臺灣博碩士論文加值系統

本論文建立阿拉比卡咖啡癒傷組織之誘導系統，首先測試種子無菌萌芽之培養條件，並藉以獲取足量之實生苗供後續試驗。切取阿拉比卡咖啡實生苗的根及葉為培植體，置於含不同種類及濃度植物生長調節劑的Murashige and Skoog（MS）培養基，以全暗培養進行癒傷組織之誘導，將誘導所得之癒傷組織進行繼代增殖，及試管內形態發生試驗。
種子萌芽試驗結果顯示，以濃度0.5%之次氯酸鈉消毒10分鐘，可獲得最高萌芽比率（63.16%）。切取萌芽十二週實生苗之根及葉做為培植體，以不同植物生長調節劑（plant growth regulators，PGRs），包括植物生長素α-naphthaleneacetic acid （NAA）或2,4-dichlorophenoxyacetic acid（2,4-D）與細胞分裂素thidiazuron（TDZ）或N6-benzyladenine（BA），搭配進行癒傷組織誘導試驗。癒傷組織誘導試驗結果顯示，根及葉培植體在不含PGR的控制組中，無法誘導出癒傷組織。然而，其餘各試驗組皆可成功誘導出癒傷組織，共獲得二十八個品系癒傷組織，這些癒傷組織可穩定生長，並以相同條件的培養基持續進行繼代，經持續觀察得知，各品系癒傷組織之外觀、顏色、形態及結構有所差異。癒傷組織增殖率試驗及形態分析結果顯示，可將癒傷組織分為四種：第一種為結構緊密、觸感堅硬之黃色癒傷組織，生長速度最快；第二種為蓬鬆顆粒狀且觸感略硬之淡黃色且透白的癒傷組織，生長速度次之；第三種為結構鬆散、觸感軟綿之白色半透明狀癒傷組織，生長速度稍慢；第四種為形態軟爛之紅褐色癒傷組織，生長速度緩慢。本論文成功建立了阿拉比卡咖啡的癒傷組織誘導及增殖系統，建議未來可以選擇本論文所篩選出之生長快速、質地緊密及外觀顏色呈淡黃色或白色之癒傷組織，進一步建立完整的植株再生體系。

The thesis attempts to establish a protocol for callus induction of Coffee arabica L. Firstly, the seeds were sown in in vitro conditions and seedlings obtained were used as donor plants for the subsequent tests. The explants were taken from these donor plants and placed on a modified Murashige and Skoog (MS) medium supplemented with combinations of plant growth regulators (PGRs) in the dark. The calli were proliferated more on the same medium, and their capacities of in vitro morphogenesis were subsequently evaluated. In the seeds germination experiment, 10 min sterilization time with 0.5% sodium hypochlorite gave the highest germination rate (63.16%). The root and leaf explants were taken from 12-week-old seedings and placed on a modified MS medium supplemented with combinations of auxins, (α-naphthaleneacetic acid : NAA or 2,4-dichlorophenoxyacetic acid : 2,4-D) and cytokinins (thidiazuron : TDZ or N6-benzyladenine : BA). In the control treatment, no response was found at all the explants. In contrast, 28 callus lines were obtained from other treatments. These calli could be subcultured on same medium and proliferated more with viable growth. Based on the proliferation rate and their morphology, the callus were classified into 4 types : 1) Yellowish compact callus with a highest proliferation rate; 2) Pale yellowish granular callus with high proliferation rate; 3) Translucent to whitish and friable callus with low proliferation rate; 4) Red to brown callus with lowest proliferation rate. Type 1 callus lines were suggested for inducing in vitro morphogenesis and subsequent plant regeneration.

謝誌 I
縮寫表 II
目錄 III
表目錄 V
圖目錄 VI
中文摘要 1
英文摘要 3
第一章 前言 5
1.1 咖啡之植物學特性 5
1.2 咖啡之商業及藥用價值 5
1.3 阿拉比卡咖啡之簡介 6
1.4 植物組織培養技術 7
1.5 咖啡組織培養之前人研究 8
1.6 研究目的 9
第二章 材料與方法 10
2.1 實驗材料 10
2.2 基礎培養基植 10
2.3 物生長調節劑 10
2.4 阿拉比卡咖啡種子萌芽試驗 10
2.5 阿拉比卡咖啡癒傷組織之誘導 10
2.6 阿拉比卡咖啡癒傷組織之增殖試驗 11
2.7 統計方式 11
第三章 結果 12
3.1 阿拉比卡咖啡種子萌芽試驗 12
3.2 阿拉比卡咖啡癒傷組織之誘導 12
3.2.1 NAA搭配BA之癒傷組織誘導 12
3.2.2 2,4-D搭配TDZ之癒傷組織誘導 13
3.2.3 不定根之形成 14
3.3 阿拉比卡咖啡癒傷組織增殖試驗 14
第四章 討論 35
第五章 參考文獻 39

表3.1 阿拉比卡咖啡種子萌芽率 18
表3.2 NAA搭配BA處理之阿拉比卡咖啡癒傷組織誘導率 23
表3.3 2,4-D搭配TDZ處理之阿拉比卡咖啡癒傷組織誘導率 28
表3.4 NAA及BA處理之阿拉比卡咖啡葉培植體不定根誘導率 31
表3.5 NAA及BA處理之阿拉比卡咖啡癒傷組織增值率與形態 32
表3.6 2,4-D及TDZ處理之阿拉比卡咖啡癒傷組織增值率與形態 33
 
圖3.1 阿拉比卡咖啡種子萌芽情形 17
圖3.2 NAA及BA對阿拉比卡咖啡根培植體癒傷組織之誘導 19
圖3.3 NAA及BA對阿拉比卡咖啡根培植體癒傷組織之誘導 20
圖3.4 NAA及BA對阿拉比卡咖啡葉培植體癒傷組織之誘導 21
圖3.5 NAA及BA對阿拉比卡咖啡葉培植體癒傷組織之誘導 22
圖3.6 2,4-D及TDZ對阿拉比卡咖啡根培植體癒傷組織之誘導 24
圖3.7 2,4-D及TDZ對阿拉比卡咖啡根培植體癒傷組織之誘導 25
圖3.8 2,4-D及TDZ對阿拉比卡咖啡葉培植體癒傷組織之誘導 26
圖3.9 2,4-D及TDZ對阿拉比卡咖啡葉培植體癒傷組織之誘導 27
圖3.10 NAA及BA對阿拉比卡咖啡葉培植不定根之誘導 29
圖3.11 NAA及BA對阿拉比卡咖啡葉培植不定根之誘導 30
圖3.12 阿拉比卡咖啡癒傷組織形態與增生能力 34

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