植物学技术

The expression of viral coat protein (CP) genes in transgenic plants can lead to different phenotypes of resistance (1). Occasionally, transgenic plants escape infection completely and do not accumulate virus or develop symptoms. In other cases, local and systemic virus accumulation a ...

This chapter describes methods to assess agronomic performance among plants of existing plant cultivars that have been transformed with viral genes for the purpose of achieving resistance to the virus (1). The principles and general guidelines for selection among plants altered by tra ...

This chapter describes methods to detect and assess commercial resistance to virus disease that may be conferred on existing plant cultivars by their transformation with genes derived from viruses (1). The principles and general guidelines for selection among plants altered by trans ...

A number of methods have been used for the quantitation of virus within infected plants. Early ones used physical techniques such as dry-wt and particle counts; more recently, serological and nucleic acid-based methods have been developed. These methods are all based on a physical aspect of the v ...

The polymerase chain reaction (PCR) is a technique that enables the specific amplification and hence detection of target DNA sequences from complex mixtures of nucleic acid. A combination of short, specific primers and thermostable DNA polymerases are used to amplify the target sequen ...

Unlike for fungal and bacterial diseases, no direct method for the control of viral diseases is yet available. The early detection of plant viruses constitutes, therefore, one of the main ways of controlling these diseases, and so sensitive detection systems are essential. Until now, methods ...

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 ...