巴西鳶尾（Neomarica gracilis）根莖組織培養及其抗氧化力與鳶尾異黃酮分析

THE RHIZOME TISSUE CULTURE AND ANALYSIS OF ANTIOXIDATION PROPERTY AND TECTORIGENIN OF NEOMARICA GRACILIS

謝幸君(Hsing-Chun Hsieh)

指導教授 ： 何錦玟

大同大學/工程學院/生物工程學系所/碩士(2006年)

摘要

巴西鳶尾(Neomarica gracilis)之試管內實生苗，於含有MS為基本鹽類添加30 g/l蔗糖、0.5 mg/l BA、0.84 mg/l NAA、0.1 mg/l 2,4-D之液態培養基中，可誘得生長快速的根莖組織，再以此根莖組織為培植體，繼代培養於相同培養基，平均每30天其組織之鮮重增加率可達4倍。
根莖組織分別繼代培養於含不同濃度茉莉酸(jasmonic acid)、苯丙胺酸(l-phenylalanine)及酵母萃取物(yeast extract)之修飾過MS液態培養基中，以探討培養基中添加有機物對於巴西鳶尾根莖組織之抗氧化力、總黃酮及鳶尾異黃酮含量之影響。在含不同濃度酵母萃取物之修飾過MS液態培養基中，可增加根莖組織之自由基清除能力(radical scavenging activity)。添加100 ~ 400 mg/l的酵母萃取物培養之根莖組織甲醇萃取液，當系統濃度為5 mg DW /ml時，其自由基清除能力可達到90 %。而在含不同濃度苯丙胺酸(l-phenylalanine)之修飾過MS液態培養基中，可促進總黃酮(total flavonoid)及鳶尾異黃酮(tectorigenin)在根莖組織中的累積量。其中以添加300 mg/l苯丙胺酸之MS修飾過液態培養基，根莖組織之甲醇萃取液的總黃酮量達到最高(1.059 % DW)。在鳶尾異黃酮方面，則以根莖組織培養於MS為基本鹽類添加200 mg/l苯丙胺酸之液態培養基，根莖組織之甲醇萃取液的鳶尾異黃酮達到最高(0.00427 % DW)。而田間種植的巴西鳶尾根莖無法測出鳶尾異黃酮含量。

關鍵字

抗氧化力 ； 根莖組織培養 ； 鳶尾異黃酮 ； 巴西鳶尾

並列摘要

Rhizomes were induced from in vitro seedlings of Neomarica gracilis cultured in MS basal medium contained with 30 g/l sucrose, 0.5 mg/l BA, 0.84 mg/l NAA and 0.1 mg/l 2,4-D. Fast growing tissue obtained from the rhizome tissues were subcultured in the same liquid medium and the multiplication of rhizome tissues were 4-fold after 30 days culture. The influence of jasmonic acid, l-phenylalanine and yeast extract addition on the radical scavenging activity, total flavonoid amount and tectorigenin accumulation in rhizome tissues were investigated.
The radical scavenging activity under system concentration 5 mg DW/ml can increase to 90 % in methanol extract from rhizome tissues subcultured in MS basal medium contained with 30 g/l sucrose, 0.5 mg/l BA, 0.84 mg/l NAA, 0.1 mg/l 2,4-D and yeast extract (100 ~ 400 mg/l).
The total flavonoid amount and tectorigenin accumulation in cell increased in methanol extract from rhizome tissues subcultured in MS basal medium contained l-phenylalanine. The maximum total flavonoid reached 1.059 % DW, when rhizome tissues subcultured in MS basal medium contained with 30 g/l sucrose, 0.5 mg/l BA, 0.84 mg/l NAA, 0.1 mg/l 2,4-D and 300 mg/l l-phenylalanine. The rhizome tissues subcultured in MS basal medium contained with 30 g/l sucrose, 0.5 mg/l BA, 0.84 mg/l NAA, 0.1 mg/l 2,4-D and 200 mg/l l-phenylalanine has the highest tectorigenin content (0.00427 % DW). The rhizome of Neomarica gracilis planted in field is unable to measure the tectorigenin.

並列關鍵字

ANTIOXIDATION PROPERTY ； NEOMARICA GRACILIS ； RHIZOME TISSUE CULTURE ； TECTORIGENIN

參考文獻

Atta, U.R., S. Nasim, I. Baig, S. Jalil, I. Orhan, B. Sener and M.I. Choudhay, 2003. Anti-inflammatory isoflavonoids from the rhizomes of Iris germanica. J. Ethnopharmcol. 86:177-180.

Duncan, D.B., 1955. Multiple range and multiple F-test. Biometrics. 11:1-42.

Filipeanu, C.M., E. Brailoiu, G. Huhurez, S. Slatineanu, O. Baltatu and D.D. Branisteanu, 1995. Multiple effect of tyrosine kinase inhibitors on vascular smooth contraction. Eur. J. pharmacol. 281:29-35.

Imoto, M., T. Yamashita, T. Sawa, S. Kurasawa, H. Naganawa, T. Takeuchi, Z. Baoquan and K. Umezwa, 1988. Inhibition of cellular phosphatidylinositol turnover by psi- tectorigenin. Eur. Biochem. Soci. 230:43-46.

Kim, H.K., B.S. Cheon, Y.H. Kim, S.Y. Kim and H.P. Kim, 1999a. Effects of naturally occurring flavonoids on nitric oxide production in the macrophage cell line RAW 264.7 and their structure-Activity relationships. Biochem. Pharmacol. 58:759-765.