植物生理

The iPhyClassifier is an internet-based research tool for quick identification and classification of diverse phytoplasmas. The iPhyClassifier simulates laboratory restriction enzyme digestions and subsequent gel electrophoresis and generates virtual restricti ...

It is no longer as difficult to determine genomic DNA sequences of uncultured bacteria as it once was, due to the development of DNA sequencing technology. It is likely that the number of whole-genome sequences of phytoplasmas will increase. In this chapter, two major strategies of whole-genome c ...

Phytoplasma identification has proved difficult due to their inability to be maintained in vitro. DNA barcoding is an identification method based on comparison of a short DNA sequence with known sequences from a database. A DNA barcoding tool has been developed for phytoplasma identific ...

Phytoplasmas are routinely detected by nucleic acid-based techniques. These approaches rely on enriched phytoplasma DNA extracts of good quality, following labor intensive and time-consuming purification protocols. Here we describe a very rapid, specific, sensitive, and rel ...

Loop-mediated isothermal amplification (LAMP) is a method for amplification and detection of target organisms which, unlike polymerase chain reaction, does not require thermal cycling. LAMP assays can be developed in the laboratory for subsequent deployment in the field, where the s ...

In this chapter, we describe a real-time PCR detection system for fast, reliable, specific, and sensitive detection and discrimination of ‘Candidatus Phytoplasma mali’, ‘Ca. P. prunorum’, and ‘Ca. P. pyri’ from the 16SrX (apple proliferation-AP) group. These phytoplasmas are causal agents ...

The real-time PCR detection system for grapevine yellows phytoplasmas described here is composed of two assays for group-specific detection of flavescence dor�e (FD) and bois noir (BN) phytoplasmas and a universal phytoplasma assay. It uses hydrolysis minor groove binder probes (Taq ...

Currently, the most efficient detection and precise quantification of phytoplasmas is by real-time PCR. Compared to nested PCR, this method is less sensitive to contamination and is less work intensive. Therefore, a universal real-time PCR method will be valuable in screening programs a ...

Conventionally, detection of phytoplasmas has been performed by PCR of the 16S rRNA gene, followed by either RFLP or DNA sequencing to determine the phytoplasma 16Sr group. This chapter demonstrates the technique of terminal restriction fragment length polymorphism (T-RFLP), a finge ...

This volume of “Methods in Molecular Biology” entitled “Phytoplasmas: Methods and Protocols” aims to provide a broad range of protocols for working with this group of plant pathogens. In this first chapter, we provide some background information about the phytoplasmas to put the protocols ...

Detection and identification of phytoplasmas is a laborious process often involving nested PCR followed by restriction enzyme analysis and fine-resolution gel electrophoresis. To improve throughput, other methods are needed. Microarray technology offers a generic assay th ...

Single-strand conformation polymorphism (SSCP) analysis is a sensitive and rapid technique for detecting DNA polymorphisms and mutations in PCR-amplified fragments. Due to its technical simplicity, it is widely used as a screening tool in various investigations, ranging from cli ...

Conventionally, diagnostics and phylogenetics of phytoplasmas have been primarily based on the 16S rRNA gene, for which “universal” primers are available that amplify from most phytoplasma 16Sr groups. However, there has been a drive in recent years to develop “universal” primers for ot ...

Tuf and secY genotyping techniques have been developed to distinguish phytoplasma strains. Tuf polymerase chain reaction sequence analyses are available for phytoplasma taxonomic groups 16SrI, 16SrV, 16SrXII-A, and XII-B. In addition to their use to confirm the taxonomic status of p ...

Differentiation and classification of phytoplasmas have been primarily based on the highly conserved 16S rRNA gene. RFLP analysis of 16S rRNA gene sequences has identified 31 16S rRNA (16Sr) groups and more than 100 16Sr subgroups. Classification of phytoplasma strains can, however, be ...

Detection of pathogen DNA by polymerase chain reaction (PCR) assays is the most widely used method for diagnosing phytoplasma diseases. Reliable and efficient detection of phytoplasmas, especially in woody perennial plants, is challenging due to the unusually low abundance and spor ...

Current phytoplasma detection and identification methods are primarily based on nested polymerase chain reaction followed by restriction fragment length polymorphism analysis and gel electrophoresis. These methods can potentially detect and differentiate all phyto ...

The method described here is a rapid, total DNA extraction procedure applicable to a large number of plant samples requiring pathogen detection. The procedure combines a simple and quick homogenization step of crude extracts with DNA extraction based upon the binding of DNA to magnetic bead ...

The use of electron microscopy, both transmission and scanning, provides reliable and accurate methods for detecting phytoplasmas in plants. Our understanding of these pathogens, their morphology, development, and intracellular location in plants and insect vectors has been gr ...

Careful examination of how explants react to experimental conditions is the first step of a successful research program. Photographing the specimen is an extremely important method that can be used to document changes of an explant throughout an experiment. This article serves to draw att ...