Conservation of plant genetic resources, especially threatened species, is an important topic in biodiversity. It is a field that requires prior knowledge of the target species, in addition to correct identification and taxonomic description. In botany, the identification of plant species traditionally relies on key morphological descriptions and anatomical features. However, in complex species and tree plants, molecular identification can facilitate identification and increase species delimitation accuracy. In the Faifa mountains of Jazan province in Saudi Arabia, 12 rarely occurring plants were recorded and identified using two DNA barcoding regions (i.e., rbcL and ITS). All the samples were successfully amplified, sequenced, and analyzed using the standard DNA barcode protocol, and this resulted in the clear and accurate identification of 11 out of the 12 sampled species. A total of five species were in agreement in terms of both morpho- and molecular-based identification. Four and two species were identified based solely on ITS and rbcL phylogenetics, respectively. The geographic distribution records of the identified species showed that some species were distributed at a distance far from their usual region, while others were reported in proximate regions and localities. Some species were found to be medicinally important and required additional conservation plans.