本研究之目的為建立觀賞鳳梨直接不定芽與癒合組織之誘導及植株再生系統，並探討auxin對促進擎天屬鳳梨Guzmania組培苗伸長生長之影響，及應用物理與化學方法誘導體細胞變異之產生。
本研究針對觀賞鳳梨組織培養再生的培植體來源及培養基成分等因子，作一系列的研究探討。試驗結果顯示，含有1.0 mg l-1 BA + 0.5-1.0 mg l-1 NAA或3.0 mg l-1 BA + 0.5 mg l-1 NAA，與50 mg l-1 arginine、50 mg l-1 aspargine、50 mg l-1 L-glutamine、0.5 mg l-1 Ca-panthothenate、200 mg l-1 citric acid及3﹪sucrose之1/3MS基本鹽類與有機成分配方，對觀賞鳳梨Aechmea fulgens var. fulgens和Guzmania 'Focus'之莖頂及側芽培植體，誘導直接不定芽發生率最佳。二品種並以最基部側芽培植體的不定芽發生率最高，分別為47.5及35 %。本研究同時發現G. 'Focus'之去頂組培苗，培養於含有3.0 mg l-1 BA + 0.5 mg l-1 NAA的培養基，第16天起，即開始形成不定芽體，但此現象並未發生在未去頂的培植體上，僅產生突起的瘤狀物。
此外，1/2MS基礎培養基添加1.0-1.5 mg l-1 2,4-D並組合1.0或0.5 mg l-1 NAA，對觀賞鳳梨A. fasciata與G. 'Hilda'之花瓣與子房培植體，可獲得較高的不定芽癒合組織誘導率；將癒合組織移植於含有1.0 mg l-1 NAA + 0.5 mg l-1 TDZ之再生培養基，可分化形成不定芽。而G. 'Hilda'花器癒合組織衍生的再生植株，培養在添加不同濃度IAA、IBA、NAA及8-azaadenine之培養基，各處理於培植體基部增殖之不定芽數只有1-2個，顯示auxin處理確可減低細胞分裂素之效應。再生植株的抽長生長以0.5 mg l-1 NAA及1.0 mg l-1 NAA的影響最大，培養4個月，分別可生長至5.73及5.62 cm，較對照組多1.95-2.22 cm，可克服Guzmania觀賞鳳梨幼苗抽長遲緩的缺點，作為產業界大量繁殖種苗之應用。
利用光強度馴化，對組培苗的生長有很大的改善效果。A. fasciata組培苗以中光強度(50 µmol m-2s-1)馴化，其成活率遠高於低光強度(1 µmol m-2s-1)的馴化苗(17.5%)，為95%，並且可產生最多的新生鬚根數(4.15條)。組培苗移植於椰纖的生長效果最好，6個月株高可達22 cm及生長19.3片葉。
本研究亦利用觀賞鳳梨癒合組織及去頂短縮莖為材料，以疊氮化鈉及γ射線進行誘變處理，以作為無性繁殖作物誘變育種之參考。G. 'Hilda'癒合組織經疊氮化鈉處理後，培植體均無法存活；G. 'Hilda'去頂短縮莖培養於1/3MS基礎培養基並添加3.0 mg l-1 BA + 0.5 mg l-1 NAA所組成的增殖培養基，並以0.5 mM之疊氮化鈉處理60分鐘，其存活率為51.3%，約達半致死劑量。觀賞鳳梨A. fasciata、G. 'Hilda'、G. 'Cherry'、G. 'Luna'及G. 'Focus'短縮莖以γ射線照射後，除G. 'Focus'之短縮莖經15 Gy照射，存活率下降至45.0%之外，其餘各處理均達74.2-100%。目前，已產生葉部具有鑲嵌條紋的變異性狀，再生植株並移出種植於溫室。

