Table 1.
List of the 11 morphological variables used in the discriminant analysis of principal components (DAPC) analysis.
Fig 1.
DAPC analysis based on 11 morphological traits recorded on 51 slide-fixed specimens.
Species identification by SSR and EF1-α: orange = M. sacchari, blue = M. sorghi.
Fig 2.
Comparison between M. sacchari and M. sorghi using the three traits showing the highest loadings in the DAPC among 51 slide-fixed specimens.
The specimens are assigned to M. sorghi or M. sacchari according to their SSR or EF1-α genotype.
Table 2.
Comparison of morphological characteristics of M. sacchari and M. sorghi apterous females (mean values, with range under brackets).
Fig 3.
The horizontal line is the limit showing the variables (morphological traits) that yield a cumulated 75% contribution to the DAPC. The individual peaks show the magnitude of the influence of each variable on separation of M. sorghi and M. sacchari.
Fig 4.
Assignment of each of the 59 MLG to the two clusters inferred by Structure.
MLG were defined using nine microsatellite markers.
Table 3.
Multi Locus Lineage (MLL) identification of the 51 slide fixed specimens morphologically assigned to M. sorghi or M. sacchari by DAPC.
Table 4.
Haplotypes defined by the concatenation of four genes and correspondence with the Multilocus Lineages (MLLs) defined using SSR.
Fig 5.
Minimum spanning network constructed using the concatenated COI, COII and EF1-α sequences.
The orange and blue boxes indicate the STRUCTURE clusters inferred from SSR data (Table 5). The eight concatenated haplotypes (cH1 to cH8) are listed in Table 5. The number of hatch marks represents the number of mutations separating the concatenated haplotypes. Circle sizes are proportional to haplotype frequencies.
Table 5.
Sequence divergences (pairwise uncorrected P-distances, %) between or within species.
Fig 6.
Molecular identification with SSRs and sequencing of COI or EF1-α of 2,332 specimens: Blue = M. sorghi, orange = M. sacchari.
Data from India is the EF1-α sequence Genbank accession KU048048.1 (exact geographical location within India not available). M. sorghi data from Brazil are EF1- α sequences (M. Kuki and C. Menezes personal communication). The map was drawn using QGIS 3.4 (www.qgis.org). The maps of the administrative boundaries of the countries and states was uploaded from the database of Global Administrative Areas GADM 3.4 (www.gadm.org).
Fig 7.
Molecular diagnosis for separation of M. sacchari and M. sorghi using sequencing of COI or EF1-α.
Fig 8.
Molecular diagnosis for separation of M. sacchari and M. sorghi using the SSR locus CIR-Ms-G01.
M. sacchari (MLL-D) are in lanes A10 (voucher # SNIB00040_0101) and A1 (voucher # SNIB00233_0102). M. sorghi (MLL-F) are in lanes A11 (voucher # SNIB00075_0101) and A5 (voucher # SNIB000237_0102). The PCR was carried out according to [14]. The migration of PCR products was carried out on a Qiagen Qiaxcel electrophoresis analyzer. The image was generated by the Qiaxcel ScreenGel 1.6.0 software (see original image in S1 Raw image).