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In animal species with separate sexes, abnormal individuals with a mix of phenotypically male and phenotypically female body parts are generally indicated as gynandromorphs, whereas individuals with intermediate sexual phenotypic traits are generally indicated as intersexes. However, this distinction, clear as it may seem, is neither universally agreed upon, nor free of critical issues. In consideration of the role of sex anomalies in understanding normal development, we reassess these phenomena of abnormal sexual development, taking into consideration the more recent advances in the study of sex determination and sexual differentiation. We argue that a distinction between gynandromorphism and intersexuality, although useful for descriptive purposes, is not always possible or sensible. We discuss the conceptual and terminological intricacies of the literature on this subject and provide reasons for largely, although not strictly, preferring a terminology based on descriptive rather than causal morphology, that is, on the observed phenotypic patterns rather on the causal process behind them.

In forensics, predicting the sex is a crucial step in identification. Many studies have aimed to find an accurate and fast technique to estimate the sex. This study was conducted to determine the accuracy of volumetric and linear measurements of three-dimensional (3D) images of the mandible obtained from cone beam computed tomography (CBCT) radiographs, using different machine-learning (ML) models for sex identification. The CBCTs of 104 males and 104 females were included in this study. The radiographs were converted to 3D images, and the volume, surface area, and ten linear measurements of the mandible were obtained. The data were evaluated using statistical analysis and five different ML algorithms. All results were considered statistically significant at p < 0.05, and the precision, recall, f1-score, training accuracy, and testing accuracy were used to evaluate the performance of the ML models. All the studied parameters showed statistically significant differences between sexes p < 0.05. The right coronoid-to-gonion linear distance had the highest discriminative power of all the parameters. Meanwhile, Gaussian Naive Bayes (GNB) showed the best performance among all the ML models. The results of this study revealed promising outcomes; the sex can be easily determined, with high accuracy (90%).

In the model system for genetics, Drosophila melanogaster, sexual differentiation and male courtship behavior are controlled by sex-specific splicing of doublesex (dsx) and fruitless (fru). In vitro and in vivo studies showed that female-specific Transformer (TRA) and the non-sex-specific Transformer 2 (TRA2) splicing factors interact, forming a complex promoting dsx and fru female-specific splicing. TRA/TRA2 complex binds to 13 nt long sequence repeats in their pre-mRNAs. In the Mediterranean fruitfly Ceratitis capitata (Medfly), a major agricultural pest, which shares with Drosophila a ~120 million years old ancestor, Cctra and Cctra2 genes seem to promote female-specific splicing of Ccdsx and Ccfru, which contain conserved TRA/TRA2 binding repeats. Unlike Drosophila tra, Cctra autoregulates its female-specific splicing through these putative regulatory repeats. Here, a yeast two-hybrid assay shows that CcTRA interacts with CcTRA2, despite its high amino acid divergence compared to Drosophila TRA. Interestingly, CcTRA2 interacts with itself, as also observed for Drosophila TRA2. We also generated a three-dimensional model of the complex formed by CcTRA and CcTRA2 using predictive approaches based on Artificial Intelligence. This structure also identified an evolutionary and highly conserved putative TRA2 recognition motif in the TRA sequence. The Y2H approach, combined with powerful predictive tools of three-dimensional protein structures, could use helpful also in this and other insect species to understand the potential links between different upstream proteins acting as primary sex-determining signals and the conserved TRA and TRA2 transducers.

Mediterranean Shag (Gulosus aristotelis desmarestii) is a seabird endemic to the Mediterranean and Black Seas, recently included in the IUCN list of threatened Species. Most of the reproductive colonies are hosted in Sardinia and surrounding islets. Bycatch in fishing nets is one of the most significant threats for this population. Our work aimed to assess alterations in the sex ratio caused by bycatch and to study the adaptive response of the population to a skewed adult sex ratio. The sex ratio of Mediterranean Shags found drowned in the gillnets near the colonies and that of the nestlings of the Corcelli (northeast Sardinia) colony was determined using the sex-linked polymorphism of the gene Chromobox-Helicase-DNA-binding 1. The data of the shags found drowned in gillnets evidenced a high mortality rate (83.3%; p < 0.001) and a larger size of males (35% heavier than females, p < 0.05) compared to females, supporting the theory that heavier individuals are able to forage at great depths. With 64.8% of the nestlings being male, the sex ratio of nestlings was statistically different from parity (p < 0.05). Furthermore, it was related to the brood size. In one- and two-chick broods, 73% and 70% of nestlings, respectively, were males, while in three-chick broods, only 33% were males. Our data identify the higher rate of male shags drowned in gillnets as a factor causing an alteration of the sex ratio in the Mediterranean Shag population. According to the Sex Allocation Theory, an adaptive adjustment of sex made by adult females restores the Mendelian sex ratio in the population.

