臺灣博碩士論文加值系統

本論文旨在建立兔眼藍莓之微體繁殖流程，試驗材料為‘Woodard’及‘Tifblue’品種，試驗目標為尋找微體繁殖法之1) 最佳培養部位; 2) 最適再生部位; 3) 最適培養基; 4) 產生細胞懸浮培養之最佳生長條件及培養基。試驗結果顯示，以當年生成熟綠枝條，截切1-2公分的單節莖段，具有汙染力低、萌芽率高且萌生之側枝較長等優點。其中單節莖段培養於添加2iP之MS培養基與WPM培養基等量混合之培養基 (MW) 於光照下培養四週，‘Woodard’枝條中部位莖段在MW培養基中添加75 μM 2iP培養，總腋芽葉片數較無添加組的0.75片可以增加至16.25片葉片；‘Tifblue’枝條下部位莖段則在添加MW培養基中50 μM 2iP，總腋芽葉片數較無添加組的0.05片增加至15.08片葉片。取‘Woodard’經單節莖段培養產生的葉片中部，於添加TDZ培養基進行四週光照培養及四週黑暗培養誘導不定芽增生，‘Woodard’於7.5 μM TDZ培養基中，芽數由較無添加組的0個增加至9.5個。而利用‘Woodard’培植體葉片產生之癒傷組織，於添加7.5 μM TDZ的培養液在90 rpm且黑暗下進行懸浮培養，培養第30天時，平均細胞體積增加17.17倍；‘Tifblue’ 經單節莖段培養產生的葉片中部，則於9.1 μM TDZ培養基中，芽數由較無添加組的0個增加至6.83個。而葉片在經過繼代於4 μM ZT培養基中，‘Woodard’可以誘導出3.25個展開的不定芽；‘Tifblue’可以誘導出2.25個展開的不定芽。由論文的試驗結果推測，兔眼藍莓‘Woodard’進行微體繁殖的合適部位為枝條中部，如於MW培養基中添加75 μM 2iP、7.5 μM TDZ及4 μM ZT，約可以生產出52.81個不定芽；‘Tifblue’則以枝條下部為較佳部位，如於培養基為添加50 μM 2iP、9.1 μM TDZ及4 μM ZT，約可以生產出33.93個不定芽。

The aim of this thesis is to establish a micropropogation protocol of rabbiteye blueberries (Vaccinium ashei). Varieties of ‘Woodard’ and ‘Tifblue’ were used as plant materials. The objectives of this study are to set up the optimal 1) position of material, 2) position for regeneration, 3) medium contents, and 4) medium for cell suspension culture. The results showed that nodal segments collected from new shoots and cut with 1-2 cm had lower rate of contamination, higher percentage of bud sprouting, and longer axillary shoots. By been cultured in the medium of mixing Murashige and Skoog medium, woody plant medium (MW) and 75μM 2iP for four weeks in light, ‘Woodard’ produced 16.25 leaves per explant and ‘Tifblue’ produced 15.08 leaves per explant. In the proliferation stage, as the leaf segments of middle section from axillary shoots were cultured in MW medium containing different TDZ concentrations for four weeks in light and another four weeks in dark condition for 4 weeks, ‘Woodard’ produced 9.5 buds in the medium containing 7.5μM TDZ and ‘Tifblue’ produced 6.83 buds in the medium containing 9.1μM TDZ. In suspension culture of treatments 7.5μM TDZ, dark condition and 90 rpm shaking, the volume of callus increased 17.17 folds after 30 day of culture. And transferred the leaves to the medium containing 4μM ZT for 4 weeks, ‘Woodard’ produced 3.25 shoots per explant and ‘Tifblue’ produced 2.25 shoots per explant. From the results, we set up a protocol that have ‘Woodard’ and ‘Tifblue’ with the ability of producing 51.81 and 33.93 shoots per explant, respectively.

縮寫對照表............................................................i
中文摘要..............................................................ii
英文摘要...........................................................iii
目次................................................................ iv
表次索引.............................................................vi
圖次索引............................................................ vii
第一章 前人研究......................................................1
一、兔眼藍莓的生長背景及價值..................................1
二、兔眼藍莓的傳統繁殖法及微體繁殖法..........................2
三、兔眼藍莓微體繁殖法系統....................................3
四、兔眼藍莓單節莖段培養試驗..................................7
五、兔眼藍莓培植體葉片癒傷組織及不定芽培養試驗................8
六、兔眼藍莓不定芽伸長試驗....................................9
七、研究目的.................................................10
第二章 材料與方法...................................................11
一、試驗材料.................................................11
二、培養基成份...............................................11
三、培養基製備...............................................11
四、培養環境條件.............................................11
五、試驗方法.................................................12
六、統計分析.................................................14
第三章 結果.........................................................15
一、初代培養—單節莖段培養試驗...............................15
二、殖培養—葉片癒傷組織及不定芽培養試驗.....................21
三、伸長培養—不定芽伸長培養試驗.............................37
第四章 討論.........................................................40
一、初代培養—單節莖段培養試驗...............................40
二、增殖培養—葉片癒傷組織及不定芽培養試驗...................41
三、伸長培養—不定芽伸長培養試驗.............................44
第五章、結論.........................................................45
參考文獻..........................................................47
附錄................................................................53

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