iTRAQ-based quantitative proteomic analysis reveals the metabolic pathways of grain chalkiness in response to nitrogen topdressing in rice.

Grain chalkiness is a highly undesirable trait that adversely affects rice quality. This chalkiness is easily influenced by the application of chemical nitrogen (N) fertilizer at the late growth stage. However, on the molecular mechanism underlying grain chalkiness caused by late N fertilization is not fully clear. In this study, proteomic differences in expression were determined in developing grains exposed to N topdressing (108 kg N ha-1, N+) and a control (0 kg N ha-1, N0), using the rice variety OM052, which has a high level of chalkiness. A total of 198 differentially expressed proteins (DEPs) were detected between the N+ and N0 treatments, including 9 up-regulated proteins and 189 down-regulated proteins. Of these DEPs, approximately half were associated with carbohydrate metabolism (glycolysis, tricarboxylic acid cycle, pentose phosphate pathway, fermentation and starch metabolism) and N metabolism (protein synthesis, folding, degradation and storage, amino acid synthesis and catabolism). A detailed pathway dissection revealed that multiple metabolic pathways during the grain filling stage were involved in the N-induced grain chalkiness. Reduced abundances of proteins associated with respiratory metabolism and energy metabolism drastically impaired the biosynthesis and deposition of starch in the developmental endosperms, which might be a crucial trigger for the increase in grain chalkiness. The disturbed N metabolism and differential expression of storage proteins up-regulated during the grain filling stage are able to partially explain the occurrence of grain chalkiness in rice.