Evaluation of Genetic Fidelity Among Micropropagated Plants Raised Through Long-term Nodal Cultures of Elite Clone of Artemisia annua L. Using DNA-based RAPD Markers

摘要

Micropropagation has been achieved employing nodal explants excised from long term cultured shoots of an elite clone of Artimisia annua L. on MS medium supplemented with different cytokinins such as 2iP, BA or Kn individually. Of the various concentrations (1, 5, 7.5 and 10 uM) tried, 10 muM 2iP proved optimum for eliciting cent percent morphogenic response with an average of 17.7 shoots per explant. The excised shoots on transfer to MS + 5 uM NAA developed an average of 10.7 roots per shoot in 100%cultures. Thus, raised plantlets were acclimatized to field and found to be phenotypically similar to the mother plants. The genetic fidelity of such plants was established using random amplified polymorphic DNA (RAPD) markers. Out of the 20 RAPD primersscreened, 18 RAPD primers produced clear, reproducible and scorable bands. In total, 91 distinct and scorable bands were generated, with an average of 5.1 bands per primer. All the banding profiles from micropropagated plants were monomorphic and similar to those of the mother plant. A similarity matrix based on Jaccard's coefficient revealed that the pair-wise value between the mother and the in vrtro-raised plantlets was 1, indicating 100% similarity. This confirmed the true-to-type nature of the invztro-raised plants even after 8th subculture. This is our first report of revealing the genetic fidelity of the micropropagated plants of a high artemisinin yielding elite clone of Artemisia annua L. employing RAPD.