H_2O_2诱变蛇足石杉培养物的后代株系变化分析

Analysis on the Progeny Diversity of H_2O_2 Induced Mutation on Huperzia serrata Culture in vitro

目的:探索H_2O_2胁迫诱导提高蛇足石杉叶状体积累石杉碱甲(huperzine A,Hup A)含量的作用。方法:对经H_2O_2诱变建立的蛇足石杉叶状体中筛选的13个株系及出发株等14份材料进行了相对增长率和Hup A含量比较及ISSR DNA分子分析。结果:3%H_2O_2胁迫诱导处理16 min的叶状体后代相对增长率和积累Hup A的含量与出发株差异显著(P<0.05),变异叶状体株系SH042相对增长率和积累Hup A含量最大,分别为4 499.28%和(261.17±0.19)μg/g DW,比出发株分别提高2.35和2.43倍;20个ISSR随机引物中有9个对14份叶状体株系遗传分子显示了多态性,聚类分析可分为4大类,说明诱变后代叶状体株系的DNA分子存在显著差异。结论:该研究结果表明H_2O_2用于诱导蛇足石杉培养物能提高Hup A含量和建立丰富的遗传资源。

Objective： To explore the effect of H2O2 stress induced to huperzine A（Hup A） accumulation of thallus of Huperzia serrata. Methods： The relative growth rate,Hup A metabolic content and ISSR DNA of fourteen materials,including variants and control thallus（thirteen lines） of Huperzia serrata were analyzed. Results： The results indicated that the relative growth rate and accumulation of Hup A were significantly different between variants and the orginal strain after 3% H2O2 stress induced to hup A with 16 min（P〈0. 05）,the relative growth rate and hup A content of strain SH042 was highest,whose growth rate and Hup A yield arrived to 4 499. 28% and（261. 17±0. 19） μg/g DW,respectively,which increased by 2. 35 times and 2. 43 times than the orginal strain; there were polymorphism for fourteen genomic DNA of thallus,nine primers in 20 arbitrary primers of ISSR were screened. Fourteen materials were clustered into four groups,which identified the genetic basis of the thallus,which indicated that there were significant differences in the DNA molecules of the thallus of Huperizia serrata. Conclusion： The results shows that H2O2 stress induced can increase the content of Hup A and the establishment of rich genetic resources.