Cytogenetic studies in Trachymyrmex holmgreni Wheeler, 1925 (Formicidae: Myrmicinae) by conventional and molecular methods

Luísa Antônia Campos Barros, Gisele Amaro Teixeira, Hilton Jeferson Alves Cardoso de Aguiar, Denilce Meneses Lopes, Silvia das Graças Pompolo


Over the past several decades, ant cytogenetic studies have focused on chromosome number and morphology; however, recently, additional information concerning heterochromatin composition and 45S rDNA location has become accessible. The fungus-growing ants are a peculiar ant group that cultivates fungus for food, and Trachymyrmex is suspected to be the sister group of leafcutter ants. Cytogenetic data are so far available for sixn Trachymyrmex species. The present study aimed to increase the knowledge about Trachymyrmex cytogenetics by the chromosomal characterization of Trachymyrmex holmgreni including the karyotyping, fluorochromes staining, 18S rDNA, and microsatellite (GA)15 fluorescence in situ hybridization (FISH). Karyotyped samples from four ant colonies showed 2n = 20 metacentric chromosomes. Centromeric heterochromatin rich in GC base pairs was detected in all chromosomes. FISH revealed the presence of rDNA clusters on the fourth chromosome pair, and an intense spreading of the microsatellite (GA)15 including exclusively euchromatic areas of the chromosomes. The GC-rich heterochromatin observed in different ant species may have a common origin and, thus, phylogenetic implication that needs to be further investigated. To the best of our knowledge, this study is the first report of the use of chromosomal physical location of repetitive DNA sequences by means of microsatellite probes in Formicidae.


heterochromatin; FISH; ribosomal genes; microsatellite repeats; ant

Full Text:



Aguiar, H. J. A. C., Barros, L. A. C., Mariano, C. S. F., Delabie, J. H. C., Pompolo, S. G. (2011). 45S rDNA localization for the giant ant Dinoponera gigantea (Perty, 1833) with evolutionary inferences for Dinoponera genus (Formicidae: Ponerinae). Sociobiology 57: 607-620.

Aguiar, H. J. A. C., Barros, L. A. C., Alves, D. R., Mariano, C. S. F., Delabie, J. H. C., Pompolo, S. G. (2017). Cytogenetic studies on populations of Camponotus rufipes (Fabricius, 1775) and Camponotus renggeri Emery, 1894 (Formicidae: Formicinae). PLoS ONE 12: e0177702.

Barros, L. A. C., Aguiar, H. J. A. C., Mariano, C. S F., Delabie, J. H. C., Pompolo, S. G. (2010). Cytogenetic Characterization of the Lower-Attine Mycocepurus goeldii (Formicidae: Myrmicinae: Attini). Sociobiology 56: 57-66.

Barros, L. A. C., Mariano, C. S. F., Pompolo, S. G., Delabie, J. H. C. (2011). Citogenética de Attini. In: Della-Lucia TMC (Ed) Formigas cortadeiras: da bioecologia ao manejo. 1st edn. Universidade Federal de Viçosa, Viçosa – MG, Brazil, pp 68-79.

Barros, L. A. C., Aguiar, H. J. A. C., Andrade-Souza, V., Mariano, C. S. F., Delabie, J. H. C., Pompolo, S. G. (2012). Occurrence of pre-nucleolar bodies and 45S rDNA location on the chromosomes of the ant Mycocepurus goeldii (Forel) (Formicidae, Myrmicinae, Attini). Hereditas 149: 50-54. doi: 10.1111/j.1601-5223.2011.02237.x

Barros, L. A C., Mariano, C. S. F., Pompolo, S. G. (2013). Cytogenetic studies of five taxa of the tribe Attini (Formicidae: Myrmicinae). Caryologia 66: 59-64. doi:

Barros, L. A. C., Aguiar, H. J. A. C., Mariano, C. S. F., Delabie, J. H. C., Pompolo, S. G. (2014a). Cytogenetic characterization of the ant Trachymyrmex fuscus Emery, 1934 (Formicidae: Myrmicinae: Attini) with the description of a chromosomal polymorphism. Annales de la Société Entomologique de France 49: 367-373. doi:

Barros, L. A. C., Teixeira, G. A., Aguiar, H. J. A. C., Mariano, C. S. F., Delabie, J. H. C., Pompolo, S. G. (2014b. Banding patterns of three leafcutter ant species of the genus Atta (Formicidae: Myrmicinae) and chromosomal inferences. Florida Entomologist 97: 1694-1701. doi:

