植物生理

Soybean, the world’s most widely grown seed legume, is an important global source of vegetable oil and protein. Though, complete draft genome sequence of soybean is now available, but functional genomics studies remain in their infancy, as this agricultural legume species exhibits genet ...

In this chapter we describe the workflow used in our laboratory to analyze rice leaf samples using label-free shotgun proteomics based on SDS-PAGE fractionation of proteins. Rice proteomics has benefitted substantially from successful execution of shotgun proteomics techniqu ...

Arabidopsis is the model plant of choice for large-scale proteome analyses, because its genome is well annotated, essentially free of sequencing errors, and relatively small with little redundancy. Furthermore, most Arabidopsis organs are susceptible to standard protein solubi ...

The question of low-abundance proteins from biological tissues is still a major issue. Technologies have been devised to improve the situation and in the last few years a method based on solid-phase combinatorial peptide ligand libraries has been extensively applied to animal extracts. T ...

Proteomic technologies have become a successful tool to provide relevant information on fungal biology. In the case of plant pathogenic fungi, this approach would allow a deeper knowledge of the interaction and the biological cycle of the pathogen, as well as the identification of pathoge ...

Plant tissues contain high levels of nonprotein contaminants such as lipids, phenolic compounds, and polysaccharides among others, which interfere with protein extraction and electrophoretic separation. Preparation of good-quality protein extracts is a critical issue for ...

Functional proteomics of plant membrane proteins is an important approach to understand the comprehensive architecture of each metabolic pathway in plants. One bottleneck in the characterization of membrane proteins is the difficulty in producing sufficient quantities of fu ...

Imaging techniques based on mass spectrometry (MS) have become powerful approaches to decipher the spatial distribution of metabolites and proteins. MS imaging (MSI) mostly relies on matrix-assisted laser desorption/ionization coupled to MS detection, but desorption electr ...

The yeast two-hybrid system is currently one of the most standardized protein interaction mapping techniques. The rationale of the yeast two-hybrid system relies on the physical separation of the DNA-binding domain from the transcriptional activation domain of several transcrip ...

Moving past the discovery phase of proteomics, the term targeted proteomics combines multiple approaches investigating a certain set of proteins in more detail. One such targeted proteomics approach is the combination of liquid chromatography and selected or multiple reaction mo ...

The Mass Western describes the absolute quantification of proteins based on stable isotope labeled integral standard peptides and liquid chromatography coupled selective reaction monitoring triple quadrupole mass spectrometry (LC-SRM/MS). Here, we present a detailed wor ...

Oil bodies, lipid-storage organelles, are stabilized by a number of specific proteins. These proteins are very hydrophobic, which complicates their identification by “classical” proteomic protocols using trypsin digestion. Due to the lack of trypsin cleavage sites, the achievab ...

As the field of proteomics shifts from qualitative identification of protein “subfractions” to quantitative comparison of proteins from complex biological samples, it is apparent that the number of approaches for quantitation can be daunting for the result-oriented biologist. T ...

Protein relative quantitation is one of the main targets in many proteomic experiments. Among the range of techniques available for both top-down and bottom-up approaches, isobaric tags for relative and absolute quantitation (iTRAQ) have gained positions within the top-rank techni ...

Although proteomists working with gel-free methods are considering the gels as coming from the past, proteomics based on gels has still a lot of opportunities to offer and acquisition of images on which thousands of spots may be resolved is still largely performed. Nowadays, two-dimensional ...

The use of analytical biochemical techniques with different separation properties allows us to better understand the proteome. To demonstrate this we have used two different methodologies to analyze embryos from a Tunisian cultivar of durum wheat (Triticum durum Desf.), variety Oum R ...

Mass spectrometry (MS) is an analytical technique that measures the mass-to-charge ratio of electrically charged gas-phase particles. All mass spectrometers combine ion formation, mass analysis, and ion detection. Although mass analyzers can be regarded as sophisticated devic ...

Two-dimensional gel electrophoresis remains the most widely used technique for protein separation in plant proteomics experiments. Despite the continuous technical advances and improvements in current 2-DE protocols, an adequate and correct experimental design and stati ...

Two-dimensional electrophoresis has nurtured the birth of proteomics. It is however no longer the exclusive setup used in proteomics, with the development of shotgun proteomics techniques that appear more fancy and fashionable nowadays. Nevertheless, 2D gel-based proteomics st ...

Proteomic approaches are useful for the identification of functional proteins. These have been enhanced not only by the development of proteomic techniques but also in concert with genome sequencing. In this chapter, 30 databases and Web sites relating to plant proteomics are reviewed a ...