Redalyc.Genetic Variability in Oligosarcus Paranensis (Teleostei
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Acta Scientiarum. Biological Sciences ISSN: 1679-9283 [email protected] Universidade Estadual de Maringá Brasil Rocha dos Santos, Michele; Renesto, Erasmo Genetic variability in Oligosarcus paranensis (Teleostei: Characiformes) from the São Francisco river, Ivaí river basin – Paraná State, Brazil Acta Scientiarum. Biological Sciences, vol. 35, núm. 3, julio-septiembre, 2013, pp. 389-394 Universidade Estadual de Maringá .png, Brasil Available in: http://www.redalyc.org/articulo.oa?id=187128703012 How to cite Complete issue Scientific Information System More information about this article Network of Scientific Journals from Latin America, the Caribbean, Spain and Portugal Journal's homepage in redalyc.org Non-profit academic project, developed under the open access initiative Acta Scientiarum http://www.uem.br/acta ISSN printed: 1679-9283 ISSN on-line: 1807-863X Doi: 10.4025/actascibiolsci.v35i3.14179 Genetic variability in Oligosarcus paranensis (Teleostei: Characiformes) from the São Francisco river, Ivaí river basin – Paraná State, Brazil Michele Rocha dos Santos* and Erasmo Renesto Departamento de Biologia Celular e Genética, Universidade Estadual de Maringá, Av. Colombo, 5790, 87020-900, Maringá, Paraná, Brazil. *Author for correspondence. E-mail: [email protected] ABSTRACT. The genetic variability of Oligosarcus paranensis was estimated from a population collected in São Francisco river, Prudentópolis county in Paraná State (Brazil) using the electrophoresis in starch gel technique. Eleven enzymatic systems were analyzed: Aspartate aminotransaminase (AAT; E. C. 2.6.1), Alcohol dehydrogenase (ADH; E. C. 1.1.1.1), Esterase (EST; E. C. 3.1.1.1), Glucose-6-phosphate isomerase (GPI; E. C. 5.3.1.9), Glycerol-3-Phosphate dehydrogenase (G3PDH; E. C. 1.1.1), Isocitrate dehydrogenase (IDH; E. C. 1.1.1.42), L-lactate dehydrogenase (LDH; E. C. 1.1.1.27), Malate dehydrogenase (MDH; E. C. 1.1.1.37 ), Malate dehydrogenase NADP (ME; E. C. 1.1.1.40), Phosphoglucomutase (PGM; E. C. 5.4.2.2) and Sorbitol dehydrogenase (SORB; E.C. 1.1.1.14). Twenty loci were identified through 15% corn starch gel electrophoresis of which nine (45%) were polymorphic. The average expected heterozygosity was estimated as 0.1229 ± 0.1728, and the observed was 0.0586 ± 0.1069, indicating high genetic variability. The average value of FIS = 0.5145 indicates homozygote excess. Keywords: allozyme, genetic diversity, heterozygosity, polymorphism, pisces. Variabilidade Genética em Oligosarcus paranensis do rio São Francisco, Bacia do rio Ivaí – Estado do Paraná, Brasil RESUMO. A variabilidade genética de Oligosarcus paranensis foi estimada a partir de uma população coletada no rio São Francisco, município de Prudentópolis no Estado do Paraná (Brasil) utilizando a técnica de eletroforese em gel de amido. Onze sistemas enzimáticos foram analisados: Aspartato aminotransaminase (AAT; E.C. 2.6.1.1), Álcool desidrogenase (ADH; E.C. 1.1.1.1), Esterase (EST; E.C. 3.1.1.1), Glicose-6-fosfato isomerase (GPI; E.C. 5.3.1.9), Glicerol-3-fosfato desidrogenase (G3PDH; E.C. 1.1.1.8), Isocitrato desidrogenase (IDH; E.C. 1.1.1.42), L-Lactato desidrogenase (LDH; E.C. 1.1.1.27), Malato desidrogenase (MDH; E.C. 1.1.1.37), Malato desidrogenase NADP+ (ME; E.C. 1.1.1.40), Fosfoglicomutase (PGM; E.C. 5.4.2.2) e Sorbitol desidrogenase (SORB; E.C. 1.1.1.14). Foram identificados vinte loci por eletroforese em gel de amido de milho 15% dos quais nove (45%) foram polimórficos. A heterozigosidade média esperada foi estimada em 0,1229 ± 0,1728, e a observada foi de 0,0586 ± 0,1069, indicando uma alta variabilidade genética. O valor médio de FIS = 0,5145 indica excesso de homozigotos. Palavras-chave: alozimas, variabilidade genética, heterozigosidade, polimorfismo, peixes. Introduction dominated by the order Characiformes, with comprises approximately 1,200 species and by the Fish are important for human consumption, Siluriformes, with 1,300 species (AGOSTINHO et al., however this importance are preventing many species 2007). Despite the prevalence of these two orders, the to go through the extinction process. Fishes are also composition and number of species among different indicators of environmental pollution (NELSON, basins varies considerably (AGOSTINHO et al., 2006). They play a key role as components of the 2007). ecosystem, participating in the nutrient cycling and The São Francisco river is a tributary of the Ivaí energy flow, such as staple food for many birds and animals. Due to their impressive diversity of species, river, that is, in turn, a tributary of the Paraná river. the fish fauna is a valuable gene bank, strategy for The fish fauna of the Paraná river basin is estimated in future applications. 