Barros, L.A. C., Aguiar, H. J. A. C., Teixeira, G. A., Mariano, C S. F., Delabie, J. H. C., Pompolo, S. G. (2015). Cytogenetic data on the threatened leafcutter ant Atta robusta Borgmeier, 1939 (Formicidae: Myrmicinae: Attini). Comptes Rendus Biologies 38:

-665. doi: 10.1016/j.crvi.2015.07.006

Barros, L. A. C., Aguiar, H. J. A. C., Mariano, C. S. F., Andrade-Souza, V., Costa, M. A., Delabie, J. H. C., Pompolo SG (2016). Cytogenetic data on six leafcutter ants of the genus Acromyrmex Mayr, 1865 (Hymenoptera, Formicidae, Myrmicinae): insights into chromosome evolution and taxonomic implications. Comparative Cytogenetics 10: 229-243. doi: 10.3897/CompCytogen.v10i2.7612

Bolton, B. (2017). An online catalog of the ants of the world. Available from Acessed 11 June 2017.

Brandão, C. R. F., Mayhé-Nunes, A., Sanhudo, C. E. D. (2011). Taxonomia e Filogenia das Formigas-Cortadeiras. In: DELLA LUCIA, T.M.C. Formigas-Cortadeiras da Bioecologia ao Manejo. Viçosa, MG: Ed. UFV. p. 27-48.

Chapela, I. H., Rehner, S. A., Schultz, T. R., Mueller, U. G. (1994). Evolutionary history of the symbiosis between fungus-growing ants and their fungi. Science 226: 1691-1694. doi: 10.1126/science.266.5191.1691

Cioffi M.B., Kejnovsky E., Bertollo L.A.C. (2010). The Chromosomal Distribution of Microsatellite Repeats in the Genome of the Wolf Fish Hoplias malabaricus, Focusing on the Sex Chromosomes. Cytogenetic and Genome Research, 132: 289–296. doi: 10.1159/000322058.

Correia, J. P. S. O., Mariano, C. S. F., Delabie, J. H. C., Lacau, S., Costa, M. A. (2016). Cytogenetic analysis of Pseudoponera stigma and Pseudoponera gilberti (Hymenoptera: Formicidae: Ponerinae): a taxonomic approach. Florida Entomologist 99: 718-721. doi:

Cristiano, M. P., Cardoso, D. C., Fernandes-Salomão, T. M. (2013). Cytogenetic and molecular analyses reveal a divergence between Acromyrmex striatus (Roger, 1863) and other congeneric species: taxonomic implications. PloS ONE 8: e59784. doi: 10.1371/journal.pone.0059784

Delabie, J. H. C., Fernandez, F., Majer, J. D. (2012). Editorial Advances in Neotropical Myrmecology. Psyche 2012: 1-3. doi: 10.1155/2012/286273

Imai, H., Taylor, R. W., Crosland, M. W., Crozier, R. H. (1988). Modes of spontaneous chromossomal mutation and karyotype evolution in ants with reference to the minimum interaction hypothesis. Japanese Journal of Genetics 63: 159-185.

Imai, H. T., Taylor, R. W., Crozier, R. H. (1994). Experimental bases for the minimum interaction theory. Chromosome evolution in ants of the Myrmecia pilosula species complex (Hymenoptera: Formicidae: Myrmeciinae). Japanese Journal of Genetics 69: 137-182.

Kubat, Z., Hobza, R., Vyskot, B., Kejnovsky, E. (2008). Microsatellite accumulation in the Y chromosome of Silene latifolia Genome, 51: 350–356. doi: 10.1139/G08-024

Levan, A., Fredga, K., Sandberg, A. (1964). Nomenclature for centromeric position on chromosomes. Hereditas 52: 201-220. doi: 10.1111/j.1601-5223.1964.tb01953.x

Mariano, C. S. F, Pompolo, S. G., Barros, L. A. C., Mariano-Neto, E., Campiolo, S., Delabie, J. H. C. (2008). A biogeographical study of the threatened ant Dinoponera lucida Emery (Hymenoptera: Formicidae: Ponerinae) using a cytogenetic approach. Insect Conservation and Diversity 1: 161-168. doi: 10.1111/j.1752-4598.2008.00022.x

Mayhé-Nunes, A. J., Jaffé, K. (1998). On the biogeography of Attini (Hymenoptera: Formicidae). Ecotrópicos 11: 45-54.

