植物学技术

Root hairs are tubular extensions from the root surface that expand by tip growth. This highly focused type of cell expansion, combined with position of root hairs on the surface of the root, makes them ideal cells for microscopic observation. This chapter describes the method that is routinely us ...

Plant organs and tissues consist of many various cell types, often in different phases of their development. Such complex structures do not allow direct studies on behavior of individual cells. In contrast, populations of in vitro-cultured plant cells represent valuable tool for studyi ...

Specific gene knockdown mediated by the antisense oligodeoxynucleotides (AODNs) strategy recently emerged as a rapid and effective tool for probing gene role in plant cells, particularly tip-growing pollen tubes. Here, we describe the protocol for the successful employment of AODN ...

Different plant cell types express unique transcriptomes, proteomes, and metabolomes. Therefore, the isolation of specific cell types prior to molecular analyses is important to understand the specification, differentiation, and function of these cells. Isolation of specif ...

A major limitation in the study of pollen tube growth has been the difficulty in providing an in vitro testing microenvironment that physically resembles the in vivo conditions. Here we describe the development of a lab-on-a-chip (LOC) for the manipulation and experimental testing of indiv ...

Optical tweezers allow noninvasive manipulation of subcellular compartments to study their physical interactions and attachments. By measuring (delay of) displacements, (semi-)quantitative force measurements within a living cell can be performed. In this chapter, we prov ...

Histochemical methods allow for identification and localization of various components within the tissue. Such information on the spatial heterogeneity is not available with biochemical methods. However, there is limitation of the specificity of such detection in context of com ...

The budding yeast (Saccharomyces cerevisiae) can serve as a unique experimental system for functional studies of heterologous genes, allowing not only complementation of readily available yeast mutations but also generation of overexpression phenotypes and in some cases also r ...

Plant cell walls are structurally diverse macromolecular composites. One of our best methodologies to determine the temporal and spatial regulation of cell wall polysaccharides in relation to development are monoclonal antibody (MAB) and carbohydrate-binding module (CBM) pr ...

This chapter gives examples of basic procedures of quantification of plant structures with the use of image analysis, which are commonly employed to describe differences among experimental treatments or phenotypes of plant material. Tasks are demonstrated with the use of ImageJ, a wid ...

Tightly controlled spatiotemporal specificity of gene expression is intrinsic to developmental and adaptation responses of living systems throughout the kingdoms. Forward genetic screens employing well-characterized reporter lines can be used to identify as yet unknown g ...

Most biochemical functions of plant cells are carried out by proteins which act at very specific places within these cells, for example, within different organelles. Identifying the subcellular localization of proteins is therefore a useful tool to narrow down the possible functions t ...

The cortical microtubules, and to some extent also the actin meshwork, play a central role in the shaping of plant cells. Transgenic plants expressing fluorescent protein markers specifically tagging the two main cytoskeletal systems are available, allowing noninvasive in vivo stud ...

Sequential replica method facilitates in vivo imaging of plant surface and provides data sufficient for detailed computation of geometry and growth. It enables obtaining a series of high-resolution images visualizing details of the examined surface. Series of molds, made in dental po ...

Analysis of shoot meristem shape and gene expression pattern has been conducted in many species over the past decades. Recent live imaging techniques have allowed an unprecedented accumulation of data on the biology of meristematic cells, as well as a better understanding of the molecular a ...

During the last 10 years, the Proteomics Standards Initiative from the Human Proteome Organization (HUPO-PSI) has worked on defining standards for proteomics data representation as well as guidelines that state the minimum information that should be included when reporting a prote ...

Numerous methods have been employed to depict the protein content of wines. Among them, two-dimensional electrophoresis (2D-E) presents a powerful resolution, but has been poorly applied to wine. Furthermore, 2D-E was coupled with various extraction methods of proteins without any re ...

Proteome analysis provides a way to identify proteins related to the quality traits of beer. A number of protein species in beer and wort have been identified by two-dimensional gel electrophoresis combined with enzyme digestion such as trypsin, followed by mass spectrometry analyses and ...

Immunoblot-coupled proteomics based on two-dimensional polyacrylamide gel electrophoresis (2D-PAGE), namely, immunoproteomics, has been used for comprehensive identification of food allergens, because it is a simple and inexpensive tool for rapid identification of sev ...

Proteomics has become a powerful tool to characterize biodiversity and natural variability in plant species, as well as to catalogue and establish phylogenetic relationships and distances among populations, provenances or ecotypes. In this chapter, we describe the standard prot ...