Low temperature is a major constraint for crop productivity. To cope with this challenge, plants have developed several mechanisms to adapt to low temperature. Developing breeding strategies to enhance cold stress tolerance in crops requires an understanding of the mechanisms by which plants perceive and transmit cold stress-related signals to their cellular machinery, thereby activating adaptive responses. Only one quantitative trait locus for tolerance to low-temperature germination, qLTG3-1, has been narrowed down to the gene level in rice. A 71 bp indel that can be used to distinguish between tolerant and susceptible parents has been identified. We tested the 71 bp indel on 65 diverse rice genotypes including those adapted to colder climates of North and Northeastern India to find evidence of the tolerant allele (insertion) and to see whether it is associated with low-temperature germinability in these genotypes. Our results show that 48% of the rice genotypes tested carried the tolerant allele. The insertion was found to be significantly associated with cold tolerance during germination. Moreover, several landraces/improved varieties known for their superior performance in other abiotic stress conditions such as drought and high salinity conditions, and which were previously never exposed to low temperature, carry the beneficial allele for qLTG3-1, suggesting an additional role of this allele in adverse climatic conditions. This study enhances current understanding of the distribution of the tolerant allele qLTG3-1 in rice germplasm, which could help in the identification of suitable donors for potential marker-assisted breeding programmes.