- 學位論文
培養條件對射干不定根培養生產類黃酮之影響
THE EFFECT OF CULTURAL FACTORS ON THE PRODUCTION OF FLAVONOIDS IN ADVENTITIOUS ROOT CULTURE OF BELAMCANDA CHINENSIS (L.) DC.
摘要
射干（Belamcanda chinensis（L.）DC.）不定根培養於MS培養基，在三種接種比例0.09、0.4與0.7 ％ 比較下，不定根在0.4 ％ 的比例下培養30天，有最佳的生長率為原接種量的11.1倍，總黃酮為101.4±5.4 eq. iridin mg/g DW，且可分析出tectoridin、iridin、tectorigenin、irigenin與isorhamnetin等指標性成份。調整MS培養配方中之有機添加物( nicotinic acid、pyridoxine HCl、thiamine HCl和glycine )之倍數濃度，將添加倍數增至4倍，可提升總黃酮與總異黃酮分別為143.78±22.46 eq. iridin mg/g DW與71.77±10.69 mg/g DW。並可提升不定根中主要產物iridin的累積，可得其最高含量63.31 ± 9.76 mg/g DW。利用化學誘變劑對射干癒合組織進行不同時間的處理，經colchicine溶液處理後，共可獲得5種不定根品系，其中1種品系具有較高之總黃酮與iridin含量，分別為103.57±8.5 eq. iridin mg/g DW和42.79±5.4 mg/g DW。經1 mgl-1 trifluralin溶液加入MS培養基中，處理60天所得之癒合組織，可得總黃酮含量78.82±6.2 eq. iridin mg/g DW與tectorigenin 2.86±0.07 mg/g DW。研究結果有助於未來射干不定根量化培養生產類黃酮條件之建立。
並列摘要
The adventitious root of Belamcanda chinensis cultured in MS liquid medium, the optimal of inoculum densities was 0.4％ which had the best growth index of 11.1 times. The total flavonoids of methanolic extract in adventitious roots was 101.4 ± 5.4 eq. iridin mg / g DW. Five kinds of flavonoids (tectoridin , iridin , tectorigenin , isorhamnetin and irigenin ) in methanolic extract were analyzed by HPLC. Total flavonoids and total isoflavonoids content reached 143.78 ± 22.46 eq. iridin mg / g DW and 71.77 ± 10.69 mg / g DW when the concentration of organic additives( nicotinic acid、pyridoxine HCl、thiamine HCl and glycine ) was raised with 4 times in MS liquid medium. Callus treated with antimitotic agent in different exposure time, five adventitious root lines were obtained and one of the lines with a higher content which total flavonoids was 103.57 ± 8.5 eq. iridin mg / g DW. Total flavonoids and iridin content reached 78.82 ± 6.2 eq. iridin mg / g DW and tectorigenin 2.86 ± 0.07 mg / g DW when callus was cultured in the medium supplemented with 1 mgl -1 trifluralin solution 60 days.