Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/89210
標題: 聖誕紅之多倍體化與種間雜交
The Polyploidization and Interspecific Hybridization of Poinsettia
作者: Ya-Fen Lu
陸雅芬
關鍵字: 聖誕紅
多倍體誘變
種間雜交
胚拯救
poinsettia
polyploidy induction
interspecific hybridization
embryo rescue
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摘要: The present research aims to obtain the mutants with fertility of novel poinsettia cultivars by colchicine treatment, then used as pollen donor to cross with E. pulcherrima. We investigated the flowering physiology and pollen viability in Euphorbia pulcherrima and its endemic relatives ex. Euphorbia cyathophora, E. formosana, E. fulgens, E. jolkini, and E. leucocephala. It also assesses the possibility of individual hybridization between E. pulcherrima and other species or colchicine mutants. Moreover, the factor of cross barriers were also clarified and applied embryo rescue technique to resolve cross barriers to create desirable poinsettia progeny containing new Euphorbia germplasm. After the treatment of different concentrations of colchicine in poinsettia interspecific hybrid cultivars, each with lanolin or with cotton, 3 polyploidy mutants were obtained from ʻDulce Rosa™', 29 polyploidy mutants were obtained from ʻPrincettiaR-Hot Pink', and 4 polyploidy mutants were obtained from ʻLuv U Pink™'.Furthermore, the fertility of ʻDulce Rosa™' mutants and ʻPrincettiaR-Hot Pink' were recovered, and the highest pollen germination rate was reached to 26.5%. It also obtained morphological difference mutants. The bract color was observed in order to investigate the difference of between the poinsettia interspecific cultivars and their mutants in the epidermal layer and longitudinal-section of bract. Pigment concentration, amount of epidermal cell & epidermal layers each with a pigment, and the arrangement of epidermal cells all were related to bract color. Moreover, the forms of bract epidermal cell with onical or papillae cells observed by scanning electron microscope were also leaded to different colors of bract in poinesettia. The result of this research shows that the anthesis of Euphorbia endemic species is as follows: E. cyathophora blossomed all year-round, E. formosana blossomed from March to October; E. fulgens blossomed from November to April of next year; E. jolkini blossomed from February to July; E. leucocephala blossomed from October to April of next year, and poinsettia commercial interspecific hybrid cultivars all blossomed from November to April of next year. The optimal sucrose concentration in Brewbaker and Kwack (1963) medium for pollen germination of Euphorbia species were 15%~20% and of poinsettia cultivars were 20%~25%. Moreover, the optimum temperature for pollen germination of Euphorbia species was 25℃ and poinsettia cultivars was 20℃. The reciprocal crossing between poinsettia and Euphorbia species were also conducted. It couldn't harvest mature seeds in the other cross combinations, only obtain progenies when E. cyathophora as maternal donor crossed with poinsettia, However, the progenies traits were similar to E. cyathophora, and the DNA content was exactly the same as maternal donor. The in vivo pollen tube was observed by fluorescence microscope, and the result indicated that the pollen tubes stopped only in the stigma surface tissue on the self-pollination of E. cyathophora, but the hybrid capsule still continued to enlarge, and also get mature seeds when the cross combinations between E. cyathophora with poinsettia as pollen donor. Observed in vivo pollen tube by fluorescence microscope, the pollen tube could enter into ovule when the crosses between Euphorbia pulcherrima with Euphorbia species or the fertile mutants. This research also evaluated the efficiency of different cross combinations and period of day after pollination in embryo rescue, the results indicated that it were easier to regenerate when the section of ovule as expants which were obtained from the capsule on 26-32 day after pollination. The real hybrids were successfully obtained in present research which confirmed the relative DNA content by flow cytometry (poinsettia ʻSparkle Star'× ʻPrincettiaR-Hot Pink'mutant P-M13).
本論文利用商業流通之新型態聖誕紅種間雜交種進行秋水仙素誘變以恢復其花粉稔性。此外調查聖誕紅近緣種如猩猩草 (Euphorbia cyathophora)、台灣大戟 (E. formosana)、羽毛花 (E. fulgens)、岩大戟 (E. jolkini) 及白雪木 (E. leucocephala) 其開花生理及花粉活力,並評估收集之大戟屬種源及具稔性之秋水仙素誘變株與聖誕紅進行雜交之可能性及釐清雜交障礙之因素,亦使用胚拯救技術解除雜交障礙以創造具新穎血緣之聖誕紅雜交後代。 聖誕紅種間雜交種以不同濃度之秋水仙素佐羊毛脂膏或棉花處理,ʻDulce Rosa™'共獲得3株多倍體誘變株;ʻPrincettiaR- Hot Pink'共獲得29株多倍體誘變株;ʻLuv U Pink™'共獲得4株多倍體誘變植株。此外ʻDulce Rosa™'和ʻPrincettiaR- Hot Pink'經秋水仙素處理後,均可成功獲得恢復稔性之誘變植株,花粉活力最高可達26.5%,且獲得多樣性植物性狀型態改變之誘變植株。 觀察聖誕紅種間雜交種及其誘變株苞葉顏色,並調查苞葉上表皮細胞與苞葉橫向切面和原品種之差異。苞葉顏色與表皮細胞色素濃度、具色素之細胞數量、具色素之表皮細胞層數,及色素細胞排列相關。此外以掃描式電子顯微鏡觀察苞葉表皮細胞形狀,圓錐狀與乳突型細胞形狀亦影響其苞葉呈色。 大戟屬原生物種自然花期,猩猩草可周年開花;台灣大戟之花期為3月到10月;羽毛花之花期為11月至翌年4月;岩大戟花期為2月至7月;而白雪木之花期為10月至翌年4月。聖誕紅商業品種與種間雜交種之花期皆為11月至翌年4月。大戟屬原生物種花粉培養於Brewbaker and Kwack (1963) 培養基中,其最適蔗糖濃度約為15%~20%,花粉發芽適宜溫度主要為25℃。而聖誕紅品種之最適蔗糖濃度約為20%~25%,其花粉發芽最適溫度為20℃。 聖誕紅與大戟屬原生物種進行正反雜交,僅於以猩猩草為母本與聖誕紅雜交後能成功獲得後代,其餘雜交組合均無法獲得成熟種子。然而該雜交後代性狀與母本猩猩草相似,且其DNA含量與母本猩猩草完全相同。使用花粉管體內螢光觀察發現猩猩草授予自體花粉及聖誕紅商業品種花粉,其花粉管均無法伸長進入花柱,僅萌發停滯於柱頭表面組織,然其雜交蒴果仍能持續膨大並獲得成熟種子。 聖誕紅被授予大戟屬近緣種及恢復稔性之誘變株花粉,其兩者之花粉管均可順利萌發並抵達胚珠。評估不同雜交組合及授粉後日數對胚拯救之影響,授粉後26-32天之胚珠切片培殖體進行培養較易再生發育。目前已獲得經流式細胞儀驗證之真實雜交後代 (聖誕紅ʻ閃亮之星' × ʻPrincettiaR- Hot Pink'誘變株P-M13)。
URI: http://hdl.handle.net/11455/89210
其他識別: U0005-1308201500330400
文章公開時間: 2018-08-25
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