Zingiber barbatum Wall. (family Zingiberaceae), is an underutilized medicinal plant and commonly known as ‘Meik tha-lin’ in Myanmar where it is used in the indigenous system of medicine. In the present study, 19 accessions of Z. barbatum from five provinces in Myanmar have been utilized to characterize and assess genetic diversity. Twenty-nine morphological characters were noted, including growth habit, leaf, pseudo-stem and rhizome characters. Fifteen primer sets of P450-based analogue (PBA) markers were used to reveal molecular characteristics. Of the 29 morphological characters, 22 showed a high degree of variation within wild ginger accessions, whereas 20 of these characters contributed significantly to morphological variation. Eleven amplified primer sets gave a total of 175 bands and exhibited 92.15% polymorphism across Z. barbatum accessions. Based on morphological characters and PBA markers, 19 accessions can be divided into two morphotype groups with comparatively higher genetic diversity. This information can be applied in future crop improvement, proper conservation and better use of this underutilized medicinal species.