篇名 | 避免黛粉葉莖頂分生組織初代培養內源感染的處理方法 |
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卷期 | 60:2 |
並列篇名 | The Methods for Avoiding Endo-contamination of Shoot-tip Explants in Initial |
作者 | 沈榮壽 、 李哖 |
頁次 | 077-088 |
關鍵字 | 分生組織大小 、 乾燥處理 、 除鱗片 、 芽體再生 、 系統性病原 、 apical meristem size 、 drought stress 、 removal of scales 、 shoot organogenesis 、 systemic pathogen |
出刊日期 | 201406 |
本研究旨在探討黛粉葉無病原培植體之準備與芽體無菌培養之建立。利用 不同大小莖頂分生組織作為起始培植體,培養結果於MS 基礎培養基含1 mg·L-1 NAA 及1 mg·L-1 TDZ 培養12 週後,最小之莖頂(0.1×0.25 mm)培植體具有20%的存 活率,較大(0.90×1.20 mm)存活之莖頂分生組織培植體,則顯現高繁殖潛力,獲得 12.3 個再生芽體;另以4 種大小黛粉葉腋芽分生組織作為無病原培植體來源,剝除 3 或4 片鱗片數的存活率可達30%-35%,剝掉1 片鱗片的分生組織培植體平均每一 個反應培植體可達11.2 個再生芽體;顯示利用黛粉葉腋芽進行初代培養可避免病原 感染,可建立其微體繁殖芽體再生。此外,母株經6 週人工乾旱處理之枝梢腋芽, 進行精細解剖再切取腋芽培養,結果顯示,剝除2-4 片鱗片莖頂培植體之存活成功 率可達80%-92.5%,可有效避免初代培養感染微生物。
In this research we described the methods of pathogen-free meristem explant preparation and the shoot proliferation of Dieffenbachia. After culturing different sizes of shoot apical meristem on solidified MS medium with 1 mg·L-1 NAA (1-Naphthylacetic acid and 1 mg·L-1 TDZ[1-phenyl-3-(1,2,3-thiadiazol-5-yl)urea] for 12 weeks, a 20% survival rate was obtained from the smallest size (0.1×0.25 mm) of shoot-tip meristem explant; while 12.3 shoots were regenerated from the largest size (0.90×1.20 mm) of shoot-tip meristem explant during initial culture. Moreover, four types of axillary meristem-tip by removing scales were used as explants for aseptic culture. Axillary meristem-tip after 3 or 4 scales removed, used as explant, had higher survival rate of 30%-35%. An axillary meristem-tip explant, with one scale excised, produced 11.2 shoots. Removal of scales from meristem-tip was ideal for the elimination of in vitro endo-contamination in Dieffenbachia. High survival rate (80%-93%) was achieved in axillary bud explants, with 2-4 scales removed, excised from the stock plants that had not been watered for 6 weeks. The methods we described here reduced in vitro endo-contamination and had high shoot multiplication of Dieffenbachia.