液体培养的蛇足石杉离体叶状体生产石杉碱甲和生化特征研究

Studies on Huperzine A Accumulation and Biochemical Characteristics in Thallus of Huperzia serrata in vitro by Liquid Culture

摘要:

为了解浅层液体培养对蛇足石杉[Huperzia serrate（Thunb.）Trev.]叶状体的影响，采用不同用量培养液进行培养，对叶状体的生长、石杉碱甲（HupA）积累、抗氧化酶活性和内源激素的变化进行了研究。结果表明，培养液用量不同，叶状体产生的HupA含量有显著差异，15 mL浅层培养的HupA生产率可达3 115.159 μg L^-1，是对照的2.20倍。浅层液体培养的叶状体的超氧化物歧化酶活性始终高于对照，过氧化物酶活性在培养前期比对照提高，过氧化氢酶活性在培养后期比对照提高；内源激素IAA与GA含量低于对照，而ABA含量高于对照。因此，浅层液体培养的蛇足石杉离体叶状体生产HupA的能力更高，且HupA含量与内源激素ABA水平相关，叶状体生长期的3种抗氧化酶活性具有协作增强抗氧化作用。

关键词:蛇足石杉;离体叶状体;石杉碱甲;液体培养;抗氧化酶;内源激素

Abstract:

In order to understand the effect of liquid cultures on thallus of Huperzia serrate (Thunb.) Trev. in vitro, the changes in thallus growth, huperzine A accumulation, antioxidant enzymes activities and endogenous hormones were studied cultured on different dosage of liquid medium. The results showed that the huperzine A contents had significant differences among different dosages of liquid medium. The huperzine A content could reach up to 3 115.159 μg L^-1 with 15 mL medium, which was 2.20 times more than the control. The superoxide dismutase activity of thallus cultured in shallow liquid was higher than that of the control, the activity of peroxidase was higher than that of the control at early stage, while did the activity of catalase at later stage. The contents of endogenous hormones IAA and GA were lower than the control, while ABA content was higher than the control. Therefore, it was suggested that the ability to produce HupA was high in thallus in vitro of H. serrata cultured with shallow liquid medium, the HupA content had relation with the content of endogenous hormone ABA, and the activities of three antioxidant enzymes in thallus had collaborative enhancement effect during growth stage.

Key words:Huperzia serrate;Thallus in vitro;Huperzine A;Liquid culture;Antioxidant enzyme;Endogenous hormone