Some sexual traits, including genitalia, have undergone coevolutionary diversification toward exaggerated states in both sexes among closely related species, but the underlying genetic mechanisms that allow correlated character evolution between the sexes are poorly understood. Here, we studied interspecific differences in gene expression timing profiles involved in the correlated evolution of corresponding male and female genital parts in three species of ground beetle in Carabus (Ohomopterus). The male and female genital parts maintain morphological matching, whereas large interspecific variation in genital part size has occurred in the genital coevolution between the sexes toward exaggeration. We analyzed differences in gene expression involved in the interspecific differences in genital morphology using whole transcriptome data from genital tissues during genital morphogenesis. We found that the gene expression variance attributed to sex was negligible for the majority of differentially expressed genes, thus exhibiting sex-concordant expression, although large variances were attributed to stage and species differences. For each sex, we obtained co-expression gene networks and hub genes from differentially expressed genes between species that might be involved in interspecific differences in genital morphology. These gene networks were common to both sexes, and both sex-discordant and sex-concordant gene expression were likely involved in species-specific genital morphology. In particular, the gene expression related to exaggerated genital size showed no significant intersexual differences, implying that the genital sizes in both sexes are controlled by the same gene network with sex-concordant expression patterns, thereby facilitating the coevolution of exaggerated genitalia between the sexes while maintaining intersexual matching.

To investigate the developmental genetics of genital formation in the carabid beetle Carabus maiyasanus, we compared gene expression patterns among five stages using transcriptomic RNA sequencing data from abdominal segments and genitalia in the third (last) larval instar (including prepupa) and pupal stages. We identified 18 839 genes, of which 10 796 were differentially expressed among stages or between sexes. There were relatively few differentially expressed genes (DEGs) between the sexes (3%). The DEGs were clustered into six groups, mainly according to stage‐specific expression patterns. Genes in clusters 1–3 showed high expression levels before pupation and low expression levels during the pupal period, whereas genes in clusters 4–6 showed high expression levels from the prepupal to the pupal stages. Genes related to the initial pupation process and differentiation of genital discs in Drosophila were involved in clusters 4 and 6 and showed low expression levels at early third instar and elevated expression levels from the early prepupal stage, suggesting that the pupation process and genital differentiation started in the prepupal stage. Clusters 4 and 5 included developmental genes related to organ size control, which may be important in the formation of internal genital structures during the pupal stage.

BackgroundWith the accelerating development of bioscience, the problem of research cost has become important. We previously devised and developed a novel concept microarray with manageable volumes (MMV) using a soft gel. It demonstrated the great potential of the MMV technology with the examples of 1024-parallel-cell culture and PCR experiments. However, its full potential failed to be expressed, owing to the nature of the material used for the MMV chip.ResultsIn the present study, by developing plastic-based MMVs and associated technologies, we introduced novel technologies such as C2D2P (in which the cells in each well are converted from DNA to protein in 1024-parallel), NGS-non-dependent microbiome analysis, and other powerful applications.ConclusionsThe reborn MMV-microarray technology has proven to be highly efficient and cost-effective (with approximately 100-fold cost reduction) and enables us to realize hitherto unattainable technologies.

Background The coevolutionary dynamics of corresponding male and female sexual traits, including genitalia, may be driven by complex genetic mechanisms. Carabus (Ohomopterus) ground beetles show correlated evolution in the size of their functionally corresponding male and female genital parts. To reveal the genetic mechanisms involved in the evolution of size, we investigated interspecific differences in gene expression profiles in four closely related species (two species each with long and short genital parts) using transcriptome data from genital tissues in the early and late pupal stages. Results We detected 1536 and 1306 differentially expressed genes (DEGs) among the species in males and 546 and 1959 DEGs in females in the two pupal stages, respectively. The DEGs were clustered by species-specific expression profiles for each stage and sex to identify candidate gene clusters for genital size based on the expression patterns among the species and gene ontology. We identified one and two gene clusters in females and males, respectively, all from the late pupal stage; one cluster of each sex showed similar expression profiles in species with similar genital size, which implies a common gene expression change associated with similar genital size in each sex. However, the remaining male cluster showed different expression profiles between species with long genital parts, which implies species-specific gene expression changes. These clusters did not show sex-concordant expression profiles for genital size differences. Conclusion Our study demonstrates that sex-independent and partly species-specific gene expression underlies the correlated evolution of male and female genital size. These results may reflect the complex evolutionary history of male and female genitalia.

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