A phylogenetic analysis of Thai Hedychium (Zingiberaceae) and development of SCAR marker for Hedychium flavescens Carey ex Roscoe

Abstract

High annealing temperature rapid amplified polymorphic DNA (HAT-RAPD) was used as the screening method for identification of 23 Thai Hedychuim species. From 16 arbitrary primers, a total of 2, 112 distinct polymorphic band patterns were generated. To understand the relationship among Hedychium samples and three other genera of Zingiberaceae, a phylogenetic tree was generated from highly polymorphic data obtained from PCR amplification of DNA samples using character state data. The important character for each clade was discussed. The unique band (519 bp) that only present in H. flavescens, a valuable species for horticulture importance and source of natural products, was obtained from the primer OPAG-19 and selected for sequencing and conversion to the more reproducible and robust sequence characterized amplified region (SCAR) marker specific to H. flavescens. These techniques for species identification and characterization provides a morphologically independent test to verify relatedness and provide species information particularly for cases where such identification was previously impossible to obtain, for instance, in case of morphologically indistinguishable rhizome or vegetative parts in off-season or dormancy period.