Advanced search
Start date
Betweenand

Development of protocols using quantitative PCR (qPCR) for molecular diagnostic of the quarantine nematode A1 Pratylenchus crenatus

Grant number: 12/19101-9
Support Opportunities:Scholarships abroad - Research
Start date: July 01, 2013
End date: January 31, 2014
Field of knowledge:Agronomical Sciences - Agronomy - Plant Health
Principal Investigator:Claudio Marcelo Gonçalves de Oliveira
Grantee:Claudio Marcelo Gonçalves de Oliveira
Host Investigator: Vivian Carol Blok
Host Institution: Instituto Biológico (IB). Agência Paulista de Tecnologia dos Agronegócios (APTA). São Paulo , SP, Brazil
Institution abroad: James Hutton Institute, Scotland  

Abstract

Molecular detection of pest and pathogens relies on rapid and dependable methods for their identification and quantification. This project describes the development and evaluation of a novel qualitative and quantitative diagnostic method for Pratylenchus crenatus Loof, 1960, consider as A1 quarantine pest to Brasil, based on a fluorogenic 5' nuclease PCR assay (TaqMan). Primer/probe sets will be designed targeting the ITS-1 region of the ribosomal cistron (rDNA). The validation experiments will be carry out using extracted DNA from P. crenatus populations from United Kingdom, Holland and a population detected recently in Brazil. In order to assess the specificity, the primer/probe sets will be tested with samples of other Pratylenchus species. Experiments with dilutions of purified plasmid standards will be used to test the sensitivity of the TaqMan assay on detection of nematode DNA to the femtogram level. Quantification of the target DNA present in field potato samples will be performed. Also, the genetic diversity between P. crenatus populations will be studied based on molecular techniques and phylogenetic analysis. Complementary to the main objective, training on identification of Globodera pallida e G. rostochiensis will be implemented based on both traditional taxonomy and modern molecular techniques. These species are on the top list of main quarantine diseases, causing damages on potato tubers cultivated in Europe and should be treated as serious pests if introduced in Brazil. The use of these tools are expected to provide an accurate identification of P. crenatus and Globodera species, enabling researchers and extension services of quarantine service from Brazil to establish protocols for detecting plant-parasitic nematodes and, consequently, avoiding their introduction.

News published in Agência FAPESP Newsletter about the scholarship:
More itemsLess items
Articles published in other media outlets ( ):
More itemsLess items
VEICULO: TITULO (DATA)
VEICULO: TITULO (DATA)

Scientific publications
(The scientific publications listed on this page originate from the Web of Science or SciELO databases. Their authors have cited FAPESP grant or fellowship project numbers awarded to Principal Investigators or Fellowship Recipients, whether or not they are among the authors. This information is collected automatically and retrieved directly from those bibliometric databases.)
OLIVEIRA, CLAUDIO M. G.; BLOK, VIVIAN; NEILSON, ROY; MROZ, TOMASZ; ROBERTS, DAVID. Hydrolysis probe-based PCR for detection of Pratylenchus crenatus, P. neglectus and P. penetrans. NEMATOLOGY, v. 19, n. 1, 1, p. 81-91, . (12/19101-9)
OLIVEIRA, CLAUDIO M. G.; BLOK, VIVIAN; NEILSON, ROY; MROZ, TOMASZ; ROBERTS, DAVID. Hydrolysis probe-based PCR for detection of Pratylenchus crenatus, P. neglectus and P. penetrans. NEMATOLOGY, v. 19, n. 1, p. 11-pg., . (12/19101-9)