臺灣博碩士論文加值系統

本試驗目的為建立水黃皮組織培養之微體繁殖及體胚誘導體系。
本試驗以水黃皮之帶節莖段為培殖體。將帶節莖段先以流水處理2 h，後以稀釋300之安期消毒液震盪30 sec，再以70％之酒精震盪30 sec，最後在5％NaOCl水溶液 (含約0.1％ (v/v) Tween 20 展著劑) 之超音波中震盪30 min，無污染率可達65％。
在器官形成方面：以MS或WPM中添加1 mg/L BA無菌苗之結果較佳。以水黃皮無菌種子苗之子葉節、帶節莖段、莖段及下胚軸為材料，體胚誘導結果以子葉節較佳，以添加1 mg/L BA前處理之子葉節培養1個月後，於MS添加2 mg/L BA及0.1 mg/L NAA中，每個培殖體平均可產生5.42個芽體。以水黃皮母樹萌蘗之枝條為培殖體，取春季正要突破休眠的春芽，芽體誘導在MS添加1-2 mg/L BA及0.1 mg/L NAA中較佳。芽體伸長結果以在MS添加1 mg/L GA3及0.2 mg/L BA中最佳(平均1.18 cm)。將單一芽體切下，培養於1/2MS或MS添加IBA之發根培養基中誘導發根，以在1/2MS添加1 mg/L IBA中之處理較佳，達發根率74.55%、平均產生2.96個根，且平均根長度約1.33 cm。以子葉為培殖體進行體胚誘導，於MS添加3 mg/L 2,4-D與0.5 mg/L Kn2個月後，體胚誘導率為20.83%，且平均體胚數為3個。
發根的小苗經馴化結果，馴化6週的成活率為85.88%，且莖伸長約0.78 cm。

The main objectives of this study are to purpose and establish the micropropagation and somatic embryogenesis system in the tissue culture of Pongamia pinnata.
Results of nodal stem segments of epicormics culture of P. pinnata were as followed:nodal stem segments were first treated with running water for two hr. , shaking in 1/300 (v/v) anticeptol solution (Benzel thyonium chloride U.S.P 10﹪ (w/v) , Alkyl aryl polyether alcohol 10﹪(w/v)） for 30 sec , then shaking in 70﹪ethanol solution for 30 seconds, then soaked in 5﹪NaOCl (supplemented with 0.1﹪ (v/v) Tween 20） and treated with ultrasonic shaking for 30 min and the nodal stem segments obtained 65﹪non contamination.
In the aspect of organ formation : the medium of 1 mg/L BA in MS or WPM gained the best results of aseptic seedlings. Micropropagation of P. pinnata from cotyledonary nodes, nodal stem, epicotyls and hypocotyls explants derived from in vitro seedlings. Multiple shoots induction from cotyledonary node explants gained the best result of that from other explants. The greatest shoots were induced from the 1 mg/L BA pretreatmenting of cotyledonary nodes explants was achieved when MS was added with 2 mg/L BA and 0.1 mg/L NAA, average of 5.42 shoots per explants were formed in one month. Spring buds at the stage of ready to burst from dormancy and shoot explants in spring were collected from the coppice shoots of P. pinnata wild trees. MS medium containing 1-2 mg/L BA and 0.1 mg/L NAA was the best for inducing more shoots. Shoot elongation of these multiple shoots was good at 1 mg/L GA3 and 0.2 mg/L BA in MS medium. Individual shoot were rooted on 1/2MS or MS supplemented with IBA. The 1 mg/L IBA in 1/2MS medium gave the highest percentage of rooting (74.55%) with root number of 2.96 roots/shoot and root length of 1.33 cm. Cotyledons were induced with the highest percentage of 20.83% and with average of 3 somatic embryo/per explant of somatic embryogenesis in media composed of MS medium supplemented with 2,4-D 3 mg/L and Kn 0.5 mg/L after two months.
Shoots with roots were suitable for acclimatization. The plantlets survival rates were 85.88% with average shoot length of 0.78 cm after six weeks of acclimatization.

目錄

誌謝 i
中文摘要 ii
英文摘要 iii
目錄 v
表目次 ix
圖目次 xi
壹、前言 ( Introduction ) 1
貳、前人研究 (Literature Review) 3
一、水黃皮之介紹 3
(一) 生質能源 3
(二) 植生復育 3
(三) 生藥功能 4
二、微體繁殖的定義 6
三、微體繁殖的步驟 6
四、水黃皮之微體繁殖 7
(一) 培養基 7
(二) 培殖體 7
(三) 器官發生 7
(四) 體胚發生 12
參、材料與方法 (Materials and Methods) 16
一、材料 16
(一) 種子 16
(二) 萌蘗枝條 16
(三) 水黃皮之微體繁殖 16
(四) 水黃皮組培苗之馴化 17
(五) 水黃皮體胚之誘導 17
二、方法 17
(一) 種子處理 17
(二) 試驗項目 18
(三) 培養基 22
(四) 培養環境 22
(五) 形態觀察 23
(六) 統計分析 23
肆、結果 (Results) 28
一、水黃皮野外帶節莖段之表面消毒 28
二、水黃皮無菌種子苗之培養與生長 30
三、水黃皮之芽體誘導試驗 34
(一) 種子苗材料 34
(二) 野外材料 49
四、水黃皮之芽體伸長試驗 53
五、水黃皮芽體誘導發根之試驗 55
六、水黃皮切片觀察結果 57
(一) 芽體試驗 57
(二) 芽體伸長試驗 57
(三) 發根試驗 57
七、水黃皮組培苗之馴化 60
八、水黃皮體胚誘導之初步探討 61
(一) 培殖體特性 61
(二) 各植物生長調節劑誘導 61
伍、討論 (Discussion) 66
一、水黃皮野外帶節莖段之表面消毒 66
二、水黃皮無菌種子苗之培養與生長 66
(一) 發芽率 66
(二) 平均根長度 67
(三) 平均莖長度 67
三、水黃皮芽體誘導試驗 68
(一) 種子苗材料 68
(二) 野外材料 70
四、水黃皮之芽體伸長試驗 71
(一) 培殖體差異 71
(二) GA3與BA兩種植物生長調節劑之影響 71
五、水黃皮芽體誘導發根之試驗 72
(一) 基礎培養基 72
(二) IBA對發根之影響 72
六、水黃皮組培苗之馴化 73
七、水黃皮體胚誘導之初步探討 73
(一) 培殖體特性 73
(二) 各植物生長調節劑誘導 74
陸、結論 (Conclusion) 75
柒、參考文獻 ( References ) 76
捌、附錄 (Appendix) 87

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