Plant reproduction is a fundamental process on Earth from the perspective of biodiversity, biomass gain, and crop productivity. It is therefore important to understand the sex determination process, and many researchers are investigating the molecular basis of this phenomenon. However, information on the influence of transcription factors (TFs), genes that encode DNA-binding proteins, on this process is limited, although cucumber is a model plant in this regard. In the present study, based on RNA-seq data for differentially expressed genes (DEGs), we aimed to investigate the regulatory TFs that may influence the metabolic processes in the shoot apex containing the forming flower buds. Therefore, the annotation of the genome of the B10 cucumber line was supplemented with the assigned families of transcription factors. By performing ontology analyses of the DEGs, the processes they participate in were identified, and TFs were located among the results. In addition, TFs that have significantly overrepresented targets among DEGs were detected, and sex-specific interactome network maps were generated, indicating the regulatory TFs based on their effects on DEGs and furthermore, on the processes leading to the formation of different-sex flowers. Among the most overrepresented TF families in the sex comparisons were the NAC, bHLH, MYB, and bZIP families. An interaction network analysis indicated the most abundant families among DEGs’ regulatory TFs were MYB, AP2/ERF, NAC, and bZIP, and those with the most significant impact on developmental processes were identified, namely the AP/ERF family, followed by DOF, MYB, MADS, and others. Thus, the networks’ central nodes and key regulators were identified with respect to male, female, and hermaphrodite forms. Here, we proposed the first model of the regulatory network of TFs that influences the metabolism of sex development in cucumber. These findings may help us to understand the molecular genetics and functional mechanisms underlying sex determination processes.

Sex determination is a key developmental process, yet it is remarkably variable across the tree of life. The dipteran family Sciaridae exhibits one of the most unusual sex determination systems in which mothers control offspring sex through selective elimination of paternal X chromosomes. Whereas in some members of the family females produce mixed-sex broods, others such as the dark-winged fungus gnatBradysia coprophilaare monogenic, with females producing single-sex broods. Female-producing females were previously found to be heterozygous for a large X-linked paracentric inversion (X’), which is maternally inherited and absent from male-producing females. Here we assembled and characterized the X’ sequence. As close sequence homology between the X and X’ made identification of the inversion challenging, we developed a k-mer-based approach to bin genomic reads before assembly. We confirmed that the inversion spans most of the X’ chromosome (approximately 55Mb) and encodes around 3500 genes. Analysis of the divergence between the inversion and the homologous region of the X revealed that it originated very recently (

Complementary sex determination (CSD) is a widespread sex determination mechanism in haplodiploid Hymenoptera. Under CSD, sex is determined by the allelic state of one or multiple CSD loci. Heterozygosity at one or more loci leads to female development, whereas hemizygosity of haploid eggs and homozygosity of diploid eggs results in male development. Sexual (arrhenotokous) reproduction normally yields haploid male and diploid female offspring. Under asexual reproduction (thelytoky), diploidized unfertilized eggs develop into females. Thelytoky is often induced by bacterial endosymbionts that achieve egg diploidization by gamete duplication. As gamete duplication leads to complete homozygosity, endosymbiont-induced thelytokous reproduction is presumed to be incompatible with CSD, which relies on heterozygosity for female development. Previously, we excluded CSD in four Asobara (Braconidae) species and proposed a two-step mechanism for Wolbachia-induced thelytoky in Asobara japonica. Here, we conclusively reject CSD in two cynipid wasp species, Leptopilina heterotoma and Leptopilina clavipes. We further show that thelytoky in L. clavipes depends on Wolbachia titer but that diploidization and feminization steps cannot be separated, unlike in A. japonica. We discuss what these results reveal about the sex determination mechanism of L. clavipes and the presumed incompatibility between CSD and endosymbiont-induced thelytoky in the Hymenoptera.

