Detection and molecular characterization of Italian Grapevine rupestris stem pitting‐associated virus isolates

Abstract

The efficiencies of two published primer pairs (RSP13-RSP14 and RSP2-RSP21) were compared in reverse transcriptionpolymerase chain reaction (RT-PCR) to improve the diagnosis of Grapevine rupestris stem pitting-associated virus (GRSPaV), which has consistently been found associated with rupestris stem pitting (RSP), one of the most common infectious diseases of grapevine. Fifty-six grapevine accessions from different locations in Northern Italy were tested. Of these, 48 were positive with primers RSP13-RSP14 and 37 with primers RSP2-RSP21, suggesting that primer pair RSP13-RSP14 has a broader detection spectrum than primers RSP2-RSP21. The coat protein (CP) gene was also amplified by RT-PCR and the amplicons analysed by single-strand conformation polymorphism (SSCP) and sequencing. The results support the existence of wide variability within this virus, independent of cultivar or geographic origin. Sequence analysis of the 17 Italian GRSPaV isolates examined showed a molecular identity ranging from 78AE8 to 99AE7%, within which, four groups of sequence variants were recognized. Flexuous rod-shaped particles, approximately 800 nm in length, were detected in plant extracts from infected grapevines and were positively decorated with the specific antiserum As7-276 in immunosorbent electron microscopy (ISEM). This is the first report of GRSPaV particles in grapevines observed by transmission electron microscopy (TEM) in Italy.

Resistance to Elsinoë Ampelina and Expression of Related Resistant Genes in Vitis Rotundifolia Michx. Grapes

“…Expression of these genes was rapid 24 h of Elsinoë inoculation. Similar validation of expression of pathogenesis-related genes has been recorded in grapevine against Uncinula necator that causes powdery mildew and rupestris stem pitting-associated virus ( GRSPaV ) [24,25]. Chalcone synthase is a phytoalexin biosynthetic enzyme [26], which is involved in defense against fungal diseases.…”

Detection of viruses associated with rugose wood in Japanese grapevines and analysis of genomic variability of Rupestris stem pitting-associated virus

“…In this study, although we also found RSPaV isolate closely related to GRSPaV-SG1 from Pione OE8, one of the eight asymptomatic grapevines (Table 2), further study is necessary to evaluate the pathogenicity of OE8 isolate. Sequencing analysis of RSPaV isolates revealed that the virus has many sequence variants (Casati et al 2003;Lima et al 2006a;Meng et al 1999bMeng et al , 2005Nolasco et al 2006;Rowhani et al 2000;Terlizzi and Credi 2003;Santos et al 2003). In our studies, the RSPaV isolates from Japanese grapevines were grouped into four phylogenetic groups supported by high bootstrap values (Fig.…”

“…In addition, a strikingly high association has been detected recently between RSPaV and vein necrosis on Vitis rupestris 9 V. berlandieri 110 Richiter (Bouyahia et al 2005). On the other hand, several studies revealed that RSPaV comprise a family of sequence variants segregated into three or four phylogenetic groups (Casati et al 2003;Lima et al 2006a;Meng et al 1999bMeng et al , 2005Nolasco et al 2006;Rowhani et al 2000;Terlizzi and Credi 2003;Santos et al 2003). The complete nucleotide sequences of three genomic variants have also been reported in addition to previously published for two other isolates (Meng et al 2005;Lima et al 2006a).…”

Grapevine Virus Diseases:economic Impact and Current Advances in Viral Prospection and Management

“…The biological indexing is time-consuming, limited for inconspicuous symptoms and latent infections, while TeM and micro-or macroarrays are low efficient or onerous (ABDULLAHI et al, 2011;MALIOGKA et al, 2015a;TERLIZZI et al, 2010;ThOMPSON et al, 2012). elISa-or PcR-based methods are limited to viruses for which antibodies or virus-specific nucleotide sequences are available, while being are more rapid and efficient (BASSO et al, 2010b;FAJARDO et al, 2007a;MALIOGKA et al, 2015a).…”