Further studies on the use of molecular probes to grapevine closterovirus A

Authors

  • A. Minafra
  • M. Russo
  • G. P. Martelli

DOI:

https://doi.org/10.5073/vitis.1992.31.87-93

Keywords:

Vitis, closterovirus, virosis, subgenomic RNAs, cDNA, molecular probes, diagnosis

Abstract

Two cloned cDNA probes to genomic RNA of grapevine closterovirus A (GVA) were utilized successfully for the detection of viral sequences in infected herbaceous hosts (Nicotiana benthamiana) and grapevines. One of the probes (pGA112) was complementary to the central part of the viral genome and gave light false positive signals with healthy grapevine extracts, whereas the other (pGA240), which is presumably colinear with the 3' terminus, was virus-specific and hybridized only with infected sample extracts. The two probes recognized smaller than genome RNAs in electrophoresed N. benthamiana extracts and hybridized differentially with the bands, thus suggesting that these represent subgenomic RNAs. Probe pGA240 may be used for GVA detection, but the preparation of samples for hybridization needs further improvement for routine testing.

Weitere Untersuchungen über die Verwendung von Molekülsonden beim Grapevine-Closterovirus A

Zwei klonierte cDNA-Sonden für genomische RNA des Grapevine-Closterovirus A (GVA} wurden mit Erfolg zum Nachweis von Virussequenzen in infizierten krautigen Wirtspflanzen (Nicotiana benthamiana) und Reben benützt. Die eine Sonde (pGA112} war komplementär zum zentralen Teil des Virusgenoms und lieferte schwache falsch-positive Nachweisreaktionen mit Extrakten aus gesunden Reben; die andere Sonde (pGA240}, die vermutlich kolinear mit dem 3'-Terminus ist, war dagegen virusspezifisch und hybridisierte nur mit Extrakten aus infizierten Proben. In elektrophoretisch aufgetrennten N. benthamiana-Extrakten "erkannten" die beiden Sonden RNAs von weniger als Genomgröße; sie hybridisierten differenziert mit den Banden, so daß diese subgenomische RNAs darstellen könnten. Die Sonde pGA240 kann für den GVA-Nachweis verwendet werden; für Routinetests muß die Vorbereitung der Proben für die Hybridisierung noch verbessert werden.

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Published

2015-10-08

Issue

Vol. 31 No. 2 (1992): Vitis

Section

Article

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