Aphid‐borne viruses infecting cultivated watermelon and squash in Spain: Characterization of a variant of cucurbit aphid‐borne yellows virus (CABYV)

Abstract

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Different RNA Elements Control Viral Protein Synthesis in Polerovirus Isolates Evolved in Separate Geographical Regions

“…To study the role in virus multiplication of the 3′-CITEs identified above, we modified two agroinfectious CABYV clones obtained from Spanish isolates, one unpublished cloned in pBIN61 and the other in pLJ89 [ 46 ], on the one hand by deleting its first 60 nt of the 3′-UTR (Δ-CITE), and, on the other hand, by exchanging its CMTE with the CXTE from CABYV-Xinjiang. The multiplication efficiencies of these clones were tested in melon cotyledons and Nicotiana benthamiana leaves.…”

“…Thus, several recombinants between poleroviruses and luteoviruses have been described, and CABYV itself has been proposed to derive from recombinations between polero- and enamo-like viruses [ 57 , 58 , 59 ]. Many CABYV recombinants have been identified, with the Brazilian isolates being the most recent ones [ 6 , 10 , 24 , 46 ].…”

“…The agro-infectious mutant clones based on the binary vector pBIN61, were obtained by digestion of these CABYV-pTOPO mutants and CABYV-pBIN61 with HpaI followed by the exchange of the excised 2460 nt fragments of the 3′-end of the viral genomes (digestion at positions 3125 and 5585), resulting in CABYV-ΔCITE-pBIN61 and CABYV-CXTE-pBIN61. The agro-infectious mutant clones based on the binary vector pJL89 were constructed directly by in vitro mutagenesis performed on the agro-infectious CABYV-pJL89 plasmid CABYV-MEC12.1 [ 46 ]. All plasmids were sequenced ( Supplementary Table S1 ).…”

“…Total RNA was extracted with TRI reagent, purified by phenol-chloroform extraction, and followed by DNAseI treatment (Sigma-Aldrich). Quantification of viral RNA was performed as described by Rabadán et al by one step RT-qPCR (NZYtech, Lisboa, Portugal) using 100 ng total RNA [ 46 ]. The results presented are the average of five independent experiments.…”

Occurrence, Distribution, and Management of Aphid-Transmitted Viruses in Cucurbits in Spain

“…Within the Mediterranean basin, Spain is among the largest producer of cucurbits, with a total of 67,500 ha cultivated in the central and southeastern areas and over 3 million tonnes harvested a year [ 31 ]. Our current observations in these crops suggest that the incidence of aphid-transmitted viruses has increased in the last few seasons [ 32 , 33 , 34 ], concurrently with an increase in the aphid population in the early stages of crop cultivation. Thus, a better understanding of the intricate epidemiology of cucurbit viral diseases will allow the improvement of the management strategies utilized for these pests and the aphid-transmitted viruses in crops [ 35 , 36 ].…”