Karyotypic mosaicism and molecular cytogenetic markers in the scleractinian coral Acropora pruinosa Brook, 1982 (Hexacorallia, Anthozoa, Cnidaria)

Abstract

Molecular cytogenetic investigation was carried out on the scleractinian coral, Acropora pruinosa (A. pruinosa). Conventional Giemsa staining techniques for karyotyping, such as G- and C-banding, were conducted. Karyotype analysis showed mosaicism of cells with two different chromosome numbers, 28 and 29, in the same embryo. The C-band positive portions appeared on the centromeres of most chromosomes and along the entire length of the unpaired chromosome 15 in cells with 29 chromosomes. Fluorescence in situ hybridization (FISH) revealed that the loci of rRNA genes (rDNA), 5S and 18S/28S, were located on chromosomes 4 and 2, respectively. Whole genome hybridization (WGH) with sperm DNA showed distinct signals not only on all centromeric regions but also on a whole unpaired chromosome 15. Comparative genomic hybridization (CGH) using DNAs from sperm and unfertilized eggs revealed that the unpaired chromosome 15 has sperm-specific DNA sequences. We therefore hypothesized the existence of a sex-related chromosome in A. pruinosa. In this study, we have presented a tentative karyotype of this coral, based on banding results, and described FISH, WGH, and CGH results. Moreover, we successfully cloned and sequenced three clones; AP-5S for 5S rDNA, AP-18S for 18S/28S rDNA, and AP-unpaired chromosome 15q for the unpaired chromosome 15-specific DNA. These molecular cytogenetic approaches will help establish a more exact karyotype of corals and promote understanding of coral genetics, including chromosome evolution. These findings will help to verify the method of classification of complex scleractinian corals.