Latent fingerprint is an important crime scene evidence, but it is not always recoverable or technically suitable for analysis with fingerprint patterns. Forensic science has shown that other information can be explored from traces using chemical compounds. Infrared spectroscopy is a nondestructive technique that is widely applied to a variety of forensic evidence. In this work, infrared spectroscopy and partial least square discriminant analysis were used to determine the human sex based on latent fingermark analysis. Fingerprint samples were taken from 42 male and female donors, then kept in either dark or light storage conditions, and the Fourier transform infrared (FTIR) spectra were measured considering a period of up to 30 days from collection. The regions from 3000 to 2800 cm-1 and 1790 to 1150 cm-1 presented the greatest differences in the peak intensities among the two sex groups. The results showed a correct discrimination rate higher than 80%.

Primordial germ cells (PGCs) are germline-restricted embryonic cells that form the functional gametes of the adult animal. The use of avian PGCs in biobanking and producing genetically modified birds has driven research on the in vitro propagation and manipulation of these embryonic cells. In avian species, PGCs are hypothesized to be sexually undetermined at an early embryonic stage and undergo differentiation into an oocyte or spermatogonial fate dictated by extrinsic factors present in the gonad. However, chicken male and female PGCs require different culture conditions, suggesting that there are sex-specific differences, even at early stages. To understand potential differences between male and female chicken PGCs during migratory stages, we studied the transcriptomes of circulatory stage male and female PGCs propagated in a serum-free medium. We found that in vitro cultured PGCs were transcriptionally similar to their in ovo counterparts, with differences in cell proliferation pathways. Our analysis also revealed sex-specific transcriptome differences between male and female cultured PGCs, with notable differences in Smad7 and NCAM2 expression. A comparison of chicken PGCs with pluripotent and somatic cell types identified a set of genes that are exclusive to germ cells, enriched in the germplasm, and associated with germ cell development.

The sexual fate of honeybees is determined by the complementary sex determination (CSD) model: heterozygosity at a single locus (the CSD locus) determines femaleness, while hemizygosity or homozygosity at the CSD locus determines maleness. The csd gene encodes a splicing factor that regulates sex-specific splicing of the downstream target gene feminizer (fem), which is required for femaleness. The female mode of fem splicing occurs only when csd is present in the heteroallelic condition. To gain insights into how Csd proteins are only activated under the heterozygous allelic composition, we developed an in vitro assay system to evaluate the activity of Csd proteins. Consistent with the CSD model, the co-expression of two csd alleles, both of which lack splicing activity under the single-allele condition, restored the splicing activity that governs the female mode of fem splicing. RNA immunoprecipitation quantitative PCR analyses demonstrated that the CSD protein was specifically enriched in several exonic regions in the fem pre-mRNA, and enrichment in exons 3a and 5 was significantly greater under the heterozygous allelic composition than the single-allelic condition. However, in most cases csd expression under the monoallelic condition was capable of inducing the female mode of fem splicing contrary to the conventional CSD model. In contrast, repression of the male mode of fem splicing was predominant under heteroallelic conditions. These results were reproduced by real-time PCR of endogenous fem expression in female and male pupae. These findings strongly suggest that the heteroallelic composition of csd may be more important for the repression of the male splicing mode than for the induction of the female splicing mode of the fem gene.

