植物学技术

By insertion into an individual xylem vessel at the root base, the multifunctional xylem probe allows the monitoring of the xylem pressure, the radial electrical gradients in the root (the so-called trans-root potential, TRP), as well as the activity of a particular ion such as K+ in the xylem sap of inta ...

Voltage clamp is one of the key techniques for the dissection, identification, and monitoring of ion transporters in plant cells. Voltage clamp-based research work on salinity stress in plants enables the characterization of many plant ATP-dependent pumps, ion channels, and ion-coupl ...

The patch-clamp technique was designed to measure any electrogenic transport across the whole cell and organelle (vacuolar) membranes and excised membrane patches. Here, we describe preparation of protoplasts and vacuoles, as well as patch-clamp assays, to detect the functional ex ...

Both nitric oxide (NO) and reactive oxygen species (ROS) are versatile molecules that mediate a variety of cellular responses in plants. In this chapter, methods for imaging NO and ROS using laser scanning confocal microscopy (LSCM) are presented. Arabidopsis roots, dyed with DAF-FM or H2DCF, ...

Purification of plasma membranes by two-phase partitioning is based on the separation of microsomal membranes, dependent on their surface hydrophobicity. Here we explain the purification of plasma membranes from a relatively small amount of material (7–30 g). The fluorescent probe A ...

Soil salinity devastates agriculture. It reduces crop yields and makes arable land unsuitable for later use. Many species have evolved highly efficient strategies to sense, transduce, and build up tolerance to high salinity and even sensitive species have endogenous mechanism for cop ...

Plant response to salt stress involves changes in intracellular ion concentrations that can be estimated by ion-selective fluorescent dyes. Conventional confocal/fluorescent imaging does not always accurately reflect the process, since it is dependent not only on the ion concen ...

Advances in the techniques required for the X-ray microanalysis of cryo-fixed, naturally hydrated plant tissues in the cryo-scanning electron microscope have reached the stage that accurate, cell-specific localization and quantification of the nutrient and toxic elements can be ...

Reactive oxygen species (ROS) participate in signaling events that regulate ion channel activity and gene expression. However, excess ROS exert adverse effects that stem from their interaction with macromolecules. Thus, the assessment of the effects of salinity on ROS changes are cen ...

Biochemical methods available for the measurement of antioxidant activity in salt-stressed tissues are reviewed, outlining the most important advantages and shortcomings of the methods. Here we consider commonly used methods for measuring total antioxidant capacity and phe ...

Plant hormones cytokinins, auxin (indole-3-acetic acid), and abscisic acid are central to regulation of plant growth and defence to abiotic stresses such as salinity. Quantification of the hormone levels and determination of their ratios can reveal different plant strategies to cope w ...

Fluorescence-activated cell sorting (FACS) provides a rapid means of isolating large numbers of fluorescently tagged cells from a heterogeneous mixture of cells. Collections of transgenic plants with cell type-specific expression of fluorescent marker genes such as green fluo ...

The chapter describes the main light microscopy techniques used for the diagnosis of phytoplasmas. Because the described techniques are generally only effective in cases where the concentration of the pathogen inside the host phloem tissue is high, they are useful as preliminary metho ...

The correct identification of the insect species involved in phytoplasma transmission is an essential condition for managing phytoplasma diseases and employing control strategies. The taxonomy of leafhoppers, planthoppers, and psyllids traditionally relies on morphol ...

Phytoplasmas are transmitted in a persistent propagative manner by phloem-feeding vectors belonging to the order Hemiptera, suborder Homoptera. Following acquisition from the infected source plant, there is a latent period before the vector can transmit, so transmission assays ...

Phytoplasmas are plant pathogens of huge economic importance due to responsibility for crop yield losses worldwide. Institutions around the world are trying to understand and control this yield loss at a time when food security is high on government agendas. In order to fully understand the m ...

Insect vectors of phytoplasmas are limited to leafhoppers, planthoppers, and psyllids. While populations can be monitored by a number of passive techniques in the field, the capture of live insects is necessary for manipulation and study. A number of physical methods for capturing these in ...

Transmission of phytoplasmas from naturally infected plant host species using the parasitic plant Cuscuta spp. (dodder) to Catharanthus roseus (Madagascar periwinkle) is an effective way to maintain a wide range of phytoplasmas for further research. Here, we describe transmissi ...

Maintenance of phytoplasma strains in tissue culture is achievable for all strains transmitted to periwinkle (Catharanthus roseus), and also for other naturally infected plant host species. Shoots of 1–3 cm length are grown in a solid medium containing Murashige and Skoog (MS) micro- and ma ...

Physical and genetic mapping of the phytoplasma chromosome can be a useful tool in a genome sequencing project in order to assemble the in silico-predicted contigs robustly. Mapping consists of four distinct steps: preparation of phytoplasma chromosomes from infected plants, single- ...