Sub-cellular proteomics provide insight into the molecular mechanisms of plant cell modulation of protein accumulation in intracellular compartments regarding various perturbations, and thus provides rectified knowledge about signal transduction in organelles. Mitochondria are important organelles for cellular respiration within the eukaryotic cell and serve many important functions including vitamin synthesis, amino acid metabolism and photorespiration for the cell as well. To define the mitochondrial proteome of the roots of wheat seedling, a systematical and targeting analysis were carried out on the mitochondrial proteome from 15 days-old wheat seedling roots material. Mitochondria were isolated by Percoll gradient centrifugation; and extracted proteins were separated and analyzed using Tricine SDS-PAGE along with LTQ-FTICR mass spectrometry. From the isolated mitochondrial proteins, a total of 140 proteins were identified. The identified proteins were functionally classified into 12 classes using ProtFun 2.2 server based on cellular roles, Proteins were shown to be involved in including amino acid biosynthesis (17.1%), biosynthesis of cofactors (6.4%), cell envelope (11.4%), central intermediary metabolism (10%), energy metabolism (20%), fatty acid metabolism (0.7%), purines and pyrimidines (5.7%), regulatory functions (0.7%), replication and transcription (1.4%), translation (22.1%), transport and binding (1.4%), and unknown (2.8%). These results indicated that many of the protein components present and functions of identified proteins are common to other profiles of mitochondrial proteomes performed to date. The data presented here will begin to reveal a better understanding the characteristics of proteins and metabolic activity in mitochondria in wheat roots.

Buckwheat sprout is used as vegetable, and also flour for making noodles, and so on. Currently, information about tissue culture in buckwheat is limited and restricted to micropropagation. We carried out somatic embryogenesis and plant regeneration using hypocotyl segments as explant of the cultivated buckwheat species Fagopyrum esculentum, differs from existing studies in the growth regulator combinations used. Maximum callus regeneration was induced on MS medium containing 2,4-D(2.0 mg/L) and benzylaminopurine BAP (1.0 mg/L) and 3% sucrose. Friable callus was transferred to solidified MS media containing BAP (1.0 mg/L) and at various concentrations for the induction of embryogensis. The optimum concentrations of additives were IAA (2 mg/L), KIN(1.0 mg/L), BAP (1.0 mg/L), and 3% (w/v) sucrose. Only 2,4-D did not show any significant effect on callus induction or embryogenesis. Regeneration of embryonic callus varied from 5 % to 20%. Whole plants were obtained at high frequencies when the embryogenic calluses with somatic embryos and organized shoot primordia were transferred to MS media with 3% sucrose. Regenerated plants after acclimation will transfer to green house. The main objective of this research was to develop a efficient protocol for plant regeneration for common buckwheat, and to apply in future for genetic transformation.

Buckwheat sprouts are a vegetable which provides health benefit with their nutritionally important substances. Buckwheat has been considered as preventive medicine in the last decade. The present study was focused on the reference maps common (Fagopyrum esculentum Möench.) and tatary(Fagopyrum tataricum Gaertn.) buckwheat leaf and stem cultured in light and dark condition. Proteins were extracted from 7-day germinated buckwheat sprout sand separated by two-dimensional electrophoresis(2-DE) with isoelectro focusing gel over pH3 to 10. A total of more than 1520 protein spots were revealed on 2-DE gel, in which 165 proteins were identified in the basis of peptide mass fingerprinting. Functional category analysis indicated that these differentially expressed proteins mainly involved in cellular process, defense responsive, energy production, metabolism, photosynthesis, DNA recombination, DNA replication, seed storage, signal transduction, stress responsive, transcription, translation, and energy transport proteins. The pattern at protein level suggested the important roles for energy and protein metabolism-related proteins in growing sprouts under dark and light condition, accompanied by the activated of the stress responsive and growth condition. The proteomic profiling of common and tatary buckwheat will give insight for understanding buckwheat physiology and application to buckwheat industry.

