2,4-D及TDZ誘導臺灣白及芽體及類原球體形成與植株再生

The Formation of Shoot and Protocorm-like Body Induced by 2,4-D and TDZ and in Vitro Plant Regeneration of Bletilla formosana (Hayata) Schltr

林葦庭(Wei-Ting Lin) ； 熊同銓(Tung-Chuan Hsiung) ； 陳麗如(Li-Ru Chen)

《作物、環境與生物資訊》 13卷3期 (2016/09) Pp. 146-152

https://doi.org/10.30061/CEB.201609_13(3).0005

摘要

本研究以建立臺灣原生藥用蘭科植物-台灣白及[Bletilla formosana (Hayata) Schltr.]的試管內植株再生途徑為目標。取無菌苗的幼葉葉尖、根尖與假球莖為培植體，培養在16 種2,4- 二氯苯氧乙酸[2,4-dichlorophenoxyaceticacid (2,4-D), 0、0.5、1、10 mgL^(-1)]與賽苯隆[thidiazuron (TDZ), 0、0.1、0.5、1.0 mg L^(-1)]濃度組合的培養基中，觀察其型態發生變化。結果顯示，葉尖及根尖培植體無進一步的型態變化，而以假球莖為培植體可誘導形成芽體及類原球體(protocorm-like body; PLB)。在1 mg L^(-1)TDZ 及0.5 mg L^(-1) 2,4-D 組合0.5 mg L^(-1) TDZ的條件下，芽體誘導率為50%，平均芽體形成數為5 個。在類原球體形成前，假球莖表面先形成透明狀的體胚，膨大轉綠形成類原球體。在10 mg L^(-1) 2,4-D 組合0.1 或1 mg L^(-1)TDZ 的條件下，誘導率為16.7%，接續類原球體繼代至原誘導培養基中可快速增殖，平均倍率133 倍，繼代至1/2 MS 基本培養基中可發育為小植株。經由芽體及類原球體途徑均可獲得健康小植株，出瓶存活率100%。

關鍵字

2,4-二氯苯氧乙酸 ； 假球莖 ； 器官發生 ； 賽苯隆 ； 類圓球體 ； 體胚

並列摘要

Medicinal orchid plant- Bletilla formosana (Hayata) Schltr. is native in Taiwan. We used B. formosana as plant materials for establishing in vitro plant regeneration pathway. Explants of leaf tip, root tip and pseudobulb derived from in vitro plantlets were cultured on 1/2 MS basal medium containing 0, 0.5, 1 and 10 mg L^(-1) 2,4- dichlorophenoxyacetic acid (2,4-D) combined with 0, 0.1, 0.5 and 1.0 mg L^(-1) thidiazuron (TDZ). The results showed that no further changes in morphology were observed in leaf tip and root tip explants. However, shoot and protocorm-like body (PLB) can be induced in pseudobulb explants. The percentage of multiple shoot formation was 50% in the conditions of 1 mg L^(-1) TDZ only and 0.5 mg L^(-1) 2,4-D combined with 0.5 mg L^(-1) TDZ, and the average shoot number was 5. In the conditions of 10 mg L^(-1) 2,4-D combined with 0.1 or 1 mg L^(-1) TDZ, transparent somatic embryos formed on the surface of pseudobulb explants, and then enlarged and turned green to form PLB with a percentage of PLB formation at 16.7%. PLBs proliferated in a significant average fold of 133 (estimated with the PLB numbers) after subcultured on the same induction medium, and further developed into plantlets after cultured on 1/2 MS basal medium. Plantlets are healthy and grow well with a survival rate of 100% after acclimation.

並列關鍵字

2,4-dichlorophenoxyacetic acid (2,4-D) ； Pseudobulbs ； Organogenesis ； Thidiazuron (TDZ) ； Protocorm-like body (PLB) ； Somatic embryo