Abstract
The genus Pterois includes nine valid species, native to the Red Sea and Indian Ocean throughout the Western Pacific. P. volitans and P. miles are native to the Indo-Pacific, and were introduced into Florida waters as a result of aquarium releases, and have been recently recognized as invaders of the Western Atlantic and Caribbean Sea (Costa Rica to Venezuela). Thus far, cytogenetic studies of the genus Pterois only cover basic aspects of three species, including P. volitans from Indo-Pacific Ocean. Considering the lack of more detailed information about cytogenetic characteristics of this invasive species, the objective of the present study was to investigate the basic and molecular cytogenetic characteristics of P. volitans in Venezuela, and compare the results with those from the original distribution area. For this, the karyotypic characteristics of four lionfish caught in Margarita Island, Venezuela, were investigated by examining metaphase chromosomes by Giemsa staining, C-banding, Ag-NOR, and two-colour-Fluorescent in situ hybridization (FISH) for mapping of 18S and 5S ribosomal genes. Comparing the sequences of the 16S gene of the specimens analyzed, with sequences already included in the Genbank, we corroborated that our specimens identified as P. volitans are in fact this species, and hence exclude the possibility of a misidentification of P. miles. The diploid number was 2n=48 (2m+10sm+36a) with FN=60. Chromosomes uniformly decreased in size, making it difficult to clearly identify the homologues except for the only metacentric pair, and the pairs number two, the largest of the submetacentric series. C-banding revealed only three pairs of chromosomes negative for C-band, whereas all remaining chromosomes presented telomeric and some interstitial C-positive blocks. Only two chromosomes were C-banding positive at the pericentromeric regions. Sequential staining revealed Ag-NOR on the tips of the short arms of chromosome pair number two and the FISH assay revealed that 18S rDNA and 5S rDNA genes are co-located on this chromosome pair. The co-localization of 5S rDNA and 45S rDNA is discussed. Both constitutive heterochromatin and NOR location detected in samples examined in this study, differ from those reported for P. volitans in previous analysis of specimens collected in Indian Ocean (Java), suggesting the occurrence of chromosome microrearrangements involving heterochromatin during the spread of P. volitans.
Mauro Nirchio1*, Nicolás Eheman1,Raquel Siccha-Ramirez3, Ernesto Ron1,Julio E. Pérez2 Anna Rita Rossi4 & Claudio Oliveira3
1Escuela de Ciencias Aplicadas del Mar, Universidad de Oriente, Isla de Margarita, Venezuela; mauro.nirchio@gmail.com
2Instituto Oceanográfico de Venezuela, Universidad de Oriente, Cumaná Venezuela; jeperezr@yahoo.com
3Departamento de Morfologia, Instituto de Biociências Universidade Estadual Paulista, 18618-970 Botucatu, São Paulo, Brazil; claudio@ibb.unesp.br
4Department of Human and Animal Biology, University of Rome “La Sapienza”, via Borelli 50, 00161 Rome, Italy; annarita.rossi@uniroma1.it
References
Aljanabi, S. M. & Martinez, I. (1997). Universal and rapid salt extraction of high quality genomic DNA for PCR based technique. Nucleic Acids Research, 25, 4692-4693.
Arai, R. (2011.) Fish Karyotypes. A Check List. Tokio: Springer.
Beaufort, L. F. & De Briggs, J. C. 1962. The fishes of the Indo-Australian archipelago. I. Leiden: Brill.
Betancur, R. R., Hines, A., Acero, A. P., Ortí, G., Wilbur, A. E., & Freshwater, D.W. (2011). Reconstructing the lionfish invasion: insights into Greater Caribbean biogeography. Journal of Biogeography, 38, 1281-1293.
Caputo, V., Nisi, C. P., Caniglia, M., & Giovannotti, M. (2003). Chromosomal studies of five tropical scorpaeniform fishes (Teleostei, Scorpaenidae). Italian Journal of Zoology, 70, 201-204.
Cioffi, M. B. & Bertollo, L. A. C. (2012). Chromosomal Distribution and Evolution of Repetitive DNAs in Fish. In M. A. Garrido-Ramos (Ed.) Repetitive DNA. (Vol. 7, pp. 197-221). Genome Dyn. Basel: Karger (DOI: 10.1159/000337950).
Courtenay, W. R. (1995) Marine fish introductions in southeastern Florida. American Fisheries Society Introduced Fish Section Newsletter, 2-3.
Dor, M. (1984). CLOFRES, Checklist of the fishes of the Red Sea. Jerusalem: Israel Academy of Science and Humanities.
Ferro, D. A., Neo, D. M., Moreira-Filho, O., & Bertollo, L. A. C. (2001). Nucleolar organizing regions, 18S and 5S rDNA in Astyanax scabripinnis (Pisces, Characidae): populations distribution and functional diversity. Genetica, 110, 55-62.
Freshwater, D. W., Hamner, R. M., Parham, S., & Wilbur, A. E. (2009). Molecular evidence that the lionfishes Pterois miles and Pterois volitans are distinct species. Journal of the North Carolina Academy of Science, 125(2), 39-46.
Fujiwara, A., Abe, S., Yamaha, E., Yamazaki, F., & Yoshida, M. C. (1998). Chromosomal localization and heterochromatin association of ribosomal RNA genes loci and silver stained nucleolar organizer regions in salmonid fishes. Chromosome Research, 6, 463-471.
González, J., Grijalba-Bendeck, M., Acero, P. A., & Betancur, R. (2009). The invasive Indo-Pacific lionfish, Pterois volitans/miles complex, in the southern Caribbean Sea. Aquatic Invasions, 4, 507-510.
