With the increase of research on establishing a lunar base, there are many challenges faced regarding sustainable living conditions. Some limitations include the cost of long-term spaceflight due to the frequent transport of food supply to the moon. The use of in-situ resources and the recycling of biomass waste can be used for sustainable crop production to decrease the costs of long-term spaceflight and make efficient use of payloads. Lunar regolith, the moon’s soil, contains all the nutrients needed for plant growth except for nitrogen. However, biomass waste, such as organic waste, can be used as a nitrogen source and a supplement providing the lunar regolith with microbial communities to sustain plant growth. In this experiment we are investigating the effects of added microbial communities in horse manure on lunar regolith simulant for optimal plant growth. The experimental design requires the testing of protocols including a watering schedule, determining an optimal ratio of manure to lunar regolith simulant, and other appropriate growth conditions. The objective of this experiment is to analyze the microbial communities in the rhizosphere for plants in different substrates in an effort to simulate the Earth’s soil that can sustain crop production for a lunar base.