The evolution of sex determination (SD) mechanisms in teleost fishes is amazingly dynamic, as reflected by the variety of different master sex-determining genes identified, even sometimes among closely related species. Pangasiids are a group of economically important catfishes in many South-Asian countries, but little is known about their sex determination system. Here, we generated novel genomic resources for 12 Pangasiid species and provided a first characterization of their SD system. Based on an Oxford Nanopore long-read chromosome-scale high quality genome assembly of the striped catfish Pangasianodon hypophthalmus, we identified a duplication of the anti-Müllerian hormone receptor type II gene (amhr2), which was further characterized as being sex-linked in males and expressed only in testicular samples. These first results point to a male-specific duplication on the Y chromosome (amhr2by) of the autosomal amhr2a. Sequence annotation revealed that the P. hypophthalmus Amhr2by is truncated in its N-terminal domain, lacking the cysteine-rich extracellular part of the receptor that is crucial for ligand binding, suggesting a potential route for its neofunctionalization. Short-read genome sequencing and reference-guided assembly of 11 additional Pangasiid species, along with sex-linkage studies, revealed that this truncated amhr2by duplication is also conserved as a male-specific gene in many Pangasiids. Reconstructions of the amhr2 phylogeny suggested that amhr2by arose from an ancient duplication / insertion event at the root of the Siluroidei radiation that is dated around 100 million years ago. Altogether these results bring multiple lines of evidence supporting that amhr2by is an ancient and conserved master sex-determining gene in Pangasiid catfishes, a finding that highlights the recurrent usage of the transforming growth factor β pathway in teleost sex determination and brings another empirical case towards the understanding of the dynamics or stability of sex determination systems.

Background and Aims The largest genus of Salicaceae sensu lato, Salix, has been shown to consist of two main clades: clade Salix, in which species have XY sex-determination systems (SDSs) on chromosome 7, and clade Vetrix including species with ZW SDSs on chromosome 15. Here, we test the utility of whole genome re-sequencing (WGR) for phylogenomic reconstructions of willows to infer changes between different SDSs. Methods We used more than 1 TB of WGR data from 70 Salix taxa to ascertain single nucleotide polymorphisms on the autosomes, the sex-linked regions (SLRs) and the chloroplast genomes, for phylogenetic and species tree analyses. To avoid bias, we chose reference genomes from both groups, Salix dunnii from clade Salix and S. purpurea from clade Vetrix. Key Results Two main largely congruent groups were recovered: the paraphyletic Salix grade and the Vetrix clade. The autosome dataset trees resolved four subclades (C1–C4) in Vetrix. C1 and C2 comprise species from the Hengduan Mountains and adjacent areas and from Eurasia, respectively. Section Longifoliae (C3) grouped within the Vetrix clade but fell into the Salix clade in trees based on the chloroplast dataset analysis. Salix triandra from Eurasia (C4) was revealed as sister to the remaining species of clade Vetrix. In Salix, the polyploid group C5 is paraphyletic to clade Vetrix and subclade C6 is consistent with Argus’s subgenus Protitea. Chloroplast datasets separated both Vetrix and Salix as monophyletic, and yielded C5 embedded within Salix. Using only diploid species, both the SLR and autosomal datasets yielded trees with Vetrix and Salix as well-supported clades. Conclusion WGR data are useful for phylogenomic analyses of willows. The different SDSs may contribute to the isolation of the two major groups, but the reproductive barrier between them needs to be studied.

