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Endophytism of Sclerotinia pseudotuberosa: PCR assay for specific detection in chestnut tissues

Published online by Cambridge University Press:  15 February 2005

Anna Maria VETTRAINO ,
Annarita PAOLACCI  and
Andrea VANNINI
Anna Maria VETTRAINO
Affiliation:
Department of Plant Protection, University of Tuscia, via S. Camillo de Lellis, I-01100 Viterbo, Italy. E-mail: [email protected]
Annarita PAOLACCI
Affiliation:
Department of Agrobiology and Agrochemistry, University of Tuscia, via S. Camillo de Lellis, I-01100 Viterbo, Italy.
Andrea VANNINI
Affiliation:
Department of Plant Protection, University of Tuscia, via S. Camillo de Lellis, I-01100 Viterbo, Italy. E-mail: [email protected]
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Abstract

The endophytic behaviour of Sclerotinia pseudotuberosa, cause of black rot of fruits of Castanea sativa, was investigated in asymptomatic tissues of the host, including nuts, buds and bark. A nested polymerase chain reaction (PCR) assay was developed for the specific detection of S. pseudotuberosa. Two specific primers, RAC1 and RAC2 were designed within the ITS1-5.8S-ITS2 region of rDNA by comparison of sequences of 24 isolates of S. pseudotuberosa and related species. Specificity of primers was tested against isolates of related and unrelated genera and the total DNA of C. sativa. The assay proved to be highly sensitive and capable of detecting picograms of S. pseudotuberosa DNA. Our study showed that S. pseudotuberosa was present asymptomatically in all the chestnut tissues tested. It is discussed that latency of S. pseudotuberosa in chestnut tissues may represent a adaptive strategy of the pathogen for rapid and massive host colonization in favourable environmental conditions.

Type
Research Article
Information
Mycological Research , Volume 109 , Issue 1 , January 2005 , pp. 96 - 102
Copyright
© The British Mycological Society 2005

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