Simultaneous detection of eight viruses and two viroids affecting stone fruit trees by using a unique polyprobe |
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Authors: | Ana Peiró Vicente Pallás Jesús ángel Sánchez-Navarro |
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Institution: | (1) Instituto de Biolog?a Molecular y Celular de Plantas, Universidad Politecnica de Valencia-CSIC, Avenida de los Naranjos s/n, 46022 Valencia, Spain; |
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Abstract: | The use of riboprobes carrying partial sequences of different plant viruses or viroids fused in tandem, has permitted the
simultaneous detection of up to six different pathogens using a non-radioactive molecular hybridization procedure. In the
present report, we describe the development of a unique polyprobe (poly10) with the capacity to detect viruses and viroids
commonly found infecting fruit trees. The poly10 covers eight viruses: Apple mosaic virus (ApMV), Prunus necrotic ringspot virus (PNRSV), Prune dwarf virus (PDV), American plum line pattern (APLPV), Plum pox virus (PPV), Apple chlorotic leaf spot virus (ACLSV), Apricot latent virus (ApLV), Plum bark necrosis and stem pitting-associated virus (PBNSPaV) and two viroids: Hop stunt viroid (HSVd) and Peach latent mosaic viroid (PLMVd). Poly10 is comparable to the individual riboprobes in terms of end-point dilution limit and specificity, allowing
the detection of up to 2.5 picograms of viral or viroidal RNA. However, the polyprobe requires a hybridization temperature
of 60°C instead of the standard 68°C. The validation of the new simultaneous detection strategy was confirmed by the analysis
of 60 field samples, which came from seven different hosts. The use of the polyprobe as an alternative to other routinely
used detection methods is discussed. |
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