Paphiopedilum callosum or Lady’s slipper orchids or “Rong Thao Nari Khang Kob” in Thai, is one of very favorite potted flower and popular in the orchids markets. Paphiopedilum spp. is regarded as endangered or even extinct from the wild because smuggling out of natural areas and illegal trade large. Therefore, they have been assigned to endanger by the Convention on International Trade in Endangered Species of Wild Fauna and Flora (Long et al., 2010). Nowadays, growing orchids is international business and orchids have high demand of consumer so large-scale multiplication of delicate and rare hybrids using tissue culture techniques has aided orchid capture a position as one of the top ten cut flower (Chugh et al., 2009). Paphiopedilum callosum is terrestrial orchid that have slow growth and low survival rate of the orchid cultivation which the relationship between mycorrhizal fungi and orchid simplify the nutrient uptake of orchid and resulting in growth stimulation, the fungus translocates sugars, vitamins and amino acids to the orchids (Arditti et al., 1990). Moreover, orchid mycorrhizal fungus was observed to prevent materials of orchid cultivation from fungal diseases and contaminant (Aewsakul et al., 2013)