Identification and characterization of a new potyvirus infecting cucurbits.

A new potyvirus, tentatively named cucurbit vein banding virus (CVBV), was identified in crops of cucurbits in San Pedro (Buenos Aires, Argentina). The complete genome sequences of two isolates of CVBV were obtained by next-generation sequencing (Illumina). The genomic RNA consisted of 9968 and 9813 nucleotides, respectively, and displayed typical potyvirus organization. The percentage identity for these two genome sequences, using BLASTn, was 77% to sweet potato virus c and 73% to tomato necrotic stunt virus. BLASTx analysis of the complete polyprotein showed that the most closely related virus is plum pox virus, with 48% amino acid sequence identity for both isolates. Sequence comparisons and phylogenetic analyses indicate that CVBV belongs to a previously undescribed species in genus Potyvirus.

Striking differences in the biological and molecular properties of onion and garlic isolates of onion yellow dwarf virus.

Complete nucleotide (nt) and deduced amino acid sequences of two onion yellow dwarf virus (OYDV) isolates showing mild and severe symptoms in onion but being unable to infect garlic were determined. The genomes consisted of 10,459 and 10,461 nt (without the 3' poly(A) tail) and were 92.2 % identical. Comparison of their whole genomes, polyproteins and P1, HC-Pro, P3, CI, VPg and NIa-Pro regions with those of garlic isolates previously identified as OYDV gave percentage values below that proposed as the molecular threshold for potyvirus species demarcation. This and the striking differences in host range between onion and garlic isolates suggest that they represent different virus species.

First Report of Shallot latent virus in Garlic in Argentina.

Because of exclusively agamic propagation, garlic is commonly infected with a virus complex mainly composed of species within the genera Potyvirus, Allexivirus, and Carlavirus. This virus complex causes leaf striping that ranges from various shades of green to yellow and results in yield losses (2,4). Onion yellow dwarf virus, Leek yellow stripe virus (potyviruses), Garlic virus A, Garlic virus C (allexiviruses), and Garlic common latent virus (carlavirus) have been detected in Argentina previously (1,2). Recently, Shallot latent virus (SLV; another carlavirus) was detected in 25 of 30 garlic plants (cv. Morado) growing in four different fields near Córdoba, Argentina by double-antibody sandwich (DAS)-ELISA using BIOREBA (Reinach, Switzerland) antibodies. To confirm the presence of the virus, DAS-ELISA-positive plants were also analyzed by one-step reverse transcription (RT)-PCR using the Access RT-PCR system (Promega, Madison, WI) with specific primers reported by Tsuneyoshi et al. (3). RNA extractions were performed from 100 mg of leaves with the Qiagen RNeasy Plant Mini Kit (Qiagen, Valencia, CA). Primers used were Car-V1 (5'-AAACCTTTTGGTTCACTTTAGG-3'); Car-V2 (5'-AGGTGCATTGTTATCATTACTGG-3'); and Car-Cp3 (5'-GCGTGCTATATTTAAGTTGCATAC-3'). Primer pairs Car-V1/Car-Cp3 and Car-V2/Car-Cp3 were used for the amplification of the coat protein (CP) gene of SLV and an isolate of SLV formerly known as Garlic latent virus, respectively. Fragments of 992 bp and 1,079 bp were amplified with these primer pairs, respectively. The RT-PCR products were cloned with the TOPO TA Cloning Kit in the 3.9-kb pCR-TOPO vector (Qiagen). The nucleotide sequences of both fragments were determined and were found to be identical (GenBank No. GU355922) showing 94.2% nt sequence identity with the CP gene of an isolate of SLV from Indonesian garlic (GenBank No. AB004686) formerly referred to as Garlic latent virus (3). Consequently, the Argentinean virus is now considered a garlic isolate of SLV. References: (1) E. Cafrune et al. Plant Dis. 90:898, 2006. (2) V. C. Conci. Virus y Fitoplasmas de Ajo. Page 267 in: 50 Temas Sobre Producción de Ajo. Vol. 3. J. L. Burba, ed. Ediciones INTA, Mendoza, Argentina. 1997. (3) T. Tsuneyoshi et al. Arch. Virol. 143:1093, 1998. (4) D. G. A. Walkey and D. N. Antill. J. Hortic. Sci. 64:53, 1989.

Yield Loss in Garlic Caused by Leek yellow stripe virus Argentinean Isolate.

Garlic plants (Allium sativum) are naturally infected by a complex of viruses in the genera Potyvirus, Carlavirus, and Allexivirus. The yield of virus-free garlic plants (noninoculated control) was compared with that of plants infected with an Argentinean isolate of Leek yellow stripe virus (LYSV; L treatment) and garlic plants infected with the virus complex (VC). Evaluations were conducted in the field and in anti-aphid cages during two crop cycles after planting three sizes of cloves (categories). The percent plant emergence in the noninoculated control and in the L treatments (between 80 and 100%) did not differ statistically, but the percent emergence for these two treatments was double that for the VC treatment (25 to 62%). Plant height and leaf number in the L treatment were lower than in the noninoculated control during the first evaluation (year 1), but they did not differ during the second evaluation (year 2). However, both treatments produced taller plants with more leaves than those of VC in both years. The L treatment decreased bulb weight up to 28% and perimeter up to 9% when compared with those in the noninoculated control maintained in the anti-aphid cages until the end of the experiment. However, differences between these treatments were higher in the field experiments where plants were exposed to infection by other viruses (up to 36% in bulb weight and 13% in perimeter). Bulbs of the VC-infected plant treatment were reduced up to 74% in weight and 37% in perimeter. In field evaluations, a high percentage of plants were infected with Onion yellow dwarf virus (58 to 100%), whereas fewer were infected with LYSV (15 to 68%). Garlic virus A infection was high in plants previously infected with LYSV (96 and 97%), but lower in the noninoculated control (12 and 68%). These results show the high impact of the virus complex on garlic yield and the effect of LYSV as a component of the garlic virus complex.

Effect of Two Allexivirus Isolates on Garlic Yield.

Garlic (Allium sativum) is infected by numerous viruses forming a viral-complex, which is widely distributed in the garlic production regions of Argentina. This work is the first report of the effect of two Allexivirus isolates, Garlic virus A (GarV-A) and Garlic virus C (GarV-C), on garlic yield. Garlic cvs. Morado-INTA and Blanco-IFFIVE were used in the experiments, and four treatments were evaluated: plants inoculated with GarV-A only, GarV-C only, virus-free plants (negative control), and plants infected with the virus-complex. Assays were performed in anti-aphid cages and in the field during 2002 and 2003. GarV-A caused significant reductions in bulb weight (14 to 32%) and diameter (6 to 11%) compared with the negative control in the two cultivars under both assay conditions. GarV-C caused less damage than GarV-A (15% in weight and 5% in diameter) with respect to the negative control in cv. Blanco-IFFIVE, and did not produce significant yield losses in cv. Morado-INTA in either year or under either assay condition.