Optimization of different DNA extraction methods for the molecular detection of resistance genes

Abstract

Tomato spotted wilt virus (TSWV) and Pepper mild mottle virus (PMMoV) not only cause economic yield losses but also limit production for pepper plants. Resistant genes are the only reliable management strategies to control such viral diseases, but these viruses are able to overcome such resistance mechanisms. Therefore, novel resistance genes should be used to control the TSWV and PMMoV diseases. For molecular detection of resistance genes, DNA quality and purity are extremely vital to obtain proper results. This study aims to identify the best extraction method using resistance gene markers from different leaves of pepper plants. PCR analyzes revealed that fresh first real leaf gave reliable DNA quality with the CTAB DNA extraction method and then the Dellaporta extraction method which yielded lower concentrations. Although the commercial DNA extraction method offered convenient results within PCR analyzes, it is cost effective for molecular breeding such as marker assisted selection programs in developing countries. The study has clearly addressed optimized DNA concentration, using different extraction methods, leaf samples from pepper plants and their storage conditions for ultimate results in breeding programs.

Keywords

• DNA quality
• Purity
• Optimization
• Extraction methods
• PCR

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