A fast and efficient method for total DNA extraction from soil filamentous fungi <span class="so-article-trans-title" dir="auto"> Translated title: Un método rápido y eficiente para la extracción de ADN total de hongos filamentosos del suelo </span>

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Abstract

Extraction of high quality of DNA from fungus can be affected because the presence of the complex cell wall, high content of polysaccharides or secondary metabolites. In this research, we adapt a simple method of DNA extraction and purification of from fungus isolated from soil for Polymerase Chain Reaction (PCR) applications. The methodology described is both rapid and cost effective.

Translated abstract

La extracción de ADN de alta calidad de hongos puede ser afectada debido a la presencia de una pared celular compleja, alto contenido de polisacáridos o metabolitos secundarios. En esta investigación se adaptó un simple método de extracción y purificación de ADN de hongos filamentosos aislados del suelo, para aplicaciones tipo Polymerase Chain Reaction (PCR). La metodología descrita es rápida y de bajo costo.

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Most cited references 11

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ITS primers with enhanced specificity for basidiomycetes--application to the identification of mycorrhizae and rusts.

M Gardes, T D Bruns (1993)

We have designed two taxon-selective primers for the internal transcribed spacer (ITS) region in the nuclear ribosomal repeat unit. These primers, ITS1-F and ITS4-B, were intended to be specific to fungi and basidiomycetes, respectively. We have tested the specificity of these primers against 13 species of ascomycetes, 14 of basidiomycetes, and 15 of plants. Our results showed that ITS4-B, when paired with either a 'universal' primer ITS1 or the fungal-specific primer ITS1-F, efficiently amplified DNA from all basidiomycetes and discriminated against ascomycete DNAs. The results with plants were not as clearcut. The ITS1-F/ITS4-B primer pair produced a small amount of PCR product for certain plant species, but the quantity was in most cases less than that produced by the 'universal' ITS primers. However, under conditions where both plant and fungal DNAs were present, the fungal DNA was amplified to the apparent exclusion of plant DNA. ITS1-F/ITS4-B preferential amplification was shown to be particularly useful for detection and analysis of the basidiomycete component in ectomycorrhizae and in rust-infected tissues. These primers can be used to study the structure of ectomycorrhizal communities or the distribution of rusts on alternate hosts.