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Cytogenetic studies on fungus-farming ants have shown remarkable karyotype diversity, suggesting different chromosomal rearrangements involved in karyotype evolution in some genera. A notable cytogenetic characteristic in this ant group is the presence of GC-rich heterochromatin in the karyotypes of some ancient and derivative species. It was hypothesized that this GC-rich heterochromatin may have a common origin in fungus-farming ants, and the increase in species studied is important for understanding this question. In addition, many genera within the subtribe  Attina  have few or no cytogenetically studied species; therefore, the processes that shaped their chromosomal evolution remain obscure. Thus, in this study, we karyotyped, through classical and molecular cytogenetic techniques, the fungus-farming ants    Cyphomyrmex   transversus    Emery, 1894,    Sericomyrmex   maravalhas    Ješovnik et Schultz, 2017, and    Mycetomoellerius   relictus    (Borgmeier, 1934), to provide insights into the chromosomal evolution in these genera and to investigate the presence the GC-rich heterochromatin in these species.    Cyphomyrmex   transversus    (2n = 18, 10m + 2sm + 6a) and    S.   maravalhas    (2n = 48, 28m + 20sm) showed karyotypes distinct from other species from their genera.    Mycetomoellerius   relictus    (2n = 20, 20m) presented the same karyotype as the colonies previously studied. Notably,    C.   transversus    presented the lowest chromosomal number for the genus and a distinct karyotype from the other two previously observed for this species, showing the existence of a possible species complex and the need for its taxonomic revision. Chromosomal banding data revealed GC-rich heterochromatin in all three species, which increased the number of genera with this characteristic, supporting the hypothesis of a common origin of GC-rich heterochromatin in  Attina . Although a single chromosomal pair carries rDNA genes in all studied species, the positions of these rDNA clusters varied. The rDNA genes were located in the intrachromosomal region in    C.   transversus    and    M.   relictus   , and in the terminal region of    S.   maravalhas   . The combination of our molecular cytogenetic data and observations from previous studies corroborates that a single rDNA site located in the intrachromosomal region is a plesiomorphic condition in  Attina . In addition, cytogenetic data obtained suggest centric fission events in    Sericomyrmex    Mayr, 1865, and the occurrence of inversions as the origin of the location of the ribosomal genes in    M.   relictus    and    S.   maravalhas   . This study provides new insights into the chromosomal evolution of fungus-farming ants.

comparative cytogenetics

Distribution of GC-rich heterochromatin and ribosomal genes in three fungus-farming ants (  Myrmicinae  ,   Attini  ,  Attina ): insights on chromosomal evolution