Spanish Melon necrotic spot virus Isolate Overcomes the Resistance Conferred by the Recessive nsv Gene of Melon

Melon necrotic spot virus (MNSV, genus Carmovirus, family Tombusviridae) is a worldwide natural pathogen that can cause significant economic losses in cucurbit crops (2). Use of commercial cultivars that carry the recessive nsv gene, the only resistance to MNSV described until now (1), is an effective means of controlling MNSV on melon crops. We report here the appearance of MNSV isolates able to overcome nsv resistance. Typical MNSV symptoms were observed in plants with an nsv/nsv genotype during a survey of melon in Almería (southeastern Spain) in 1999. The presence of MNSV in symptomatic plants was confirmed by standard double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) using commercial antisera (Loewe Biochemica GmbH, Sauerlach, Germany). MNSV was isolated from these samples by three serial single-lesion inoculation passages in melon (cv. Bola de Oro). The isolate obtained (MNSV-264) was characterized by sequencing a fragment of its RNA genome corresponding to most of the capsid protein (CP) gene. Briefly, total RNA extracts (TRI reagent, Sigma Chemical, St. Louis, MO) of MNSV-264-infected plants were used in a reverse transcription polymerase chain reaction (RT-PCR) in which the oligonucleotide for the reverse transcription was 5'-TTAGGCGAGGTAAGCAGTTTC-3' (MA34), and the oligonucleotides for the PCR were MA34 and 5'-ATGGCGATGGTTAAACGC-3' (MA32). MA32 and MA34 were designed based on an alignment of nucleotide sequences of CP genes of diverse MNSV isolates. A DNA product of approximately 1.1 kbp was obtained, purified, and sequenced (GenBank Accession No. AF488692). Sequence comparisons (GCG Software Package, Madison, WI) of MNSV-264 and MNSV-Dutch (2) showed a 93% nucleotide sequence identity and a 96% similarity of the deduced amino acid sequence. Therefore, both isolates appear to be closely related. MNSV-264 was inoculated on melon accessions known to be susceptible (cvs. Panal and Bola de Oro) or resistant (cvs. Primal F1, Planters Jumbo, and PI161375) to common MNSV strains. Inoculations with MNSV-Dutch were used as a control for this experiment. The results showed that MNSV-264 was able to infect plants of all genotypes, whereas MNSV-Dutch infected only plants of the susceptible genotypes. Therefore, MNSV-264 is an isolate able to overcome the resistance conferred by nsv. The genetic determinant of the MNSV-264 virulence on resistant genotypes is currently under investigation. From 55 MNSV isolates obtained from fields in southeastern Spain during the last 3 years, only one could overcome nsv, like MNSV-264. Therefore, resistance-breaking isolates are not frequent in the MNSV populations in Spain. It is plausible that the situation may change as a consequence of the widespread use of commercial melon cultivars with the nsv genotype. A search for new sources of genetic resistance to MNSV isolates like MNSV-264 is in progress in our laboratory. References: (1) D. L. Coudriet et al. J. Am. Soc. Hortic. Sci. 106:789, 1981. (2) C. J. Riviere et al. J. Gen. Virol. 70:3033, 1989.