植物学技术

Non-cell-autonomous (NCA) control of plant development is an emerging field. Transcription factors (TFs) are the most important plant proteins involved in development and cell fate determination. In plants specialized intercellular symplastic channels, called plasmodes ...

Similar to the activities of transcription factors (TFs) in other eukaryotes, activities of many plant TFs are determined via regulated proteolysis by the ubiquitin/26S proteasome system. Thus, to fully understand the function of a TF, it is important to determine the fate of the active TF prot ...

Defining protein complexes is critical to virtually all aspects of cell biology because most cellular processes are regulated by stable or more dynamic protein interactions. Elucidation of the protein–protein interaction network around transcription factors is essential to f ...

Protein–protein interactions are an important aspect of the gene regulation process. The expression of a gene in response to certain stimuli, within a specific cell type or at a particular developmental stage, involves a complex network of interactions between different regulatory pr ...

Microarray analysis is widely used to identify transcriptional changes associated with genetic perturbation or signaling events. Here we describe its application in the identification of plant transcription factor target genes with emphasis on the design of suitable DNA const ...

Regulation of gene expression by transcription factors is a fundamental mechanism in essentially all aspects of cellular processes. Transient expression assay of a reporter plasmid containing a reporter gene driven by a promoter of interest and an effector plasmid expressing a tran ...

Activation tagging is a powerful strategy to find new gene functions, especially from genes that are redundant or show lethal knock-out phenotypes. It has been applied using T-DNA or transposons. En/Spm-I/dSpm engineered transposons are efficient activation tags in Arabidopsis. An im ...

The yeast one-hybrid (Y1H) system is a powerful tool for the identification and isolation of cDNAs of transcription factors using promoter segments or regulatory elements as baits. Here we propose an adaptation of the Y1H system for identification and cloning of transcription factors us ...

WRKY transcription factors are key regulators of numerous biological processes in plant growth and development, as well as plant responses to abiotic and biotic stresses. Research on biological functions of plant WRKY genes has focused in the past on model plant species or species with larg ...

All major processes of life depend on differential gene expression, which is largely controlled by the activity of transcription factors (TFs). In plants many TFs are encoded by members of multigene families that expanded much more dramatically during land plant evolution than during the ...

Physical interactions between transcription factors and specific DNA sites are essential for gene regulation. Recent progress in genome-wide in vivo techniques, like chromatin immunoprecipitation followed by high-throughput sequencing (ChIP-SEQ), enables plant rese ...

DamID (DNA adenine methylation identification) is an adenine methylation-based tagging method designed to map protein–DNA interactions in vivo. DamID, an alternative method to chromatin immunoprecipitation (ChIP), is based on the covalent linking of a “fingerprint” in the vicin ...

Unlike rational protein engineering, directed evolution provides an a priori approach toward the engineering of improved proteins and novel promoters. This minimally recursive technique builds upon small improvements by selecting and combining the best changes. Protein–pr ...

Stromules are thin stroma-filled tubules that extend from all plastid types in all multicellular plants examined. They are most easily visualised by epifluorescence or confocal microscopy of plastids containing green fluorescent protein (GFP) or other fluorescent proteins. Tr ...

Although chloroplasts have their own genome, most chloroplast proteins are encoded in the nuclear genome and are targeted to chloroplasts posttranslationally. In vitro import studies with isolated chloroplasts have been widely used and have helped to elucidate the complex mechan ...

Immunofluorescence microscopy reveals localization of proteins in cells and tissues by means of highly specific, fluorescently labeled antibodies. This technique is an important complement to localization methods that use genetically encoded fluorescent tags. This chap ...

Methods are described which allow one to observe chloroplasts in mesophyll cells from leaves of Arabidopsis, determine their number per cell, measure their area, and determine a value for chloroplast coverage inside mesophyll cells. Non-green plastids can also be imaged either by using s ...

The power of Arabidopsis as a model organism lies in the depth and breadth of genetic tools available for its study. This also applies to the study of chloroplast biology. Although vast numbers of mutants have been identified in Arabidopsis, the continued use of forward-genetic screening appro ...

The ubiquitous signaling molecule nitric oxide (NO) plays an important role in seed biology. Experiments with this biologically important gas require special provisions because NO in aerobic environments is readily converted into other oxides of nitrogen. In this chapter, we descri ...

Analysis of gene expression data sets is a potent tool for gene function prediction, cis-element discovery, and hypothesis generation for the model plant Arabidopsis thaliana, and more recently for other agriculturally relevant species. In the case of Arabidopsis thaliana, experim ...