A novel tomato spotted wilt virus isolate encoding a noncanonical NSm C118F substitution associated with Sw-5 tomato gene resistance breaking

The nonstructural protein NSm of tomato spotted wilt virus (TSWV) has been identified as the avirulence determinant of the tomato single dominant Sw-5 resistance gene. Although Sw-5 effectiveness has been shown for most TSWV isolates, the emergence of resistance-breaking (RB) isolates has been observed. It is strongly associated with two point mutations (C118Y or T120N) in the NSm viral protein. TSWV-like symptoms were observed in tomato crop cultivars (+Sw-5) in the Baja California peninsula, Mexico, and molecular methods confirmed the presence of TSWV. Sequence analysis of the NSm 118–120 motif and three-dimensional protein modelling exhibited a noncanonical C118F substitution in seven isolates, suggesting that this substitution could emulate the C118Y-related RB phenotype. Furthermore, phylogenetic and molecular analysis of the full-length genome (TSWV-MX) revealed its reassortment-related evolution and confirmed that putative RB-related features are restricted to the NSm protein. Biological and mutational NSm 118 residue assays in tomato (+Sw-5) confirmed the RB nature of TSWV-MX isolate, and the F118 residue plays a critical role in the RB phenotype. The discovery of a novel TSWV-RB Mexican isolate with the presence of C118F substitution highlights a not previously described viral adaptation in the genus Orthotospovirus, and hence, the necessity of further crop monitoring to alert the establishment of novel RB isolates in cultivated tomatoes.     

Genetic diversity,population structure and validation of SSR markers linked to Sw-5 and I-2 genes in tomato germplasm

Tomato is the world’s second largest cultivated vegetable crop. Tomato spotted wilt virus (TSWV) and fusarium wilt (FW) are the two major biotic stresses in India limiting tomato production. Identification and utilization of resistant lines to realize the full genetic potential of varieties for yield gain is an eco-friendly approach. The present research work involved genetic diversity study of 48 genotypes, augmented from different exotic, and indigenous sources belonging to three species using SSR markers. A total of 195 alleles were generated by employing 84 polymorphic markers. The PIC value was ranged from 0.12 to 0.93. Two sub-populations (K = 2) were revealed by model based structure analysis. The cluster analysis using the UPGMA method classified the genotypes into 6 clusters. Pusa Ruby, EC-310310 and EC-620452 were found to be highly diverse. Molecular characterization of 48 genotypes with SSR markers divulged seven genotypes with Sw-5 gene and nine genotypes with I-2 gene showing resistance to TSWV and FW, respectively and further, on artificial screening, they were found to be phenotypically resistant. Out of 195 alleles generated from 84 polymorphic SSR markers, 43 alleles from 26 SSR markers were identified with positive average allele effect distributed across nine chromosomes and positive average allele effect was identified for the average weight of the fruit, the number of fruits formed per plant, and fusarium wilt PDI score. Fruit weight and fruit yield per plant registered a significant and positive correlations. The identified genotypes with varied backgrounds and performances will be very useful as diversified sources in resistant breeding programs of tomato.Supplementary InformationThe online version contains supplementary material available at 10.1007/s12298-021-01037-8.     

Construction of a bacterial artificial chromosome (BAC) library of Lycopersicon esculentum cv. Stevens and its application to physically map the Sw-5 locus

The Sw-5 gene is a dominantly inherited resistance gene in tomato and functional against a number of tospovirus species. The gene has been mapped on chromosome 9, tightly linked to RFLP markers CT220 and SCAR421. To analyse the Sw-5 locus, a BAC genomic library was constructed of tomato cv. Stevens, homozygous for the Sw-5 gene. The library comprised 18 816 clones with an average insert size of 100 kb, corresponding to two genome equivalents. The library was screened by PCR using primers designed for the CT220 and SCAR421 sequences, resulting in a 250 kb contig of known orientation on the long arm of chromosome 9. Using degenerate primers based on homologous sequences in the nucleotide binding site of resistance gene sequences, three discrete PCR fragments obtained from this contig were cloned and sequenced. Analysis of these fragments revealed a high similarity with numerous resistance genes or resistance gene like sequences. The present data indicate that at least three different resistance gene candidate (RGC) sequences are present in the vicinity of marker CT220, supporting the view that a resistance gene family may be responsible for the unusually broad resistance to tospoviruses conferred by the Sw-5 locus. This revised version was published online in June 2006 with corrections to the Cover Date.