In addition to increasing grain yield, improving rice (Oryza sativa L.) quality has received increasing attention recently. The cooking and eating quality (CEQ) is an important indicator of rice quality. Chalkiness and floury endosperm have a significant impact on the CEQ of rice, resulting in noticeable changes. Due to the easily observable phenotype of floury endosperm, cloning single gene mutations that cause floury endosperm and indirectly evaluating changes in CEQs facilitates the exploration of minor genes controlling CEQ. In this study, a stable genetic allele variant of flo4, named flo4-7, was obtained through EMS mutagenesis. The flo4-7 allele variant carries the cyOsPPDKB mutation. flo4-7 showed a significant reduction in compound starch granules and a significant increase in single starch granules in endosperm cells, indicating the involvement of cyOsPPDKB in the synthesis of endosperm starch. Additionally, flo4-7 exhibited a significant decrease in gel consistency (GC) compared to the wild type. Through the analysis of GC data from 166 rice germplasm resources, a C-T variation in the 18th exon of cyOsPPDKB was found to be a crucial site, causing a significant difference in GC between indica and japonica rice. This locus can be used in the future to develop molecular markers for molecular marker-assisted breeding. This study provides a good research foundation for improving the CEQ of rice.