(65(4):384-394) Influence of Explant, Plant Growth Regulator and Illumination on Adventitious Shoot Induction of In Vitro Cultured Salvia miltiorrhiza

本研究利用丹參 (Salvia miltiorrhiza) 組織培養苗之葉柄和葉片培植體進行不定芽的誘導試驗。葉柄培植體於含有1 mg L-1 芐腺嘌呤 (N6-benzyladenine; BA) 與0.5 mg L-1 奈乙酸 (α-naphthaleneacetic acid; NAA) 之MS (Murashige & Skoog 1962) 培養基中培養6 wk 後不定芽形成率達100%，平均每個培植體可形成3.7 個不定芽。將葉柄和葉片培植體預培養於含有1–2 mg L-1 2,4-二氯苯氧乙酸 (2,4-dichlorophenoxyacetic acid; 2,4-D) 之MS培養基2 或3 wk 後，再分別繼代培養於不含植物生長調節劑之MS 培養基中培養4 或3 wk，可誘導癒合組織和不定根形成。葉片癒合組織於含有0.25 mg L-1 BA 與0.2 mg L-1 2,4-D 之MS 培養基於黑暗中培養8 wk，顯示除了癒合組織增殖外，亦可形成少數不定芽。將癒合組織繼代培養於含有相同2,4-D 濃度但不同BA 濃度之培養基中，於光照或黑暗環境培養8 wk 後，其中以2 mg L-1 BA 配合光照處理可產生最多不定芽，每接種0.2 g 癒合組織平均可形成14.1 個不定芽。本研究利用丹參組織培養苗建立直接與間接不定芽再生大量繁殖體系，除可供生產丹參種苗所需外，亦可應用於誘變與轉基因之研究。 Petiole and leaf explants derived from in vitro Salvia miltiorrhiza were used for shoot regeneration in this study. The highest adventitious shoot formation rate of 100% with 3.7 shoots/explant in average was obtained from petiole segments cultured on Murashige and Skoog’s (MS) medium containing 1 mg L-1 N6-benzyladenine (BA) and 0.5 mg L-1 α-naphthaleneacetic acid (NAA) for 6 wk of culture. Callus and adventitious roots were induced from petiole and leaf segments pre-cultured on the MS medium supplemented with 1–2 mg L-1 2,4-dichlorophenoxyacetic acid (2,4-D) for 2–3 wk followed by transferring on a hormone-free MS basal medium for a total 6 wk of culture. Calli were proliferated on the MS basal medium containing 0.25 mg L-1 BA and 0.2 mg L-1 2,4-D under darkness for 8 wk of culture along with few adventitious shoots were found. Proliferated calli were subcultured to the medium containing with same concentration of 2,4-D in combination with various BA concentration under light and dark condition for shoot regeneration. The highest induction number of adventitious shoots was 14.1 shoots 0.2 g-1 callus from the medium containing 2 mg L-1 BA under light condition. An efficient micropropagation system of Salvia miltiorrhiza by direct and indirect adventitious shoot regeneration systems were established in this study which would not only supply for plantlet production but also apply on mutation and genetic transformation studies