FIG 3.

Molecular characterization of Tomato spotted wilt virus resistance-breaking isolates showing a recovery phenotype obtained in controlled conditions from defined wild-type field isolates. (A and B) Nucleotide alignments of portions of the S genomic segment; mutations occurring in each of the derived isolates are indicated by arrows. (C) Amino acid sequence alignment of the nonstructural protein of isolates p202/3WT and p202/3RB. A single deletion in the nucleotide sequence of p202/3RB resulted in a frameshift and a truncated protein of 443 aa. (D) Amino acid sequence alignment of all the different NSs alleles present in isolates obtained from the WT isolate p105. In isolate p105RBMar, a nucleotide deletion resulted in a frameshift and a truncated protein of 443 aa. A nucleotide mutation in isolate p105RBMaxI resulted in a stop codon, whereas a further mutation resulted in a full-length NSs in isolate p105WTMaxII with an E-to-Q substitution at position 324. Two single nucleotide deletions in p105-803RB caused a frameshift and a truncated protein of 382 aa. (E) Western blot analysis of total protein leaf extracts from Nicotiana benthamiana infected by different isolates, using antinucleocapsid and anti-NSs antibodies.