Abstract
Urtica dioica is an ethnobotanically and medicinally important Complementary and Alternative Medicine (CAM) plant worldwide and in Turkey; 90 % of herbal CAM applications depend on it in Turkey. It has a wide range of habitats in nearly all continents. It is found in all three phytogeographical regions in Turkey (Euro-Siberian, Irano-Turanian, Mediterranean) with high adaptivity to heterogeneous geographies such as climate, soil types and altitudes. This fact in relation to the assessment of chemical constituents of the plant and combining with further genetic and morphological variation data can assist and enhance the works for the utility and reliability of CAM applications in effect and activity of this plant species. In this work we have made some preliminary experiments with novel approaches to reveal the ecotypes and genetic variation of mighty ecotypes of Urtica dioica from different phytogeographical regions of Turkey (Euro-Siberian and Mediterranean). The ecotypes have heterogeneity in both its parts (leaf, stem, root) as revealed by Random Amplified Polymorphic DNA-Polymerase Chain Reaction (RAPD-PCR) using random primers and High-resolution Melt (HRM) analysis using Urtica dioica specific primers and universal chloroplast DNA (cpDNA) primers and morphological traits such as phenolic contents and antioxidant capacities of plants' leaf infusions as used in medicinal applications in Turkey. This work will contribute a lot for the development of molecular markers to detect the genetic variation and heterogeneity of Urtica dioica to further relate with expected phenotypes that are most useful and relevant in CAM applications.
Highlights
It is becoming increasingly interesting and popular to work with medicinal plants because the use of herbal materials for treatment and health maintenance as part of complementary and alternative medicine applications is on the rise and there is a trend of living in harmony with nature
To overcome various complications and difficulties in diagnosis of this species and to get rid of the need for personal expertise and experiences we designed a set of species specific primers for the U. dioica agglutinin isolectin VII precursor gene, exon 3 (UDA) which encodes UDA protein that is a single-chain peptide found in roots and rhizome of U. dioica (Van Damme and Peumans, 1987) and used it for both molecular diagnostics and further to assess genetic variation using real-time PCR and High-resolution Melt (HRM) analysis in combination with universal, conserved chloroplast DNA sequence 16S rRNA gene
Various environmental factors like rainfall, temperature, altitude, dosage of the UV rays, soil type, climate, environmental pollution can be responsible for appreciable variation in the species, ecotypes should be determined in this respect with careful observations
Summary
It is becoming increasingly interesting and popular to work with medicinal plants because the use of herbal materials for treatment and health maintenance as part of complementary and alternative medicine applications is on the rise and there is a trend of living in harmony with nature. Cultivation experiments carried out by Pollard & Briggs (1982) with individuals from nine populations covering a range of habitat provide evidence that phenotypic plasticity is important in U. dioica; for example, fewer hairs were produced on plant grown in the shade than on those in full sun These experiments support the conclusion that much of the variation (including the polymorphism in stinging-hair density) is genetically based and heritable (Pollard and Briggs 1982; Bharmauria, et al, 2009; Taylor, 2009). Taxonomy of U. dioica for ethnobotanical, especially complementary and alternative medicinal usages might have some complications due to wide range of habitats, and the heterogeneity of the morphological traits such as phenolic contents and antioxidant capacities of plants. Levels of intra-specific and inter-specific variation and heterogeneity in U. dioica accessions and their parts by RAPD-PCR and real-time PCR HRM analysis, sequence matching function have been detected with dPCR’s basic replicative dilution idea using gene sequences belonging to chloroplast genome, and U. dioica isolectin VII precursor gene (Uzonur et al, 2004a, 2004b, 2011). This work can expand the detectability of variation that can not be detected by conventional methods or much more expensive with the others
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