以體外誘殖葉片生產兔眼藍莓組織培養苗之流程

The Establishment of the Plantlet Production of Rabbiteye Blueberry through Tissue Culture from ＂In Vitro＂ Derived Leaf Explants

For rapid and mass-propagation of rabbiteye blueberry (”Vaccinium ashei”) plants, a protocol for tissue culture was required. The objectives of this study were 1) to investigate the optimal tissue culture condition for rapid proliferation of rabbiteye blueberry plants, and 2) to establish standard operating procedure of tissue culture for rabbiteye blueberry. ”In vitro” derived leaf explants of 'Blueshower', 'Woodard' and 'Tifblue' rabbiteye blueberry were used as plant materials to identify the optimal medium for tissue culture and ”ex vitro” rooting. The results showed that segments from the middle section of a leaf from regenerated axillary shoots were suitable for culture in the medium containing 2.3 μM TDZ. After incubation in the dark for one month and then in the light for another one month, the number of adventitious buds per explant was 15.89 in 'Blueshower', 2.5 in 'Woodard', and 0.17 in 'Tifblue'. The bud clumps of 'Blueshower' were transferred to medium containing 4 μM zeatin and cultured for 1 month. On average, a bud clump generated 2.25 shoots longer than 1 cm. The regenerated shoots were used for ex vitro rooting test. The shoots were dipped in 2000 ppm IBA and inserted in moist rooting mixture of 3 peat moss and 2 perlite (by volume), 80%-90% shoot cuttings rooted after 40 days. 建立組織培養標準流程是快速大量繁殖兔眼藍莓苗的重要步驟之一。本研究之目標為1）探討快速增殖兔眼藍莓之最佳組培條件，2）建立標準組培操作流程。本研究以‘Blueshower　’、‘Woodard　’及‘Tifblue　’三品種體外誘殖之葉片為材料，探討最適合兔眼藍莓苗組織培養及瓶外發根之條件。初代培養所獲得之葉片中段培養於含2.3μM TDZ之培養基，經一個月黑暗與一個月照光培養，‘Blueshower　’平均每培植體可再生15.89個不定芽，‘Woodar　’可再生2.5個芽，‘Tifblue　’則可再生0.17個芽。再生之‘Blueshower　’芽叢移至含4μM玉米素的培養基培養一個月，每培植體可得2.25個大於1 cm的枝條。此外，生長於4μM玉米素的大於1 cm枝條適合作為瓶外發根之培植體，將其沾2000 ppm之IBA後插入泥炭苔：珍珠石體積比為3：2的濕潤發根介質，40天後可獲得80%-90%的已發根苗。