臺灣博碩士論文加值系統

銀柳（Salix garcilistyla Miq.）為楊柳科多年生落葉灌木，喜愛溫暖潮濕、日照充足之氣候，為宜蘭三星地區重要的花卉作物。本研究是利用不同培殖體如頂芽、莖段、葉片培養癒傷組織探討植株再生之條件及方法，並利用石蠟切片技術觀察生長調節劑誘導葉片組織之癒傷組織分化、體胚形成及芽體再生之過程，建立一套完整的銀柳組織培養之再生系統。本試驗中銀柳‘蘭陽一號’頂芽長度小於0.5 cm者，培養於不同濃度TDZ之固體培養基，芽體不易生長且易褐化。取約0.5-1 cm左右之頂芽培養於光環境下之濾紙橋液體培養基，於無添加TDZ之培養基生長較佳且無褐化，而芽體褐化率隨濃度增加而增加，以0.5 μM TDZ之芽體褐化情形最為嚴重。葉片培殖體分別培養於添加27μM 2,4-D之全量MS液體及固體培養基中，液體培養之葉培殖體之癒傷組織形成速率較固體培養快，但癒傷組織之形成量無明顯差異。
葉培殖體於光環境（45±5 μmol m-2 s-1）較暗環境（0 μmol m-2 s-1）下培養容易褐化，顯示不同光度會影響培殖體褐化速率。癒傷組織分別繼代培養於添加不同濃度之TDZ、Kinetin、BA及不添加生長調節劑之全量MS培養基，試驗結果均以不添加生長調節劑之全量MS培養基癒傷組織增殖量為最高。癒傷組織於添加不同濃度之細胞分裂素（TDZ、Kinetin、BA）癒傷組織增殖量會增加，但不會誘導芽體產生。培養於0.1μM NAA及0.5μM Kinetin之MS全量培養基有芽體組織產生。

Salix garcilistyla Miq. is perennial deciduous shrub of Saliaceae. It is important for flower of crop in area of Sunshin of Ilan, like warm, humid and abundant sunshine of climate. This study establish complete regeneration system of Salix garcilistyla Miq. by tissue culture to probe into conditions and method of plant regeneration used different explants such as shoot apex, nodal segments and leaves, also used paraffin section technique to observe process of growth regulator to induce callus differentiation, somatic embryogenesis and shoot regeneration. This study are length less than 0.5 cm of shoot apex of Salix garcilistyla Miq. ‘Lanyang No. 1’ , bud are not growth and easy to browning when culture in add different concentration of TDZ of solid medium. In light condition, the length about 0.5-1 cm of shoot apex culture in filter paper bridge in liquid medium, the medium no add TDZ have better growth and without browning, and bud browning rate raise follow with TDZ concentration increase, highest of browning rate of bud in add 0.5 μM TDZ medium. Leaf explants separately culture in add 27 μM 2,4-D of liquid or solid of MS medium, the callus form rate in liquid medium is rapid than solid medium, but amount of callus are not different.
Leaf explants are easier to brownng in light condition（45±5 μmol m-2 s-1）than dark condition（0 μmol m-2 s-1）. That exhibit different luminosity Effects explant browning rate. Callus separately subculture in add different concentration of TDZ, BA, Kinetin and no add plant growth regulators of MS medium. The results are all of the highest amount of callus multiplication in no add plant growth regulators of MS medium. Callus at different concentrations of Cytokinins（TDZ、Kinetin、BA）medium will increase the amount of callus multiplication, but don’t induce shoot regeneration. Shoot regeneration in add 1μM NAA and 0.5μM kinetin of MS medium.

中文摘要.....................................................i
英文摘要.....................................................ii
表目錄.......................................................iii
圖目錄.......................................................iv
壹、前言.......................................................1
貳、前人研究...................................................2
一、銀柳型態介紹及栽培管理.......................................2
二、木本植物之組織培養..........................................2
三、體胚形成...................................................4
四、體胚形成與芽體再生之影響因子.................................6
（一）培殖體種類...............................................6
（二）培養基成分...............................................7
參、材料與方法.................................................13
一、試驗材料...................................................13
二、培養基成分及調配............................................13
三、培養室環境條件 ..............................................14
四、試驗方法...................................................14
（一）莖頂培養.................................................14
（二）莖段培養.................................................15
（三）葉片培養.................................................16
（四）組織型態檢定 .............................................18
（五）統計分析.................................................18
肆、結果......................................................19
一、莖頂培養之芽體誘導..........................................19
二、莖段培養之癒傷組織誘導.......................................19
三、葉片培養...................................................20
（一）癒傷組織誘導 .............................................20
（二）體胚形成.................................................21
伍、討論......................................................25
一、莖頂培養..................................................25
二、莖段培養..................................................26
三、葉片培養..................................................27
四、體胚形成..................................................29
五、組織形態..................................................31
陸、參考文獻..................................................58
附錄.........................................................67

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