Caroline Regina Dias Machado, Matheus Azambuja, Camila Domit, Gabriel Fraga da Fonseca, Larissa Glugoski, Camilla Borges Gazolla, Rafael Bonfim de Almeida, Marcela Baer Pucci, Thais Torres Pires, Viviane Nogaroto and Marcelo Ricardo Vicari

Correspondence: vicarimr@uepg.br

This supporting information includes:

• Table S1
• Table S2
• Table S3
• Table S4
• Fig. S1
• Fig. S2
• Fig. S3
• Fig. S4
• Fig. S5

Table S1
To perform this study we used biological samples obtained from four sea turtle species that nidificate in different places on the Brazilian coast (Chelonia mydas, N = 27: 1 female and 26 juveniles that is, sex unidentified; Caretta caretta, N = 12: 2 males, 5 females, and 5 juveniles; Eretmochelys imbricata, N = 6: 2 females and 4 juveniles; and, Lepidochelys olivacea, N = 6: 1 male and 5 juveniles). The blood samples from these individuals were used in the methods. The sea turtle samples were obtained from the different places that work with sea turtle rehabilitation or from wildlife by-catch expeditions and during the process of nidification. The place where each sea turtle was sampled was summarized in Table S1.

Table S2
Sea turtles species-specific alleles (pure lines) corresponding to mitochondrial DNA sequences (mtDNA) and nuclear DNA (nucDNA) were mined from the GenBank database. Thus, the accession number of the mtDNA 12S and 16S, and nucDNA of brain-derived neurotrophic factor - BDNF, oocyte maturation factor - Cmos, and oRNA fingerprint protein 35 gene - R35 were provided in Table S2. These sequences were used in our molecular data analysis comparatively to the gene sequences obtained from the hybrids in the study.

Table S3
Table S3 summarizes the diversity indices for each analyzed gene analyzed in this study (12S, 16S, BDNF, Cmos, and R35). Thus, the table shows detailed data on the number of sequences, haplotypes, polymorphic sites, haplotype diversity, and nucleotide diversity.

Table S4
Partial 5S rDNA sequences were obtained from the pure species of C. mydas, C. caretta, E. imbricata, and L. olivacea. The sequences identity were checked in different databases, including BLASTn algorithm and the Rfam. The data confirm the sequences belonging to the 5S rDNA gene in each sea turtle species.

Fig. S1
In this file, we provide representative idiograms from G-band method for Chelonia mydas, Caretta caretta, Eretmochelys imbricata, and Lepidochelys olivacea. The readers can use these images to verify the interspecific differences in chromosome morphology among chromosome pairs 4, 5, 7, and 12. All data used to draw the ideogram were according to images published by Machado et al. (2020a).

Fig. S2
In this file, we provide representative idiograms from repetitive DNA units chromosomal locations for Chelonia mydas, Caretta caretta, Eretmochelys imbricata, and Lepidochelys olivacea. The readers can use these images to verify the interspecific differences in the number and chromosome location of the rDNA and microsatellite sites. All data used to draw the ideogram were according to images published by Machado et al. (2020b).

Fig. S3
Metaphases used in Figure 4 edition are provided in this file. Here, the reader can find the metaphases from C. mydas (CMY, images a-f), C. caretta (CCA, images g-l), E. imbricata (EIM, images m-r), and L. olivacea (LOL, images s-x) pure lineages subjected to FISH using 5S (red signals) and 18S rDNAs (green signals) probes. The images are sequentially presented, i.e., DAPI, rDNA probe, and merged image.

Fig. S4
Metaphases from the Hybrid A used in Figure 5 karyotyping. The images are ordered according to what appears in Figure 5. For fluorescence in situ images, all layers are provided: DAPI, red channel, green channel (exclusively to 18S rDNA probing), and merged images. Excessive background may have been deleted in the final image for clearance.

Fig. S5
Metaphases from the Hybrid B used in Figure 6 karyotyping. The images are ordered according to what appears in Figure 6. For fluorescence in situ images, all layers are provided: DAPI, red channel, green channel (exclusively to 18S rDNA probing), and merged images. Excessive background may have been deleted in the final image for clearance.

References

• Machado CRD, Glugoski L, Domit C, Pucci MB, Goldberg DW, Marinho LA et al. (2020a) Comparative cytogenetics of four sea turtle species (Cheloniidae): G-banding pattern and in situ localization of repetitive DNA units. Cytogenet Genome Res 160:531-538
• Machado CRD, Domit C, Pucci MB, Gazolla CB, Glugoski L, Nogaroto V et al. (2020b) Heterochromatin and microsatellites detection in karyotypes of four sea turtle species: Interspecific chromosomal differences. Genet Mol Biol 43:e20200213