| 摘要: |
| 以石牌广藿香悬浮细胞为材料,对影响其原生质体分离和培养的酶浓度、作用时间、溶液渗透压和材料的生理状态等因素进行了研究。结果表明:以0.5%的果胶酶、0.2%离析酶和0.8%的纤维素酶组合处理继代培养3～11 d的悬浮细胞8 h,渗透压调节剂为9%甘露醇,原生质体产量达1.65×106 protoplasts·mL-1 PCV,活力超过86%。在原生质体的液体浅层培养中,细胞分裂频率为13.5%。 |
| 关键词: 石牌广藿香 原生质体分离 原生质体培养 |
| DOI：10.3969/j.issn.1000-3142.2012.05.019 |
| 分类号:Q943.1 |
| Fund project: |
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| The protoplasts isolation and culture of Pogostemon cablin cv.Shipaiensis |
|
MO Xiao-Lu, ZENG Qing-Qian, HUANG Shan-Shan,
CHEN Yu-Zhen, YAN Zhen*
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Guangdong Research Institude of TCM, Guangzhou, 510520, China
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| Abstract: |
| A protocol for protoplasts isolation and culture from the cell suspension of Polgostemon cablin cv.Shipaiensis were developed. The suspention cells grew 3-11 days after subculture was incubated in the enzyme solution consisting of 0.5%(w/v)pectolyase Y-23,0.2%(w/v)macerozyme R-10 and 0.8%(w/v)cellulase R-10 in 9%(w/v)mannitol buffered with 0.1% MES and 0.02% CaCl2,for 8 h,the yield was 1.65×106 protoplasts·mL-1 PCV(Packed Cell Voloume)and the viability was above 86%. In thin layer culture of protoplasts,observed division rate was 13.5%. |
| Key words: Polgostemon cablin cv.Shipaiensis protoplast isolation protoplast culture |
