菌物学报 15 July 2009, 28(4):512～520
黄檗根围丛枝菌根真菌菌群组成
蔡柏岩1,2 葛菁萍1 接伟光2 阎秀峰3﹡
1黑龙江大学生命科学学院 黑龙江省普通高等学校微生物重点实验室 哈尔滨 150080
2黑龙江东方学院食品与环境工程学部 哈尔滨 150086
3东北林业大学生命科学学院 哈尔滨 150040
摘 要：从东北林业大学林场采集黄檗Phellodendron amurense根系及根围土壤，采用Nested-PCR技术扩增黄檗菌根及根围土壤AM真菌18S rDNA NS31/Glol区域，利用该产物进行DGGE分析，并结合DNA测序、系统发育分析及DGGE图谱分析技术对黄檗AM真菌菌群组成进行分析。结果表明：Nested-PCR技术具有较高的灵敏性，可有效地从微量DNA中扩增出约230bp的目的片段；黄檗根系及根围土壤具有不同的DGGE指纹图谱特征，DGGE带谱在条带的数量、亮度、优势度等方面均存在较大差异，全部序列可分为3类菌群，即球囊霉属Glomus、盾孢囊霉属Scutellospora及植物病原菌黄杨亚赤壳Hyponectria buxi，其中Glomus sp.（EF177624）和Glomus sp.（DQ085205）分别为黄檗根系和根围土壤样品中最具优势的AM真菌。
关键词：18S rDNA，Nested-PCR技术，变性梯度凝胶电泳，系统发育分析
The community composition of the arbuscular mycorrhizal fungi in the rhizosphere of Phellodendron amurense
CAI Bai-Yan1, 2 GE Jing-Ping1 JIE Wei-Guang2 YAN Xiu-Feng3*
1Key Laboratory of Microbiology, College of Life Sciences, Heilongjiang University, Harbin 150080, China
2Insitute of Food and Environment Engineering, Heilongjiang Oriental College, Harbin 150086, China
3College of Life Sciences, Northeast Forestry University, Harbin 150040, China
Abstract: The rhizospheric soil and root samples of Phellodendron amurense were collected in the Logging Station of Northeast Forestry University. Nested-PCR was conducted to specifically amplify NS31/Glol domain sequences of the small-subunit (18S) rDNA from the AM fungi in the root and rhizospheric soil samples of P. amurense, respectively. The Nested-PCR products were applied to denaturing gradient gel electrophoresis (DGGE) analysis. Then, the community composition of the AM fungi was analyzed by DGGE, sequencing, and phylogenetic analysis. The result indicated that the targeted product (230bp) was successfully amplified from trace DNA by Nested-PCR. There were differences in DGGE profiles such as band numbers, densities and dominance between the rhizospheric soil and root samples. All sequences were divided into three groups, viz. Glomus, Scutellospora and Hyponectria buxi. Glomus sp. (EF177624) and Glomus sp. (DQ085205) were the prevalent AM fungi in the root and rhizospheric soil, respectively.
Key words: 18S rDNA, Nested-PCR, denaturing gradient gel electrophoresis, phylogenetic analysis
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