Studies on Inheritance of Leaf Margin Spine in Pineapple and Mutation Breeding of 'Tainung No. 4' Pineapple

In this study, use different pineapple cultivars for hybridization tests. The pollen germination rate of 'Tainung No. 4' was 4.1%; 'Tainung No. 11', 'Tainung No. 13' and 'Tainung No. 20' were 53.3%, 36.4% and 45.1%, respectively, all were significantly higher than that of 'Tainung No. 4'. Progenies from crosses between spiny 'Tainung No. 4', spiny tip 'Tainung No. 11', spiny tip 'Tainung No. 13' and spineless 'Tainung No. 20' pineapple were analyzed for leaf margin types to evaluate the genotypes of these cultivars. The leaf type ratio in progenies of 'Tainung No. 4' ×'Tainung No. 20' was spiny : spineless = 1 : 1; 'Tainung No. 11' × 'Tainung No. 20' was spiny + spiny tip : spineless = 1 : 1 and spine : spiny tip = 1 : 1. 'Tainung No. 13' ×'Tainung No. 20' was spiny + spiny tip : spineless = 1 : 1 and spine : spiny tip = 1 : 1. According to the hypothesis of Collins and Kerns (1946), the genotype of'Tainung No. 4' is ppss, 'Tainung No. 11' is ppSs, 'Tainung No. 13' is ppSs and 'Tainung No. 20' is Ppss. The results could be used in the future pineapple breeding programs. The effects of using EMS (ethyl methanesulphonate) as a mutagen on the survival rate of 'Tainung No. 4' pineapple plantlets cultured in vitro were examined. The survival rate of plantlets (5-8 mm) in the treatments of 0.6% EMS 12hr, 14 hr and 16 hr were 96.7%, 76.7% and 63.3%, respectively. However, in the treatment of 0.6% EMS 18 hr, the survival rate decreased to 20%. The LD50 treatment should be EMS 0.6% and duration between 16 hr and 18 hr. The growth vigor recovered slowly with increased treatment duration by evaluating the rooting rate of plantlets. Adding 1% DMSO (dimethyl sulfoxide) into the EMS solution increased the survival rate of plantlets. Large plantlets (4-5 cm) were treated with EMS followed by dark treatment to induce etiolated plantlets. The survival rate of large plantlets decreased with increased EMS concentration and treatment duration. The survival rate of large plantlets increased when adjusting the pH value of EMS solution to 7.0. The survival rate of etiolated plantlets in the treatment of 0.01% 1hr was 20.0%, and that of the control was 36.7%. There were significant difference between two treatments. In summary, the survival rate of different plantlets decreased when EMS concentration and treatment duration increased.本研究以不同品種鳳梨進行雜交試驗，觀察所得之雜交子代其葉緣形態，藉以了解台灣重要鳳梨品種之葉緣形態表現基因型。