Premise Plants synthesize information from multiple environmental stimuli when determining their direction of growth. Gravity, being ubiquitous on Earth, plays a major role in determining the direction of growth and overall architecture of the plant. Here, we utilized the microgravity environment on board the International Space Station (ISS) to identify genes involved influencing growth and development of phototropically stimulated seedlings of Arabidopsis thaliana. Methods Seedlings were grown on the ISS, and RNA was extracted from 7 samples (pools of 10–15 plants) grown in microgravity (μg) or Earth gravity conditions (1‐g). Transcriptomic analyses via RNA sequencing (RNA‐seq) of differential gene expression was performed using the HISAT2‐Stringtie‐DESeq2 RNASeq pipeline. Differentially expressed genes were further characterized by using Pathway Analysis and enrichment for Gene Ontology classifications. Results For 296 genes that were found significantly differentially expressed between plants in microgravity compared to 1‐g controls, Pathway Analysis identified eight molecular pathways that were significantly affected by reduced gravity conditions. Specifically, light‐associated pathways (e.g., photosynthesis‐antenna proteins, photosynthesis, porphyrin, and chlorophyll metabolism) were significantly downregulated in microgravity. Conclusions Gene expression in A. thaliana seedlings grown in microgravity was significantly altered compared to that of the 1‐g control. Understanding how plants grow in conditions of microgravity not only aids in our understanding of how plants grow and respond to the environment but will also help to efficiently grow plants during long‐range space missions.