植物生理

The phenotypic response of plants to high external levels of NaCl is very dynamic so the methods used for phenotyping should be able to capture those dynamics. Non-destructive high-throughput plant imaging enables researchers to measure salt stress-induced changes in plant growth over ...

Radioisotopes (particularly 22Na, 24Na, 42K, and 86Rb) have been used for many decades to trace the fluxes and accumulation of sodium and potassium ions in plant tissues. In this article, standard procedures for the tracing of ion fluxes are described, with emphasis on special problems encoun ...

The expression of plant transporters in simple well-characterized cell systems is an irreplaceable technique for gaining insights into the kinetic and energetic features of plant transporters. Among all the available expression systems, yeast cells offer the highest simplici ...

Plant vacuoles have multifaceted roles including turgor maintenance, cytosolic pH and ionic homeostasis, plant protection against environmental stress, detoxification, pigmentation, and cellular signaling. These roles are achieved through the coordinated activiti ...

Abiotic stresses such as drought, salinity, and extreme temperatures are major limiting factors in plant growth and development and pose serious threat to global agricultural production. Here we describe a procedure, using a tobacco plastid transformation vector, to generate tran ...

Interrogating the cell-specific transcriptome forms an important component of understanding the role that specific cells play in assisting a plant to overcome abiotic stress. Among the challenges arising when extracting RNA from individual plant cells are: the isolation of pure ce ...

Marker-assisted selection (MAS) is the process of using morphological, biochemical, or DNA markers as indirect selection criteria for selecting agriculturally important traits in crop breeding. This process is used to improve the effectiveness or efficiency of selection for the t ...

Exposure to high ambient levels of NaCl affects plant water relations and creates ionic stress. To a large extent, responses to such stress depend on the action of membrane transporters, particularly those that move cations such as Na+ and K+. A genomics approach can greatly help with the identific ...

The recent advances made in the use of infrared thermal imaging (thermography) as a non-invasive, high-throughput technique for the screening of salinity tolerance in plants is reviewed. Taking wheat seedlings as an example, the methods and protocols used to impose a homogeneous salt stre ...

The controlled cDNA overexpression system (COS) was developed to identify novel regulatory genes in model plants as well as in other species that might have a particular valuable trait. The COS system (Papdi et al. Plant Physiol 147:528–542, 2008) is composed of a random cDNA library prepared in a T-D ...

Salinity and oxidative stress can transiently elevate cytosolic free Ca2+ (cyt) of plant cells. The cyt increase may be part of a signaling cascade or cell death, depending on cell type, the magnitude and the duration of stress exposure. Several approaches for determining cyt responses are avai ...

Knowledge of the fluxes of ions and neutral molecules across the outer membrane or boundary of living tissues and cells is an important strand of applied molecular biology. Such fluxes can be measured non-invasively with good resolution in time and space. Two systems (MIFE™ and SIET) have been dev ...

Single-cell sampling and analysis allows the determination of solute concentrations in individual cells and tissues. This is particularly important when studying a stress such as salinity, where the cell- and tissue-specific distribution of sodium and chloride may decide a plant’s f ...

Ion-selective microelectrodes can be used to measure intracellular ion concentrations. The use of multi-barrelled electrodes enables the identification of the cellular compartment. For example, the inclusion of a pH-selective electrode enables the cytoplasm and vacuole to be d ...

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 ...