Wheat amino acid transporters highly expressed in grain cells regulate amino acid accumulation in grain

Amino acids are delivered into developing wheat grains to support the accumulation of storage proteins in the starchy endosperm, and transporters play important roles in regulating this process. RNA-seq, RT-qPCR, and promoter-GUS assays showed that three amino acid transporters (TaAAP2, TaAAP13, and TaAAP21) are differentially expressed in the endosperm transfer cells, starchy endosperm cells, aleurone cells and the embryo of the developing grain, respectively. Yeast complementation revealed that all three transporters can transport a broad spectrum of amino acids. RNAi-mediated suppression of TaAAP13 expression in the starchy endosperm did not reduce the total nitrogen content of the whole grain, but significantly altered the composition and distribution of metabolites in the starchy endosperm, with increasing concentrations of some amino acids (especially glutamine and glycine) from the outer to inner starchy endosperm cells. Overexpression of TaAAP13 under the endosperm-specific HMW-GS promoter significantly increased grain size, grain nitrogen concentration, and thousand grain weight, indicating that the sink strength for nitrogen improvement was increased by manipulation of amino acid transporters. However, the total grain number was reduced, suggesting that source nitrogen remobilized from leaves is a limiting factor for productivity. Therefore, simultaneously increasing loading of amino acids into the phloem and delivery to the spike would be required to increase protein content while maintaining grain yield.