植物生理

Enzyme-linked immunosorbent assay (ELISA), since its use with plants was described by Clark and Adams (1), has become the most popular method for detection of viruses in plants, because of its simplicity and wide applicability. Used mainly to confirm presence or absence of infection, it can be ada ...

Western blotting combines the resolving power of protein electrophoresis and the specificity of immunology in a rapid and sensitive format for the identification of proteins in complex mixtures. Proteins resolved by electrophoresis are transferred to a solid support, which is norm ...

Southern analysis (1) is routinely carried out to determine whether a plant regenerated from tissue-culture has been transformed with foreign DNA. The technique of Southern analysis begins with the extraction of genomic DNA from the plant, digestion of the DNA with diagnostic-restric ...

The detection of RNA transcript expressed from transgenes is often one of the first steps in the analysis of transgenic plants. Such analysis might include confirmation that RNA transcripts are of the expected size and the quantification of transcript in different transgenic lines. Until ...

Although the display of antibiotic resistance can be a good indication that a regenerated plant is transformed with introduced DNA, escapes that represent partial or no transfer may be present within the population of primary transformants. The polymerase chain reaction (PCR) provid ...

Geminiviruses, named for their unique geminate capsid morphology, have small single-stranded (ss) circular DNA genomes that replicate in the nuclei of infected cells via a double stranded (ds) DNA intermediate. They are responsible for economically devastating diseases in a wide var ...

It makes little sense to invest time and effort in assaying biological activities of transgenic plants if the plants do not contain a functional gene of interest. Although this appears to be self-evident, many investigators attempt to use biological assays for the presence of the gene of intere ...

Oryza sativa L. has three subspecies: indica, japonica, and javanica. Subsp. indica grows well in southern temperate and tropical regions; subsp. japonica grows well in cooler climates such as Japan; and subsp. javanica is grown mostly in the Americas and Europe. Several viruses infect rice, cau ...

Genetic transformation of plants is a technique by which foreign DNA is introduced into plant cells, leading to regeneration of transgenic plants, with new features resulting from integration and expression of the foreign DNA. This technique has become an indispensable tool, both for pla ...

The generation of genetically transformed plants is central to, and has indeed revolutionized, plant molecular biology. This is true for studies at both the fundamental and more applied levels of research. For researchers interested in unraveling the roles of specific genes in particul ...

Since 1986, when Beachy and coworkers (1) first published a protection of transgenic tobacco plants expressing the tobacco mosaic virus (TMV) coat protein (CP) against TMV infection, transgenic plants have become an important tool in plant virology. Coat protein transgenic plants have b ...

Potato is the world’s fourth most important food crop, being surpassed in total production only by wheat, corn, and rice. Improved resistance to disease and pests during growth and storage is therefore of significant economic importance.

Transgenic plants expressing the coat protein (CP), and/or the RNA transcript of the CP gene of many RNA viruses, may be protected against the virus from which the CP gene was derived, and, in some cases, against related viruses (for further details, see Chapter 3). Since it is not possible to predict from the l ...

Genome sequences are of minimal use without an adequate interpretation of the sequences of putative protein products, which is only possible on the basis of detailed computer analysis. The approaches to amino acid sequence analysis can be roughly divided into those that explore intrinsic ...

Transient gene expression studies of plant virus genes or parts of genes can be studied using plant protoplasts. Such studies are dependent on a number of important factors: 1. The method for isolating plant cells or protoplasts. Met

Methods developed for plant virology have been of central importance to other branches of plant pathology. Fungal and bacterial pathogens were recognized and characterized in some detail by 1880, but it was not until after the turn of the century that viral pathogens were identified, and cons ...

Large amounts of active mRNA can be synthesized in vitro. In vitro-transcribed RNA molecules may subsequently be used for various purposes, e.g., in vitro translation. Several plasmids are available that are used to produce transcript molecules from cloned DNA inserts. These contain one or m ...

When trying to identify a clone within a cDNA library, which may contain a coat protein (CP) gene, one useful technique may by immunological screening, using antibodies raised against either purified virus or isolated CP. Antibody screening can be carried out on a cDNA cloned into a wide range of vect ...

Antibodies are produced by the immune systems of animals in response to the presence of foreign substances. Antibodies raised against regions of the viral coat protein (CP) (epitopes) can be exploited for virus detection; in 1977 Clark and Adams described the use of antibodies in ELISA to rapidly ...

The polymerase chain reaction (PCR) is used to amplify DNA between two oligonucleotide primers, of which one is complimentary to a sequence on the (+)-strand and the other to a downstream sequence on the (−)-strand. Amplification between the two primers is achieved by reiterative cycles of templ ...