The 14‐3‐3 protein GF 14f negatively affects grain filling of inferior spikelets of rice ( Oryza sativa L.)

Summary In rice ( Oryza sativa L.), later flowering inferior spikelets ( IS ), which are located on proximal secondary branches, fill slowly and produce smaller and lighter grains than earlier flowering superior spikelets ( SS ). Many genes have been reported to be involved in poor grain filling of IS , however the underlying molecular mechanisms remain unclear. The present study determined that GF 14f, a member of the 14‐3‐3 protein family, showed temporal and spatial differences in expression patterns between SS and IS . Using GF 14f – RNA i plants, we observed that a reduction in GF 14f expression in the endosperm resulted in a significant increase in both grain length and weight, which in turn improved grain yield. Furthermore, pull‐down assays indicated that GF 14f interacts with enzymes that are involved in sucrose breakdown, starch synthesis, tricarboxylic acid ( TCA ) cycle and glycolysis. At the same time, an increase in the activity of sucrose synthase (SuSase), adenosine diphosphate‐glucose pyrophosphorylase ( AGP ase), and starch synthase (StSase) was observed in the GF 14f – RNA i grains. Comprehensive analysis of the proteome and metabolite profiling revealed that the abundance of proteins related to the TCA cycle, and glycolysis increased in the GF 14f – RNA i grains together with several carbohydrate intermediates. These results suggested that GF 14f negatively affected grain development and filling, and the observed higher abundance of the GF 14f protein in IS compared with SS may be responsible for poor IS grain filling. The study provides insights into the molecular mechanisms underlying poor grain filling of IS and suggests that GF 14f could serve as a potential tool for improving rice grain filling.