盤固草A254(Digitaria decumbens)細胞懸浮培養與植株再生

Plant Regeneration from Cell Suspension Culture of Pangolagrss A254 (Digitaria decumbens)

施意敏(Yih-Min Shy) ； 廖成康(Cherng-Kang Liao) ； 盧虎生(Huu-Sheng Lur) ； 朱鈞(Chun Chu)

《台灣農學會報》 9卷5期 (2008/10) Pp. 433-445

https://doi.org/10.6730/JAAT.200810_9(5).0002

摘要

盤固草A254 (Digitaria decumbens)是台灣地區適應性相當廣的熱帶牧草，由於盤固草遺傳組成為三倍體，花粉不稔。為加速傳統育種效率，期利用生物技術提供遺傳改良的另一個途徑。因此，本研究主要建立具植株再生能力的細胞懸浮培養方法，以因應基因轉殖或突變育種之需。將盤固草A254未成熟花穗，接種於含有2 mg/l 2,4-D (2,4-dichlorophenoxyacetic acid)及0.5 mg/l BA (N^6-benzyladenine)的MS (Murashige and Skoog's)固體培養基，誘導的癒合組織幾乎全部為白色結構緊密的胚性癒合組織，BA顯著影響癒合組織的型態。一些透明鬆軟的癒合組織，可持續繼代培養於含有2 mg/l 2,4-D的MS液體培養基，至少六個月以上。一旦需要植株再生時，可於原懸浮細胞培養基添加0.5 mg/l BA，培養一週後，再移至含有2 mg/l 2,4-D及0.5 mg/l BA的固體培養基，以誘導白色緊密的胚性癒合組織。將這些胚性癒合組織移至含有0.5 mg/l BA或0.05 mg/l TDZ的MS培養基，分別含88.9%或87.7%的癒合組織可再生為完整植株。本研究結果對盤固草基因轉殖非常重要且相當有益，因為可以在人為的控制下，精準地促進細胞持續增殖，並保有高頻度的植株再生能力。

關鍵字

植株再生 ； 癒合組織形態 ； 盤固草

並列摘要

Pangolagrass A254 (Digitaria decumbens) is a triploid forage grass with wide adaptation in Taiwan. The objective of this study was to develop an efficient plant regeneration system from cell suspension culture of pangolagrass to accelerate genetic improvement of this species by gene transformation or mutation. The embryogenic callus was induced from immature inflorescences of pangolagrass on MS medium with 2 mg/l 2,4-D and 0.5 mg/l BA. The type of callus induction was influenced by BA. The induced transparent and friable calli were used to establish and maintain suspension cultures with 2 mg/l 2,4-D for at least 6 months. For plant regeneration, the embryogenic calli were induced by adding 0.5 mg/l BA in suspension medium for one week and then transferred to a solid medium with 2 mg/l 2,4-D and 0.5 mg/l BA. The addition of BA in the suspension medium enhanced embryogenic callus formation and the ability of plant regeneration. The plant regeneration frequency of the embryogenic calli derived from cell suspension system reached up to 88.9% and 87.8% cultured with 0.5 mg/l BA and 0.05 mg/l TDZ, respectively. The results showed that the calli could keep proliferating and regenerating into the plantlets with high frequency under control. The results show potential for the genetic transformation of pangolagrass.

並列關鍵字

Plant regeneration Callus type Pangolagrass