Mushroom cultivation is one of the technology for the efficient conversion of energy utilising the agro residues. In the present study we have evaluated the various substrates for the spawn production for the cultivation of shiitake mushroom. Spawn was prepared using wheat, sorghum and pearl millet grains solely and in combination with their respective grain powder at 9:1 proportion as substrate. The fastest coverage of mycelia was observed on the sorghum grains (20 days), whereas slowest mycelial run on the grain surface was observed on the pearl millet grains (45 days). There was a significant difference between all the spawning substrates in their pure form and their combinations except for the pearl millet and its grain combination. The study documented the best substrate for the shiitake mushroom spawn production.