ABSTRACT
The molecular characterization of a tomato leaf curl New Delhi virus (ToLCNDV) isolate, denoted ToLCNDV-Le, is reported. The virus was associated with severe and recurrent outbreaks in protected crops of zucchini squash grown in the Province of Lecce (Apulia, southern Italy). The fully sequenced genome of ToLCNDV-Le consists of two genomic components named DNA-A and DNA-B of 2738 and 2683 nt in size, respectively. Like other ToLCNDV isolates, ToLCNDV-Le DNA-A contains the AV2 and AV1 open reading frames (ORFs) in the virion-sense orientation and five additional ORFs named AC1, AC2, AC3, AC4 and AC5 in the complementary-sense orientation. The DNA-B contains BV1 ORF in the virion-sense orientation and BC1 ORF in the complementary-sense orientation. No DNA betasatellites were found associated with ToLCNDV-Le in naturally infected samples. Phylogenetic analysis clustered ToLCNDV-Le with the ToLCNDV-ES strain of western Mediterranean Basin isolates. Consequently, the ToLCNDV-ES-[IT-Zu-Le18] name is proposed as the descriptor for ToLCNDV-Le. Using recombination detection program RDP4, one putative recombination breakpoint (Rbp) was identified close to nucleotide positions 2197-2727, covering approximately half of the AC1 region, including the AC4 ORF and the 3' UTR. RDP4 indicated the event represents an Rbp of an isolate similar to ToLCNDV [Pk-06] (Acc. No. EF620534) found in Luffa acutangula in Pakistan and identified as putative minor parent into the background of ToLCNDV [BG-Jes-Svr-05] (Acc. No. AJ875157), found in tomato in Bangladesh, and identified as putative major parent. To the best of our knowledge, this is the first report of a ToLCNDV-ES recombinant isolate in the AC1-AC4 region in Italy.
ABSTRACT
Tomato leaf curl New Delhi virus (ToLCNDV) is an emerging begomovirus (Geminiviridae family) listed in the EPPO Alert List 2, present in the Mediterranean area and in Italy, where it was reported in 2015 in Sicilian courgette. The virus is widespread in cucurbits where it causes up to 100% production losses. In 2018, ToLCNDV was isolated in Apulia (southern Italy) in commercial fields of zucchini squash and since then its recurrent outbreaks generated justified concern among growers. Thus, a sustainable and environmentally friendly approach must be adopted. Genetic resistances have been identified in Cucurbita moschata and Luffa cylindrica but, compared to genetic resistance, grafting could provide a faster and more flexible solution because the graft wounding induces tolerance rather than resistance against airborne virus infection. Compared to tolerance, the up-regulation of resistance genes requires energy resources mobilized at the expense of primary metabolism, plant growth, and development. Results of screening among twenty-one local cucurbit cvs. ecotypes and accessions to evaluate tolerance levels against rub-inoculation of ToLCNDV led to the identification of potential rootstocks to attain suitable levels of tolerance against the virus in commercial cucurbit varieties. Cucurbit plants were challenged by a ToLCNDV isolated in Apulia denoted ToLCNDV-Le and evaluated for disease symptoms development and viral DNA accumulation up to 28 days after inoculation. On the basis of disease symptoms developed, plants were classified as tolerant, moderately tolerant, moderately susceptible, and susceptible. Cucumis melo cv. Barattiere did not show any detectable disease symptoms and very low levels of viral DNA accumulation was recorded; thus, it was used as rootstock for some of the remaining cucurbit genotypes that were used as scions. The tolerance trait was transmitted to the otherwise susceptible and moderately susceptible cucurbit genotypes grafted onto the cv. Barattiere. The results of this study suggest practical implications of the approach described.
