Single seeds of common buckwheat cultivar Suwon No. 1 when subjected to SDS-PAGE revealed very high polymorphism. High variation existed for protein or protein subunits with molecular weight 54-47kDa, 45-25kDa and 16-11kDa. The electrophoregram showed variation for globulin as well as other protein fractions. About 300 proteins were separated by two-dimensional electrophoresis in common buckwheat (Fagopyrum esculentum Moench.) seed. Seed maturation is a dynamic and temporally regulated phase of seed development that determines the composition of storage proteins reserves in mature seeds. Buckwheat seeds from 5, 10, 15, 20, and 25 days after pollination and matured stage were used for the analysis. This led to the establishment of high-resolution proteome reference maps, expression profiles of 48 spots. It was identified 48 proteins from MALDI-TOF/MS analysis of wild buckwheat seed storage proteins. The 48 proteins were found identical or similar to those of proteins reported in buckwheat and other plants; it is belonging to 9 major functional categories including seed storage proteins, stress/defense response, protein synthesis, photosynthesis, allergy proteins, amino acid, enzyme, metabolism, and miscellaneous. It appears that the major allergenic storage protein separated played the important role in buckwheat breeding and biochemical characterization.

Buckwheat is one of the traditional crops and has become a renewed target of interest or a popular crop as a healthy foodstuff, because it is a good source of cereal protein which is rich with essential amino acids. However, what is critical to our health is that buckwheat contains proteins which cause a allergy. Buckwheat allergy resulting from ingestion is caused by the storage proteins in the grain with molecular weights ranging from 15, 22, 35, 39 and 50 kDa proteins of the inner fractions to low, and there were clear differences in the protein compositions between the inner and outer buckwheat flour fractions. A major allergenic protein of buckwheat is Fag e 1 with molecular weight 22 kDa (BW22KD). Buckwheat allergy is an immunoglobulin E (IgE)-mediated hypersensitive response capable of causing anaphylactic shock. Buckwheat seeds were dissected to endosperm and embryo. From each fraction we extracted proteins and analyzed extracts by SDS-PAGE and 2-DE. On electrophoregrams of endosperm proteins, 6 intense bands were detected. The most intense corresponded to molecular weights ranging from 54 to 65 kDa. These proteins have been reported not to be allergenic. We show here that the allergenic buckwheat seed proteins are found only among embryo proteins. No allergenic proteins were found in the buckwheat endosperm. The results presented here lead to the proposal that patients with hypersensitivity to buckwheat flour should use only fine flour from buckwheat endosperm, as this fraction contains no allergenic proteins. At present, specific protein spots will be selected and in-gel digested for MALDI-TOF-TOF/MS analysis.

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.