植物生理

A large body of evidence has lead to the suggestive proposal of a potential interplay between viroid-induced pathogenesis and RNA silencing regulatory mechanisms. A variety of techniques have been used to examine this interaction. This chapter outlines the use of a green fluorescent pro ...

Our increasing understanding of virus–host interactions is revealing a complex role for host factors during virus replication. Besides the role of some host proteins in defense against viruses, it is becoming clear that viruses also hijack several host functions to utilize them for their ...

The identification of small molecule inhibitors of plant virus RNA replication is useful to develop antiviral drugs and to dissect various steps in the replication process. Moreover, small molecule inhibitors could be effective tools to study similarities in replication strategi ...

Given the widespread impact of RNA silencing on the Arabidopsis thaliana genome, it is indeed remarkable that this means of gene regulation went undiscovered for so long. Since the publication of landmark papers in 1998 (Fire et al., Nature 391:806–811, 1998; Waterhouse et al., Proc Natl Acad Sci U S A 95: ...

RNA interference (RNAi) plays multiple biological roles in eukaryotic organisms to regulate gene expression. RNAi also operates as a conserved adaptive molecular immune mechanism against invading viruses. The antiviral RNAi pathway is initiated with the generation of virus-de ...

Virus-induced gene silencing (VIGS) has emerged as a powerful method for studying gene function. VIGS is induced by infecting a plant with a plant virus that has had its genome modified to include a sequence from the host gene to be silenced. DNAβ and DNA1 are satellite and single-stranded DNA molecul ...

Agroinfiltration assay using green fluorescent protein (GFP)-expressing Nicotiana benthamiana line 16c is a powerful method for screening of putative plant virus-encoded gene silencing suppressors. This method allows the investigator to know whether the putative viral sup ...

Virus-induced gene silencing (VIGS) is a useful functional genomics tool for rapidly creating plant gene knockout phenotypes that can be used to infer gene function. Until recently, VIGS has only been possible in dicotyledonous plants. However, the development of cloning vectors based on ...

One of the most effective forms of plant defense against viruses is posttranscriptional gene silencing (PTGS). This process implies that a virus can trigger the host plant’s RNA-silencing machinery and ultimately become the target of RNA silencing. PTGS is, therefore, an attractive endo ...

Viral cross protection in plants is known as an acquired immunity phenomenon, where a mild virus isolate/strain can protect plants against economic damage caused by a severe challenge strain/isolate of the same virus. Mild strain cross protection (MSCP) has been used extensively to contr ...

Small interfering RNAs (siRNAs) and microRNAs (miRNAs) are non-coding regulatory RNAs that play an important role in development and genome stability in plants. Conventional cloning and sequencing approaches have identified hundreds of miRNAs and a large number of siRNAs, but are no lo ...

The following chapter describes a PCR method for the identification of the raspberry root rot pathogen Phytophthora fragariae var. rubi. Furthermore, a nested PCR suitable for the detection of the pathogen in infected raspberry roots and validated against the “Duncan bait test” (EPPO Bull ...

Traditional methods for the isolation and identification of fungal spores can be time-consuming and laborious. DNA-based methods for fungal detection can be used to detect the spores of plant-pathogenic fungi. Air borne spores can be collected and identified by PCR allowing identific ...

FISH is a widely used technique in many laboratories not only for cytogenetic studies, but also in other biological fields. It requires a combination of skills in molecular biology, cytogenetics, immunocytochemistry, microscopy and cellular imaging analysis.

Erwinia amylovora is the causative agent of fire blight, a very destructive disease of numerous members of the rosaceae. The primary route of infection for host species, including commercially grown apple and pear, is the newly opened blossom. Susceptibility of flowers to infection for only a ...

Ring rot, brown rot and blackleg represent major bacterial pathogens of potato. The methods described below are aimed at basic identification of ring rot, brown rot or blackleg in a tuber sample.

Immunofluorescence microscopy is a very sensitive serological test which harnesses both the power of antibodies to bind to targets along with the use of the flourescence microscope to visualise the structures to which they bind. Antibody binding is visualised by the fluorescent emiss ...

Enzyme-linked immunosorbent assay (ELISA) is the most commonly used serological diagnostic technique. A number of different ELISA formats can be used for the detection of bacterial plant pathogens and in particular Erwinia amylovora and Clavibacter michiganensis subsp. sepedo ...

The use of monoclonal antibodies in plant pathology has improved the quality and specificity of detection methods for diseases. Hybridoma technology allows the limitless production of highly specific antibodies which can be used to identify pathogens to the species or even sub-speci ...

Outbreaks of bacterial wilt caused by Ralstonia solanacearum in South Africa in the 1980s necessitated the development of a sensitive assay for its detection. In this chapter, the development of an enzyme-linked immunosorbent assay (ELISA) for the detection of Ralstonia solanacearum ...