Assessment of EST-SSR and RAPD markers for genetic analysis on Artemisia species

Artemisia species has various medicinal properties, constituting the main source artemisinin, an antimalarial drug which has widespread importance to mankind. The production of artemisinin either in cell/tissue/the whole plant of Artemisia species is therefore highly desirable. This artemisinin varies in different species of Artemisia based on their genetic diversity. Thus in the present study an attempt has been made successfully to assess the genetic diversity among Artemisia species using EST- derived SSR marker as well as RAPD marker. The five species of Artemisia were collected from different regions of Karnataka, India and used for the DNA isolation using CTAB method and DNA quantified using 260/280 in UV-Vis spectrophotometer. The Isolated DNA was amplified using five EST derived Simple Sequence Repeat (SSR) markers and 10 RAPD marker. Further the amplified products were separated by Gel Electrophoresis. The bands were scored for the presence or absence of bands. SSR produced a total of 25 alleles with an average of 5 alleles per locus. The polymorphism information content (PIC) reflections of allele diversity, frequency among the varieties ranged from 0.03 to 0.33, with an average of 0.18. The 10 decamer-RAPD primers generated 352 RAPD fragments. Most of the RAPD markers studied showed different level of genetic polymorphism. Pairwise Nei and Li’s similarity coefficient value ranged from 0.62-0.83 for 5 species of medicinal plants. A dendrogram was constructed using Unweight Pair Group Method with Arithmetic Mean (UPGMA) revealing 5 genotype with 4 clusters (SSR marker) and 3 clusters (RAPD marker), wide range of dissimilarity values which showed a high degree of diversity among the cultivars. The Artemisia species showed rich allelic diversity, indicating that there is great potential for identification of associations with present genotypes. Thus this method of analysis can be a helpful tool in selecting diverse parents and give broadness to the germ plasm base of medicinal plant breeding programs for the future, which can be utilized to develop new varieties with traits of interest.