Induction of multiple shoots and proliferation of Acorus macrospadiceus (Yamamoto) F.N.Wei et Y.K.Li

Abstract

摘要 本試驗以茴香菖蒲(Acorus macrospadiceus)之葉片和匍匐莖上側芽為培植材料,經由多芽體的誘導建立快速大量繁殖再生植株的方法。試驗結果摘述如下: 以匍匐莖上側芽為培植體接種於添加1 mg/L TDZ之處理培養基中,可誘導多芽體,每個培植體平均可獲得3個芽體。但以葉片為培植材料,培養於添加不同auxin和cytokinin之16種組合培養基中,均無法誘導多芽體形成。 不同生長調節劑對多芽體增殖之影響,以添加4 mg/l TDZ之處理培養基可獲得最多芽體數,平均為3.5個;BA之處理中以添加8 mg/l BA之處理可得最多芽體數,平均為2.7個;添加BA之處理在多芽體形成率和平均芽體形成數均較TDZ處理低。另外發現,單獨添加TDZ和BA之處理在多芽體増殖效果較NAA和2,4-D組合BA之處理為佳。 基本鹽類濃度方面,以全量MS鹽類對多芽體増殖效果最好,多芽體形成率為100﹪,每個培植體平均可誘導3個芽體。蔗糖濃度則以添加50 g/l 蔗糖之處理可得最多芽體,每個培植體平均可誘導3.4芽體形成,而10 g/l蔗糖之處理則會抑制多芽體形成。胺基酸添加物方面,添加CH之處理對多芽體増殖之效果優於添加peptone之處理,以添加100 mg/l CH培養基有最好的增殖效果,芽體數平均為3.3,但高濃度之CH處理則會抑制芽體形成。 以2和4 mg/L TDZ二種培養基所誘導出之芽體進行連續4次繼代培養後,所誘導芽體數分別介於2.9~ 3.0和3.4 ~ 3.6個,顯示各世代處理間無顯著差異,多芽體在添加TDZ之培養基可穩定增殖。 再生芽體的發根試驗中,添加CH處理之發根率介於83.3~100﹪,而peptone處理之發根率介於76.7~ 96.7﹪,添加50和100 mg/l CH之培養基有最高的發根率(100﹪),優於添加抗壞血酸之培養基(發根率介於56.7~76.7﹪),及添加NAA和IBA之培養基(發根率<45﹪)。Summary The leaves and lateral buds of Acorus macrospadiceus were used as materials in this study to establish the optimal protocol for rapid multiplication through induction of multiple shoot. The results were summarized as follows: However, when leaf explant was used no multiple shoot was induced in all of medium containing different levels of auxin and cytokinin. The lateral bud explants cultured on the media containing 1 mg/L TDZ were induced and averaged 3 shoots per explant. The regenerated shoot cultured on media with 4 mg/L TDZ had highest proliferation efficiency and averaged 3.51 per explant. The optimal concentration of BA to induced and multiply shoot was at 8 mg/L, averaged 2.73 shoots per explant. However addition of TDZ was found to be better than BA in the rate and number of shoot formation. The results also showed that addition of TDZ or BA alone proliferated more shoots than the combined media with NAA or 2, 4-D and BA. The highest proliferation efficiency was found in the medium with full MS salt, reached 100% and averaged 3 shoots per explant . Media containing 50 g /L sucrose produced more multiple shoots, averaged 3.37 shoots per explant when sucrose more than 50 g/L inhibited shoot proliferations. Media containing CH proliferated shoot more than peptone and 100 mg/L CH had the best performance averaged 3.27 shoots per explant. Shoot proliferation was inhibited under high CH concentration level. The averaged shoots induced from shoot explant through four times subculture on media with 2 and 4 mg/L TDZ were ranged 2.86 to 3.03 and ranged 3.43 to 3.56, respectively. The result indicated that shoots can be stable multiplied on medium containing TDZ. Rooting efficiency of media containing CH and peptone were ranged 83.3 to 100﹪and ranged 76.7 to 96.7﹪, respectively. Regenerated shoot cultured on media with 50 and 100 mg/L CH rooted batter than media containing ascorbic acid (56.7 to 76.7﹪) or NAA and IBA (<45%).目 錄 章節 頁次 中文摘要 -------------------------------------------------------------- i 英文摘要 -------------------------------------------------------------- iii 目錄 ------------------------------------------ ------------------------- iv 表目錄 ----------------------------------------------------------------- v 圖目錄 ----------------------------------------------------------------- v 前言 -------------------------------------------------------------------- 1 前人研究 -------------------------------------------------------------- 2 材料與方法 ---------------------------------------------------------- 13 結果 ------------------------------------------------------------------- 21 討論 ------------------------------------------------------------------- 65 參考文獻 ------------------------------------------------------------- 7

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