Molecular identification and in vitro propagation of the endangered medicinal Cleome droserifolia plant of Saint Katherine

Abstract

The protectorate of Saint Katherine considers one of the biggest protected areas in Egypt characterized by a unique location and environment. It supports a high number of rare and endangered species. Cleome droserifolia (Rorida droserifolia) is an endangered plant found in Saint Katherine, Sinai, Egypt. It has been known for its medicinal uses so it becomes rare due to over-grazing and over-collection for use in traditional medicine, research and trade home use. DNA barcoding analysis and in vitro culture tools were conducted for saving the plant. This research supports the goal number 15 of the United Nations Sustainable Development (SDGs); aimed at preserving, restore and reinstate sustainable usage of terrestrial ecosystems and halting biodiversity deterioration. Two pairs primers of three DNA markers: [ribulose-1, 5-bisphosphate carboxylase/oxygenase large subunit (rbcL), maturase K (matK) and trnH-psbA] were used for PCR amplification and sequencing. BLAST results and phylogenetic analysis proved a correct authentication of Cleome droserifolia on both; species and genus levels and confirms the involvement of Cleome droserifolia in Cleome genus. For in vitro propagation, cotyledonary node explants of germinated seedlings were cultured on Murashige and Skoog (MS) medium fortified with 0.27 µM α-naphthaleneacetic acid (NAA), and 2.32 µM of kinetin (Kin). The highest mean number of axillary shoots was 8.16 shoots/explant. 85% of multiple shoots were successfully rooted on ¼ MS medium fortified with 7.42 µM indole-3-butyric acid (IBA). The successfully rooted plants were transferred to a mixture of sand, soil, and peat moss (1: 1: 1) and efficiently acclimatized in the greenhouse.