https://scholars.lib.ntu.edu.tw/handle/123456789/91738
標題: | 鐵線蕨孢子無菌播種與部分組織培養繁殖系統之建立 | 作者: | 葉德銘 | 關鍵字: | 脆鐵線蕨;孢子;原葉體;無菌繁殖;乙烯;激勃素;Adiantum tenerum ‘Scutum Roseum’;spore;prothallium;in vitro propagation ethylene;GA3 | 公開日期: | 2002 | 出版社: | 臺北市:國立臺灣大學園藝學系暨研究所 | 摘要: | 脆鐵線蕨(Adiantum tenerum ‘Scutum Roseum’)孢子播於無土介質及試管內,比 較其原葉體生長及發育。孢子播於泥炭 苔、蛭石與珍珠砂之無土混合介質下,原 葉體長度約1.2 mm、寬度約1.3 mm 以上 時,即有藏精器的分化;原葉體長度約2.0 mm、寬度約2.5 mm,方觀察到藏卵器之 形成。將孢子播於試管內,經增殖而來的 原葉體僅分裂數個細胞後即有藏精器的形 成。且於無菌撒播20 天後,自翼片下側近 假根處,開始有原葉體的增殖與鮮重之增 加。於60 天後,原葉體的鮮重即無明顯增 加,而葉綠素含量下降。適當的繼代時期 為無菌撒播後50-60 天,於第一次繼代6-8 週後需再行第二次繼代,使原葉體大量增 生。若將無菌撒播60 天之原葉體團,以全 量強生氏營養液浸泡5 秒鐘後,再置於無 土介質中,其孢子體形成速率,較前人所 推薦之½ MS 為佳。本研究建立以部分組 織培養方式(partial tissue culture system),可 兼顧原葉體培育、增殖並可獲得更多的孢 子體。此外由迴歸分析得知: 撒播後每瓶原 葉體鮮重增加至8-10 g 時,瓶內乙烯含量 亦近呈直線增加至0.5-0.6 µL/L。於含 0.1-10 mg/L GA3 之培養基中,原葉體發育 階段在未形成頂端分生組織前,即有藏精 器的分化。若培養基中添加0.01-10 mg/L 之GA3 抑制脆鐵線蕨原葉體的生長;100 mg/L 之GA3 處理者抑制孢子發芽。 Comparative growth and development of gametophyes and sporophytes were determined in Adiantum tenerum ‘Scutum Roseum’ sown in soilless mix and in vitro. Prothallia in soilless mix were competent to initiate antheridia provided that these prothallia had reached a critical size at length > 1.2 mm and width > 1.3 mm, while archegonia were observed at prothallia with length > 2.0 mm and width > 2.5 mm. In contrast, antheridia were observed in the young adventitious prothallia cultured in vitro, with only a few cells. The young adventitious prothallia were differentiated from the lower region of the wing cells of the prothallia after spores were sown in vitro for 20 days. Within sowing for 60 days, the fresh weight of prothallia increased due to adventitious outgrowths. The fresh weight of prothallia increased little and the chlorophyll content started to decrease after sowing in vitro for 60 days. The first optimum in vitro transferring duration was approximately 50 to 60 days after sowing, and the second subculture was 6 to 8 weeks after the former transferring. Before transferring from in vitro to soilless mix, the prothallia soaked with Johnson’s solution for 5 seconds resulted in a faster sporophyte formation than those soaked with 1/2 MS as recommended previously. The established partial tissue culture system could facilitate both mass-propagation of the prothallia and production of more sporophytes. Regression analysis revealed that the ethylene production increased almost linearly up to 0.5-0.6 µL/L with increasing prothallial fresh weight up to 8-10g per vase. Supplementary GA3 at 0.1-10 mg/L reduced the length and width of prothallia but promoted the formation of antheridia, which were observed in the young gametophytes prior to apical meristem initiation stage. Increasing GA3 at 100 mg/L prevented spore germination. |
URI: | http://ntur.lib.ntu.edu.tw//handle/246246/18224 | 其他識別: | 902313B002269 | Rights: | 國立臺灣大學園藝學系暨研究所 |
顯示於: | 園藝暨景觀學系 |
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902313B002269.pdf | 103.81 kB | Adobe PDF | 檢視/開啟 |
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