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A Non-targeted Approach for Extended Liquid Chromatography-Mass Spectrometry Profiling of Free and Esterified Jasmonates After Wounding

Upon wounding or herbivory, plants quickly react by activating various defense mechanisms. A major part of these defenses is thought to be regulated by the jasmonate pathway through the induction of jasmonic acid and its biologically active jasmonoyl-isoleucine conjugate. Yet, these ...

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Jasmonate Signaling in the Field, Part II: Insect-Guided Characterization of Genetic Variations in Jasmonate-Dependent Defenses of Transgenic and Natu

The introduction of genetically modified plants into natural habitats represents a valuable means to determine organismic level functions of a gene and its effects on a plant’s interaction with other organisms. Nicotiana attenuata, a wild tobacco species native of the southwestern U ...

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Jasmonate Signaling in the Field, Part I: Elicited Changes in Jasmonate Pools of Transgenic Nicotiana attenuata Populations

Nicotiana attenuata, a wild tobacco species native of the southwestern USA that grows in the immediate postfire environment, is one of the important host plants for herbivore populations recolonizing recently burned habitats in the Great Basin Desert. Based on more than 20 years of field re ...

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Pseudomonas syringae Infection Assays in Arabidopsis

Pseudomonas syringae pv. tomato DC30000 (Pst DC3000) infection of Arabidopsis thaliana has been widely used to elucidate many of the general principles underlying the plant immune response and bacterial pathogenesis. Study of Pst DC3000 virulence factors has also proven useful in the ...

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Elicitation of Jasmonate-Mediated Defense Responses by Mechanical Wounding and Insect Herbivory

Many plant immune responses to biotic stress are mediated by the wound hormone jasmonate (JA). Functional and mechanistic studies of the JA signaling pathway often involve plant manipulations that elicit JA production and subsequent changes in gene expression in local and systemic tis ...

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Bioassays for Assessing Jasmonate-Dependent Defenses Triggered by Pathogens, Herbivorous Insects, or Beneficial Rhizobacteria

Jasmonates, together with other plant hormones, are important orchestrators of the plant immune system. The different hormone-controlled signaling pathways cross-communicate in an antagonistic or a synergistic manner, providing the plant with a powerful capacity to finely re ...

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Phenotyping Jasmonate Regulation of Root Growth

Root architecture is a complex and highly plastic feature of higher plants. Direct treatments with jasmonates and alterations in jasmonate signaling have been shown to elicit a range of root phenotypes. Here, we describe a fast, noninvasive, and semiautomatic method to monitor root archi ...

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Characterizing Jasmonate Regulation of Male Fertility in Arabidopsis

Coordination of events leading to fertilization of Arabidopsis flowers is tightly regulated, with an essential developmental cue from jasmonates (JAs). JAs coordinate stamen filament elongation, anther dehiscence, and pollen viability at stage 12 of flower development, the sta ...

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Phenotyping Jasmonate Regulation of Senescence

Osmotic stress induces several senescence-like processes in leaves, such as specific changes in gene expression and yellowing. These processes are dependent on the accumulation of jasmonates and on intact jasmonate signaling. This chapter describes the treatment of Arabidopsis ...

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Identifying Meiotic Mutants in Arabidopsis thaliana

Arabidopsis is a very powerful tool for understanding meiosis in plants with genetic approaches. We provide here a simple summary of the techniques used to test if a candidate gene has an essential meiotic function. These protocols require no specific prior knowledge and help eliminate easi ...

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Analysis of Meiotic Protein Complexes from Arabidopsis and Brassica Using Affinity-Based Proteomics

The application of proteomics techniques to the study of plant meiosis has the potential to make a valuable contribution to our understanding of the molecular events underpinning meiotic processes. Here we describe the preparation of meiotic protein complexes from Arabidopsis tha ...

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Analyzing the Meiotic Transcriptome Using Isolated Meiocytes of Arabidopsis thaliana

Improved transcriptome sequencing technologies (RNA-seq) have advanced our understanding of the tissue-specific transcriptome landscapes, including those of messenger RNAs, noncoding RNAs and small RNAs. However, transcriptome profiles of plant meiocytes remain cha ...

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Chromatin Immunoprecipitation for Studying Chromosomal Localization of Meiotic Proteins in Maize

Chromatin immunoprecipitation (ChIP) is a method that allows identification of chromosomal sites occupied by specific proteins. In this technique, chromatin is extracted from cells, sheared, and, using a specific antibody, enriched in fragments that contain a protein of interest. G ...

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Characterization of Meiotic Non-crossover Molecules from Arabidopsis thaliana Pollen

Meiotic recombination is essential for proper segregation of homologous chromosomes and thus for formation of viable gametes. Recombination generates either crossovers (COs), which are reciprocal exchanges between chromosome segments, or gene conversion not associated w ...

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Preparing Thin Sections of Meiotic Nuclei for Transmission Electron Microscopy

Transmission electron microscopy (TEM) of thin sections of meiocytes embedded in plastic resin allows analysis of structures in meiotic nuclei, such as the synaptonemal complex, at very high resolution while preserving the spatial organization of chromosomes.

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Analysis of the Synaptonemal Complex in Brassica Using TEM

Much of meiosis research is focussed on Arabidopsis thaliana, largely due to the significant advantages it brings, having a small sequenced genome with comparatively little repetitive DNA, the ease of forward and reverse genetics, and a short life cycle. On the other hand, due the small genome si ...

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Preparing SC Spreads with RNs for EM Analysis

Recombination nodules (RNs) are associated with synaptonemal complexes (SCs) during early prophase I of meiosis. RNs are too small to be resolved by light microscopy and can be observed directly only by electron microscopy. The patterns of RNs on SCs can be analyzed using three-dimensional r ...

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Immunolocalization of Meiotic Proteins in Brassicaceae: Method 1

Plant meiosis studies have enjoyed a fantastic boom in recent years with the use of Arabidopsis thaliana as an important model species for developmental studies because of its small genome, short life cycle, and large mutant collections. Unlike other eukaryotic models, plant meiosis does n ...

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Live Imaging of Chromosome Dynamics

Progression of meiosis has been traditionally reconstructed from microscopic images collected from fixed cells. However, studies conducted in a number of species, including plants, indicate that this approach has clear shortcomings in accurately portraying the dynamic nature ...

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Analyzing Maize Meiotic Chromosomes with Super-Resolution Structured Illumination Microscopy

The success of meiosis depends on intricate coordination of a series of unique cellular processes to ensure proper chromosome segregation. Many proteins involved in these cellular events are directly or indirectly associated with chromosomes, especially those required for homo ...

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