结构与功能

Oocytes of the South African clawed toad, Xenopus laevis, have proved a versatile and powerful heterologous expression system for eukaryotic genes (1). Xenopus oocytes were first used to express plant genetic material in 1979 (2), and in the past decade it has become increasingly evident that t ...

In Chapter 28, a method for the isolation of intact chloroplasts was described. The availability of this technique has allowed researchers the opportunity to investigate the mechanisms of chloroplast biogenesis. The chloroplast is a complex organelle, being made up of six subcompartm ...

The isolation of intact chloroplasts from plant tissue allows the study of processes carried out in this subcellular compartment. Such processes may be enzymatic, e.g., photosynthetic oxygen-evolution, or mechanistic, like the import and targeting of precursor proteins synthesi ...

The chloroplast is the product of two genomes with the majority of its polypeptide complement encoded by nuclear DNA and synthesized on cytosolic ribosomes. Proteins destined for the chloroplast must therefore contain targeting information to allow their correct localization. It h ...

During the last years the structure and gene content of the mitochondrial genome of land plants have been subject of intensive molecular investigations. Because of the large size (200–2500 kb), and the complex, multipartite organization, the actual configuration of the mitochondrial g ...

Protein synthesis by isolated plant mitochondria under in vitro conditions is a direct method to study the translational products of this organellar genetic system. Compared to fungal and animal mitochondrial systems, a considerably higher number of proteins is expected to be encoded ...

Direct identification of the product of a transferred gene is clearly an important part of the characterization of a transgenic plant. It is possible to detect proteins using highly sensitive ELISA techniques (Chapter 36, this volume), and to determine their precise histological or subce ...

The immunodetection of proteins bound to a membrane has widespread applications in plant biochemistry and molecular biology, including the identification and semiquantitative determination of foreign proteins expressed in transgenic plants. The approach is usually appl ...

Immunoassays were first developed over 30 yr ago. The radioimmunoassay for insulin described by Yalow and Berson (1) heralded a new era in the use of antibody reagents for the quantification of proteins and peptides. The Nobel Prize-winning research revolutionized analysis by virtue of much ...

The localization of proteins within plant tissues is readily accomplished using the techniques of immunocytochemistry: the identification of a cell-bound antigen in situ by means of a specific antigen-antibody reaction, tagged microscopically by a visible label (1–3).

Electroporation is now a standard method of transfection and cell loading. There is a variety of commercial electroporation equipment, and many published and manufacturer-supplied protocols. Many of these protocols are results of trial and error. These empirical protocols are val ...

Direct DNA transformation is currently the method of choice for obtaining transformed cereals, such as Zea mays, owing to the inherent host limitations of Agrobacterium tumefaciens. This soil bacterium, which naturally acts by transferring its own genes into a host plant, is commonly eng ...

The techniques of electroporation and electrofusion require that cells be subjected to brief pulses of electric fields of the appropriate amplitude, duration, and wave form. In this chapter, the term electro cell manipulation (ECM) shall describe both techniques. ECM is a quite universal ...

The introduction of DNA into plant cells, protoplasts, or intact tissues has been accomplished by a variety of mechanisms, including electroporation, electrofusion, particle bombardment, liposome transfer, the use of bacterial vectors, polyethylene glycol treatment, and oth ...

Agrobacterium tumefaciens is a soil bacterium that causes tumors on dicotyledonous plants. Virulent strains harbor a large plasmid, the Ti (tumor-inducing) plasmid, which is involved in tumorigenesis. A small segment of this plasmid, the T-DNA, is transferred to the plant cell and becomes ...

Molecular characterization of complex cellular processes depends on the use of model systems in which a classical genetic approach can be combined with molecular techniques of gene cloning and DNA transfer. Several features of the green alga Chlamydomonas reinhardtii make it a suitab ...

Micropropagation techniques are being used by an increasing number of research workers and commercial firms. The main use has been that of mass production of plants ranging from nursery stock species (such as rhododendron or rose), through ornamentals (such as fuchsia or carnation) to frui ...

The essence of meristem-tip culture is the excision of the organized apex of the shoot from a selected donor plant for subsequent in vitro culture. The conditions of culture are regulated to allow only for organized outgrowth of the apex directly into a shoot, without the intervention of any advent ...

Regeneration of buds, shoots, and roots in cells and explants in vitro has provided useful developmental systems to analyze the processes of cell differentiation and morphogenesis. Interest in these studies was greatly stimulated by the demonstration of chemical regulation of morp ...

Plant cells can be totipotent, i.e., each cell may be capable of developing into an entire plant when provided with the correct environmental stimuli. Research during the last 30 yr has demonstrated that successful organogenesis in callus cultures can be achieved by the correct choice of medium ...