Chalkiness is an important grain quality trait in rice. Chalk5, encoding a vacuolar H⁺-translocating pyrophosphatase, is a major gene affecting both the percentage of grains with chalkiness (PGWC) and chalkiness degree (DEC) in rice. Reducing its expression can decrease both PGEC and DEC. In this study, the first exon of Chalk5 was edited in the elite restorer line 9311 using the CRISPR/Cas9 system and two knockout mutants were obtained, one of which did not contain the exogenous Cas9 cassette. PGWC and DEC were both significantly reduced in both mutants, while the seed setting ratio (SSR) was also significantly decreased. Staggered sowing experiments showed that the chalkiness of the mutants was insensitive to temperature during the grain-filling stage, and the head milled rice rate (HMRR) could be improved even under high-temperature conditions. Finally, in the hybrid background, the mutants showed significantly reduced PGWC and DEC without changes in other agronomic traits. The results provide important germplasm and allele resources for breeding high-yield rice varieties with superior quality, especially for high-yield indica hybrid rice varieties with superior quality in high-temperature conditions.

Ratooning ability is a key factor that influences ratoon rice yield, in the area where light and temperature are not enough for second season rice. In the present study, an introgression line population derived from Minghui 63 as the recipient parent and 02428 as the donor parent was developed, and a high-density bin map containing 4568 bins was constructed. Nine ratooning-ability-related traits were measured, including maximum tiller number, panicle number, and grain yield per plant in the first season and ratoon season, as well as three secondary traits, maximum tiller number ratio, panicle number ratio, and grain yield ratio. A total of 22 main-effect QTLs were identified and explained for 3.26–18.63% of the phenotypic variations in the introgression line population. Three genomic regions, including 14.12–14.65 Mb on chromosome 5, 4.64–5.76 Mb on chromosome 8, and 10.64–15.52 Mb on chromosome 11, were identified to simultaneously control different ratooning-ability-related traits. Among them, qRA5 in the region of 14.12–14.65 Mb on chromosome 5 was validated for its pleiotropic effects on maximum tiller number and panicle number in the first season, as well as its maximum tiller number ratio, panicle number ratio, and grain yield ratio. Moreover, qRA5 was independent of genetic background and delimited into a 311.16 kb region by a substitution mapping approach. These results will help us better understand the genetic basis of rice ratooning ability and provide a valuable gene resource for breeding high-yield ratoon rice varieties.

Rice yield is a complex quantitative trait. Although a lot of genes for yield have been cloned, their genetic basis remains unknown. In the present study, a set of chromosome segment substitution line population (CSSL) was developed, derived from the indica variety Huanghuazhan as the recipient parent and the Aus variety N22 as the donor parent, and a high-density bin map containing 609 bins was constructed by resequencing. The CSSL population comprised 155 families with an average background recovery rate of 93.02%. Nine yield-related traits, including plant height, panicle number, panicle length, primary branch number, spikelet number per panicle, grain number per panicle, seed setting rate, 1000-grain weight, and grain yield per plant, were evaluated across four environments. The results showed significant differences in yield-related traits between the two parents across four environments. All nine traits showed continuous distribution with transgressive segregation. Spikelet number per panicle, grain number per panicle and 1000-grain weight showed strong correlations with each other, whereas panicle number had weak correlations with them. A total of 80 main-effect quantitative trait loci (QTLs) affecting yield-related traits were identified, among which 13 QTLs were repeatedly detected in multiple environments, 45 QTLs were located in 8 pleiotropic QTL regions, and 47 QTLs showed significant interactions with environments. In addition, 260 pairs of epistatic QTLs underlying yield-related traits were identified, of which 2 pairs stably expressed across different environments, and 11 pairs controlled more than two traits. These findings provide a theoretical basis for clarifying the genetic differentiation between indica and Aus and cloning yield-related genes, and offer valuable gene resources for molecular breeding of high-yield rice varieties.