Articles
DETECTION AND QUANTIFICATION OF MRNA BY REVERSE TRANSCRIPTION REAL TIME PCR AS INDICATOR OF VIABILITY OF PHYTOPHTHORA CAMBIVORA IN SOIL
Article number
844_50
Pages
361 – 366
Language
English
Abstract
Detection of introduced invasive Phytophthoras is a main issue in order to forecast the risk of Ink disease and evaluate the efficacy of mitigation and eradication plans.
Diagnosis of these pathogens is based on analysis of symptoms and/or isolation in pure culture from different substrates; recently PCR techniques based on DNA template were widely introduced.
However biological detection is scarcely sensible resulting in false negatives especially from difficult substrates; furthermore PCR based on DNA templates does not answer the question of viability of the inoculum.
In this study mRNA was used as a viability marker of Phytophthora cambivora one of the causal agents of Ink disease of chestnut.
Cytochrome oxidase gene encoding subunits I and II was chosen to detect changes in gene transcription.
The goal is to develop a differential display RT-PCR technique able to detect changes in transcription of COXI and COXII cytochrome oxidase subunits in Phytophthora spp. following exposition of samples to different chemical and physical tretments.
The technique would make available a larger amount of transcripts as templates for molecular diagnosis with RT-PCR and would offer an additional tool to assess the vitality of the pathogen through the differential display analysis of subsamples differently treated.
Diagnosis of these pathogens is based on analysis of symptoms and/or isolation in pure culture from different substrates; recently PCR techniques based on DNA template were widely introduced.
However biological detection is scarcely sensible resulting in false negatives especially from difficult substrates; furthermore PCR based on DNA templates does not answer the question of viability of the inoculum.
In this study mRNA was used as a viability marker of Phytophthora cambivora one of the causal agents of Ink disease of chestnut.
Cytochrome oxidase gene encoding subunits I and II was chosen to detect changes in gene transcription.
The goal is to develop a differential display RT-PCR technique able to detect changes in transcription of COXI and COXII cytochrome oxidase subunits in Phytophthora spp. following exposition of samples to different chemical and physical tretments.
The technique would make available a larger amount of transcripts as templates for molecular diagnosis with RT-PCR and would offer an additional tool to assess the vitality of the pathogen through the differential display analysis of subsamples differently treated.
Publication
Authors
A.M. Vettraino, A. Tomassini, A. Vannini
Keywords
chestnut, cytochrome oxydase gene, Phytophthora cambivora, real-time RT-PCR, viability
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