Abstract

The pollen wall is a specialized extracellular cell wall that protects male gametophytes from various environmental stresses and facilitates pollination. Here, we reported that bHLH010 and bHLH089 together are required for the development of the pollen wall by regulating their specific downstream transcriptional and metabolic networks. Both the exine and intine structures ofbhlh010 bhlh089pollen grains were severely defective. Further untargeted metabolomic and transcriptomic analyses revealed that the accumulation of pollen wall morphogenesis-related metabolites, including polysaccharides, glyceryl derivatives, and flavonols, were significantly changed, and the expression of such metabolic enzyme-encoding genes and transporter-encoding genes related to pollen wall morphogenesis was downregulated inbhlh010 bhlh089mutants. Among these downstream target genes,CSLB03is a novel target with no biological function being reported yet. We found that bHLH010 interacted with the two E-box sequences at the promoter ofCSLB03and directly activated the expression ofCSLB03.The cslb03 mutant alleles showedbhlh010 bhlh089–like pollen developmental defects, with most of the pollen grains exhibiting defective pollen wall structures.Keywords:anther;pollen wall development;bHLH010;CSLB03;Arabidopsis thaliana