植物学技术

Lipid modifications play a key role in protein targeting and function. The two Arabidopsis Gγ subunits, AGG1 and AGG2, have been shown to undergo prenylation (AGG1) and S-acylation (AGG2). Prenylation involves covalent nonreversible attachment of either farnesyl (15 carbons) or geran ...

Myristoylation is a lipid modification conserved among eukaryotes and involves the addition of a 14-carbon myristoyl moiety to a glycine at the N-terminus of cargo proteins. Since not every protein with an N-terminal glycine is myristoylated, experimental verification is necessary ...

S-acylation is increasingly being recognized as an important posttranslational modification of proteins controlling activity, subcellular localization, microdomain residence, and stability. Heterotrimeric G-proteins and GPCRs are particularly well studied S-a ...

Protein prenylation, like other lipid posttranslational modifications of eukaryotic proteins, plays important roles in protein–membrane association and protein–protein interactions. In Arabidopsis, hundreds of proteins involved in a great variety of biological proc ...

Heterotrimeric G-proteins are important signaling intermediates in all eukaryotes. These proteins link signal perception by a cell surface localized receptor to the downstream effectors of a given signaling pathways. The minimal core of the heterotrimeric G-protein complex co ...

Genomic sequencing has provided a vast resource for identifying interesting genes, but often an exact “gene-of-interest” is unknown and is only described as putatively present in a genome by an observed phenotype, or by the known presence of a conserved signaling cascade, such as that facilit ...

The heterotrimeric G-proteins form classical signal transduction complexes conserved in all eukaryotes. The repertoire of G-protein signaling complex is much simpler in plants than in metazoans. One of the best understood functions for the plant G-protein complex is its modulation ...

Phospholipase Ds (PLDs) play diverse roles in plant lipid metabolism and cellular signaling processes. The sole canonical G-protein α-subunit (Gα) in Arabidopsis also plays multiple roles in plant growth and cellular signaling processes. Interestingly, overlapping functions ...

Heterotrimeric GTP-binding proteins (G-proteins) and G-protein-coupled receptors are important signaling components in eukaryotes. In plants, the G-proteins are involved in diverse physiological processes, some of which are exerted via changes in the level of cytosolic free c ...

The unfolded protein response (UPR) is fundamental for development and adaption in eukaryotic cells. Arabidopsis has become one of the best model systems to uncover conserved mechanisms of the UPR in multicellular eukaryotes as well as organism-specific regulation of the UPR in plants. M ...

Increased susceptibility to Fusarium oxysporum is one of the most conspicuous characteristics of the Arabidopsis mutants lacking the heterotrimeric G protein β and γ1 subunits. The molecular mechanisms placing these G proteins in the plant innate immunity network are yet to be discov ...

Functional analysis of G-proteins has been extensively carried out using their over-expressing lines and knockout mutants in plants. Since α subunit exists in an active or inactive form, overexpressing α subunit does not mean that G-protein signaling pathways are activated in the trans ...

There are various preparatory techniques for light microscopy permitting access to the inner structure of plant body and its development. Minute objects might be processed as whole-mount preparations, while voluminous ones should be separated into smaller pieces. Hereby we summar ...

Confocal microscopy is a technique widely used to live-image plant tissue. Cells can be visualized by using fluorescent probes that mark the cell wall or plasma membrane. This enables the confocal microscope to be used as a 3D scanner with submicron precision. Here we present a protocol using the 3D i ...

Use of electron tomography methods improves image resolution of transmission electron microscopy especially in the z-direction, enabling determination of complicated 3D structures of organelles and cytoskeleton arrays. The increase in resolution necessitates preser ...

Indentation methods on the micro- and nanoscale are increasingly used to assess mechanical properties of living plant tissues. These techniques rely on recording the force resulting from indenting the cell surface with a small probe. Depending on the scale of indentation and the indenter ...

In micrographs acquired with a transmission electron microscope, 3-dimensional (3D) objects are superimposed onto a 2D screen. This reduction in dimension necessarily leads to a degradation of image resolution. To overcome this problem, 3D microscopy techniques, such as tomograp ...

Scanning electron microscopy (SEM) is a powerful technique that can image exposed surfaces in 3D. Modern scanning electron microscopes, with field emission electron sources and in-lens specimen chambers, achieve resolutions of better than 0.5 nm and thus offer views of ultrastructur ...

Knowledge about the spatio-temporal distribution patterns of proteins and other molecules of the cell is essential for understanding their function. A widely used technique is immunolabeling which uses specific antibodies to reveal the distribution of molecular components at v ...

This protocol describes a quantitative analysis of the morphology of small plants from the moss Physcomitrella patens. The protocol can be used for the analysis of growth phenotypes produced by transient RNA interference or for the analysis of stable mutant plants. Information is presen ...