The red brocket deer Mazama americana is a neotropical species that exhibits extensive karyotype variation under an unvarying morphotype. In order to deduce red brocket deer genetic units for conservation, gene flow between populations, and genetic variation, we initiated a cytogenetic and molecular genetic study based on representative samples from throughout their Brazilian geographic range. These data represent the first cytotaxonomical and molecular systematics, and although sample sizes are limited, our results clearly suggest that red brocket deer populations are significantly differentiated with respect to karyotypes and the mitochondrial sequences analyzed. We clearly recognized 2 independent species, and we will be focusing further research in analyzing the meiotic dynamic to determine the existence of other evolutionarily significant units under the red brocket complex.

The Brazilian dwarf brocket deer (Mazama nana) is the smallest deer species in Brazil and is considered threatened due to the reduction and alteration of its habitat, the Atlantic Rainforest. Moreover, previous work suggested the presence of intraspecific chromosome polymorphisms which may contribute to further population instability because of the reduced fertility arising from the deleterious effects of chromosome rearrangements during meiosis. We used G- and C-banding, and nucleolus organizer regions localization by silver-nitrate staining (Ag-NOR) to investigate the causes of this variation. Mazama nana exhibited eight different karyotypes (2n = 36 through 39 and FN = 58) resulting from centric fusions and from inter and intraindividual variation in the number of B chromosomes (one to six). Most of the animals were heterozygous for a single fusion, suggesting one or several of the following: a) genetic instability in a species that has not reached its optimal karyotypic evolutionary state yet; b) negative selective pressure acting on accumulated rearrangements; and c) probable positive selection pressure for heterozygous individuals which maintains the polymorphism in the population (in contrast with the negative selection for many rearrangements within a single individual)

BackgroundThe red brocket deer, Mazama americana, has at least six distinct karyotypes in different regions of South America that suggest the existence of various species that are today all referred to as M. americana. From an evolutionary perspective, the red brockets are a relatively recent clade that has gone through intense diversification. This study sought to prove the existence of post-zygotic reproductive isolation in deer offspring between distinct chromosome lineages. To achieve this, inter-cytotype and intra-cytotype crosses were performed, which resulted in both F1 hybrid (n = 5) and pure offspring (n = 3) in captivity.ResultsF1 females were analyzed in terms of their karyotypes, ovarian histology, estrous cycles and in vitro embryo production. Pure females presented parameters that were similar to those previously reported for M. Americana; however, the parameters for hybrid females were different. Two hybrids were determined to be sterile, while the remaining hybrids presented characteristics of subfertility.ConclusionsThe results support the existence of well-established reproductive isolation among the most distant karyotype lineages and elucidates the need to define all karyotype variants and their geographical ranges in order to define the number of species of red brocket.

The red brocket (Mazama americana) is a South American deer with a wide geographical distribution that presents different chromosomal variants depending on their location. At least six different cytotypes belonging to two distinct evolutionary lineages have been described. This study aimed to verify the existence of postzygotic reproductive isolation between cytotypes of M. americana by comparative evaluation of pure and hybrid males. Seven 18-month-old bucks were submitted to seminal collection and evaluation and testicle histological evaluations. The pure males showed normal parameters for sperm quality and testicular histology. Hybrids from the same evolutionary lineage (≤3 chromosomes different from the progenitors) showed similar results to pure males, except for the reduced ratio of round spermatids to pachytene spermatocytes. Hybrids between cytotypes of different evolutionary lineages (≥10 chromosomes different from progenitors) presented azoospermia and evidence of testicular degeneration. Despite the striking morphological similarities, we can conclude that populations with more distinct karyotypes possess an effective reproductive barrier; moreover, there is evidence that reproductive isolation mechanisms exist between some closer karyotypes, corroborating the hypothesis that M. americana is best characterized as a superspecies. Thus, the future description of several new species for this taxon is expected, since the tendency is to establish efficient mechanisms of postzygotic reproductive isolation, preventing the introgression and fusion of genomes from different populations through chromosome variation.

ABSTRACT. Deer species of the genus Mazama show significant inter-and intraspecific chromosomal variation due to the occurrence of rearrangements and B chromosomes. Given that carriers of aneuploidies and structural rearrangements often show anomalous chromosome pairings, we here performed a synaptonemal complex analysis to study chromosome pairing behavior in a red brocket deer (Mazama americana) individual that is heterozygous for a Robertsonian translocation, is a B chromosome carrier, and has a multiple sex chromosome system (XY 1 Y 2 ). The synaptonemal complex in spermatocytes showed normal chromosome pairings for all chromosomes, including the autosomal and sex trivalents. The electromicrographs showed homology among B chromosomes since they formed bivalents, but they also appeared as univalents, indicating their anomalous behavior and non-Mendelian Meiotic pairing in a red brocket deer segregation. Thus, synaptonemal complex analysis is a useful tool to evaluate the role of B chromosomes and rearrangements during meiosis on the intraspecific chromosomal variation that is observed in the majority of Mazama species.