Authors
Marcelo
Helguera
,
Instituto de Investigaciones en Ingeniería Genética y Biología Molecular (INGEBI-CONICET), Vuelta de Obligado 2490, 1428, Buenos Aires, Argentina, and Instituto de Fitopatología y Fisiología Vegetal (IFFIVE-INTA), Camino 60 Cuadras Km 5 1/2, CP 5014, Córdoba, Argentina
;
Fernando
Bravo-Almonacid
,
INGEBI-CONICET and FCEyN-UBA
;
Ken
Kobayashi
,
INGEBI-CONICET and FCEyN-UBA
;
Pablo D.
Rabinowicz
,
INGEBI-CONICET and FCEyN-UBA
;
Vilma
Conci
,
IFFIVE-INTA
; and
Alejandro
Mentaberry
,
INGEBI-CONICET and FCEyN-UBA
ABSTRACT
A cDNA library representing the genomes of several garlic viruses was generated using a viral RNA mixture as template. Analysis of randomly selected clones allowed the identification of different viral genomic sequences. On the basis of amino acid and nucleotide sequence comparisons, one of them was assigned to garlic virus A (GarV-A), a novel flexuous, rod-shaped virus recently reported in Japan. The coat protein (CP) of this virus was expressed in Escherichia coli cells and used as immunogen to produce polyclonal antibodies. The expression protein was recognized by an antiserum detecting garlic mite-borne filamentous virus, but did not react with antibodies specific for other garlic viruses. Antibodies raised against the viral CP reacted with extracts infected with garlic mite-borne viruses and fail to recognize preparations of onion yellow dwarf potyvirus, leek yellow stripe potyvirus, and carnation latent carlavirus. The same antibodies decorated viral particles exhibiting a modal length of 586 nm in immuno electron microscopy with decoration assays. Double-antibody enzyme-linked immunosorbent assays and tissue printing assays performed with these antibodies allowed detection of GarV-A in most garlic cultivars used in Argentina.
