Diseases and Management of Crops under Protected Cultivation

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(Diseases and Management of Crops under Protected Cultivation) Data Analysis: score all the gels twice manually and independently. Indicate band presence by 1 and its absence by 0. Score all unique bands and include in the analysis. Presence or absence of unique and shared, polymorphic band should be used to generate similarity coefficients. The similarity coefficients then used to construct a dendogram manually by UPGMA (unweighted pairgroup method with arithmetical averages). Application of PCR in Plant Pathology - PCR technology has become an essential research and diagnostic tool for improving knowledge regarding identification, characterization, detection and diagnosis of plant pathogens. PCR technology allows scientists to take a specimen of genetic material, even from just one cell, copy its genetic sequence over and over, and generate a test sample sufficient to detect the presence or absence of a specific virus, bacterium or any particular sequence of genetic material. Therefore, it is hard to exaggerate the impact of the polymerase chain reaction. PCR, the quick, easy method for generating unlimited copies of any fragment of DNA, is one of those scientific developments that actually deserve timeworn superlatives like "revolutionary" and "breakthrough." Diagnosis and characterization of the pathogen- PCR utilises specific oligonucleotide primers, which are designed based on nucleic acid sequences that are diagnostic for the pathogen. These may be sequences identified by genomic sequencing or techniques such as DNA fingerprinting of pathogens. To improve the sensitivity of these techniques, it is sometimes an advantage to use nested PCR methods, in which the products from the initial PCR amplification are diluted and reamplified with a second set of primers internal to the original primer set in the pathogen sequence. These conventional PCR-based techniques have proved to be useful for diagnosis of fungal, bacterial and phytoplasma-associated diseases with a number of good taxon-specific primers developed for example from the rRNA subunit genes. Because of the sequence variation between these regions in different isolates, it has sometimes been possible to identify restriction endonucleases that give different restriction patterns upon digestion of the PCR products depending upon the isolate. Primers have also been developed based on more specific sequences such as the argk-tox gene of P. syringae pv. phaseolicola which encodes a gene involved in phaseolotoxin biosynthesis and can be used to identify bacteria that possess this trait, or the aflR gene of Aspergillus flavus which regulates aflatoxin production in these fungi. In addition, PCR-based techniques have proved useful for identifying the vectors for insect-transmitted diseases. For example, DNA extracted from leafhoppers that are potential vectors for phytoplasma diseases can be PCR amplified using phytoplasma-specific primers to identify which species are the true vectors. Single-strand RNA viruses can be detected by modifying the PCR to include a reverse transcriptase step (RT-PCR). In this, the reverse transcriptase uses a viral specific primer to make a cDNA copy of the viral RNA which is then amplified using Taq polymerase through conventional PCR. RT-PCR has the potential for use in diagnostics of bacterial and fungal pathogens. Through - 253 -
(Diseases and Management of Crops under Protected Cultivation) identification of particular genes that are expressed during pathogen growth, or at specific stages of development, it may be possible to use RT-PCR to identify the developmental stage of the pathogen in infected material. It has also been possible to produce ‘multiplex’ PCR kits capable of detecting more than one pathogen present in a particular plant or soil sample. Kits are commercially available to unravel the cereal stem-based complex of fungi comprising Tapesia yallundae and T. acuformis (eyespot fungi), Fusarium culmorum, F. avenaceum, F. graminearum and F. poae (ear blights), and Microdochium nivale (snowmould of cereals). REFERENCES • Dickinson. M. 2005. Molecular Plant Pathology. BIOS Scientific Publishers, Taylor & Francis Group. London and New York. • Schaad, N. W. and Frederick, R. D. 2002. Real-time PCR and its application for rapid plant disease diagnostics. Can. J. Plant Pathol. 24: 250–258. • Gelsomino, G., Faedda, R., Rizza, C., Petrone, G. and Cacciola, S.O. 2011. New platforms for the diagnosis and identification of fungal and bacterial pathogens. In: Science against microbial pathogens: communicating current research and technological advances .( E_d_.) A. Méndez Vilas. 622-630. - 254 -
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CENTRE OF ADVANCED FACULTY TRAINING
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CONTENTS Sl. No. Title Speaker Page
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REMARKS by Dr. K.S. Dubey Director
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and extension literature that have
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discussions. No doubt the course is
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Introduction (Diseases and Manageme
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Loads For Greenhouse: (Diseases and
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Chemical Control Chemical Abamectin
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Water Requirement (lpd/plant) (Dise
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clear (Boulard et al, 1989). (Disea
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Page 211 and 212: Risks of biocontrol agents: (Diseas
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