New insight into puroindoline function inferred from their subcellular localization in developing hard and soft near-isogenic endosperm and their relationship with polymer size of storage proteins

Abstract

Wheat endosperm texture is correlated with one major locus, Ha, located on the short arm of chromosome 5D, which comprises several genes among which are two puroindoline genes, Pina and Pinb. In this study, we used two near-isogenic lines, the hard-textured line lacking Pina and the soft-textured line containing both Pina and Pinb wild-type genes. Hard and soft endosperms were observed at four kernel developmental stages, from 180 °Cd to 750 °Cd. Puroindolines were located within protein bodies at the onset of prolamin accumulation by transmission electron microscopy and immunolabelling. Ab initio modeling showed a closer structural relationship between puroindolines and 2S storage proteins from dicots than between puroindolines and other cysteine-rich wheat proteins, i.e. LTP and amylase inhibitors. Compared to the soft line, storage protein polymers in the hard line exhibited higher molecular mass (increase of from 6 to 93%) and polydispersity indices (increase of from 26 to 63%) over the course of the 4-year experiment. This suggests that puroindolines might impact the aggregation of storage proteins. Finally, these data pave the way for investigation of the role of protein–protein interactions in the texture of wheat endosperm.