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研究生:李佾玲
研究生(外文):Yi-Ling Li
論文名稱:台灣土黨參(Cyclocodon lancifolius (Roxb.) Kurz) 組織培養及植株再生
論文名稱(外文):Tissue culture and plant regeneration of Cyclocodon lancifolius (Roxb.) Kurz
指導教授:何錦玟
指導教授(外文):Chin-wen Ho
口試委員:何錦玟
口試委員(外文):Chin-wen Ho
口試日期:2015-07-29
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:2015
畢業學年度:103
語文別:中文
論文頁數:81
中文關鍵詞:植株再生台灣土黨參組織培養
外文關鍵詞:plant regenerationCyclocodon lancifolius (Roxb.) Kurztissue culture
相關次數:
  • 被引用被引用:3
  • 點閱點閱:170
  • 評分評分:
  • 下載下載:41
  • 收藏至我的研究室書目清單書目收藏:0
本研究建立台灣土黨參(Cyclocodon lancifolius(Roxb.)Kurz)組織培養的流程,田間來源培植體易產生乳汁不利試管內植株生長。從未成熟果實取得的種子萌發的實生苗作為培植體來源可以避免乳汁造成的不良影響。使用葉片培植體進行癒傷組織的誘導及器官發生,在Murashige and Skoog (1962)培養基添加0.5 mg/l Zeatin 組合0.1mg/l 之Gibberellic acid (GA3)和α-Naphthalene acetic acid(NAA) 可誘導出不定芽及癒傷組織。而添加0.5mg/l 6-Benzyl-aminopurine (BA)組合0.1mg/l 的GA3、NAA及2,4-Dichlorophenoxy acetic acid (2,4-D)的MS培養基能產生最多癒傷組織,每個培植體平均鮮重量2.89 ± 1.12g。液態培養中以幼芽作為培植體進行芽體增殖,培養於添加1 mg/l BA和0.1 mg/l NAA的MS培養基每個培植體有最多平均芽數2.92 ± 1.05。莖段培植體,則在不添加生長調節劑的MS培養基每個培植體能得到最多平均芽數2.2 ± 0.76。液態培養中所產生之玻璃質化植株,繼代至固態培養15天後可改善此現象。再生植株以1/2MS培養基繼代培養能有良好的發根反應,且經出瓶馴化後有正常的外觀。抗氧化能力分析再生植株測得14.783 ± 1.629 eq. quercetin μg / mg DW總黃酮含量,相近於田間植株的含量。未成熟果實測得的總黃酮含量為7.533 ± 0.236 eq. quercetin μg / mg DW及總酚含量為8.554 ± 2.147 eq. gallic acid μg / mg DW。
In present study, we established the tissue culture process of Cyclocodon lancifolius (Roxb.) Kurz. Explants from field were easy to exudate latex which was adverse to grown of in vitro plant. Shoots from immature seeds of fruits as a source of explants could avoid latex causing adverse effects. To induce callus and organogenesis, explants of leaf blades cultured in Murashige and Skoog (1962) medium supplemented with 0.5 mg/l Zeatin combined 0.1mg/l of Gibberellic acid (GA3) and α-Naphthalene acetic acid (NAA) could quickly induce adventitious shoots and more callus. The maximum average fresh weight of callus was 2.89 ± 1.12g per explant when cultured in MS medium supplemented with 0.5 mg/l 6-Benzyl-aminopurine (BA) combined 0.1mg/l GA3, NAA and 2,4-Dichlorophenoxy acetic acid (2,4-D). Proliferation of shoots in liquid culture, the maximum average number of shoots was 2.92 ± 1.05 per explant cultured in MS medium supplemented with 1 mg/l BA and 0.1 mg/l NAA when using shoots as explants. However, using stem segments as explants, the maximum average number of shootswas 2.2 ± 0.76 per explant cultured in MS medium without plant growth regulator. Subculture the hyperhydric plantlets produced from liquid culture to solid culture become healthy after 15 days. Plantlets developed roots cultured in one half MS medium. After acclimatized successfully, Plantlets have normal pherotype. The total flavonoid content in plantlets was 14.783 ± 1.629 eq. quercetin μg / mg which was similar to plant from field. The total flavonoids and phenols content in immature fruit were 7.533 ± 0.236 eq. quercetin μg / mg DW and 8.554 ± 2.147 eq. gallic acid μg / mg DW respectively.
致 謝i
中文摘要ii
Abstractiv
目錄vi
表目錄x
圖目錄xi
縮寫字xii
一、前人研究1
1.1 台灣土黨參植株特性及藥用1
1.2 桔梗科植物之利用3
1.3 桔梗科植物組織培養之相關研究4
1.3.1 植株材料4
1.3.2 培植體種類4
1.3.3 培養基與生長調節劑配方10
1.4 植物分泌乳汁的特性11
1.5 研究目的12
二、材料與方法13
2.1 材料來源13
2.2 培養基13
2.3 培養環境15
2.4 莖段培養15
2.5 未成熟果實培養15
2.6 生長調節劑組合對葉片培植體癒傷組織培養及器官發生之影響17
2.7 培植體來源及BA濃度對芽體增殖之影響20
2.7.1 不同BA濃度對幼芽芽體增殖之影響20
2.7.2 不同BA濃度對莖段芽體增殖之影響20
2.8 培養方式對再生植株生長之影響20
2.9 甜肉桂植物萃取液對再生植株生長之影響22
2.10 植株出瓶馴化22
2.11 抗氧化能力分析23
2.11.1 乾燥及研磨23
2.11.2 甲醇萃取23
2.11.3總黃酮含量測定23
2.11.4 總酚含量測定24
2.12 統計分析24
三、結果25
3.1莖段培養25
3.2 未成熟果實培養25
3.3 生長調節劑組合對葉片培植體癒傷組織誘導及器官發生之影響28
3.4 2,4-D與不同Cytokinin組合對葉片培植體癒傷組織誘導及器官發生之影響31
3.5 培植體來源及BA濃度對芽體增殖之影響35
3.5.1 不同BA濃度對幼芽芽體增殖之影響35
3.5.2 不同BA濃度對莖段芽體增殖之影響35
3.6 培養方式對再生植株生長之影響41
3.7 甜肉桂植物萃取液對再生植株生長之影響41
3.8 植株出瓶馴化45
3.9 抗氧化能力分析45
四、討論48
4.1莖段及未成熟果實培養48
4.2 乳汁對台灣土黨參植株生長之影響48
4.3生長調節劑組合對葉片培植體癒傷組織誘導及器官發生之影響49
4.4 2,4-D與不同Cytokinin組合對葉片培植體癒傷組織誘導及器官發生之影響50
4.5培植體來源及BA濃度對芽體增殖之影響51
4.6 培養方式對再生植株生長之影響51
4.7 甜肉桂植物萃取液對再生植株之影響52
4.8 出瓶馴化及植株生長53
4.9 抗氧化能力分析53
五、結論55
六、參考文獻57
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