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研究生:季亞垠
研究生(外文):Ya Yin Ji
論文名稱:金門保育植物寬葉毛氈苔的組織培養繁殖
論文名稱(外文):Tissue culture propagation of Drosera burmannii Vahl: a conserved plant species in Kinmen
指導教授:廖宇賡
指導教授(外文):Yue Ken Liao
學位類別:碩士
校院名稱:國立嘉義大學
系所名稱:森林暨自然資源學系研究所
學門:農業科學學門
學類:林業學類
論文種類:學術論文
畢業學年度:100
語文別:中文
中文關鍵詞:細胞分裂素寬葉毛氈苔綴化金門徒長
外文關鍵詞:cytokininDrosera burmanniifasciationKinmenspindling
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本研究利用組織培養技術誘導金門保育類植物寬葉毛氈苔(Drosera burmannii Vahl) 發生綴化 (fasciation),再透過綴化多頂端生長點增殖之性狀建立其大量繁殖的再生系統。首先在誘導莖頂培殖體生成芽體時,以20 μM 6-furfurylaminopurine (kinetin) 處理所獲得的芽體有50%會轉變成綴化植株 (fasciated plant, FP),經石蠟切片形態觀察得知FP 莖頂縱切面上具有橫向排列的多數頂端生長點。FP 在1/2 MS 培養基中繼代時會持續地延展其寬幅,切取寬幅長度0.5 cm進行30 天 (1 代) 繼代培養,則以20 μM 6-benzylaminopurine (BAP) 誘導而來之FP 有最大的延展寬幅長度,在一次繼代後可增長至4.38 ±0.11 cm,紀錄FP 這種綴化現象已持續了270 天 (9 代)。若終止FP之繼代,並將FP 黑暗處理60 天使其徒長後,再以解剖刀切取1 cm徒長後之寬幅長度進行密集切碎培養,約45 天可獲得恢復正常形態之寬葉毛氈苔,其中來自20 μM BAP 誘導所得之FP 獲得恢復正常的單株達20.1 ± 1.8 個。目前推估經270 天繼代並進行60 天的黑暗徒長與45 天的正常形態單株回復培養後,起始0.5 cm 寬幅的FP 可獲得的正常寬葉毛氈苔單株數量達3.2 × 10 的10 次方。
The present study has developed a tissue culture protocol incorporating the induction of shoot fasciation in Drosera burmannii Vahl for mass propagation of this plant species conserved in Kinmen. The explants (shoot tips) exhibited 50% of fasciated plant (FP) formation after being induced by 20 µM 6-furfurylaminopurine (kinetin). The anatomical analysis revealed that there are multiple apical meristems arranged perpendicularly to the longitude section of FP apical stem. When a 0.5 cm wide piece of the FP was excised and cultured on the cytokinin-free 1/2 MS medium, it proliferated with its width continuously extended every 30 days (one subculture). However, the greatest width increase (4.38 ± 0.11 cm per subculture) was obtained from 20 µM 6-benzylaminopurine (BAP) induced FP. Such events of FP to proliferate and maintain fasciated characteristics during subcultures prolonged for 270 days (nine subcultures) without apparent changes. The termination of FP subculture was possible when dark incubation was applied for 60 days followed by fine mincing of the spindling FP. Morphologically normal plants were recovered from these operations and the maximal production of normal plants (20.1 ± 1.8 plants per 1.0 cm width of spindling FP) was obtained from 20 µM BAP induced FP. An estimated production of normal plants, initiated from a 0.5 cm piece of FP, was thus achieved to the amount of 3.2 × 1010 within one year (270-day FP subculture plus 60-day dark spindling and 45-day normal plant recovery cultures).
目錄......................................I
圖目錄...................................III
表目錄....................................IV
英文縮寫表.................................V
I、前言..................................1
II、前人研究...............................4
(I)、茅膏菜屬植物之特性....................4
(II)、茅膏菜屬植物之藥用價值.................6
(III)、茅膏菜屬植物之組織培養.................6
(IV)、綴化現象之外觀形態.....................9
(V)、影響植物體綴化發生之因子...............11
1.生理因子.................................11
2.基因調控.................................14
(VI)、正常植株與綴化植株內部構造之差異.........15
III、材料與方法.............................17
(I)、植物材料之準備........................17
(II)、誘導芽體生成與植株綴化.................17
(III)、組織解剖觀察..........................19
(IV)、綴化現象之維持........................19
(V)、綴化現象之終止及正常形態植株之回復........20
(VI)、正常形態植株總數量之推估................21
(VII)、培養環境.............................22
IV、結果...................................24
(I)、芽體生成與植株綴化.....................24
(II)、綴化現象之維持.........................30
(III)、終止綴化現象與推估單株數量..............33
V、討論....................................39
VI、引用文獻................................48
附錄一.....................................61

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