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Identification of HR-Inducing cDNAs from Plant Pathogens via a Gateway-Compatible Binary Potato Virus X-Expression Vector

Identification of pathogen effectors that elicit a hypersensitive response (HR) in resistant plant hosts is essential to study disease resistance. In this method, it is described how to generate a cDNA library, how to transfer the library into a binary PVX-expression vector, and finally how to ...

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Proteomic Techniques for PlantFungal Interactions

Proteomics is a key technique that is helping elucidate many complex biological processes. The analysis of plant–pathogen interactions using proteomics is complicated by the presence of the proteomes of two species, but is benefiting from the developing maturity and power of these tec ...

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The Use of Agroinfiltration for Transient Expression of Plant Resistance and Fungal Effector Proteins in Nicotiana benthamiana Leaves

Agroinfiltration is a versatile, rapid and simple technique that is widely used for transient gene expression in plants. In this chapter we focus on its use in molecular plant pathology, and especially for the expression of plant resistance (R) and fungal avirulence (Avr) (effector) genes in le ...

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Freeze-Fracture of Infected Plant Leaves in Ethanol for Scanning Electron Microscopic Study of Fungal Pathogens

Fungi often are found within plant tissues where they cannot be visualized with the scanning electron microscope (SEM). We present a simple way to reveal cell interiors while avoiding many common causes of artifact. Freeze-fracture of leaf tissue using liquid nitrogen during the 100% ethan ...

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Real-Time PCR for the Quantification of Fungi In Planta

Methods enabling quantification of fungi in planta can be useful for a variety of applications. In combination with information on plant disease severity, indirect quantification of fungi in planta offers an additional tool in the screening of plants that are resistant to fungal disease ...

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Using Non-homologous End-Joining-Deficient Strains for Functional Gene Analyses in Filamentous Fungi

Fungal strains deficient in the non-homologous end-joining (NHEJ) pathway are excellent recipient strains for gene targeting approaches. In addition, NHEJ-deficiency can facilitate the formation of heterokaryons which allows rapid identification of essential genes. How ...

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ISSR: A Reliable and Cost-Effective Technique for Detection of DNA Polymorphism

With the emergence of more and more molecular markers as useful tools in plethora of population genetic and phylogenetic studies, choice of marker system for a particular study has become mind boggling. These marker systems differ in their advantages and disadvantages, so it is imperative to ...

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Chromatin Immunoprecipitation to Identify Global Targets of WRKY Transcription Factor Family Members Involved in Plant Immunity

The completion of the alfalfa, Arabidopsis, papaya, poplar, and rice genome sequences along with ongoing sequencing projects of various crop species, offers an excellent opportunity to study gene expression at the whole genome level and to unravel the complexity of gene networks underl ...

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Restriction Fragment Length Polymorphism of the 5S-rRNA-NTS Region: A Rapid and Precise Method for Plant Identification

Molecular genetic methods have several advantages over classical morphological and chemical analyses. The genetic method requires genotype instead than phenotype, therefore PCR-based techniques have been widely used for a rapid identification of plant species, varieties a ...

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Random Amplified Marker Technique for Plants Rich in Polyphenols

More than 10,000 publications using the random amplified polymorphic DNA (RAPD) or related arbitrary marker techniques have been published in two decades of its inception in 1990. Despite extensive use, RAPD technique has also attracted some criticisms, mainly for lack of reproducibi ...

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Amplified Fragment Length Polymorphism: An Invaluable Fingerprinting Technique for Genomic, Transcriptomic, and Epigenetic Studies

Amplified fragment length polymorphism (AFLP) is a PCR-based technique that uses selective amplification of a subset of digested DNA fragments to generate and compare unique fingerprints for genomes of interest. The power of this method relies mainly in that it does not require prior info ...

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DNA Purification from Multiple Sources in Plant Research with Homemade Silica Resins

DNA purification is a routine procedure in most plant laboratories. Although different kits are available in the market allowing convenient DNA purification, the cumulative cost of purchasing multiple kits for a laboratory can be staggering. Here, we describe a protocol using homemade ...

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Plant Genetics for Forensic Applications

An emerging application for plant DNA fingerprinting and barcoding involves forensic investigations. Examples of DNA analysis of botanical evidence include crime scene analysis, identifying the source of commercial plant products, and investigation of trade in illicit drugs. ...

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Challenges in the DNA Barcoding of Plant Material

DNA barcoding, using a short gene sequence from a standardized region of the genome, is a species identification tool which would not only aid species discovery but would also have applications ranging from large-scale biodiversity surveys through to identification of a single fragment ...

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DNA Fingerprinting, DNA Barcoding, and Next Generation Sequencing Technology in Plants

DNA fingerprinting of plants has become an invaluable tool in forensic, scientific, and industrial laboratories all over the world. PCR has become part of virtually every variation of the plethora of approaches used for DNA fingerprinting today. DNA sequencing is increasingly used eith ...

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A Taxonomists View on Genomic Authentication

A brief history of taxonomy, for the most part plant oriented, is provided, which demonstrates the use of morphology early on, through the stages when different technologies became available at different times until the present use of genomic tools. Genomic authentication facilitates w ...

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Using GenBank for Genomic Authentication: A Tutorial

The GenBank� database is perhaps one of the most important repositories of genetic information. A researcher working in the field of genomic authentication must therefore be equipped with the skills needed to competently access the required information from this database whilst ulti ...

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Genomic DNA Extraction and Barcoding of Endophytic Fungi

Endophytes live inter- and/or intracellularly inside healthy aboveground tissues of plants without causing disease. Endophytic fungi are found in virtually every vascular plant species examined. The origins of this symbiotic relationship between endophytes go back to the emer ...

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Loop-Mediated Isothermal Amplification for the Detection of Plant Pathogens

Loop-mediated isothermal amplification (LAMP) is a technique involving the use of four to six primers (two inner primers, two outer primers, and two loop primers) and the strand displacement activity of Bacillus subtilis-derived (Bst) DNA polymerase. The end result of strand displaceme ...

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Multiplex PCR Method to Discriminate Artemisia iwayomogi from Other Artemisia Plants

Some plants in the genus Artemisia have been used for medicinal purposes. Among them, Artemisia iwayomogi, commonly referred to as “Haninjin,” is one of the major medicinal materials used in traditional Korean medicine. By contrast, Artemisia capillaris and both Artemisia argyi and Arte ...

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