Hamner, R. M., Freshwater, D. W., & Whitfield, P. E. (2007). Mitochondrial cytochrome b analysis reveals two invasive species with strong founder effects in the western Atlantic. Journal of Fish Biology, 71 (Sup B), 214-222.
Hett, A. K., Nirchio, M., Oliveira, C., Siccha, Z. R., Rossi, A. R., & Sola, L. (2011). Karyotype characterization of Mugil incilis Hancock, 1830 (Mugiliformes: Mugilidae), including a description of an unusual co-localization of major and minor ribosomal genes in the family. Neotropical Ichthyology, 9(1), 107-112.
Howell, W. & Black, D. (1980). Controlled silver staining of nucleolus organizar regions with a protective colloidal developer: a one-step method. Experientia, 36, 1014-1015.
Inafuku, J., Nabeyama, M., Kikuma, Y., Saitoh, J., Kubota, S., & Kohno, S. (2000). Chromosomal location and nucleotide sequences of 5S ribosomal DNA of two cyprinid species (Osteichthyes, Pisces). Chromosome Research, 8, 193-199.
Kocher, T. D., Thomas, W. K., Meyer, A., Edwards, S. V., Pääbo, S., Villablanca, F. X., & Wilson, A. C. (1989). Dynamics of mitochondrial DNA evolution in animals: Amplification and sequencing with conserved primers. Proceedings of the National Academy of Sciences, 86, 6196-6200.
Lasso-Alcalá, O. M. & Posada, J. M. (2010). Presence of the invasive red lionfish, Pterois volitans (Linnaeus, 1758), on the coast of Venezuela, southeastern Caribbean Sea. Aquatic Invasions, 5, S53-59.
Lee, M. R. & Elder, F. F. B. (1980). Yeast stimulation of bone marrow mitosis for cytogenetic investigations. Cytogenetics and Cell Genetics, 26, 36-40.
Levan, A., Fredga, K., & Sandberg, A. (1964). Nomenclature for centromeric position on chromosomes. Hereditas, 52, 201-220.
Martínez, J. L, Móran, P., Garcia-Vásquez, E., & Pendás, A. M. (1996). Chromosomal localization of the major and minor 5S rRNA genes in the European eel (Anguilla anguilla). Cytogenetics and Cell Genetics, 73,149-152.
Moran, P., Martinez, J., Garcia-Vazquez, E., & Pendas, A. (1996). Sex linkage of 5 S rDNA in rainbow trout (Oncorhynchus mykiss). Cytogenetics and Cell Genetics, 75, 145-150.
Morris, J. A. Jr. & Green, S. J. (2012). Lionfish Research: Current Findings and Remaining Questions. In J.A. Morris Jr. (Ed.) Invasive Lionfish: A Guide to Control and Management (pp. 3-14). Gulf and Caribbean Fisheries Institute Special Publication Series Number 1, Marathon, Florida, USA.
Nirchio, M. & Oliveira, C. (2006). Citogenética de peces. Nueva Esparta, Venezuela: Coordinación de publicaciones del rectorado de la Universidad de Oriente.
Nirchio, M., Oliveira, C., Ferreira, D. C., Rondón, R., Pérez, J. E., Hett, A. K., Rossi, A. R., & Sola, L. (2009). Cytogenetic characterization of Rhomboplites aurorubens and Ocyurus chrysurus, two monotypic genera of Lutjaninae from Cubagua Island, Venezuela, with a review of the cytogenetics of Lutjanidae (Teleostei: Perciformes). Neotropical Ichthyology, 7, 587-594.
Nishikawa, S., Honda, M., & Wakatsuki, A. (1977) Comparative studies on the chromosomes in Japanese fishes–II. Chromosomes of eight species in scorpionfishes. Journal of the Shimonoseki University of Fisheries, 25, 187-191. (In Japanese with English abstract).
Pendas, A. M., Moran, P., Freije, J. P., & Garcia-Vazquez, E. (1994). Chromosome mapping and nucleotide sequence of two tandem repeats of Atlantic salmon 5S rDNA. Cytogenetics and Cell Genetics, 63, 31-36.
Schultz, E. T. (1986). Pterois volitans and Pterois miles: two valid species. Copeia, 3, 686-690.
Semmens, B. X., Buhle, E. R., Salomon, A. K., & Pattengill-Semmens, C. V. (2004). A hotspot of non-native marine fishes: evidence for the aquarium trade as an invasion pathway. Marine Ecology Progress Series, 266, 239-244.
Sumner, A. (1972). A simple technique for demostrating centromeric heterochromatin. Experimental Cell Research, 75, 304-306.
Whitfield, P. E., Gardner, T., Vives, S. P., Gilligan, M. R., Courtenay, W. R., Ray, G. C., & Hare, J. A. (2002). Biological invasion of the Indo-Pacific lionfish Pterois volitans along the Atlantic coast of North America. Marine Ecology Progress Series, 235, 289-297.
Whitfield, P. E., Hare, J. A., David, A. W., Harter, S. L., Muñoz, R. C., & Addison, C. M. (2007). Abundance estimates of the Indo-Pacific lionfish Pterois volitans/miles complex in the Western North Atlantic. Biological Invasions, 9, 53-64.
Zenzes, M. T. & Voiculescu, I. (1975). C-banding pattern in Salmo salar, a species of tetraploid origin. Genetica, 45, 531-536.
Comments
This work is licensed under a Creative Commons Attribution 4.0 International License.
Copyright (c) 2014 Revista de Biología Tropical