Molecular analysis of Phyllanthus spp. in Thailand based on rapd and dna sequencing

Abstract

The genus Phyllanthus (Euphorbiaceae) is distributed in tropical and subtropical regions, and its members are widely used as medicinal plants in many countries. Plants in this genus are important source of natural products and bioactive compounds. The medicinal effects of Phyllanthus are diverse including antipyretic, analgesic, anti-inflammatory, antihepatotoxic and antiviral activity. In this study the genetic variation of twenty-three Phyllanthus species existing in Thailand were analyzed. Random amplified polymorphic DNA (RAPD) markers were used to discriminate between Phyllanthus species. Nine out of eighty arbitrary primers were screened for DNA polymorphism. Primer OPS-01, OPS-03, OPS-07, OPS-08, OPS-12, OPS-19, OPD-02, OPD-04, and OPD-07 produced unique DNA fingerprints that individuated the twelve Phyllanthus species. In this study, we also analyzed the nucleotide sequences of the internal transcribed spacers (ITS) of nuclear ribosomal DNA. Furthermore, a simple protocol to discriminate three important medicinal Phyllanthus species, P. amarus, P. debilis, and P. urinaria, was developed using a Polymerase Chain Reaction- Restriction Fragment Length Polymorphism (PCR-RFLP) method based on ITS sequences and successfully applied to the crude drug samples obtained in Thai markets.