Cell-type proteomic and metabolomic resolution of early and late grain filling stages of wheat endosperm.

The nutritional value of wheat grains, particularly their protein and metabolite composition, is a result of the grain-filling process, especially in the endosperm. Here, we employ laser microdissection (LMD) combined with shotgun proteomics and metabolomics to generate a cell type-specific proteome and metabolome inventory of developing wheat endosperm at the early (15 DAA) and late (26 DAA) grain-filling stages. We identified 1803 proteins and 41 metabolites from four different cell types (aleurone (AL), sub-aleurone (SA), starchy endosperm (SE) and endosperm transfer cells (ETCs). Differentially expressed proteins were detected, 67 in the AL, 31 in the SA, 27 in the SE and 50 in the ETCs between these two-time points. Cell-type accumulation of specific SUT and GLUT transporters, sucrose converting and starch biosynthesis enzymes correlate well with the respective sugar metabolites, suggesting sugar upload and starch accumulation via nucellar projection and ETC at 15 DAA in contrast to the later stage at 26 DAA. Changes in various protein levels between AL, SA and ETC support this metabolic switch from 15 to 26 DAA. The distinct spatial and temporal abundances of proteins and metabolites revealed a contrasting activity of nitrogen assimilation pathways, e.g. for GOGAT, GDH and glutamic acid, in the different cell types from 15 to 26 DAA, which can be correlated with specific protein accumulation in the endosperm. The integration of cell-type specific proteome and metabolome data revealed a complex metabolic interplay of the different cell types and a functional switch during grain development and grain-filling processes.

Effects of Cysteine and Inorganic Sulfur Applications at Different Growth Stages on Grain Protein and End-Use Quality in Wheat.

The aim of this study was to test the significant effects of inorganic sulfur and cysteine on grain protein and flour quality in wheat and to provide a theoretical basis of wheat cultivation techniques with high yield and quality. In the field experiment, a winter wheat cultivar, Yangmai 16, was used, and five treatments were established, i.e., S0 (no sulfur fertilizer application during the whole wheat growth period), S(B)60 (60 kg ha-1 inorganic sulfur fertilizer was applied as the basal fertilizer), Cys(B)60 (60 kg ha-1 cysteine sulfur fertilizer was applied as the basal fertilizer), S(J)60 (60 kg ha-1 inorganic sulfur fertilizer was applied as the jointing fertilizer), and Cys(J)60 (60 kg ha-1 cysteine sulfur fertilizer was applied as the jointing fertilizer). The fertilizer application at jointing stage showed a better influence than basal fertilizer application on protein quality; for the content of albumin, gliadin, and high molecular weight glutenin (HMW-GS), Cys(J)60 was the best among these treatments. An increase of 7.9%, 24.4%, 43.5%, 22.7% and 36.4% was found in grain yield, glutenin content, glutenin macro-polymer (GMP), low molecular weight glutenin (LMW-GS), and S content under Cys(J)60, in relation to the control, respectively. A similar trend was found in the end-use quality, as exemplified by an increase of 38.6%, 10.9%, 60.5%, and 109.8% in wet gluten content, dry gluten content, sedimentation volume, and bread-specific volume, respectively; a decrease of 69.3% and 69.1% in bread hardness and bread chewiness was found under Cys(J)60. In terms of application period, topdressing at jointing stage is compared with base fertilizer, the sulfur fertilizer application at jointing stage showed larger effects on grain protein and flour quality, from the different types of sulfur fertilizer, the application of cysteine performed better than the use of inorganic sulfur. The Cys(J)60 exhibited the best effects on protein and flour quality. It was suggested that sufficient sulfur application at jointing stage has the potential to enhance the grain protein and flour quality.