Ex-Situ conservation of Haplophyllum Gilesii: a critically endangered and narrow endemic species of the Karakoram range

摘要

The present study aimed to develop effective In vitro methods by means of callus and axillary buds for ex-situ conservation and rapid micro-propagation of a critically endangered, narrow endemic species Haplophyllum gilesii (Hemsel) C.C. Townsend, present in Pakistan confined to Gilgit Baltistan region. Media used was MS basal with various combinations of growth regulators (kinetin, GA 3 , BAP, IBA and NAA) for shoot multiplication, callus induction, regeneration and root induction. Supplementation of 2.5mgL -1 BAP for 35 days of culture subsequently resulted in maximum number (6.8) of shoots. Healthy roots were induced effectively with addition of GA 3 (1.5mgL -1 ) and (0.4mgL -1 ) kinetin in half strength MS media. 2.5mgL -1 NAA, 0.25mgL -1 kinetin and 2.5mgL -1 BAP exhibited effective initiation and proliferation of callus while best regeneration was obtained at the medium having 1mgL -1 IBA. Acclimatized plantlets exhibited 100% survival rate for one week, after one month 60%, while survival rate after 5 months in pots was 45%. The protocol developed here can be used effectively for in-situ and ex-situ conservation as well as mass multiplication for production of H. gilesii plantlets and callus to assess its advantages without disturbing wild populations of this plant. There is still need to investigate secondary metabolic and molecular composition of this plant.