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Pure breeding multiplication Shitake mushrooms generally use Potato Dekstrose Agar (PDA) media. However, the cost of basic media is relatively higher, so it needs alternative media to multiply pure broodstock for shitake mushrooms to reduce the cost of producing pure broodstock such as rice wastewater. Goal of the research was to find out ability of rice wastewater medium and determine the concentration of the best rice wastewater medium on the growth rate of shitake mushroom mycelium. The study used an experimental method consisting of 11 treatments of rice wastewater concentration 50%, 55%, 60%, 65%, 70%, 80%, 85%, 90%, 95%, 100% and Potato Dekstrose Agar (PDA) as the repeated control three replication. The results showed that influence of various concentrations of rice wastewater on the growth rate of shitake mushroom mycelium. The concentration of 100% rice wastewater produces the best growth and quality of mycelium compared to other treatments. Therefore Rice wastewater can be used as an alternative medium to multiply pure broodstock of shitake mushrooms.