菌物學(xué)報 15 July 2009, 28(4):512～520
黃檗根圍叢枝菌根真菌菌群組成
蔡柏巖1,2 葛菁萍1 接偉光2 閻秀峰3﹡
1黑龍江大學(xué)生命科學(xué)學(xué)院 黑龍江省普通高等學(xué)校微生物重點實驗室 哈爾濱 150080
2黑龍江東方學(xué)院食品與環(huán)境工程學(xué)部 哈爾濱 150086
3東北林業(yè)大學(xué)生命科學(xué)學(xué)院 哈爾濱 150040
摘 要：從東北林業(yè)大學(xué)林場采集黃檗Phellodendron amurense根系及根圍土壤,，采用Nested-PCR技術(shù)擴增黃檗菌根及根圍土壤AM真菌18S rDNA NS31/Glol區(qū)域，利用該產(chǎn)物進(jìn)行DGGE分析,，并結(jié)合DNA測序,、系統(tǒng)發(fā)育分析及DGGE圖譜分析技術(shù)對黃檗AM真菌菌群組成進(jìn)行分析。結(jié)果表明：Nested-PCR技術(shù)具有較高的靈敏性,，可有效地從微量DNA中擴增出約230bp的目的片段,；黃檗根系及根圍土壤具有不同的DGGE指紋圖譜特征，DGGE帶譜在條帶的數(shù)量,、亮度,、優(yōu)勢度等方面均存在較大差異，全部序列可分為3類菌群,，即球囊霉屬Glomus,、盾孢囊霉屬Scutellospora及植物病原菌黃楊亞赤殼Hyponectria buxi，其中Glomus sp.（EF177624）和Glomus sp.（DQ085205）分別為黃檗根系和根圍土壤樣品中最具優(yōu)勢的AM真菌,。
關(guān)鍵詞：18S rDNA,，Nested-PCR技術(shù)，變性梯度凝膠電泳,，系統(tǒng)發(fā)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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