Supplementary Material for: Cytogenetic Analysis of Lagria villosa (Coleoptera, Tenebrionidae): Emphasis on the Mechanism of Association of the Xyp Sex Chromosomes

The Xy<sub>p</sub> sex determination mechanism is the system most frequent and ancestral to Coleoptera. Moreover, the presence of argyrophilous material associated with the sex bivalent is described as being responsible for the maintenance and association of these chromosomes. There are no karyotype data available regarding the genus <i>Lagria</i> and no consensus in the literature regarding the argyrophilous material present in the lumen of sex bivalent. Therefore, the aim of this work was to investigate the mechanism of sex chromosome bivalent association in <i>Lagria villosa</i> by analyzing the argyrophilous nature of the material present in the Xy<sub>p</sub> lumen. It was also intended to characterize <i>L. villosa</i> cytogenetically. The analysis of meiotic cells showed 2n = 18 = 16+Xy<sub>p</sub> for males and 2n = 18 = 16+XX in females and the meiotic formula was 2n = 8<sup>II</sup>+Xy<sub>p</sub>. The C-banding showed blocks of pericentromeric heterochromatin in all chromosomes except in the y<sub>p</sub> chromosome. In these regions, the use of fluorochromes revealed the presence of heterochromatin containing GC rich DNA sequences. The study of synaptonemal complex showed a gradual increase in the electron-density of the axial elements of the sex chromosomes and their association with strongly electron-dense material. The pepsin pretreatment revealed that the material impregnated by silver is protein.