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Title: | 台灣相思樹人工誘導多倍體的研究 Artificial polyploidy induction of Acacia confusa |
Authors: | Tsun-Yu Pan 潘遵宇 |
Advisor: | 王亞男 |
Keyword: | 臺灣相思樹,誘導多倍體,秋水仙素,染色體,流式細胞儀,寶為細胞, Acacia confusa,induced polyploidy,colchicine,flow cytometer,guard cell, |
Publication Year : | 2011 |
Degree: | 碩士 |
Abstract: | 本研究以採自林務局台南甲仙工作站之台灣相思樹(Acacia confuse Merr.)種子為材料,藉由不同濃度的秋水仙素以不同時間處理,以誘導其產生多倍體化的現象。期望可藉由本研究尋找以秋水仙素誘導多倍體發生的最佳方式,做為未來台灣相思樹育種的一個參考依據。
將浸泡於 100℃熱水中,任其自然冷卻 24 小時的台灣相思樹種子,以重量百分率濃度 0%、0.01%、0.05%、0.1%、1%、2%、4%之秋水仙素浸泡 12 與 24 小時,觀察其發芽率以及根部膨大的比例。其中以 1%秋水仙素浸泡 12 至 24 小時的結果最佳。再經過染色體觀察與流式細胞儀分析之後,確定芽體具膨大現象之幼苗確 實為多倍體化的幼苗。除了直接鑑定之外,觀察初生葉的葉背氣孔之後,發現多倍體幼苗葉背氣孔的平均面積較二倍體幼苗大 64.97μm2,單位面積內的氣孔數也較少,二倍體在單位面積(1mm2)下的平均數量為 131 個,多倍體則僅有 53.4 個。 然而,以人工方式誘導的多倍體幼苗,由於生長點受到抑制的因素,生長速度遠較未處理的幼苗慢,常常因無法長出幼葉或是生長點受微生物感染而死亡。此外,由流式細胞儀分析假葉及根部的結果也顯示,以秋水仙素誘導種子所產生的人工多倍體,常常會有混倍體的現象,且多倍體細胞亦容易被二倍體細胞所取代,使幼苗逐漸由混倍體轉回為二倍體,可見得人工誘導多倍體具有不穩定的缺點。未來可將研究方向轉為以誘導培植體,或是將多倍體化的組織進行組織培養,來提高誘導多倍體的穩定性以及存活率。 The Acacia confusa Merr. seeds collected from Tainan Chia Shian Work Station of Forest Bureau were used for the study. Different concentrations of colchicine and different time were dealed with in the experimant. It was expected to find a best way to induce the polyploidy of Acacia confusa. The seeds of Acacia confuse were first soaked in hot water at 100 ℃. Then let them cool down for 24 hours. The seeds were put into colchicine of mass percentage by 0%, 0.01%, 0.05%, 0.1%, 1%, 2% and 4% for 12 and 24 hours. The germination rate and the ratio of the swollen roots of each treatment were recored. The result indicated that the seeds soaked in mass percentage by 1% for 12 to 24 hour, was the best concentration to induce polyploidy of Acacia confusa. Polyploidy was proved by counting the chromosome numbers from the tissue of swollen roots, and analysis of the cells took from roots and phyllodes by flow cytometer. It was confirmed that the germinating seeds with swollen roots are the polyploidy of Acacia confusa. The stoma and guard cells of polyploids are different from diploids by size and number by unit area. The average size of stoma and guard cells from polyploids was larger than those from diploids by 64.97μm2. The average number of stoma from poliploids were 131﹐ and those from diploid were 53.4. vi However, the polyploids induced by colchicine grew slowly and were easily killed by microbe caused by the restrant of meristem. Besides, the analysis cells of roots and phyllodes by flow cytometer showed that most of polyploidy seedlings were mixoploid. It is possible that most of polyploidy cells would be replaced by diploidy cells. Therefore, it is important to find a way to make the polyploidy more stable and improve survival in the future, such as tissue culture or use colchicine to treat the callus of culture. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/7045 |
Fulltext Rights: | 同意授權(全球公開) |
Appears in Collections: | 森林環境暨資源學系 |
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