The wild relative’s diploid species, which are reproductively isolated from one another, compromise populations with marked morphological variation, wide climatic tolerance, and adaptation to diverse habitats, and also vary genetically in biotic, abiotic stresses, and in seed protein content and quality. Large-scale proteomic analysis of three wild relatives of wheat grain (AA, BB, and DD genome) using matrix assisted laser desorption/ionization- time of flight (MALDI-TOF-MS), multi-dimensional protein identification technology (MudPIT), allowed the detection and classification of 213, 255 unique proteins (peptide match ≥ 2), which represents the most wide-ranging proteome exploitation to date. Development of standard proteomes exhibiting all of the proteins involved in normal physiology will facilitate the delineation of disease/defense (no. of unique protein (n) =33, 51), metabolism (n=15, 32), energy metabolism (n= 21, 27), protein synthesis (n=16, 22), folding/stability (n=17, 18), transcription (n=6, 18), cell growth/division (n=17, 17), signal transduction (n=16, 15), cellular organization (n=11, 12), development (n=9, 9), storage protein (n= 30, 7), transport facilitation (n=8, 6), and unclear classification (n= 14, 21), which is identification by using MALDI-TOF and LCQ DECA mass spectrometry couple to mascot database search, respectively. For instance, ABA inducible protein PHVA1 (HVA1), which can be induced by drought, cold, heat and salinity condition, and also basic endochitinase (RSCC, RSCA) showed defense against chitin containing fungal pathogens. Gluten (glutenin and gliadin), which is very important determinant for making high quality bread, noodles, and also associated with visco-elasticity. By using MALDI-TOF, we identified abundant disease related protein such as NBS-LRR involves in response to the presence of a foreign body or the occurrence of an injury, which result in restriction of damage to the organism attacked or prevention/recovery from the infection caused by the attack, puroindoline (a & b) and grain softness protein represents the molecular-genetic basis of grain texture. In addition, the PIN A and PIN B gene products have anti microbial properties with potential role in plant defense. Recent advances in mass spectrometry and bioinformatics have provided the means to characterize complex protein landscapes from a wide variety of organisms. Hierarchical clustering could be applied to protein information from different samples using Gene Pattern and NCSS software. Here we report also genome specific protein interaction network using Cytoscape software, which provides further insight into the molecular mechanism of biochemical pathways. By integrating shotgun proteomics with statistical and computation alanalyses, we developed promising understand about expressed protein and protein functions. Our approach should be applicable for marker assisted breeding or genetransfer for quality and stress research of cultivated wheat.

To better understanding the function of the luminal sub-organelles within the thylakoid network, we have carried out a systematical analysis and identification of the lumenal proteins in the thylakoid of wheat by using Tricine / 1D-PAGE, and LTQ-ESI-FTICR mass spectrometry followed by SWISS-PROT database searching. We isolation and fractionation these membrane from fully developed wheat leaves using a combination of differential and gradient centrifugation couple to high speed ultra-centrifuge. After collecting all proteins to eliminate possible same proteins, we estimated that there are 407 different proteins including chloroplast, chloroplast stroma, lumenal, and thylakoid membrane proteins excluding 20 proteins, which were identified in nucleus, cytoplasm and mitochondria. A combination of these three programs (PSORT, TargetP, and TMHMM) was found to provide a useful tool for evaluating chloroplast localization, transit peptide, transmembranes, and also could reveal possible alternative processing sites and dual targeting. Finally, we report also sub-cellular location specific protein interaction network using Cytoscape software, which provides further insight into the biochemical pathways of photosynthesis. The present work helps understanding photosynthesis process in wheat at the molecular level and provides a new overview of the biochemical machinery of the thylakoid in wheat.

Wheat (Triticum aestivum L.) grain texture is an important determinant of milling properties and end product use. Two linked genes, puroindoline a (PINA) and puroindoline b (PINB), control most of the genetic variation in wheat grain texture. Wheat seed proteins were examined to identify PINA and PINB gene using two pre-harvest sprouting wheat cultivars; Jinpum (resistant) and Keumgang (susceptible).Wheat seed proteins were separated by two-dimensional electrophoresis with IEF gels over pH ranges: pH 3-10. A total of 73 spots were digested with trypsin resulting peptide fragmentation were analyzed by matrix assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF/MS). Mass spectra were automatically processed and searched through NCBInr, SWISS-PORT and MSDB database with mono isotopic masses and complete gene sequence were found by UniProt database. Puroindoline a and puroindoline b that is responsible for grain texture related with baking performance and roughness. Two spots were found Pin b (16.7 kDa) and Pin a (16.3 kDa) in Jinpum compare to seven spots were identified Pin a (16.1 kDa, 16.3 kDa) and Pin b (16.7 kDa, 9.5 kDa and 14.4 kDa) in Keumgang. Some selected spots were identified puroindoline like grain softness protein (16.9 kDa, 17 kDa and 18.1 kDa) in Keumgang. Moreover, to gain a better inferring the identification of puroindoline related proteins using proteomics, we accomplished a complete gene sequence of PINA and PINB gene in pre-harvesting sprouting wheat seeds between resistant (Jinpum) and susceptible (Keumgang).