A novel rice grain size gene OsSNB was identified by genome-wide association study in natural population

Increasing agricultural productivity is one of the most important goals of plant science research and imperative to meet the needs of a rapidly growing population. Rice (Oryza sativa L.) is one of the most important staple crops worldwide. Grain size is both a major determinant of grain yield in rice and a target trait for domestication and artificial breeding. Here, a genome-wide association study of grain length and grain width was performed using 996,722 SNP markers in 270 rice accessions. Five and four quantitative trait loci were identified for grain length and grain width, respectively. In particular, the novel grain size gene OsSNB was identified from qGW7, and further results showed that OsSNB negatively regulated grain size. Most notably, knockout mutant plants by CRISPR/Cas9 technology showed increased grain length, width, and weight, while overexpression of OsSNB yielded the opposite. Sequencing of this gene from the promoter to the 3'-untranslated region in 168 rice accessions from a wide geographic range identified eight haplotypes. Furthermore, Hap 3 has the highest grain width discovered in japonica subspecies. Compared to other haplotypes, Hap 3 has a 225 bp insertion in the promoter. Based on the difference between Hap 3 and other haplotypes, OsSNB_Indel2 was designed as a functional marker for the improvement of rice grain width. This could be directly used to assist selection toward an improvement of grain width. These findings suggest OsSNB as useful for further improvements in yield characteristics in most cultivars. Author summary Grain weight, including grain length and grain width, is a complex trait, and hundreds of quantitative trait loci (QTLs) were detected in different genetic rice populations. However, only about 10 genes have been isolated and characterized until now. Nine QTLs for grain size were identified by genome-wide association study in a natural rice population. The novel grain size gene OsSNB was identified from qGW7 based on the difference of expression levels between two different varieties with significantly different grain width. OsSNB is an AP2 transcription factor that is negatively regulated grain size. However, OsSNB was found to regulate the transition from the spikelet meristem to the floral meristem and the floral organ development in previous study. Compared to other haplotypes, Hap 3 has a 225 bp insertion in the promoter. Based on the difference between Hap 3 and other haplotypes, OsSNB_Indel2 was designed as a functional marker for the improvement of rice grain width. This can be directly used to assist selection for grain width improvement.