In vitro generation of high artemisinin yielding salt tolerant somaclonal variant and development of SCAR marker in Artemisia annua L

Abstract

Low levels of artemisinin in Artemisia annua is posing serious limitation in sustainable supply of this drug and ultimately affecting global struggle to cure malaria. The present study explores new ways for higher artemisinin production through generating somaclonal variant tolerant against salt stress. Here, through gamma-rays irradiation a total of 13 somaclonal variants (ASV1 to ASV13) were identified. They were characterized on the basis of morphological traits and further molecular characterization was carried out through RAPD analysis. Dendrogram, constructed on the basis of RAPD scores, showed six major clusters in which ASV12 was most distant from control to fallen alone in last cluster. Based on 16 metric traits examined among 13 somaclonal variants and wild type, correlation analysis was carried out. In addition, principle component analysis was conducted to attest the results. All statistical analysis suggested ASV12 as stable salt tolerant somaclonal variant of A. annua. ASV12 had higher artemisinin content as compared to wild type and under salt stress; expression of key artemisinin biosynthetic genes (ADS, CYP71AV1, DBR2 and ALDH1) was more in ASV12 as compared to wild type. Further SCAR marker (570 bp) linked with high artemisinin was developed and further tested on the low and high artemisinin yielding lines, F1 and F2 progenies. A single band was amplified in ASV12 and all high artemisinin-yielding plants. The study provides ample possibility of cultivation of ASV12 variant of A. annua on saline soil for dual benefit: high artemisinin production and utilization of waste saline land. © Springer Science+Business Media Dordrecht 2016.