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Utilizing Bimolecular Fluorescence Complementation (BiFC) to Assay ProteinProtein Interaction in Plants

Protein function is often mediated by the formation of stable or transient complexes. Here we present a method for testing protein–protein interactions in plants designated bimolecular fluorescence complementation (BiFC). The advantages of BiFC are its simplicity, reliabili ...

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Laser Microdissection of Paraffin-Embedded Plant Tissues for Transcript Profiling

High-resolution cellular analysis will help answer many important questions in plant biology including how genetic information is differentially used to enable the formation and development of the plant body. By comparing transcriptome data from distinct cell types during vari ...

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Fluorescence-Activated Cell Sorting in Plant Developmental Biology

Understanding the development of an organ requires knowledge of gene, protein, and metabolite expression in the specific cell types and tissues that comprise the organ. Fluorescence-activated cell sorting (FACS) is an efficient method to isolate specific cells of interest, and the inf ...

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Luciferase and Green Fluorescent Protein Reporter Genes as Tools to Determine Protein Abundance and Intracellular Dynamics

To get insight into molecular mechanisms governing plant development, the dynamics of abundance and cellular localisation of signalling components need to be understood. Luciferase and green fluorescent protein (GFP)-derived reporters are suitable markers to determine dyn ...

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Growth Protocols for Model Plants in Developmental Biology

Arabidopsis is the dominating model species for plant developmental biology, but other species serve as models for processes that cannot be studied in Arabidopsis, such as compound leaf or wood formation, or to test the universality of developmental mechanisms initially identified in ...

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Quantitative Real Time PCR in Plant Developmental Biology

Gene expression patterns are important determinants of a cell’s state, and changes in the expression profile indicate adaptation processes as a response to developmental transitions or environmental changes. Assaying gene expression can, therefore, help to elucidate mechani ...

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Detection of Small Non-coding RNAs

Gene expression is regulated at several levels in plants, and one of the most recently discovered regulatory layers involve short RNAs. Short RNAs are produced through several pathways and target either mRNAs or genomic DNA. Different classes of short RNAs have slightly different sizes and ...

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Immunolocalization of Proteins in Plants

Rapid advances in the field of plant biology, especially in plant cell biology, have created the need for methods that allow the localization of proteins in situ at subcellular resolution. Although in many cases recombinant proteins with fluorescent proteins can fulfill this task, antibo ...

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mRNA Detection by Whole Mount In Situ Hybridization (WISH) or Sectioned Tissue In Situ Hybridization (SISH) in Arabidopsis

Gene expression can be analyzed at high spatial resolution via RNA in situ detection methods. For many tissues and species, these will be performed on sections of embedded and fixed plant material. When very small or fragile tissues, such as embryos or roots are being investigated, whole mount met ...

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Flowering Time Control

A dramatic change in the life cycle of plants is the transition to flowering, which is triggered by both environmental signals, such as temperature and photoperiod, and endogenous stimuli. The dicotyledonous annual plant Arabidopsis thaliana is widely used as a model organism to study how th ...

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Kinematic Analysis of Cell Division and Expansion

Plant growth is readily analysed at the macroscopic level by measuring size and/or mass. Although it is commonly known that the rate of growth is determined by cell division and subsequent cell expansion, relatively few studies describing growth phenotypes include studies of the dynamics ...

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Phenotypic Characterization of Photomorphogenic Responses During Plant Development

Light is one of the most important exogenous factors regulating plant development throughout the entire life cycle. Light is involved in the breaking of seed dormancy, the regulation of photomorphogenic seedling development, the adaptation of plant morphology toward spectral com ...

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Analysis of Root Meristem Size Development

Plant post-embryonic development takes place in the meristems. In the root of the model plant Arabidopsis thaliana, stem cells organized in a stem-cell niche in the apex of the root meristem generate transit-amplifying cells, which undergo additional division in the proximal meristem and ...

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Grafting as a Research Tool

Grafting as a means to connect different plant tissues has been enormously useful in many studies of long-distance signalling and transport in relation to regulation of development and physiology. There is an almost infinite number of pairwise graft combinations that can be tested, typi ...

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Virus-Induced Gene Silencing as a Reverse Genetics Tool to Study Gene Function

Reverse genetics has proven to be a powerful approach to elucidating gene function in plants, particularly in Arabidopsis. Virus-induced gene silencing (VIGS) is one such method and achieves reductions in target gene expression as the vector moves into newly formed tissues of inoculated ...

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The CRE/lox System as a Tool for Developmental Studies at the Cell and Tissue Level

Targeted gene manipulation has been used in the last few decades for better understanding of gene function. Most often mutant or overexpression genotypes are analyzed, but in many cases these are not sufficient to obtain a detailed picture on the mode of action of the corresponding protein. For e ...

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Inducible Gene Expression Systems for Plants

Several systems for induction of transgene expression in plants have been described recently. Inducible systems were used mainly in tobacco, rice, Arabidopsis, tomato, and maize. Inducible systems offer researchers the possibility to deregulate gene expression levels at partic ...

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Trichome Development in Arabidopsis

Arabidopsis trichomes are giant single epidermal cells that are easily accessible for genetic, genomic and cell-biological analysis. They have therefore become a convenient model system to study developmental and physiological processes. Trichome studies are greatly facil ...

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Phenotyping the Development of Leaf Area in Arabidopsis thaliana

The study of leaf expansion began decades ago and has covered the comparison of a wide range of species, genotypes of a same species and environmental conditions or treatments. This has given rise to a large number of potential protocols for today’s leaf development biologists. The final size of the l ...

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Analyzing Shoot Apical Meristem Development

The shoot apical meristem of Arabidopsis thaliana contains a reservoir of pluripotent stem cells that functions as a continuous source of new cells for organ formation during development. The SAM forms during embryogenesis, when it becomes stratified into specific cell layers and zones ...

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