Standardization of DNA extraction from invasive alien weed Parthenium hysterophorus

Abstract

DNA isolation from the weed, Parthenium hysterophorus is complicated due to the presence of high amount of allelochemicals in the form of secondary metabolites that causes hindrance in extraction and enzymatic reactions. A modified and efficient DNA extraction from P. hysterophorus leaf has been developed. The present protocol is a modified version of cetyltrimethylammonium bromide (CTAB) method constituting high salt concentration to remove polysaccharides. The increased concentration of β-mercaptoethanol, polyvinylpolypyrrolidone (PVPP), and phenol/chloroform/isoamyl alcohol extractions eliminated protein and phenolic compounds well. Good amount and quality DNA was obtained by this method. The resulted genomic DNA showed fine random amplified polymorphic DNA (RAPD), inter-simple sequence repeat (ISSR) banding pattern, polymerase chain reaction (PCR) amplification of actin gene and restriction digestion confirm the efficiency of modified procedure.

Keywords: Allelochemicals, DNA extraction, Parthenium hysterophorus, stress tolerance