http://www.cnr.it/ontology/cnr/individuo/prodotto/ID88386

Application of molecular procedures for detection of Pochonia chlamydosporia from soil (Abstract in rivista)

Type

Prodotto della ricerca (Classe)
Abstract in rivista (Classe)

Label

Application of molecular procedures for detection of Pochonia chlamydosporia from soil (Abstract in rivista) (literal)

Anno

2006-01-01T00:00:00+01:00 (literal)

Alternative label

Rosso L., Ciancio A., Finetti-Sialer M. (2006)
Application of molecular procedures for detection of Pochonia chlamydosporia from soil
(literal)

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Rosso L., Ciancio A., Finetti-Sialer M. (literal)

Pagina inizio

145 (literal)

Pagina fine

145 (literal)

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36 (literal)

Rivista

Nematropica (Rivista)

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Nematropica, 36, 145. (literal)

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1 (literal)

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We compared two procedures for detection of the nematode parasitic fungus Pochonia chlamydosporia, directly from soil. Since DNA amplification from soil is often limited by organic contaminants purified together with nucleic acids, we tested two DNA extraction and detection procedures. The methods relied on the extraction of DNA through a traditional, phenol-chloroform based extraction protocol, or on the use of magnetic beads binding to nucleic acids in the suspension. Tests for sensitivity and specificity were developed using DNA extracted from a sandy soil treated with a commercial product based on P. chlamydosporia, or from pure cultures of the same isolate as control. The detection of the fungus DNA was performed through molecular hybridization (Dot-blot) and Real-time PCR amplification. Two probes of 500 and 600 nucleotides, respectively, were synthesized for Dot-blot detection. They recognized an internal region complementary to an alkaline serineprotease (VCP1) gene, or a fragment of the rRNA mitochondrial (rDNA) gene. For Real-time PCR, a molecular beacon was used to detect a specific sequence of 23bp in the VCP1 gene. The DNA obtained from the conventional procedure was not sufficiently pure to allow PCR amplification, but it enabled identification through the Dot-blot procedure. This protocol, although more time consuming, allowed the testing of several samples with a high sensitivity and specificity with a lower detection limit of 100ng of total DNA from soil. The use of magnetic beads in the DNA extraction from soil appeared as a simple procedure allowing the collection in a few minutes of large amounts of high quality DNA, ready for use in Real Time PCR. (literal)

Note

ISI Web of Science (WOS) (literal)

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Istituto per la Protezione delle Piante, CNR, Bari; Dipartimento di Protezione delle Piante e Microbiologia Applicata, Università degli Studi di Bari (literal)

Titolo