尼羅草（Acroceras macrum Stapf）細胞懸浮培養與植株再生

Plant regeneration from cell suspension culture of nilegrass (Acroceras macrum Stapf)

施意敏(Yih-Min Shy) ； 張珈錡(Jia-Ci Chang) ； 廖成康(Cherng-Kang Liao)

《畜產研究》 48卷2期 (2015/06) Pp. 135-142

摘要

本研究主要建立尼羅草臺畜草一號（Acroceras macrum Stapf NLcv.TS1）細胞懸浮培養及植株再生的方法。將尼羅草臺畜草一號未成熟花穗消毒後，培養於含有2.0 mg L^(-1) 2, 4-D與0.5 mg L^(-1) BA的MS培養基，五週後形成癒合組織。將癒合組織壓碎後，懸浮培養於添加1.0 mg L^(-1) 2,4-D和50 mg L^(-1)水解酪蛋白（casein hydrolysate）之MS液體培養基，進行震盪懸浮培養，每隔2週繼代一次。將細胞懸浮液增生之細胞團，移至添加2.0 mg L^(-1) 2, 4-D和0.5 mg L^(-1) BA之MS固體培養基，培養4週後形成結構緊實的胚性癒合組織。將癒合組織移至添加1.0 mg L^(-1) BA的MS固體培養基，僅4.7%的植株再生，再將未分化之癒合組織移至0.5 mg L^(-1) NAA與0.05 mg L^(-1) TDZ的二次分化培養基，可獲得75.0%的植株再生，再生之綠色植株移至盆栽種植存活率達100%。因此，根據本試驗結果，在適當的生長調節劑處理下，尼羅草臺畜草一號的懸浮細胞經生長分化，可發育至完整植株並於田間正常生長。

關鍵字

細胞懸浮培養 ； 植株再生 ； 尼羅草

並列摘要

The objective of this study was to develop an efficient system for plant regeneration from cell suspension culture of nilegrass (Acroceras macrum Stapf NLcv.TS1). The callus used for cell suspension culture was induced from immature inflorescences cultured on MS medium with 2.0 mg L^(-1) 2,4-D and 0.5 mg L^(-1) BA for 5 weeks. For establishing and maintaining the suspension culture system, the callus was cut to small pieces and sub-cultured on MS liquid medium with 1.0 mg L^(-1) 2,4-D and 50 mg L^(-1) casein hydrolysate every 2 weeks for 3 months. When the cell clumps were proliferated, they were transferred to MS solid medium with 2.0 mg L^(-1) 2,4-D and 0.5 mg L^(-1) BA for inducing white and compact callus that was beneficial for shoot regeneration. Plant regeneration from callus cultured on MS medium with 1.0 mg L^(-1) BA was sub cultured on MS medium with 0.5 mg L^(-1) NAA and 0.05 mg L^(-1) TDZ. The frequency of plant regeneration increased from 4.7% to 75%. The plantlets grew normally in the field with 100% survival. The results showed that a successful culture system for plant regeneration from cell suspension culture of nilegrass could be established.

並列關鍵字

Cell suspension culture ； Plant regeneration ； Nilegrass