서 풀어헤치는 방법을 병행하였으며, 배양 시 메스로 자르는 방법의 작업소요시간은 17∼19분이고, 집게로 찢고 풀어헤치는 방법의 작업소요시간은 10∼12분이어서 집게로 찢고 풀어헤치는 방법이 메스로 절단하는 방법보다 7분 가량 시간이 단축되었고, 미생물 오염도도 30% 보다 적은 17%로 감소 하였지만 성장 후 산삼배양근의 수거량에 있어서는 10∼12배로 현격한 차이가 나타났다. Cultivation is a combination of cutting and tearing and releasing, and the working time of cutting with a scalpel during culture is 17-19 minutes, and the working time of tearing and releasing with forceps is 10-12 minutes. The time was 7 minutes shorter than the method of cutting with scalpel, and microbial contamination was reduced to 17%, less than 30%.

<표2> 산삼근 배양시 자르는 방법과 찢어 풀어헤치는 방법의 생산량 비교<Table 2> Production Comparison of Cutting and Tearing Methods in Cultured Wild Ginseng Roots

절단방법Cutting method 실험수량(g)Experimental quantity (g) 작업소요시간(분)Work time required (minutes) 오염도(%)Pollution degree (%) 성장 후 수거량(g)Harvest after growth (g) 메스로 자르는 방법How to cut with a scalpel 5050 17-1917-19 3030 600-700(12배)600-700 (12x) 집게로 찢고 풀어헤치는 방법How to tear and loosen with tongs 5050 10-1210-12 1717 500-600(10배)500-600 (10x)

식물생장조절물질인 IBA 1ppm + 0.05g/ℓ, 0.1g/ℓ, 0.5g/ℓ의 키토산(키토올리고당) 첨가방법, IBA 1ppm + 0.05g/ℓ, 0.1g/ℓ의 크로렐라 첨가방법, IBA 1ppm + 0.1g/ℓ키토산 + 0.05g/ℓ 크로렐라 + 0.5g/ℓ 코코넛 밀크 첨가방법으로 급속배양통에 함께 넣고 배양하였다. IBA 1ppm + 0.05g / l, 0.1g / l, 0.5g / l chitosan (chitooligosaccharide) addition method, IBA 1ppm + 0.05g / l, 0.1g / l croella addition method, IBA 1ppm + 0.1 g / l chitosan + 0.05 g / l chlorella + 0.5 g / l coconut milk was added together and cultured in a rapid culture vessel.

IBA 1ppm + 0.05g/ℓ, 0.1g/ℓ의 크로렐라 첨가방법은 평균 생산수거량이 750과 780이었고 향은 약하고 엷은 녹색을 띠었으며, IBA 1ppm + 0.1g/ℓ키토산 + 0.05g/ℓ 크로렐라 + 0.5g/ℓ 코코넛 밀크 첨가방법의 산삼배양근 평균 생산수거량은 850이었고 향은 약하고 엷은 녹색을 띈 반면, IBA 1ppm + 0.1g/ℓ의 키토산 첨가방법은 산삼배양근의 평균 생산수거량이 920으로서 아주 높았고 향도 진한데다 색상도 진한 밤색으로 나타나 품질이 가장 우수한 것으로 나타났다. Cropped IBA 1ppm + 0.05g / l, 0.1g / l, the average production yield was 750 and 780, the fragrance was weak and pale green, IBA 1ppm + 0.1g / l chitosan + 0.05g / l chlorella + 0.5 The ginseng root production of ginseng roots was 850, and the fragrance was weak and light green. In addition, the color is also dark brown, the highest quality.

<표3> 4/5MS배지에 영양원 첨가시 생산량 비교 <Table 3> Production Comparison of Nutrient Added to 4 / 5MS Medium

배지badge 기본basic 첨가영양원Additive Nutrition 실험수량 (개)Experimental quantity (pcs) 수거 후 평균생산량(g)Average yield after collection (g) 특징 Characteristic 4/5배지4/5 badge IBA 1ppmIBA 1 ppm +0.05g/l 키토산 +0.05 g / l chitosan 1010 890890 조금 약한 향, 발색 Slightly weak scent, color development +0.01g/l 키토산 +0.01 g / l chitosan 1010 920920 진한 향, 진한 밤색 Dark scent, dark brown +0.5g/l 키토산 +0.5 g / l chitosan 1010 900900 진한 향, 진한 밤색 Dark scent, dark brown +0.05g/l 크로렐라 +0.05 g / l chlorella 1010 750750 약한 향, 녹색 Weak fragrance, green +0.1g/l 크로렐라 +0.1 g / l chlorella 1010 780780 약한 향, 녹색 Weak fragrance, green +0.1g/l 키토산 +0.05g/l 크로렐라 +0.5g/l 코코넛밀크 + 0.1g / l Chitosan + 0.05g / l Chlorella + 0.5g / l Coconut Milk 1010 850850 다소 약한 향, 엷은 녹색 Somewhat weak scent, pale green