The objectives of this study were to develop an in vitro direct adventitious bud induction and an organogenic callus induction and shoot regeneration system via floral organ segments culture for bromeliads, moreover, explore the effect of auxin on plantlet elongation of Guzmania. And further, apply the above micropropagation system to physical and chemical methods to induce somaclonal variances of bromeliad plantlets in vitro for mutation breeding.
The explant sources of bromeliads and the components of culture medium were studied to develop a micropropagation system for bromeliads. The results indicated that the 1/3MS basal medium supplemented with a combination of 1.0 mg l-1 BA + 0.5-1.0 mg l-1 NAA, or a combination of 3.0 mg l-1 BA + 0.5 mg l-1 NAA, showed the highest frequency of direct initiation of adventitious buds derived from shoot apex and lateral bud explants of Aechmea fulgens var. fulgens and Guzmania 'Focus'. The best results of adventitious buds induction of the both species were found in the lower lateral bud explants, at 47.5% and 35%, respectively. In addition, the adventitious buds began to form on day 16 after the G. 'Focus' decapitated plantlets had been cultured in medium supplemented with 3.0 mg l-1 BA + 0.5 mg l-1 NAA. However, this phenomenon did not occur in case of undecapitated explants, where only protruding nodules appeared.
Petal- and ovary-derived calli of A. fasciata and G. 'Hilda' were induced on 1/2MS basal medium supplemented with 1.0-1.5 mg l-1 2,4-D in combination with 1.0 or 0.5 mg l-1 NAA. Organogenic calli were cultured on medium with 1.0 mg l-1 NAA and 0.5 mg l-1 TDZ could be induced to differentiate and regenerate the adventitious buds. Furthermore, the number of adventitious buds proliferating at the base of the plantlets derived from G. 'Hilda' floral organs, cultured in media with different concentrations of IAA, IBA, NAA, and 8-azaadenine, was only 1-2 adventitious buds individually. This result shows that auxin can indeed suppress cytokinin-effects. The influence on plantlet elongation was greatest in the treatments using 0.5 mg l-1 NAA and 1.0 mg l-1 NAA. After 4 months culture, plantlets grew to 5.73 and 5.62 cm in height, that was 2.22 and 1.95 cm higher than the control, respectively.
Plantlets of A. fasciata hardened under the middle (50 μmol m-2s-1) light intensity condition had a higher survival rate, 95%, than that hardened at a low light intensity (1 μmol m-2s-1; 17.5%). The maximum number of newly developing roots, up to 4.15 per shoot, was also observed at the same light intensity treatment. During transplantation, plantlets growing in coir fiber showed the best results in terms of plant growth within 6 months ex vitro culture. The average length of the plantlets was 22.0 cm, and an average of 19.3 leaves per plantlet was achieved.
When calli of G. 'Hilda' treated by sodium azide, the survival rate was 0%. The survival rate of decapitated plantlet explants treated with 0.5 mM sodium azide for 60 minutes was 51.3%, about half-lethal dose. In addition to the survival rates of decapitated plantlet explants of A. fasciata, G. 'Hilda', G. 'Cherry', G. 'Luna' and G. 'Focus' irradiated by γ-ray showed 74.2-100% with the exception of the G. 'Focus' irradiated by 15 Gy, which dropped to 45.0%. At present, mutant plantlets showed a great deal of chimeras in leaf and were transplanted to potting media.

論文審定書
謝誌
摘要………………………………………………………………… i
Abstract……………………………………………………………… iii
目錄………………………………………………………………… v
圖次………………………………………………………………… vii
表次………………………………………………………………… x
緒言………………………………………………………………… 1
第一章 前人研究………………………………………………… 3
第二章 觀賞鳳梨直接不定芽誘導及植株再生系統之建立…… 15
中文摘要………………………………………………………… 15
Abstract…………………………………………………………… 16
前言……………………………………………………………… 17
材料與方法……………………………………………………… 19
結果……………………………………………………………… 21
討論……………………………………………………………… 23
第三章 觀賞鳳梨花器癒合組織誘導與植株再生系統之建立… 35
中文摘要………………………………………………………… 35
Abstract…………………………………………………………… 36
前言……………………………………………………………… 37
材料與方法……………………………………………………… 39
結果……………………………………………………………… 42
討論……………………………………………………………… 45
第四章 Auxin對擎天屬觀賞鳳梨組培苗生長之影響………… 58
中文摘要………………………………………………………… 58
Abstract…………………………………………………………… 59
前言……………………………………………………………… 60
材料與方法……………………………………………………… 62
結果……………………………………………………………… 64
討論……………………………………………………………… 66
第五章 光強度及介質對觀賞鳳梨組培苗馴化及生長之影響… 77
中文摘要………………………………………………………… 77
Abstract…………………………………………………………… 78
前言……………………………………………………………… 79
材料與方法……………………………………………………… 81
結果……………………………………………………………… 82
討論……………………………………………………………… 83
第六章 觀賞鳳梨疊氮化鈉與γ射線誘變之研究……………… 88
中文摘要………………………………………………………… 88
Abstract…………………………………………………………… 89
前言……………………………………………………………… 90
材料與方法……………………………………………………… 92
結果……………………………………………………………… 94
討論……………………………………………………………… 96
結論……………………………………………………………… 110
參考文獻………………………………………………………… 112
附錄……………………………………………………………… 132

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