Improving rice quality remains a crucial breeding objective, second only to enhancing yield, yet progress in quality improvement lags behind yield. The high temperature and ripening conditions in Southern China often result in poor rice quality, impacting hybrid rice production and utilization. Therefore, to address this challenge, analyzing the molecular basis of high-quality traits is essential for molecular design breeding of high-quality hybrid rice varieties. In this study, we investigated the molecular basis of grain shape, amylose content, gel consistency, gelatinization temperature, and aroma, which influence rice quality. We discovered that quality related alleles gs3, GW7^TFA, gw8, chalk5, Wx^b, ALK^TT, and fgr can enhance rice quality when applied in breeding programs. Polymerization of gs3, GW7^TFA, gw8, and chalk5 genes improves rice appearance quality. The gs3 and GW7^TFA allele polymerization increasing the grain’s length-width ratio, adding the aggregation of gw8 allele can further reducing grain width. The chalk5 gene regulates low chalkiness, but low correlation to chalkiness was exhibited with grain widths below 2.0 mm, with minimal differences between Chalk5 and chalk5 alleles. Enhancing rice cooking and eating quality is achieved through Wx^b and ALK^TT gene polymerization, while introducing the fgr(E7) gene significantly improved rice aroma. Using molecular marker-assisted technology, we aggregated these genes to develop a batch of indica hybrid rice parents with improved rice quality are obtained. Cross-combining these enhanced parents can generate new, high-quality hybrid rice varieties suitable for cultivation in Southern China. Therefore, our findings contribute to a molecular breeding model for grain quality improvement in high-quality indica hybrid rice. This study, along with others, highlights the potential of molecular design breeding for enhancing complex traits, particularly rice grain quality.

Rice (Oryza sativa L.) is an important staple food crop for more than half of the world’s population. Enhancing the grain quality and yield of rice to meet growing demand remains a major challenge. Here, we show that OsMKK3 encode a MAP kinase kinase that controls grain size and chalkiness by affecting cell proliferation in spikelet hulls. We showed that OsSPL16, GS5, and GIF1 have a substantial effect on the OsMKK3-regulated grain size pathway. OsMKK3 has experienced strong directional selection in indica and japonica. Wild rice accessions contained four OsMKK3 haplotypes, suggesting that the OsMKK3 haplotypes present in cultivated rice likely originated from different wild rice accessions during rice domestication. OsMKK3-Hap1, gs3, and gw8 were polymerized to enhance the grain length. Polymerization of beneficial alleles, such as OsMKK3-Hap1, gs3, gw8, fgr, alk, chalk5, and wx, also improved the quality of hybrid rice. Overall, the results indicated that beneficial OsMKK3 alleles could be used for genomic-assisted breeding for rice cultivar improvement and be polymerized with other beneficial alleles.

Starch branching enzyme IIb (BEIIb) and soluble starch synthase IIa (SSIIa) play important roles in starch biosynthesis in cereals. Deficiency in the BEIIb gene produces the amylose extender (ae) mutant rice strain with increased amylose content (AC) and changes in the amylopectin structure. The SSIIa gene is responsible for the genetic control of gelatinization temperature (GT). The combined effects of BEIIb and SSIIa alleles on the AC, fine structures, and physicochemical properties of starches from 12 rice accessions including 10 recombinant inbred lines (RIL) and their two parents were examined in this study. Under the active BEIIb background, starches with the SSIIa-GC allele showed a higher GT than those with the SSIIa-TT allele, resulting from a lower proportion of A chain and a larger proportion of B1 chains in the amylopectin of SSIIa-GC. However, starch with the BEIIb mutant allele (be2b) in combination with any SSIIa genotype displayed more amylose long chains, higher amylose content, B2 and B3 chains, and molecular order, but smaller relative crystallinity and proportion of amylopectin A and B1 chains than those with BEIIb, leading to a higher GT and lower paste viscosities. These results suggest that BEIIb is more important in determining the structural and physicochemical properties than SSIIa. These results provide additional insights into the structure-function relationship in indica rice rather than that in japonica rice and are useful for breeding rice with high amylose content and high resistant starch.

Although great progress has been made in the development of hybrid rice with increased yield, challenges for the improvement of grain quality still remain. In this study, the textural properties of cooked rice and physicochemical characteristics of starch were investigated for 29 new hybrid rice derived from 5 sterile and 11 restorer rice lines. Except for one sterile line Te A (P1) with high apparent amylose content (AAC) (26.9%), all other parents exhibited a low AAC. Gui 263 demonstrated the highest AAC (20.6%) among the restorer lines, so the Te A/Gui 263 hybrid displayed the highest AAC (23.1%) among all the hybrid rice. The mean AAC was similar between sterile, restorer lines and hybrid rice. However, the mean hardness of cooked rice and gels of sterile lines were significantly higher than that of restorer lines and hybrid rice (p < 0.05). Pasting temperature and gelatinization temperatures were significantly higher in the hybrids than in the restorer lines (p < 0.05). Cluster analysis based on the physicochemical properties divided the parents and hybrid rice into two major groups. One group included P1 (Te A), P12 and P14 and three hybrid rice derived from P1, while the other group, including 39 rice varieties, could be further divided into three subgroups. AAC showed significant correlation with many parameters, including peak viscosity, hot peak viscosity, cold peak viscosity, breakdown, setback, onset temperature, peak temperature, conclusion temperature, enthalpy of gelatinization, gel hardness and cooked rice hardness (p < 0.05). Principal component analysis revealed that the first component, comprised of the AAC, peak viscosity, breakdown, setback, onset temperature, peak temperature, conclusion temperature and gel hardness, explained 44.1% of variance, suggesting AAC is the most important factor affecting the grain quality of hybrid rice. Overall, this study enables targeted improvements to key rice grain quality attributes, particularly AAC and textural properties, that will help to develop superior rice varieties.