또한 1BA 1ppm 첨가와 IBA 1ppm + 0.1g/ℓ 키토산 첨가방법으로 배양하여 생산한 후 인삼조사포닌 함량성분을 분석한 결과, IBA 1ppm 첨가방법으로 배양한 산삼배양근의 인삼조사포닌의 평균 함량은 75㎎/g으로서 5% 이상 증가추세를 나타냈다. The content of ginseng irradiated ponson ginseng cultured by the addition of 1ppm 1BA and 1ppm + 0.1g / ℓ chitosan was analyzed, and the average content of ginseng irradiated root cultured by the addition of IBA 1ppm was 75mg. / g increased more than 5%.

<표4> 영양원 첨가후 인삼조사포닌 함량비교 <Table 4> Comparison of Ginseng Irradiated Phonine Contents after Addition of Nutrition Sources

배지       badge 첨가영양원Additive Nutrition 인삼조사포닌 함량(㎎/g)Ginseng Survey Ponine Content (mg / g) 4/5 MS 배지4/5 MS Badge IBA 1ppmIBA 1 ppm 68∼82(75)68-82 (75) 4/% MS 배지4 /% MS medium IBA 1ppm + 0.1g/l 키토산IBA 1 ppm + 0.1 g / l chitosan 74∼83(78.2)74 to 83 (78.2)

급속배양통에서 산소발생기를 이용하여 산소농도 35% 이상 정제된 O2를 배양통 내부 하단에 1주일 간격으로 25분 주입하는 방법과, 초음파 기계로 1주일 간격으로 배양통을 60∼80 KHz로 25분 이상 진동하였다.Method of injecting O 2 purified at least 35% oxygen concentration into the bottom of the culture vessel for 25 minutes at a weekly interval by using an oxygen generator in a rapid culture vessel and 60-80 KHz at a weekly interval with an ultrasonic machine. It vibrated more than 25 minutes.

배양 30일 경과 후 O2 주입방법과 초음파 진동방법을 확인 후 비교한 결과, O2 주입에 의한 산삼배양근의 성장은 평균 420g이었고 초음파 진동에 의한 산삼배양근의 성장은 평균 510g으로 나타나서 초음파 진동쪽이 O2 주입보다 90g 많아 17.3%의 중량증가사실을 확인하였다. After 30 days of cultivation, after comparing O 2 injection method and ultrasonic vibration method, the growth rate of wild ginseng cultured roots by O 2 injection was 420g and the growth of wild ginseng cultured roots by ultrasonic vibration was 510g. It was confirmed that the weight increase of 17.3% more than 90g than O 2 injection.

<표5> O2 주입과 초음파 진동의 비교Table 5 Comparison of O 2 Injection and Ultrasonic Vibration

주입원Injection 실험수량 (개)Experimental quantity (pcs) 위치별 방법Location-specific method 조건방법Condition method O2함유량 (%)O 2 content (%) 주입시간 (분)Injection time (minutes) 성장무게 (g)Growth weight (g) O2 O 2 5050 배양통 내부 하단에 주입Injection into bottom of culture vessel 1∼8(ℓ/min)1 to 8 (ℓ / min) 3535 2525 420420 초음파ultrasonic wave 5050 배양통 주위에 진동방법Vibration method around culture vessel 68∼80KHz68 ~ 80KHz 00 2525 510510

성장과정에서는 배양통내에서 산삼배양근의 위치에 따라 성장도에 큰 차이가 나타나게 되므로 공기량을 조절하여 배양액의 하단부에서 산삼배양근이 성장하게 하는 방법과 배양액 속 상단부에서 산삼배양근이 성장하게 하는 방법을 비교 실험하였다.In the growth process, there is a big difference in the growth rate according to the location of wild ginseng roots in the culture vessel. It was.

