摘要
从东北林业大学林场采集黄檗根际土壤及菌根，采用Nested—PCR技术分别扩增黄檗菌根及根际土壤AM真菌18SrDNANS31／Glol区域，利用该扩增产物对AM真菌DGGE条件进行优化。表明Nested—PCR技术具有较高的灵敏性，可有效的从微量DNA中扩增出约230bp的目标片段；经DGGE条件优化，确定AM真菌最佳DGGE条件为丙烯酰胺浓度8．0％，纯化后的PCR产物作为DGGE上样样品，凝胶变性梯度范围30％～60％，电泳电压130V，电泳时间8h，电泳温度60℃。
DGGE is used to investigate bacterial community composition, but this approach to community analysis of AM fungi in the rhizospheric soil and root samples of Phellodendren amurense Rupr. has not been reported. The rhizospherie soil and root samples of Phellodendren amurense Rupr. were collected in the Logging Station of Northeast Forestry University. The Nested- PCR was conducted to specifically amplify NS31/Glol domain sequences of the small - subunit （18S） rDNA from the AM fungi in the rhizospheric soil and root samples of Phellodendren amurense Rupr. , respectively. The conditions of DGGE were optimized by the Nested - PCR products. The result indicated the targeted product （230 bp） successfully amplified from trace DNA by Nested - PCR. The optimum condition of DGGE was finally confirmed. DGGE was performed on polyaerylamide gels with the denaturing gradient from 30% to 60%. The purified PCR products were used for DGGE analysis. The electrophoresis was running at a fixed voltage of 130 V for 8 h at 60℃.
出处
《黑龙江大学自然科学学报》
CAS
北大核心
2008年第4期534-538,共5页
Journal of Natural Science of Heilongjiang University
基金
黑龙江省博士后基金资助项目(LBH-Z05013)
黑龙江大学博士启动基金资助项目
关键词
黄檗
丛枝菌根真菌
Nested—PCR
变性梯度凝胶电泳
PheUodendren amurense Rupr.
arbuscular mycorrhizal fungi
Nested -PCR
denaturing gradient gelelectrophoresis