310 species (LANGEANI et al., 2007), but this The fish that have a great diversity of shapes, sizes number may be much higher (AGOSTINHO et al., and habitats. Of the 54,711 known species of 2007). Although many species are not yet described, vertebrates, fish are 27,977 of which 11,952 are the antropogenic changes in aquatic environments in freshwater ones (NELSON, 2006). South America is recent decades have been threatening the perpetuation Acta Scientiarum. Biological Sciences Maringá, v. 35, n. 3, p. 389-394, July-Sept., 2013 390 Santos and Renesto of natural populations. To accomplish its conservation, the true value of the fish fauna needs to be urgently better appreciated in the economic, scientific and ecological fields (AGOSTINHO et al., 2007). The Genus Oligosarcus represents 16 species, characterized as agile predators, preying on small fish and insects (RUBERT; MARGARIDO, 2007). They Figure 1. Oligosarcus paranensis collected in the São Francisco can be found in the river basins of the Paraná and river, located at Prudentópolis (Paraná state). Standard Length = Paraguay rivers and in the coast of south and 120.10 mm. southeastern Brazil (MENEZES, 1969). The species Oligosarcus paranensis Menezes and The starch gel was prepared by making a solution of 15% corn starch (Penetrose 50) in buffer solution Géry, 1983, reaches the standard length of 20.5 cm (VAL et al., 1981). The solution was heated to boiling (OLIVEIRA et al., 2002), and has a diploid number of and then poured into a glass plate with 0.6 cm edges. 2n = 50 (RUBERT; MARGARIDO, 2007). Despite After cooling, the solution was consistent with 0.7 cm of its small size and low commercial value it is of great thick gel. The gel remained in the refrigerator for 24 ecological importance because it is a predator of smaller hours. animals. Samples from each fish were loaded onto the gel Abundance is a way to measure the species through strips of paper filter 3mm and subjected to adaptedness - the state of being adapted to their horizontal electrophoresis with a continuous current of environment (FISCH-MULLER, 2003), as well as 200V and 50V at source, measured at the ends of the adaptability (ability to survive and reproduce in gel. The starch gels were sliced horizontally and different environments), which are closely linked to revealed to the desired enzyme. Tissues of gills, white genetic variability of the species. The future of a species muscle, heart, liver and kidney were subjected to diversity depends upon the genetic diversity of that electrophoresis in Tris 0.0135 M/0.043 M citrate, pH species (VIDA, 1994). The greater the genetic diversity 7.0 (diluted 15 times in the gel) and 0.18 M Tris / 0.1 maintained the greater adaptability, and thus, the M borate / EDTA 0.004 M, pH 8.7 (diluted 4 times in probability of survival in an unstable environment. It is the gel) to visualize the expression patterns of enzyme important to estimate the genetic diversity degree in and choose the most suitable tissue for the analysis this species to support conservation programs. population. Since there were no major differences in The objective of this study was to estimate the the expression of enzymes in the two buffers, we used genetic variability of Oligosarcus paranensis in São only Tris / citrate for the population analysis. The loci Francisco river of the Paraná river basin and compare it and alleles were named according to Murphy et al. with other species from the upper Paraná river basin to (1996). The data were analyzed by PopGene 1.31 support preservation programs. (YEH et al., 1997). Material and methods Results Thirty specimens of Oligosarcus paranensis (Figure 1) Through the starch gel electrophoresis technique, were collected just above the São Francisco falls (25° 8’ 11 enzyme systems of Oligosarcus paranensis, collected in São Francisco ri 21.27’’ S/51° 18’4.81’’ W) an approximately 190 meters ver, were analyzed. Twenty loci were detected, high waterfall, in São Francisco river, located at with a total of thirty alleles. Prudentópolis (Paraná State) as shown in Figure 2. The enzyme systems tested were aspartate The São Francisco river is a tributary of the Ivaí river, aminotransaminase (AAT, EC 2.6.1.1), Alcohol that is, in turn, a tributary of the Paraná river. The dehydrogenase (ADH, EC 1.1.1.1), Esterase (EST, EC individuals were collected in February 2007 and were 3.1.1.1), Glucose-6-phosphate isomerase (GPI, EC 5.3. immediately frozen in liquid nitrogen, transported to 1.9), Glycerol-3-phosphate dehydrogenase (G3PDH, the laboratory and stored in liquid nitrogen. EC 1.1.1.8), Isocitrate dehydrogenase (IDH, EC Samples were homogenized with plastic sticks in 1.1.1.42), L-lactate dehydrogenase (LDH, EC 1.1.1.27), 1.5 mL Eppendorf tubes with 100 μL of Tris / HCl Malate dehydrogenase (MDH, EC 1.1. 1.37), Malate 0.02 M, pH 7.5 buffer, in order to be analyzed. Since dehydrogenase NADP + (ME, EC 1.1.1.40), the liver has a lot of fat, carbon tetrachloride was added Phosphoglucomutase (PGM, EC 5.4.2.2), and Sorbitol to liver samples at a ratio of 1:2 (tissue: tetrachloride).