| Abstract: | 本研究利用植物組織培養法對紅心萊菔 ( Raphanus sativus )及紫蘇 ( Perilla frutescens ) 探討癒合組織誘發、細胞生長及花青素生產之的 影響，並且對植物體及癒合組織所含色素進行分析及安定性之測試。不同 植物生長調節劑對紅心萊菔塊根及葉片培植體癒合組織誘發程度以添加 1μM 2,4-D 及 0.1 μM kinetin 最佳；塊根培植體中，以 4 號表現最 好。在細胞生長方面，以添加 3% 蔗糖、30 mM 氮源最好。塊根花青素於 95℃ 加熱 60 分鐘時，以 1 號有最好的熱安定性，殘留率為 117%；5 號具有良好的紫外光安定度，照射 5 小時後，殘留率為 107%。葉片培植 體癒合組織花青素 95℃ 加熱 60 分鐘時，以 1、4 及 5 號仍有超過 80% 的殘留；紫外光安定度以 1 號最佳。紫蘇組織培養方面，葉片及莖 培植體以添加 1μM 2,4-D 及 1μM kinetin 可得到最大癒合組織誘發程 度。癒合組織細胞的生長最佳條件為 3% 蔗糖，氮源濃度為 30 mM，溫度 為 25 ~ 30℃ 之間；培養基起始 pH 值於 4.7 ~ 6.7 對紫蘇細胞生長影 響不明顯。紫蘇花青素在加熱 15、30 及 60 分鐘後，殘留率分別為 71.5%、69.9% 及 64.6%；在紫外光照射 1、2、3 及 5 小時後，殘留率 分別為 93%、88.7%、 83.2% 及 70.8%。
The initiation and growth of calli and the production of anthocyanins by the tissue culture of red- tuber Raphanua sativus and Perilla frutescens were studied. The stability, quality and content of the red pigments in the explant and calli initiated from the explant were also investigated.The addition of 1μM 2,4-D and 0.1μM kinetin induced more calli than other combination from the tuber and leaf explant of Raphanus sativus. The best result was found in tuber explant of strain No. 4. Subcultured calli grew well in the MS medium containing 3% sucrose and 30 mM nitrogen. After being heated under 95℃ for 60 minutes, the anthocyanins extracted from the tuber explant of strain No. 1showed the best stability with a retention of 117%, while those from strain No. 5 showed the best UV stability with a retention of 107% after being exposed under UV for 5 hours. The retention of anthocyanins of calli initiated from leaf explant of No. 4, 5, 1 was over 80% after being heated for 60 minutes at 95℃. The anthocyanins of calli from strain No.1 showed the best UV stability.For the tissue culture of Perilla frutescens, the addition of 1μM 2,4-D and 1μM kinetin induced more calli than other combination on leaf and stem explant. Well-growth calli were obtained in the MS medium containing 3% and 30 mM nitrogen, and the culturing temperature was among 25~30℃. The initial pH in the range of pH 4.7~6.7 not significantly affected the cell growth. The retention of anthocyanins from leaves was 71.5%, 69.9% and 64.6% after 15, 30 and 60 minutes at 95℃, and 93%, 88.7%, 83.2%, 70.8% after being exposed for 1, 2, 3 and 5 hours under UV. |
