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The population of Prochilodus lineatus found in the Mogi-Guaçu River is karyotypically polymorphic, carrying acrocentric, metacentric, and submetacentric B chromosomes. The analysis of each B chromosome frequency in this species revealed a variation in the distribution pattern, with the metacentric type having the highest frequency (73.30%), followed by submetacentric (25.22%) and acrocentric B chromosomes (1.48%). The transmission pattern of the supernumerary chromosomes was identified by controlled crosses, and it was shown that the acro- and submetacentric B chromosomes have a transmission pattern below the Mendelian rate (kB = 0.333 and kB = 0.385, respectively), but the metacentric variant has a cumulative transmission pattern (kB = 0.587). These results indicate that the acro- and submetacentric B chromosomes are undergoing an extinction process, while the metacentric B chromosomes appear to be accumulating in frequency with each generation., (© 2016 S. Karger AG, Basel.)

Cytogenetic studies were developed in Prochilodus lineatus (Valenciennes 1836), describing an interesting system of small supernumerary chromosomes. The purpose of this work is to study the frequency and morphology of B chromosomes in individuals from the parental line and the inheritance patterns of these elements in individuals obtained from controlled crosses in the species P. lineatus. The transmission rate of B chromosomes revealed a k(B)=0.388 for the acrocentric type, a kB=0.507 for the metacentric type and a k(B)=0.526 for the submetacentric type. The obtained results raise hypothesis that B-acrocentric chromosomes are involved in an extinction process in this species, while the metacentric and submetacentric supernumerary elements comprises a neutral mechanism and follows a Mendelian transmission rate.

To ascertain the origin of B chromosomes in 2 fish species of the genus Prochilodus, i.e. P. lineatus and P. nigricans, we microdissected them and generated B-specific DNA probes. These probes were used to perform chromosome painting in both species and in 3 further ones belonging to the same genus (P. argenteus, P. brevis and P. costatus). Both probes hybridized with the B chromosomes in P. lineatus and P. nigricans, but with none of the chromosomes in the 5 species. This indicates that the B chromosomes have low similarity with DNAs located in the A chromosomes and suggests the possibility that the B chromosomes in the 2 species have a common origin. The most parsimonious explanation would imply intergeneric hybridization in an ancestor of P. lineatus and P. nigricans yielding the B chromosome as a byproduct, which remained in these 2 species after their phylogenetic origin, but was perhaps lost in other Prochilodus species. This hypothesis predicts that B chromosomes are old genomic elements in this genus, and this could be tested once a species from a relative genus would be found showing homology of its A chromosomes with the B-probes employed here, through a comparison of B chromosome DNA sequences with those in the A chromosomes of this other species.

In Prochilodus lineatus B-chromosomes are visualized as reduced size extra elements identified as microchromosomes and are variable in morphology and number. We describe the specific total probe (B-chromosome probe) in P. lineatus obtained by chromosome microdissection and a whole genomic probe (genomic probe) from an individual without B-chromosome. The specific B-chromosome was scraped and processed to obtain DNA with amplification by DOP-PCR, and so did the genomic probe DNA. Fluorescence in situ hybridization using the B-chromosome probe labeled with dUTP-Tetramethyl-rhodamine and the genomic probe labeled with digoxigenin-FITC permitted to establish that in this species supernumerary chromosomes with varying number and morphology had different structure of chromatin when compared to that of the regular chromosomes or A complement, since only these extra elements were labeled in the metaphases. The present findings suggest that modifications in the chromatin structure of B-chromosomes to differentiate them from the A chromosomes could occur along their dispersion in the individuals of the population.

Prochilodus lineatus is widely used in pisciculture projects, mainly in the south of Brazil. It shows a basic karyotype composed of 2n = 54 chromosomes, in addition to as many as seven supernumerary chromosomes, also known as B-chromosomes. These additional small chromosomes are frequently heterochromatic, vary in number and morphology, and generally have no homology with the A complement. Intensive studies have investigated the function, origin and inheritance of these supernumerary chromosomes. The present study aimed to determine the inheritance pattern of the B-chromosomes resulting from directed cross-fertilization of P. lineatus, Prochilodontidae isolated from the Mogi-Guaçu River, Pirassununga, SP. These cross-fertilization experiments were performed at CEPTA/ICMBio in Pirassununga. The transmission patterns of these micro-chromosomes agreed (K(B)= 0.48) with the expectation of regular meiotic behavior following a Mendelian transmission model (K(B)= 0.5). A non-accumulation process was observed for these B-chromosomes in filial generations. Together, our results indicate that the supernumerary chromosomes in P. lineatus samples from the Mogi-Guaçu River exist in a neutral state and follow a Mendelian inheritance pattern.

The genus Prochilodus includes individuals ranging in size from medium to large, being highly relevant for commercial and subsistence fishing. Prochilodus species have a diploid number of 2n = 54 chromosomes and up to seven supernumerary (B) microchromosomes. Previous research has shown that B frequency increased drastically in the Mogi-Guaçu river population of Prochilodus lineatus in the early 1980s, whereas it remained about constant in the 1990s. Here we analyses B frequency in this population during the 2003-2007 period and have found that frequency has not changed significantly since 1987, and that these B chromosomes do not show the intra-individual variation in number that characterized them in the 1980s. This indicates that these B chromosomes have been neutralized, after their invasion, through their mitotic stabilization.