Seed priming with biofertilizers offers a sustainable and eco-friendly approach to improving rice seed germination and early seedling growth. This study investigated the effects of biofertilizer treatments on germination speed, vigor index, plant height, and root length in three rice varieties (Nipponbare, Koshihikari, and NU1). Treatments included seeds soaked or unsoaked in biofertilizer, compared to control and autoclaved biofertilizer under controlled conditions. Results showed that biofertilizer significantly enhanced germination speed, with NU1 demonstrating a 9% improvement compared to the control. The vigor index increased by 34% in Nipponbare, 61% in Koshihikari, and 87% in NU1. Biofertilizer-soaked seeds also recorded a substantial increase in plant height, with Nipponbare showing a 114% improvement, while root length improved markedly, with NU1 exhibiting a 25.8% increase. Statistical analysis using ANOVA and Tukey’s HSD test confirmed that these improvements were significant (p < 0.05). The autoclaved biofertilizer showed limited effects, emphasizing the necessity of active microbial components. These findings illustrate the potential of biofertilizers to accelerate germination, improve early growth parameters, and enhance rice productivity, supporting sustainable agricultural practices and contributing to global food security efforts.