Agrobacterium tumefaciens-mediated transformation as a tool for insertional mutagenesis in the symbiotic ectomycorrhizal fungus Hebeloma cylindrosporum

被引：172

作者：

Combier, JP ^{[1
]}

Melayah, D ^{[1
]}

Raffier, C ^{[1
]}

Gay, G ^{[1
]}

Marmeisse, R ^{[1
]}

机构：

[1] Univ Lyon 1, CNRS, UMR Ecol Microbienne, F-69622 Villeurbanne, France

来源：

FEMS MICROBIOLOGY LETTERS | 2003年 / 220卷 / 01期

关键词：

ectomycorrhizal fungus; Hebeloma cylindrosporum; Agrobacterium tumefaciens; genetic transformation; insertional mutagenesis;

D O I：

10.1016/S0378-1097(03)00089-2

中图分类号：

Q93 [微生物学];

学科分类号：

071005 ; 100705 ;

摘要：

We transformed haploid mycelium of Hebeloma cylindrosporum via Agrobacterium tumefaciens and optimised the procedure to develop a new tool for insertional mutagenesis in this fungus. Southern blot analysis of 83 randomly selected transformants showed that they all contained plasmid inserts. Each of them showed a unique hybridisation pattern. suggesting that integration was random in the fungal genome. Sixth percent of transformants obtained in the presence of bacteria pre-treated with acetosyringone integrated a single transferred DNA copy. Thermal asymmetric interlaced polymerase chain reaction allowed us to recover the left border and the right border junctions in 85% and 15% of transformants analysed, respectively. Results show that A. tumefaciens-mediated transformation may be a powerful tool for insertional mutagenesis in H. cylindrosporum. (C) 2003 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.

引用

页码：141 / 148

页数：8

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