Production of Anthocyanins by Vitis Hybrid Cell Culture

Vitis Hybrid 세포배양에 의한 Anthocyanin의 생산

The induction of calli from tissues of a grape, Vitis hybrid, and their suspension cultures were performed and various factors were investigated on cell growth and anthocyanin production. It was shown that light intensity and inorganic nitrogen concentration played an important role on anthocyanin production.1:he contents of anthocyanin produced under 10,000 Iux light irradiation were about twice as much as under the dark. The reduction of inorganic nitrogen concentration of MS medium to one to twenty brought about the increase of approximately five to six-fold in total anthocyanin or sixteenfold in anthocyanin content per dry cell weight and addition of nitrate only as inorganic nitrogen source was shown to be the best for anthocyanin production. Miller medium and Gamborg medium were suitable for the anthocyanin production, as well as high concentrations of Co$^{2+}$ and Fe$^{2+}$. And high yield of 40mg anthocyanins per 200m1 flask was obtained by two stage culture using MS medium for the first stage and the modified MS medium for the second stage.

착색용 포도품종(Vitis hybrid)의 조직으로부터 callus를 유도하고 액체 진탕배양을 행하여 세포 생육 및 anthocyanin의 생성에 미치는 각종 인자들에 대하여 조사한 결과 광조사량 및 무기질소원 농도가 가장 큰 영향을 미치는 것으로 나타났다. 즉, 광조사량이 10,000lux 였을 때는 암소에서 배양했을 때보다 anthocyanin 생성량은 약 2배 증가하였다. 또한 기존 MS 배지의 무기질소원 농도를 20배로 감소시켜 배양했을 때 전체 anthocyanin 생성량은 약 5-6배, 건조 중량당 함량은 약 16배 증가함을 알 수 있었다. 또한 무기질소원으로 nitrate만을 첨가했을 때 가장 높은 수율을 나타냈다. 고농도의 Co$^{2+}$, Fe$^{2+}$와 Miller 배지, Gamborg 배지 등이 anthocyanin 생산에 적합한 것으로 나타났으며, MS medium 및 modified MS medium을 이용한 2단계 배양을 시도하여 약 40mg/flask의 높은 anthocyanin 수율을 얻을 수 있었다.