Breeding Rice to Increase Anthocyanin Yield Per Area through Small, Black Grain Size and Three Grains per Spikelet

Rice varieties with high anthocyanin content are often recognized for their vibrant colors and health benefits. The demand for rice with high anthocyanin is increasing domestically and internationally due to consumers becoming more health-conscious. However, the current increase in yield might not raise the anthocyanin content due to its location in the grain pericarp and seed coat, which are relative to the grain surface area. This study aims to develop rice lines to increase anthocyanin yield per production area by improving rice varieties with small, black, and three grains per spikelet. Accordingly, six rice recombinant inbred lines (RILs) were bred by crossing Niaw Dam Chaw Mai Pai 49 (NDCMP49) with Khao Nok (LLR059). The grain color, size, and number of grains per spikelet were selected from the F₁ to the F₄ population through the pedigree selection method. Six RILs and their parents were assigned in a randomized complete block design (RCBD) with three replications under field conditions during the rainy season of 2021 and 2022 in four locations. The results showed statistically significant differences in environmental conditions, affecting productivity and the yield components of rice lines. Consequently, the rice lines adapted to a specific environment, and there were significant differences in genotype. This study identified three RILs with higher yield performance (13-1 (3842 kg/ha), 374-1 (3699 kg/ha), and 903-3 (3550 kg/ha)) compared with the parent NDCMP49 (1996 kg/ha). However, the grain yields were unstable in the three top-yielding RILs due to varying environmental conditions, indicating that selective breeding requires a specific, narrow environment. Based on grain yield and grain size, the RILs performed better in the grain surface area than in the parent NDCMP49. Moreover, only two RILs (374-1 and 903-3) produced the highest anthocyanin content and yield, although this was lower than in the parent NDCMP49. However, the 374-1 and 903-3 RILs produced more grains, black grains, and three grains per spikelet with high yield and moderate anthocyanin content. They can, therefore, be backcrossed to the parent NDCMP49 to increase the accumulated anthocyanin content with a stable, high yield. This work provides a resource of small grains, black grains, and three grains per spikelet in the rice breeding line for breeding programs in the future.