人参根际土壤微生物PCR-DGGE反应体系优化

摘要

[目的]优选人参根际土壤微生物的PCR-DGGE反应体系.[方法]以种植1年的人参土壤为材料,利用PCR-DGGE技术,对反应体系中的几种重要参数不同梯度进行了优化研究,包括模板、dNTPs、引物及Mg2+的用量.[结果]最适宜的反应体系为:模板DNA浓度为0.8μg/μl,dNTPs浓度为0.2 mmol/L,引物浓度为0.4 μmol/L,Mg2+浓度为1.5 mmol/L.[结论]该方法简便快捷,为进一步研究人参根际土壤微生物多样性奠定了基础.%[ Objective] To explore the PCR-DGGE reaction system on rhizosphere soil microorganism for Panax ginseng C. A. Mey. . [ Method] Some important factors for PCR-DGGE amplification system including the concentration of DNA template,Mg2+ ,dNTPs and primers were optimized with 1-year-old P. ginseng soil as materials. [ Result] An optimal syslem (25 μl) was obtained as 0. 8 μg/μl DNA template,0. 2 mmol/L dNTPs,0.4 μmol/L primer,and 1.5 mmol/L Mg2+ [Conclusion] The method is simple and fast. This research can laid basis for further analyzing the genetic diversity of rhizosphere soil microorganism for P. ginseng.