Our study confined to observe the effect of season of explant collection on in vitro survival and rooting. Dalbergia sissoo is an important timber yielding species in India. From last decade, heavy mortality rate and slow propagation has threatened its existence. So rapid multiplication is the urgent need of hour. Season of explant collection plays a crucial role for shoot proliferation and establishment of cultures. Nodal explants were excised from five superior clones of Dalbergia sissoo and cultured in vitro using MS media supplemented with auxin and cytokinin. Explants were collected in two seasons i.e., S1 (April to June) and S2 (July to September). Maximum survival, bud response and rooting were observed in explants collected in the months of April to June. A significant variation of explant collection season was observed in establishment and rooting of cultures. Explants collected in month of July to September exhibited maximum contamination.