在花粉平均發芽力方面，'台農4號'僅4.1%，而'台農11號'、'台農13號'、'台農20號'鳳梨則分別為53.5%、36.3%、45.1%，均顯著大於'台農4號'。以葉緣皆有刺之'台農4號'及葉緣少刺之'台農11號'、'台農13號'鳳梨為母本與葉片全緣無刺之'台農20號'鳳梨為父本進行雜交，觀察所得之雜交子代其葉緣形態，藉以了解各品種控制葉緣形態表現之可能基因型。'台農4號'雜交'台農20號'得到之後代葉緣全刺：葉緣無刺之比為1：1。'台農11號'雜交'台農20號'得到之後代葉緣有刺(全刺+少刺)：葉緣無刺為1：1；葉緣全刺：葉緣少刺亦為1：1。以'台農13號'雜交'台農20號'得到之後代葉緣有刺(全刺+少刺)：葉緣無刺為1：1；葉緣全刺：葉緣少刺亦為1：1。故以CollinsandKerns(1946)之假說推測，'台農4號'鳳梨品種之葉緣刺基因型為ppss、'台農11號'之葉緣刺基因型為ppSs、'台農13號'之葉緣刺基因型亦為ppSs、'台農20號'鳳梨品種之葉緣刺基因型為Ppss，此結果可作為日後鳳梨雜交育種親本選擇之參考。 另以'台農 4 號'鳳梨組培苗為材料，探討 EMS (ethyl methanesulphonate)誘變處理之最適條件。以組培小苗(5-8 mm)做為誘變材料，0.6% EMS 處理 12、14、16小時，存活率分別為 96.7%、76.7%、63.3%，然而若浸泡 18 小時，其存活率則僅有 20%，半致死率(LD50)之處理時間應界於 16 至 18 小時。發根率方面，結果顯示誘變處理時間愈長，植株恢復生長勢所需時間愈長。而添加 1% DMSO (dimethylsulfoxide)處理之組培小苗其存活率有增加之趨勢。以組培大苗(4-5 cm)為誘變材料，經 EMS 處理後，進行暗期處理誘變誘導白化苗，植株存活率隨著 EMS 濃度與處理時間增加而降低。而誘變溶液調整 pH 值為 7.0 之組培大苗植株存活率高於未調整之處理。組培白化苗方面，以 0.01%之 EMS 溶液，浸泡 1 小時植株存活率為 20.0%；對照組為 36.7%，具顯著差異。整體而言，經誘變處理後之各種植株存活率均隨著 EMS 濃度與處理時間增加而降低。中文摘要..........i Abstract..........ii 表目次..........v 圖目次..........vi 一、前言..........1 二、前人研 究..........2 (一)鳳梨概述..........2 (二)鳳梨分類..........2 (三)植物刺的種類..........4 (四)植物刺之遺傳..........4 (五)鳳梨葉緣性狀之遺傳..........6 (六)鳳梨育種應用..........8 1.雜交育種..........8 2.誘變育種..........9 三、材料與方法..........11 試驗一、不同品種鳳梨花粉體外發芽試驗..........11 (一)材料..........11 (二)方法..........11 (三)花粉發芽調查..........11 試驗二、不同品種鳳梨雜交試驗..........12 (一)'台農 4 號'與'台農 20 號'雜交試驗..........12 (二)'台農 11 號'與'台農 20 號'雜交試驗..........12 (三)'台農 13 號'與'台農 20 號'雜交試驗..........13 試驗三 '台農 4 號'鳳梨組培小苗 EMS 誘變..........14、 (一)不同濃度 EMS 與處理時間組合之誘變處理..........14 (二)EMS 添加 DMSO 及調整 pH 值與不同處理時間組合之誘變處理….15 試驗四 '台農 4 號'鳳梨組培大苗 EMS 誘變..........16、 (一)不同濃度 EMS 與處理時間組合之誘變處理..........16 (二)不同濃度 EMS 添加 DMSO 與不同處理時間組合之誘變處理.....16 (三)EMS 添加 DMSO 及調整 pH 值與不同處理時間組合之誘變處理.....17 試驗五 '台農 4 號'鳳梨組培白化苗 EMS 誘變..........18、 (一)不同濃度 EMS 與處理時間組合之誘變處理..........18 (二) EMS 添加 DMSO 及調整 pH 值與不同處理時間組合之誘變處理..18 四、結果 ..........20 (一)不同品種鳳梨花粉體外發芽試驗..........20 1.花粉體外發芽試驗..........20 2.雌雄蕊形態觀察..........20 (二)不同品種鳳梨雜交試驗..........25 1.'台農 4 號'與'台農 20 號'雜交試驗..........25 2.'台農 11 號'與'台農 20 號'雜交試驗..........25 3.'台農 13 號'與'台農 20 號'雜交試驗..........25 (三)'台農 4 號'鳳梨組培小苗 EMS 誘變..........32 1.不同濃度 EMS 與處理時間組合之誘變處理..........32 2.EMS 添加 DMSO 及調整 pH 值與不同處理時間組合之誘變處理...32 (四)'台農 4 號'鳳梨組培大苗 EMS 誘變..........38 1.不同濃度 EMS 與處理時間組合之誘變處理..........38 2.不同濃度 EMS 添加 DMSO 與不同處理時間組合之誘變處理....38 3.EMS 添加 DMSO 及調整 pH 值與不同處理時間組合之誘變處理...38 (五)'台農 4 號'鳳梨組培白化苗 EMS 誘變..........42 1.不同濃度 EMS 與處理時間組合之誘變處理..........42 2.EMS 添加 DMSO 及調整 pH 值與不同處理時間組合之誘變處理...42 五、討論 ..........46 (一)不同品種鳳梨雜交之探討..........46 (二)鳳梨組培苗 EMS 誘變條件之探討..........47 (三)總結..........50 六、參考文獻..........5