An efficient method of in vitro shoot propagation of six elite accessions of Withania somnifera collected throughout India was developed. Maximum numbers of shoots in all accessions were achieved from axillary explant on Murashige and Skoog (MS) medium supplemented with 1 mgL~(-1) BAP and 1 mgL~(-1) kinetin. Inclusion of kinetin increased shoot numbers in a shorter time-period and was effective on all the elite accessions. The highest number of shoots (60 +- 1.82 and 60.05 +- 2.03) was observed in two lines developed by CIMAP, Lucknow, while the lowest number of shoots (19.4 +- 1.63) occurred on a wild collected accession with bushy growth. The in vitro raised shoots of all the accessions could be easily rooted on MS medium supplemented with 2 mgL~(-1) IB A. Rooted shoots were successfully established in a garden soil-vermi-compost (3:1, w:w) medium in a glasshouse. Stable production of withanolides from in vitro regenerated shoots was comparable to the yields from field grown mother plants, indicating the in vitro methodology could be used successfully for the true-to-type plant regeneration of Withania somnifera accessions.
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