In Vitro Propagation of Platycerium Bifurcatum (Cav.) C. Chr. via Green Globular Body Initiation

建立二歧鹿角蕨組織培養綠球體之繁殖系統

廖宇賡(Yue Ken Liao) ； 吳怡慧(Yi Hui Wu)

《Botanical Studies》 52卷4期 (2011/10) Pp. 455-463

摘要

本研究描述取用二歧鹿角蕨(Platycerium bifurcatum (Cav.) C. Chr.) 之幼葉培殖體，經由誘導綠球體 (GGB) 的方式進行組織培養繁殖方法。在添加α-naphthalene acetic acid (NAA) 及2-isopentenyl adenine (2ip) 或 6-benzylaminopurine (BA) 的 MS 培養基中可誘導GGB 發生，NAA 的濃度以維持在5.37 μM 為佳，而增加細胞分裂素(cytokinin) 的濃度則抑制GGB 的發生。 單一GGB 可以在添加5.37 μM NAA 及 2.22 μM 2ip 之培養基中進行增殖，此時GGB 展現較少的孢子體增生而有較快的直徑生長。增殖的GGB 接著可以在不添加植物生長調節劑(PGR) 的MS 培養基中形成多數孢子體。個別分離且無根的孢子體可以直接轉移到試管外的四種栽培介質( 濕水苔、蛭石、泥炭土以及等體積混合蛭石及泥炭土)中生長，在健化的前四週，植物體出現自發性的發根現象，在沒有使用不定根誘導的處理之下，植物體健化成活率高於88%。在不同介質之間植物存活率並無顯著差異。採用本研究建議的繁殖步驟，推估在32 週之內可以從一個葉片培殖體(≤ 2.0 cm 的葉子；取近葉基1/2 之部分) 產生170 個孢子體。

關鍵字

綠球體 ； 二歧鹿角蕨 ； 微體繁殖 ； 鹿角蕨

並列摘要

This study describes a Platycerium bifurcatum (Cav.) C. Chr. propagation method through initiation of green globular bodies (GGBs) from juvenile leaf explants of in vitro grown sporophytes. The GGB initiation was first obtained by incubating the explants on MS medium supplemented with α-naphthalene acetic acid (NAA) and two other cytokinins, 2-isopentenyl adenine (2ip) or 6-benzylaminopruine (BA). NAA at 5.37 μM was determined a better concentration for the initiation. However, as the cytokinin concentration in the medium increased, it inhibited GGB production. Individually harvested GGBs proliferated well on a medium supplemented with 5.37 μM NAA and 2.22 μM 2ip; they displayed less sporophyte formation but a faster increase in diameter. Multiple sporophytes were subsequently regenerated by culturing GGBs on a hormone-free MS medium at the regeneration stage. Non-rooted and separated sporophytes were directly transplanted to four different types of potting media, namely sphagnum, vermiculite, peat, and a mixed medium (vermiculite:peat = 1:1). Spontaneous rooting was observed during the first 4-week acclimatization period. Even with the omission of an adventitious root induction step, the survival rate of the plants was 88% or higher. No significant difference in plant survival was detected among the different media. Using our proposed propagation procedures, it was possible to obtain an estimated production of about 170 sporophytes per initial leaf explant (proximal section; 1/2 of the leaf ≤ 2.0 cm in length) during a 32-week culturing period.

並列關鍵字

Green globular body GGB) ； Platycerium bifurcatum ； Micropropagation ； Staghorn fern