植物学技术

Protein–protein interactions play important roles in many cellular processes, including the regulation of phytohormone signaling pathways. Identification of interacting partners of key proteins involved in the cellular signaling control can provide potentially unex ...

The jasmonate (JA) ZIM-domain (JAZ) proteins of Arabidopsis thaliana repress JA signaling and negatively regulate the JA responses. Recently, JAZ proteins have been found to inhibit the transcriptional function of several transcription factors, among which the basic helix-loop- ...

Protein–protein interaction studies are crucial to unravel how jasmonate (JA) signals are transduced. Among the different techniques available, yeast two-hybrid (Y2H) is commonly used within the JA research community to identify proteins belonging to the core JA signaling module. ...

Pull-down assays are key tools to test specific protein–protein interactions and have been particularly fruitful in jasmonate signaling research. Here, we describe a standard protocol in which a matrix-bound “bait” protein, expressed in Escherichia coli, pulls down a “prey” protein t ...

Jasmonic acid (JA) is activated for signaling by its conjugation to isoleucine (Ile) through an amide linkage. The Arabidopsis thaliana JASMONIC ACID RESISTANT1 (JAR1) enzyme carries out this Mg-ATP-dependent reaction in two steps, adenylation of the free carboxyl of JA, followed by cond ...

To determine the location of specific molecules within tissues or cells, immunological techniques are frequently used. However, immunolocalization of small molecules, such as jasmonic acid (JA) and its bioactive amino acid conjugate, JA-isoleucine, requires proper fixation and ...

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 ...

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 ...

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 ...

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 ...

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 ...

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 ...

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 ...

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 ...

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 ...

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 ...

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 ...

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 ...

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 ...

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 ...