兔眼藍莓組織培養苗生產流程之建立

Abstract

本研究的目的為找出兔眼藍莓單節莖段體外培養之最佳培植體、基礎培養基、枝條再生之最佳培植體、植物生長調節劑組合與濃度與瓶外發根，建立快速大量繁殖兔眼藍莓種苗之組織培養生產流程。試驗材料為‘Blueshower’、‘Woodard’及‘Tifblue’三品種。試驗結果顯示以綠熟枝條上部2/3，截切成2公分的帶節莖段是初代培養最好的培植體，其汙染率低、萌芽率高且萌生之側枝長。帶節莖段培養於Murashige and Skoog medium與woody plant medium等量混和並添加50 μM 2iP之培養基，於照光下培養一個月後，‘Blueshower’平均每培植體可產生7.33片葉片，‘Woodard’可產生5.49片葉片，‘Tifblue’則可產生3.85片葉片。於增殖期，初代培養所獲得之葉片中段培養於含2.3 μM TDZ之培養基，經一個月黑暗與一個月照光培養後，‘Blueshower’平均每培植體可再生15.89個不定芽，‘Woodard’可再生2.5個芽，‘Tifblue’可再生0.17個芽。再生之‘Blueshower’芽叢移至含4 μM玉米素的培養基培養一個月後，每培植體可得2.25長於1cm的枝條。以此程序獲得之枝條，切口沾2000 ppm之IBA後，以泥炭苔：珍珠石體積比為3：2的濕潤發根介質，進行瓶外扦插，40天後可得80%至90%之發根率。

To fast propagate rabbiteye blueberry (Vaccinium ashei) plants, a protocol for tissue culture was required. The objectives of this study are 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 blueberries. ‘Blueshower’, ‘Woodard’ and ‘Tifblue’ rabbiteye blueberries were used as plant materials. The results showed that in the culture establishment stage, nodal segments collected from the upper 2/3 of a mature green shoot and adjusted to 2 cm long have a low rate of contamination, high percentage of bud sprouting, and produced long axillary shoots. Nodal segments of ‘Blueshower’, ‘Woodard’, and ‘Tifblue’ cultured in medium mixed of equal parts of Murashige and Skoog medium and woody plant medium and contained 50 μM 2iP in light conditions for 1month produced 7.33, 5.49, and 3.85 leaves per explant, respectively. In the proliferation stage, segments from the middle section of a leaf from regenerated axillary shoots were recommended to be cultured 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 shoot was dipped in 2000 ppm IBA and inserted in moist rooting mixture of peat moss and perlite at 3:2 ratio (v/v), 80%-90% shoot cuttings rooted after 40 days.