彩葉屬（Neoregelia）、鶯歌屬(Vriesea)及空氣屬(Tillandsia)觀賞鳳梨組織培養器官發生與體胚發生之研究

Abstract

紅羽鶯歌鳳梨（Vriesea erecta）吸芽的莖頂分切以及幼葉培養，接種於1/2 MS培養基添加NAA 1.0 mg/L + TDZ 1.0mg/L，兩個月後分別可由葉腋及葉基誘導出多量的不定芽，但芽體叢生不易抽長，需分切並移至含有NAA 0.5 mg/L的培養基以助芽體生長與不定根形成。 以牡丹彩葉鳳梨（Neoregelia cv. Peony）之花序作為培植體誘導不定芽，於2個月後有不定芽體的再生，移至1/2 MS添加NAA 1.0 mg/L + 2ip 1.0 mg/L的培養基中進行芽體的增殖，在一年內可再生小植株並達出瓶階段。紅羽鶯歌鳳梨與牡丹彩葉鳳梨之花器培植體，培養於1/2 MS添加2,4-D的培養基，於2 ~ 3個月可誘導粒狀癒合組織發生。紅羽鶯歌鳳梨誘導發生率較低，僅花托與花藥分別有6.6與20％的發生率；牡丹彩葉鳳梨以子房與花柱誘導發生率較高，分別為71.4％與60％。 玉扇空氣鳳梨（Tillandsia cyanea）葉片液體培養以1/2 MS培養基添加NAA 0.1 mg/L + BA 0. 1 mg/L效果較好，在3週後由葉片基部產生芽體，每1小植株8 ~ 10葉片可誘導出3.2個芽體。 玉扇空氣鳳梨之芽體健化以添加IAA 0.2 ~ 2.0 mg/L有較高的發根率，而添加NAA會造成毒害。牡丹彩葉鳳梨則以添加NAA 1.0 mg/L有較好的健化及發根效果，在移出溫室後的存活率可達100％。 以玉扇空氣鳳梨之胚狀體進行細胞懸浮培養，以MS為基礎培養基添加2,4-D 1.5 mg/L，在3 ~ 4個月後可得到均質的細胞系。每繼代週期內細胞增生量約為原來之1倍，提高蔗糖濃度到9.0％，對於細胞生長相沒有影響，但可能不利胚性細胞的釋放。取30 ~ 60目大小之細胞團，於含有NAA 0.2 mg/L、2ip 0.2 mg/L、kinetin 0.1 mg/L、zeatin 0.05 mg/L及ABA 0.1 mg/L的MS培養基，2個月可觀察到體胚的再生，然而體胚發生模式為間接體胚再生，尚未能同步化與個別化

The compressed shoot tip and young leaf from suckers of Vriesea erecta were excised as explants. The regeneration medium used was 1/2 MS supplemented with NAA 1.0 mg/L + TDZ 1.0 mg/L, showing high efficiency in stimulation of caulogenesis. Numerously adventitious buds appeared in the leaf axil and the base of leaf explants, but the cluster buds sprouting was restricted. Better growth could be approached by transferring the adventitious buds to medium supplemented with NAA 0.5 mg/L after separation. Adventitious buds were obtained 2 months after excising inflorescence of Neoergelia cv. Peony as explants；those buds were then transferred to 1/2 MS supplemented with NAA 1.0 mg/L + 2ip 1.0 mg/L for multiplication. Regeneration of Neoergelia cv. Peony via inflorescence tip culture from one stock plant can provide mass plantlets transfer to greenhouse in one year. The florets of Vriesea erecta and Neoregelia cv. Peony were excised as explants and inoculated on 1/2 MS medium supplemented with 2,4-D. The granular calli could be induced after 2 ~ 3 months of culture. However, the frequency of callogenesis of Vriesea erecta was low, only 6.6% and 20% in anther and receptacle explants. The frequency were 71.4％ and 60％ in ovary and style explants of Neoregelia cv. Peony. The leaves of Tillandsia cyanea were separated from the base of plantet for liquid culture. The axillary buds were sprouting from leaf basal after 3 weeks of culture. The treatment of 1/2 MS supplemented with NAA 0.1 mg/L + BA 0. 1 mg/L has better result, an average of 3.2 buds was obtained from each plantlet with 8 ~ 10 leaves. For Tillandsia cyanea, a medium supplemented with IAA 0.2 ~ 2.0 mg/L was suitable of root development, while NAA was toxic. Neoregelia cv. Peony showed better response to NAA 1.0 mg/L, high concentration could enhance growth in plantlets. The survival in exvitro transplant of the plantlets approached to 100％ in greenhouse. Embryoids were used as initiative material for cell suspension of Tillandsia cyanea .The medium was MS supplemented with 2,4-D 1.5 mg/L, which achieved homogenous cell clusters after 3 ~ 4 months of culture. The PCV（Packed cell volume）doubled during one subculture period. High sucrose concentration up to 9.0％ did not change the cell line type, but inhibited the releasing of embryogenic cell. Two months after plating suspension cells by 30 ~ 60 mesh scale on MS supplemented with NAA 0.2 mg/L、2ip 0.2 mg/L、kinetin 0.1 mg/L、zeatin 0.05 mg/L and ABA 0.1 mg/L, somatic embryos were developed via indirect somatic embryogenesis, but not synchronized and individualized.