Abstract Plant starch is naturally phosphorylated at a fraction of the C6 and the C3 hydroxyl groups during its biosynthesis in plastids. Starch phosphate esters are important in starch metabolism and they also generate specific industrial functionality. Cereal grains starch contains little starch bound phosphate compared with potato tuber starch and in order to investigate the effect of increased endosperm starch phosphate, the potato starch phosphorylating enzyme glucan water dikinase (StGWD) was overexpressed specifically in the developing barley endosperm. StGWD overexpressors showed wild-type phenotype. Transgenic cereal grains synthesized starch with higher starch bound phosphate content (7.5 (±0.67) nmol/mg) compared to control lines (0.8 (±0.05) nmol/mg) with starch granules showing altered morphology and lower melting enthalpy. Our data indicate specific action of GWD during starch biosynthesis and demonstrates the possibility for in planta production of highly phosphorylated cereal starch.