A rapid method for isolation of total DNA from pathogenic filamentous plant fungi

D. Gonzlez-Mendoza, R. Argumedo-Delira, A. Morales-Trejo, A. Pulido-Herrera, L. Cervantes-Daz, O. Grimaldo-Juarez, A. Alarcn
2010 Genetics and Molecular Research  
DNA isolation from some fungal organisms of agronomic importance is difficult because they have cell walls or capsules that are relatively unsusceptible to lysis. We have developed a fast DNA isolation protocol for Fusarium oxysporum, which causes fusarium wilt disease in more than 100 plant species, and for Pyrenochaeta terrestris, which causes pink root in onions. This protocol was based on the sodium dodecyl sulfate/ phe nol method, without β-mercaptoethanol and without maceration in liquid
more » ... ceration in liquid nitrogen; it uses phenol/chloroform extraction to remove proteins and co-precipitated polysaccharides. The A 260/280 absorbance ratios of isolated DNA were around 1.9, suggesting that the DNA fraction was pure and may be used for further analysis. Additionally, the A 260/230 values were higher than 1.8, suggesting negligible contamination by polysaccharides. The DNA isolated by this protocol is of sufficient quality for molecular applications; this tech nique could be applied to other organisms that have similar substances that hinder DNA extraction.
doi:10.4238/vol9-1gmr680 pmid:20198572 fatcat:kaeds7uicvcfrbw7pxnew4d6nu