Molecular detection of a Closterovirus associated with apricot stem pitting in Southern Italy [Prunus armeniaca L. - Apulia]

Cortical and leaf tissue extracts from apricot trees of cv Tyrinthos from Apulia (Southern Italy) affected by stem pitting contained multiple double-stranded RNA (dsRNA) species, the largest with an estimated size of 15 kbp. A segment of 590 nucleotides, showing sequence homology with the HSP70 homo...

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Bibliographic Details
Published in:Journal of plant pathology 2001-07, Vol.83 (2), p.125-131
Main Authors: Abou Ghanem-Sabanadzovic, N, Mahboubi, M, Di Terlizzi, B, Sabanadzovic, S. (Istituto Agronomico Mediterraneo, Valenzano, Bari (Italy)), Savino, V, Martelli, G.P. (Bari Univ. (Italy). Dipartimento di Protezione delle Piante e Microbiologia Applicata), Uyemoto, J.K. (California Univ., Davis (USA). Dept. of Plant Pathology)
Format: Article
Language:English
Subjects:
ADN
APULIA
ARN
Bark
CLONACION MOLECULAR
CLONAGE MOLECULAIRE
CLONAZIONE MOLECOLARE
CLOSTEROVIRUS
CLOSTEROVIRUSES
Complementary DNA
DNA
Double stranded RNA
ENFERMEDADES DE LAS PLANTAS
EXPERIMENTACION IN VITRO
EXPERIMENTATION IN VITRO
IDENTIFICACION
IDENTIFICATION
IDENTIFICAZIONE
IN VITRO EXPERIMENTATION
MALADIE DES PLANTES
MALATTIE DELLE PIANTE
MOLECULAR CLONING
NUCLEOTIDE SEQUENCE
PCR
Phytopathology
Pitting
PLANT DISEASES
Polymerase chain reaction
POUILLES
PRUNUS ARMENIACA
PUGLIA
Reverse transcriptase polymerase chain reaction
RNA
SECUENCIA NUCLEOTIDICA
SEQUENCE NUCLEOTIDIQUE
SEQUENZA NUCLEOTIDICA
SINTOMAS
SINTOMI
SPERIMENTAZIONE IN VITRO
SYMPTOME
SYMPTOMS
Viruses
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Description
Summary:Cortical and leaf tissue extracts from apricot trees of cv Tyrinthos from Apulia (Southern Italy) affected by stem pitting contained multiple double-stranded RNA (dsRNA) species, the largest with an estimated size of 15 kbp. A segment of 590 nucleotides, showing sequence homology with the HSP70 homologue gene of members of the family Closteroroviridae was amplified by RT-PCR from symptomatic trees using degenerated primers designed on the conserved phosphate 1 and 2 motifs of the HSP70 gene sequence. Computer-assisted analysis showed that the 590 nt fragment from Italian apricot trees had 97% sequence homology with Plum bark necrosis stem pitting-associated virus (PBNSPaV), a recently reported closterovirus infecting Black Beaut plum (Prunus salicina) in California. A set of primers that amplified a 290 nt virus-specific fragment was designed and a cRNA probe was generated. This riboprobe and RT-PCR assays with these primers detected PBNSPaV in diseased but not in healthy apricot trees [Estratti di tessuti della corteccia e delle foglie di piante di albicocco della cv Tyrinthos coltivata in Puglia (Italia Meridionale) colpite da butteratura del fusto contenevano specie multiple di RNA a doppia elica (dsRNA), la piu' grande con una dimensione stimata di 15 kbp. Un segmento di 590 nucleotidi, presentante un'omologia di sequenza con il gene omologo HSP70 dei membri della famiglia closteroviridae, e' stato amplificato, mediante RT-PCR, da piante sintomatiche utilizzando primer degenerati ottenuti sui motivi fosfatici 1 e 2 conservati della sequenza genica di HSP70. L'analisi assistita da computer ha dimostrato che il frammento da 590 nt delle piante di albicocco italiane presentava un'omologia di sequenza del 97% con il Virus associato alla butteratura del fusto e necrosi della scorza del susino (PBNSPaV), un closterovirus di segnalazione recente che infetta il susino Black Beaut (Prunus salicina) in California. E' stato messo a punto un gruppo di primer che amplificavano un frammento virus-specifico di 290 nt ed e' stata generata una sonda a cRNA. Questa ribosonda e i saggi RT-PCR con questi primer hanno consentito di individuare PBNSPaV nelle piante malate, ma non in quelle sane, di albicocco]
ISSN:1125-4653
2239-7264