비교 실험결과, 산삼배양근의 생산량은 상단부에서 자라는 방법의 경우 성장 후 산삼배양근의 수거량은 720g였고, 하단부에서 자라게 한 방법의 경우 성장 후의 산삼배양근 수거량은 780g으로 하단부에서 자라는 방법이 12%의 증대효과가 나타났다. 이는 As a result of the comparative experiments, the production of wild ginseng roots was 720g in the case of the method of growing in the upper part, and the harvest of wild ginseng roots in the method of growing in the lower part was 780g. Appeared. this is

<표6> 배양통속의 배양근의 위치별 생산량 비교<Table 6> Comparison of production volume by location of culture roots

배양 위치Culture location 실험 수량(통)Experiment quantity (bucks) 성장후 수거량(g/통)Post-growth harvest (g / bag) 증대효과(%)Increase effect (%) 상단 부분Upper part 5050 720720 -- 하단 부분Bottom part 5050 780780 1212

금속배양통에서 배양할 때, pH증가와 영양분의 감소로 인하여 5∼6주 중에 pH 6.5 이상 상승하여 성장이 중단되는 경우가 있다. 그래서 식용목초액 또는 식용초산을 첨가 주입하여 pH 5.8 내외로 조절하고, 영양분도 4/5 MS 배지를 20% 투입하여 8주 경과 후의 상태를 비교 실험하였다.When cultured in metal culture vessels, growth may be stopped due to an increase of pH 6.5 or more in 5-6 weeks due to the increase of pH and nutrients. Thus, the mixture was added with edible vinegar or edible acetic acid to adjust the pH to around 5.8, and 20% of nutrient content was added to 4/5 MS medium.

비교 실험 후의 산삼배양근의 생산량은 pH와 영양분을 조절하지 않은 경우의Production of Wild Ginseng Roots after Comparative Experiments without pH and Nutrients

산삼배양근 생산수거량은 750g이었고, pH 조절과 영양분을 투입한 경우의 산삼배양근 생산수거량은 850g으로서, pH 조절 및 영양분 투입방법이 13.3% 증가추세를 보였다. The production of wild ginseng roots was 750g, and the production of wild ginseng roots was 850g when pH control and nutrients were added.

<표7> 5∼6주 배양 후 pH조절한 경우의 수거된 산삼배양근의 생산량 비교<Table 7> Production of harvested wild ginseng cultured roots after pH adjustment after 5-6 weeks of culture

pHpH 실험수량(통)Experiment volume 성장기간(주)Growth period 성장방법How to grow 생산수량(g/통)Production quantity (g / bag) 비고(%)Remarks (%) 조절하지 않음Do not adjust 3030 88 6주까지 성장후 멈춤Growing up to 6 weeks 750750 -- pH조절, 영양분투입pH control, nutrition 3030 88 8주까지 지속적으로 성장Continuous growth up to 8 weeks 850850 13.3%13.3%

급속배양통에서 8주 배양한 후에 산삼배양근을 꺼내어 수세하고 배양배지를 제거하여 포장단위별 용기에 포장하고 건조기에서 동결건조 또는 열풍건조한다. After 8 weeks of incubation in a rapid culture vessel, take out wild ginseng roots, wash them, remove the culture medium, pack them in containers for each packaging unit, and lyophilize or dry with hot air in a dryer.

이상 설명한대로 산삼배양근을 배양함에 있어서 본 발명은 산삼시료의 철저한 오염방지와 배양 중 신선한 O2 및 영양분의 적기 보충을 통하여 인삼사포닌 함량이 높고 품질도 우수한 양질의 산삼배양근을 대량으로 생산할 수 있다. As described above, in cultivating wild ginseng culture root, the present invention can produce high quality ginseng saponin and high quality wild ginseng culture root through thorough prevention of wild ginseng sample and timely supplementation of fresh O 2 and nutrients during cultivation.

Claims (3)

삭제delete 엄선된 산삼시료의 무균화, MS배지를 이용한 캘러스의 유도, IBA가 함유된 4/5배지상에서의 캘러스의 배양, 배양 중의 산소공급, 배양 5~6주의 pH5.8 조절로 산삼배양근을 배양함에 있어서, 상기 4/5배지에 키토산 0.05~0.5g/l, 크롤렐라 0.05~0.1g/l, 코코넛 밀크 0.5g/l 중의 어느 1종으로 된 인삼조사포닌 함량증가 촉진제를 첨가하여 배양하는 것을 특징으로 하는 고품질 산삼배양근 대량 생산방법. Sterilization of selected ginseng samples, induction of callus using MS medium, culture of callus on 4/5 medium containing IBA, oxygen supply during culture, and cultivation of wild ginseng root with pH5.8 control of culture for 5-6 weeks In the 4/5 medium, chitosan 0.05 ~ 0.5g / l, crawlella 0.05 ~ 0.1g / l, Coconut milk 0.5 g / l of any one of the ginseng irradiated increase in phosphorus content is added to culture Production method of high quality wild ginseng root. 삭제delete
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KR100318091B1 (en) * 2001-06-02 2001-12-22 대한민국(관리청:특허청장, 승계청:산림청 임업연구원장) Mass propagation method of adventitious root mountain grown ginseng using the bioreactor culture
KR20040050317A (en) * 2002-12-10 2004-06-16 주식회사 씨비엔바이오텍 Method of bioreactor culture of adventitious roots

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