Fine-Tuning of the Grain Size by Alternative Splicing of GS3 in Rice

Grain size is subtly regulated by multiple signaling pathways in rice. Alternative splicing is a general mechanism that regulates gene expression at the post-transcriptional level. However, to our knowledge, the molecular mechanism underlying grain size regulation by alternative splicing is largely unknown. GS3, the first identified QTL for grain size in rice, is regulated at the transcriptional and post-translational level. In this study, we identified that GS3 is subject to alternative splicing. GS3.1 and GS3.2, two dominant isoforms, accounts for about 50% and 40% of total transcripts, respectively. GS3.1 encodes the full-length protein, while GS3.2 generated a truncated proteins only containing OSR domain due to a 14 bp intronic sequence retention. Genetic analysis revealed that GS3.1 overexpressors decreased grain size, but GS3.2 showed no significant effect on grain size. Furthermore, we demonstrated that GS3.2 disrupts GS3.1 signaling by competitive occupation of RGB1. Therefore, we draw a conclusion that the alternative splicing of GS3 decreases the amount of GS3.1 and GS3.2 disrupts the GS3.1 signaling to inhibit the negative effects of GS3.1 to fine-tune grain size. Moreover, the mechanism is conserved in cereals rather than in Cruciferae, which is associated with its effects on grain size. The results provide a novel, conserved and important mechanism underlying grain size regulation at the post-transcriptional level in cereals.

Automated summary

This study reveals the molecular mechanism underlying grain size regulation in rice. GS3 produces two isoforms, GS3.1 and GS3.2, through alternative splicing, with GS3.1 encoding a full-length protein and GS3.2 generating a truncated protein due to intronic sequence retention. Overexpression of GS3.1 reduces grain size, while GS3.2 has no significant effect on grain size. Furthermore, GS3.2 disrupts the signaling of GS3.1 by competitively occupying RGB1. This mechanism is conserved in cereals.