蛇足石杉离体培养物形态变化与石杉碱甲积累的研究

In vitro-cultured Morphological Changes in Huperzia serrata and Accumulation of Huperzine A

通过蛇足石杉(Huperzia serrata)离体培养研究不同培养基对培养物细胞特征、总DNA特征、生物增长量和石杉碱甲含量的影响。结果表明,Sh、W和Shx三种培养基中,培养物分化成叶状体、根状体和苔藓状体3种不同的形态,三者的叶细胞拟由单细胞出芽成串丛生发育成组织,苔藓状体和根状体比叶状体的细胞表面有更多的根和假根生长;总DNA也存在差异,35、45和55天的苔藓状体均出现DNA梯带,表明形态变异可能与细胞凋亡相关;叶状体、根状体和苔藓状体的生物量增长分别为(1 788±31)%、(833±27)%和(1 963±52)%,石杉碱甲的含量分别为71.7±1.54、20.1±0.82和0μg·L–1(P<0.01)。上述结果表明,培养基成分变化可以使相同的基因型材料生成不同的形态,且生长速度和代谢产生石杉碱甲的量等都发生明显变化,叶状体是产生石杉碱甲的最好形态。

To study the effect of different culture media on cell morphological characteristics, DNA, biomass growth and huperzine A content in in vitro cultures of Huperzia serrata. In the different culture media, named Sh, W, and Shx, H. serrata differentiated into thallus, rhizomorph and mossy culture, respectively. The single cell developed into clusters and split to form tissue. Roots and rhizoids developed on rhizomorph and mossy cell surface more than on thallus. The total DNA content differed. The morphologic changes may be related to apoptosis because mossy culture showed a DNA ladder after 35, 45 and 55 days. Biomass increase was measured for thallus（1 788±31）%, rhizomorph（833±27）%, and mossy culture（1 963±52）%, and huperzine A content was 71.7±1.54, 20.1±0.82 and 0 μg·L–1, respectively, with significant differences（P〈0.01） among the 3 morphologies. Thallus grew slower than did mossy culture, but huperzine A content was the highest, and mossy growth was the fastest but produced no huperzine A content. The medium can lead to different forms for the same genotype in H. serrata, with changes in growth rate and huperzine A content. Thallus is the best form for producing huperzine A.