赤松根际土壤微生物PCR—DGGE反应体系优化

为建立赤松根际土壤微生物的PCR-DGGE反应体系，以赤松根际土壤为材料，利用PCR—DGGE技术，对反应体系中的几个重要因素不同梯度进行优化，包括模板DNA、dNTPs、引物及Mg2＋的用量。在25μL PCR反应体系中，模板DNA浓度为10ng，dNTPs浓度为0．15mM，引物浓度为0．32μM，Mg2＋浓度为1．0mM时，其PCR效果最佳。用该反应体系扩增出来的PCR产物，在DGGE凝胶上能够清晰地分辨出赤松根际土壤细菌和真菌多样性。这为进一步研究赤松根际土壤微生物，特别是研究赤松外生菌根奠定了基础。

Optimization of PCR-DGGE reaction system on rhizosphere soil microorganism for Pinus densiflora Sieb. et Zucc

To explore the PCR-DGGE reaction system of rhizosphere soil microorganism for P. densiflora. Some important factors for PCR-DGGE amplification system including the concentration of DNA tem- plate,dNTPs, primers and Mg2＋ were optimized with P. densiflora soil as materials. An optimal system （25 gL） was obtained as 10 ng DNA template,0.15 mM dNTPs,0.32 μM primer,and Mg2＋ 1.0 mM. The method simple and fast. Amplification of PCR products in the reaction system on the DGGE gel can be clearly distinguished P. densiflora rhizosphere bacterial and fungal diversity. For the further research of P. densiflora rhizosphere microbes, especially research P. densiflora ectotrophic mycorrhiza laid a foundation.