TabHLH95-TaNF-YB1 module promotes grain starch synthesis in bread wheat

Starch is the most abundant substance in wheat(Triticum aestivum L.)endosperm and provides the major carbohydrate energy for human daily life.Starch synthesis-related(SSR)genes are believed to be spatiotemporally specific,but their transcriptional regulation remains unclear in wheat.Here,we investigate the role of the basic helix-loop-helix(bHLH)transcription factor TabHLH95 in starch synthesis.TabHLH95 is preferentially expressed in the developing grains in wheat and encodes a nucleus localized protein without autoactivation activity.The Tabhlh95 knockout mutants display smaller grain size and less starch content than wild type,whereas overexpression of TabHLH95 enhances starch accumulation and significantly improves thousand grain weight.Transcriptome analysis reveals that the expression of multiple SSR genes is significantly reduced in the Tabhlh95 mutants.TabHLH95 binds to the promoters of ADP-glucose pyrophosphorylase large subunit 1(AGPL1-1D/-1B),AGPL2-5D,and isoamylase(ISA1-7D)and enhances their transcription.Intriguingly,TabHLH95 interacts with the nuclear factor Y(NF-Y)family transcription factor TaNF-YB1,thereby synergistically regulating starch synthesis.These results suggest that the TabHLH95-TaNF-YB1 complex positively modulates starch synthesis and grain weight by regulating the expression of a subset of SSR genes,thus providing a good potential approach for genetic improvement of grain productivity in wheat.