Low-Stringency Single-Specific-Primer PCR as a tool for detection of mutations in the matK gene of Phaseolus vulgaris exposed to paranitrophenol

摘要

Low-stringency single specific primer polymerase chain reaction (LSSP)-PCR was assessed for its suitability in detecting the genotoxic effect of paranitrophenol (PNP) in the dwarf bean (Phaseolus vulgaris) exposed to different concentrations of PNP. DNA was extracted from both PNP-treated and non-treated shoots that was amplified by specific PCR, using universal primers of maturase K chloroplast DNA. PCR products of approximately 776 bp were subsequently used as a template for LSSP-PCR analysis. We detected the genotoxic effect based on LSSP-PCR profiles of the DNA generated in PNP-treated over the non-treated control of bean shoots. A complex electrophoretic pattern consisting of many bands was obtained from control and treated samples. Surprisingly, DNA sequencing data revealed that the homology among the maturase gene amplified from PNP-treated vs. non-treated samples of dwarf beans are comparable. These results showed that the use of LSSP-PCR analysis is not a proper tool to detect genotoxic effect in bean, at least in bean shoots that were exposed to PNP.