Fig 4. Phylogenetic analysis.

The relationship between LmDEFs with other insect defensin was inferred using the Neighbor-Joining method. Ixodes ricinus defensin 2 (tick) was used as the out-group. LmDEFs are marked with solid squares. Accession numbers are written next to insect species. Only representatives for defensin-2 were used. More closely related insect defensins within this group were removed to facilitate phylogenetic analysis and representation; therefore, only different lineages of the defensin2 family were shown. Bootstrap (1000 replicates) values are indicated for each root. The evolutionary distances were computed using the p-distance method and are in the units of the number of amino acid differences per site. The analysis involved 26 amino acid sequences that were aligned by Kalign. All ambiguous positions were removed for each sequence pair. The evolutionary analysis was conducted with MEGA6.