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研究生:顏志宇
研究生(外文):Jhih-Yu Yan
論文名稱:培養條件對射干不定根培養生產類黃酮之影響
論文名稱(外文):THE EFFECT OF CULTURAL FACTORS ON THE PRODUCTION OF FLAVONOIDS IN ADVENTITIOUS ROOT CULTURE OF BELAMCANDA CHINENSIS (L.) DC.
指導教授:何錦玟
指導教授(外文):Chin-Wen Ho
學位類別:碩士
校院名稱:大同大學
系所名稱:生物工程學系(所)
學門:工程學門
學類:生醫工程學類
論文種類:學術論文
論文出版年:2010
畢業學年度:98
語文別:中文
論文頁數:87
中文關鍵詞:不定根總黃酮總異黃酮化學誘變劑射干
外文關鍵詞:Belamcanda chinensisadventitious roottotal flavonoidstotal isoflavonoidsantimitotic agent
相關次數:
  • 被引用被引用:3
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  • 收藏至我的研究室書目清單書目收藏:1
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。
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.
目錄
中文摘要 I
Abstract II
目錄 III
表目錄 VII
圖目錄 IX
縮寫字 XI
一、前人研究 1
1.1 射干植株特性 1
1.2射干之成份及藥理作用 1
1.3培養條件對細胞生長及二次代謝產物生產之影響 8
1.3.1細胞分化程度 8
1.3.2接種比例 11
1.3.3生長調節劑 12
1.3.4誘導物質之添加 14
1.3.5 改變有機添加物 15
1.3.6化學誘變劑 colchicine、oryzalin及 trifluralin 17
1.4研究目的 21
二、材料與方法 23
2.1. 材料來源 23
2.2 培養基製備 26
2.3 誘變劑製備 28
2.4培養環境 28
2.5實驗架構流程 29
2.6不定根液態培養及生長曲線 30
2.7培養條件對不定根及癒合組織生產類黃酮之影響 30
2.7.1 接種比例 30
2.7.2 改變有機添加物 31
2.7.3 化學誘變劑colchicine、oryzalin及 trifluralin處理 32
2.7.3.1 colchicine水溶液進行處理 32
2.7.3.2 oryzalin溶液進行處理 32
2.7.3.3 oryzalin 或 trifluralin溶液添加入培養基中進行培養 33
2.8樣品乾燥製備 33
2.9萃取方式 33
2.10 總黃酮(total flavonoid)測定 34
2.11標準品 34
2.12 HPLC儀器系統 35
2.13生長率 ( Growth index ) 計算 37
2.14觀察及拍照記錄 37
2.15統計分析 37
三、結果 38
3.1不定根液態培養及生長曲線 38
3.2培養條件對不定根及癒合組織其生產類黃酮之影響 40
3.2.1接種比例 40
3.2.2改變有機添加物 43
3.2.3化學誘變劑( colchicine、oryzalin及 trifluralin) 45
3.2.3.1以colchicine水溶液進行短時間處理 45
3.2.3.2以colchicine水溶液進行長時間處理 51
3.2.3.3以oryzalin溶液進行短時間處理 54
3.2.3.4以oryzalin溶液進行長時間處理 59
3.2.3.5 oryzalin或trifluralin溶液添加入培養基中進行培養 62
四、討論 65
4.1不定根液態培養及生長曲線 65
4.2培養條件對不定根及癒合組織其生產類黃酮之影響 66
4.2.1接種比例 66
4.2.2改變有機添加物 67
4.2.3化學誘變劑 colchicine、oryzalin及 trifluralin 69
4.2.3.1以colchicine進行短時間處理 69
4.2.3.2以colchicine進行長時間處理 70
4.2.3.3以oryzalin溶液進行短時間處理 72
4.2.3.4以oryzalin溶液進行長時間處理 73
4.2.3.5 oryzalin 或 trifluralin添加入培養基中進行培養 74
五、結論 77
六、參考文獻 79
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