Development and Characterization of SSR Marker in Artemisia annua utilizing Glandular Secretory Tricomes (GST) derived EST Sequences

Antimalarial drug artemisinin is a bioactive compound of Artemisia annua, a traditional Chinese medicinal plant. Low and inconsistent production with higher commercial demand of artemisinin alarms improvement in the A. annua plant in which molecular marker especially Expressed Sequence Taq (EST) sequences derived Simple Sequence Repeat (SSR) marker is very useful in the identification of elite plants as well as in identification of gene (s). A total of 24963 Glandular Secretory Tricomes (GST) derived EST sequences were searched for the presence of SSR motifs with criteria of minimum five repeat units and 1942 SSR (7.78%) was detected in 1666 (6.67%) EST sequences with relative abundance of 9.01 kb/SSR. The di-nucleotide repeats had highest frequency of 61.4% followed with 31.3% tri-nucleotide. Fifty primer pairs were designed for SSR containing EST sequences and amplification was performed with five genotypes of A. Annua out of which 29 primers amplified the genomic DNA and 26 primers were found to be specific. In the present study, 26 EST-SSR markers in A. annua derived from GST was developed which will be useful to understand the molecular biology of A. annua.