Wheat-rye translocation lines were developed to produce a main crop resistant to biological and physical stress. 'Chaupon' rye contains 2RL chromatin to harbor resistance genes for powdery mildew and leaf rust. In order to identify chromosome 2RL-derived rye proteins and 2RL-perturbed proteins in wheat-rye translocation lines, the gel-based proteomics was employed with 'Coker797' (non-2RL), 'Hamlet' (2RL) and 'near-isogenic line' (stabilized 2RL). The leaf proteome was resolved on 2D-gel, resulting in 216 spots in a final selection. A total of 90 proteins were identified with the identification success rate of 42%. The identified proteins were classified by functional annotation: metabolism (64%), cellular process (5%), translation (2%), regulatory function (1%) and hypothetical (28%). The proteins belonged to metabolism were subdivided into carbohydrate metabolism (36%), energy metabolism (35%), metabolism of lipid, amino acid, other amino acid and biosynthesis of secondary metabolites (each 6%) and others (5%). A total of 53 proteins were differentially expressed, in which β-glucosidase, in particular, originated from the chromosome 2RL of rye, was exclusively appeared in NIL. In addition, small Ras-related GTP binding-protein assigned to wheat was predominantly found in 2RL rye chromatin-possessing NIL. These results suggest that the acquired genetic traits obtained from rye 2RL enhance the resistance to biotic and abiotic stress in wheat-rye translocation lines by altered the proteome expression. In leaf metabolome analysis, 11 predominant metabolites containing trans-aconitate, glutamate, and betaine were identified by 1H-NMR-based metabolite fingerprinting. The overall metabolites pattern of NIH appears to be closer to Coker797 rather than Hamlet. Thus, the metabolic phenotype of NIL was not so much lineated from Hamlet contrast to proteomic phenotyping.

Seed color is an important trait affecting flour yield and quality in wheat. Seed color also is either tightly linked to or pleiotropically controls seed dormancy in wheat, because most of the red-seeded wheats are tolerant to pre-harvest sprouting in comparison to white-seeded wheats. Recently, metabolomics approaches have recently been used to assess the natural variance in metabolite content between individual plants, an approach with great potential for the improvement of the compositional quality of crops. Basically, in the study here, the simultaneous proteomic and metablomic approaches are being investigated to identify the expressed proteins of genes and specific metabolism responsible for the expression of red and white colors of seed. Red seed “Jinpum” and white seed “Kumkang” cultivars were used in this study to identify the storage proteins use of 2-DE, MALDI-TOF/MS. Here we optimize tissue extraction methods compatible with high-throughput, reproducible nuclear magnetic resonance (NMR) spectroscopy based metabolomics. It appears that the proteins expressed were different each other according to two different cultivars from the seeds of hexaploid wheat. Some selected protein spots were identified as follows: B3-hordein, Gamma-hordein-3,bifunctionalalpha amylase/subtilisin Inhibitor. To monitor metabolic profile, wheat grain was ground in liquid nitrogen, ensuring a homogeneous mix of the tissue, solution samples extracted from seed grains of two wheat cultivars were conducted to measurement of metabolite using 1H-1D NMR method. Representative 1H-1D NMR spectra showing the metabolic fingerprints of wheat grain extracted and presented in Fig. The different peaks, observed at 3.4 and 4.3 ppm, were detected and difference in each two cultivars. The metabolic fingerprint of each two wheat cultivars by 1H-1D NMR were analysed using partial least squares (PLS) in mutivariate analysis to confirm metabolic profiling between different cultivars and to screen chemical shift spectrum corresponding to metabolite specifically abundant in each cultivars. Profiling using 1H-1D NMR was applied to measure of abundance of major metabolite. In total metabolites were compared between “Jinpum” and “Kumkang” cultivars. Therefore, NMR based on the metabolic-phenotyping should be mostly applicable to systematic exploration of plant genetic resources as well as to metabolite based on the breeding program involved in crops productivity.

The wheat-rye translocation lines have been agriculturally developed for the resistance to the biotypes of Hessian fly as a major insect pest of wheat. In order to compare the proteomic profiles between ‘Coker797’ (non-2RL), ‘Hamlet’ (2RL), and near-isogenic line (NIL) carrying 2RL, we evaluated the protein extraction and preparation methods for two-dimensional gel electrophoresis approach. The tissues such as leaves, stems, and roots from three wheat-rye lines were extracted by following trichloroacetic acid (TCA)/acetone precipitation. In a preliminary proteome analysis, a commonly expressed protein in Hamlet and NIL strain was identified as methionine synthase annotated in Hordeum vulgare subsp. The present study will provide the experimental guideline for the proteomic study of other useful crop plant tissues.