利用組織培養繁殖臺灣一葉蘭

Tissue Culture of Pleione formosana Hayata

盧美君(Mei-Chun Lu)

《植物種苗》 6卷1期 (2004/03) Pp. 60-71

https://doi.org/10.30077/SN.200403.0006

摘要

臺灣一葉蘭(Pleione formosana Hayata)為臺灣之原生蘭，因其自然繁殖倍率低，種源日漸匱乏。本試驗探討無菌播種和癒合組織再生之影響因子，種子發芽時無機鹽類濃度以1/4MS到1/8MS顯著較佳；0.5mg/L菸鹼酸可促進種子的發芽與生育，高於4mg/L反而有抑制效果；花寶培養基中添加NAA3mg/L及BA1mg/L可顯著增加小苗鮮重；Tryptone3g/L可增加兩倍的小苗鮮重。由播種到長根出瓶約80天，馴化率可達95%以上；培養一季後可得直徑4到5mm的小球。此外，於含有2,4-D0~5mg/L及TDZ 0~0.5mg/L之1/2Murshige-Skoog培養基中進行癒合組織誘導，篩選出了12個營養系，繼代後每月增殖4到5倍。培養2.5到3個月可經由擬原球體(protocorm-like body)再生芽體，每0.01g胚性癒合組織在含有TDZ0.5mg/L的1/2MS培養基中最多再生出134個芽。

關鍵字

臺灣一葉蘭 ； 無菌播種 ； 側芽培養 ； 癒合組織

並列摘要

Pleione formosana Hayata, a native orchid in Taiwan, is endangered due to its low proliferation rate. The optimum medium compositions for sowing and the totipotent callus lines were studied in this report. Direct seed sterilization resulted in the formation of protocorm-like bodies (19.6%) and the browning of seeds (37%). A mineral concentration of 1/4MS or 1/8MS was significantly superior than MS or 1/2 MS for seed germination. Nicotinic acid of 0.5mg/L enhanced seed germination and development, while concentrations higher than 4mg/L showed negative effects. Tryptone of 3g/L promoted a 2-fold fresh weight increase. The Hyonex medium containing 3mg/L NAA and 1 mg/L BA was effective for increasing fresh weight of seedlings. The addition of charcoal enhanced the seed germination and development. The duration for in vitro seedling growth was 80 days and the ex vitro survival rate was above 95%. Twelve totipotent callus lines of Pleione formosana Hayata were established from protocorm-derived callus cultured on half-strength of Murashige-Skoog medium with 2,4-dichlorophenoxyacetic acid (2,4-D, 1-5mg/L) and 1-phenyl-3-(1,2,3-thiadiazol-5-yl) urea (TDZ, 0-0.5mg/L) in the dark. The proliferation rate of totipotent calli was 4 to 5 folds in fresh weight after 30 days of culture on basal medium containing 5 mg/L 2,4-D and 0.5mg/L TDZ in the dark. These calli were regenerated to plantlets via protocorm-like bodies (PLBs) on medium containing 0.5mg/L TDZ after 75 to 150 days of culture. The maximum number of shoot buds generated by 0.01 g callus explant was 134 after 4 months of culture.

並列關鍵字

Pleione formosana Hayata ； embryo culture ； bud culture ； callus