Detection and Identification of <Emphasis Type="Italic">Phytophthora</Emphasis> Species in Southern Italy by RFLP and Sequence Analysis of PCR-amplified Nuclear Ribosomal DNA |
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Authors: | Email author" target="_blank">Ippolito?CameleEmail author Carmine?Marcone Gennaro?Cristinzio |
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Institution: | (1) Dipartimento di Biologia, Difesa e Biotecnologie Agro-Forestali, Università degli Studi della Basilicata, Viale dell’Ateneo Lucano 10, 85100, Potenza;(2) Dipartimento di Arboricoltura, Botanica e Patologia Vegetale, Sez. di Patologia Vegetale, Università degli Studi di Napoli (Portici), Via Università 100, 80055 Portici, NA |
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Abstract: | In four neighbouring regions of southern Italy, Basilicata, Campania, Apulia and Calabria, pepper and zucchini plants showing
Phytophthora blight symptoms, tomato plants with either late blight or buckeye rot symptoms, plants of strawberry showing
crown rot symptoms and declining clementine trees with root and fruit rot were examined for Phytophthora infections by means of polymerase chain reaction (PCR) assays, using primers directed to nuclear ribosomal DNA (rDNA) repeat
sequences. All diseased plants and trees examined tested positive. The detected fungal-like organisms were differentiated
and characterized on the basis of primer specificity as well as through extensive restriction fragment length polymorphism
(RFLP) and sequence analysis of PCR-amplified rDNA. Phytophthora capsici was identified in diseased pepper and zucchini plants, P. infestans was identified in tomato with late blight symptoms whereas buckeye rot-affected tomatoes and diseased strawberry plants proved
to be infected by P. nicotianae and P. cactorum, respectively. Declining clementine trees were infected with P. citrophthora and P. nicotianae in about the same proportion. Also, thirty-one pure culture-maintained isolates of Phytophthora which had previously been identified in southern Italy by traditional methods but were never examined molecularly, were examined
by RFLP and sequence analysis of PCR-amplified nuclear rDNA. Among these, an isolate from gerbera which had previously been
identified by traditional methods only at genus level, was assigned to P. tentaculata. For the remaining pure culture-maintained isolates examined, the molecular identification data obtained corresponded with
those delineated by traditional methods. Most of the diseases examined were already known to occur in southern Italy but the
pathogens were molecularly detected and fully characterized at nuclear rDNA repeat level only from other geographic areas,
very often outside Italy. A new disease to southern Italy was the Phytophthora blight of zucchini. This is also the first
report on the presence and molecular identification of P. tentaculata from Italy. |
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Keywords: | DNA-based methods fungal-like organism taxonomy ITS regions phylogenetic analysis Phytophthora diseases restriction endonucleases |
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