Milani, D., Cabral-de-Mello, D. C. (2014). Microsatellite organization in the grasshopper Abracris flavolineata (Orthoptera: Acrididae) revealed by FISH mapping: remarkable spreading in the A and B chromosomes. PLoS One 9: e97956. doi:

Murakami, T., Fujiwara, A., Yoshida, M. C. (1998) Cytogenetics of ten ant species of the tribe Attini (Hymenoptera, Formicidae) in Barro Colorado Island, Panama. Chromosome Science 2: 135-139.

Palacios-Gimenez, O. M., Carvalho, C. R., Soares, F. A. F., Cabral-de-Mello, D. C. (2015) Contrasting the chromosomal organization of repetitive DNAs in two Gryllidae crickets with highly divergent karyotypes. PLoS ONE 10: e0143540. doi:

Palacios-Gimenez, O. M., Cabral-de-Mello, D. C. (2015) Repetitive DNA chromosomal organization in the cricket Cycloptiloides americanus: a case of the unusual X1X20 sex chromosome system in Orthoptera. Molecular Genetics and Genomics 290: 623-631. doi: 10.1007/s00438-014-0947-9.

Pereira, J. O. P. (2006). Diversidade genética da abelha sem ferrão Melipona quinquefasciata baseada no sequenciamento das regiões ITS1 parcial e 18S do DNA ribossômico nuclear. Thesis, Universidade Federal do Ceará, Brazil.

Pinkel, D., Straume, T., Gray, J. W. (1986). Cytogenetic analysis using quantitative, high-sensitivity, fluorescence hybridization. Proceedings of the National Academy of Sciences 83: 2934-2938.

Schweizer, D. (1980) Simultaneous fluorescent staining of R bands and specific heterocromatic regions (DA/DAPI-bands) in human chromosomes. Cytogenetics and Cell Genetics 27: 190-193. PMID: 6156801

Santos, I. S., Costa, M. A., Mariano, C. S., Delabie, J. H. C., Andrade-Souza, V., Silva, J. G. (2010). A cytogenetic approach to the study of Neotropical Odontomachus and Anochetus ants (Hymenoptera: Formicidae). Annals of the Entomological Society of America 103: 424-429. doi: 10.1603/AN09101

Santos, I. S., Mariano, C. S. F., Delabie, J. H. C., Costa, M. A.,

Carvalho, A. F., Silva, J. G. (2016). "Much more than a neck": karyotype differentiation between Dolichoderus attelaboides (Fabricius, 1775) and Dolichoderus decollates F. Smith, 1858 (Hymenoptera: Formicidae) and karyotypic diversity of five other Neotropical species of Dolichoderus Lund, 1831. Myrmecological News 23:61-69.

Schultz, T. R., Meier, R. (1995). A phylogenetic analysis of the fungus-growing ants (Hymenoptera: Formicidae: Attini) based on morphological characters of the larvae. Systematic Entomology 20(4):337-370. doi: 10.1111/j.1365-3113.1995.tb00100.x

Schultz TR, Brady SG (2008). Major evolutionary transitions in ant agriculture. Proceedings of the National Academy of Sciences 105: 5435-5440.

Schweizer, D. (1980). Simultaneous fluorescent staining of R bands and specific heterocromatic regions (DA/DAPI-bands) in human chromosomes. Cytogenetics and Cell Genetics 27: 190-193. doi: 10.1159/000131482

Sumner, A. T. (1972). A simple technique for demonstrating centromeric heterochromatin. Experimental Cell Research 83: 438-442.

Sumner, A. T. (2003). Chromosomes: Organization and Function. Blackwell Bublishing, North Berwick – United Kingdom.

Teixeira, G. A., Barros, L. A. C., Aguiar, H. J. A. C., Pompolo, S. G. (2017) Comparative physical mapping of 18S rDNA in the karyotypes of six leafcutter ant species of the genera Atta and Acromnyrmex (Formicidae: Myrmicinae). Genetica 145: 351-357. doi 10.1007/s10709-017-9970-1



  • There are currently no refbacks.

JCR Impact Factor 2018: 0.604