Despite molting is an important and expensive activity in the life cycle of birds, this process is poorly known for neotropical species and even less so for species inhabiting urban zones. We studied molt patterns, aging and sexing criteria, and morphometrics for seven common resident species in an urban area of Lima city, Peru. All species evidenced complete prebasic molts. Partial to eccentric incomplete preformative molts were evidenced for six species (Camptostoma obsoletum, Pyrocephalus rubinus, Catamenia analis, Poospiza hispaniolensis, Coereba flaveola, and Thraupis episcopus); whereas Columbina cruziana exhibited a complete molt. All thraupid species exhibited partial prealternate molts. Preformative and prebasic molts were recorded mostly during the dry season, and the few records of prealternate molts might suggest its occurrence soon after the wet season. Our species also exhibited low molt synchrony, with some of them (C. cruziana, P. rubinus, and C. flaveola) exhibiting extensive molt periods likely explained by its potential to reproduce throughout the year and/or the existence of suspended/ protracted molts. Morphometric analysis evidenced significant differences according to sex for C. cruziana, P. rubinus, and C. analis, with the wing chord being the most common characteristic for distinguishing between sexes. Our study covers an important gap in the knowledge of the natural history of urban birds and provides information to test hypotheses of molt pattern variation on species with widespread distribution throughout the Americas. RESUMEN A pesar de que la muda constituye una actividad importante y costosa en el ciclo de vida de aves, este proceso es poco conocido en especies neotropicales y lo es aún menos para especies que habitan ambientes urbanos. Estudiamos los patrones de muda, criterios de determinación de edad y sexo y datos mor-fométricos correspondientes a siete especies de aves residentes comunes en un área urbana de la ciudad de Lima, Perú. Todas las especies evidenciaron muda prebásica completa. Mudas preformativa parcial a incompleta excéntrica fueron evidenciadas para Camptostoma obsoletum, Pyrocephalus rubinus, Cata-mema analis, Poospiza hispaniolensis, Coereba flaveola y Thraupis episcopus; mientras que Columbina cruziana evidenció una muda completa. Todos los traúpidos exhibieron mudas prealterna parcial. Las mudas preformativa y prebásica fueron registradas principalmente durante la temporada seca, y los pocos registros de muda prealterna sugieren su ocurrencia poco después de la temporada húmeda. Nuestras especies también exhibieron una baja sincronía de muda, y algunas de ellas (C. cruziana, P. rubinus y C. flaveola) exhibieron períodos extensos de muda probablemente explicados por su potencial para reproducirse durante el año o la existencia de mudas suspendida / prolongada. El análisis morfométrico evidenció diferencias significativas según sexo para C. cruziana, P. rubinus y C. analis, siendo la cuerda alar la característica más común para distinguir sexos. Nuestro estudio cubre un vacío importante en conocimiento de la historia natural de aves urbanas y proporciona información para probar hipótesis sobre la variación de patrones de muda en especies con amplia distribución en las Américas.

Pheromones are pivotal to sexual communication in insects. These chemical signals are processed by sexually dimorphic circuitries in the antennal lobe (AL) of the insect brain. However, there is limited understanding of how these circuitries form during AL development. Our review addresses this issue by comparing how circuitries develop throughout the growth processes of peripheral and deutocerebral neurons in various insect orders. Olfactory sensory neurons (OSNs) expressing novel pheromone receptors are eligible candidates to initiate new sexually-dimorphic circuitries when these OSNs survive programmed cell death and match the physiological properties of pheromone-sensing sensilla. The probability of these OSNs forming new glomeruli is largely determined by the degree of glia-OSN interactions and projection neuron (PN) prepatterning. The relative contribution of either of these processes determines the degree of evolutionary neuroplasticity, which is particularly prevalent in those species with complex ALs lacking specific macroglomerular structures. The extent of sexual dimorphism is determined by sex-determination genes, such as Doublesex and Fruitless, that regulate factors inducing OSN programmed cell death. Currently, these mechanisms are largely unexplored. This review, therefore, aims to provide a solid foundation for ongoing research into the evolution of AL sexual dimorphism and formation of pheromone circuitries in the light of insect sex determination.

Des modifications dans le gène codant pour la sous-unité II du cytochrome c oxydase du génome mâle (Mcox2) ont été recensées chez des espèces de bivalves présentant un mode unique de transmission mitochondriale nommé double transmission uniparentale (DUI). Dans la DUI, les mitochondries paternelles (et leur ADNmt mâle) ainsi que les mitochondries maternelles (et leur ADNmt femelle) sont transmises aux descendants mâles. Scrobicularia plana, une espèce de bivalves présentant ce modèle d'hérédité, possède une insertion importante d'environ 4,8 kb dans son gène Mcox2 qui ne change pas le cadre de lecture et qui est traduite en un polypeptide de 1 892 acides aminés, ce qui en fait la plus grande protéine COX2 connue à ce jour chez les métazoaires. L’objectif de cette étude était de caractériser l'évolution et la fonction potentielle de l'insertion dans Mcox2 chez S. plana par RT-PCR, tests immunologiques et analyses bio-informatiques. L'insertion est présente parmi les individus de différentes populations, contient des variations dans la longueur de sa séquence, est riche en zones de désordre intrinsèque et évolue sous sélection purificatrice. La longue insertion pourrait modifier la structure 3D du complexe IV de la chaîne de transport d'électrons (CTE), affectant sa fonction dans la phosphorylation oxydative (OXPHOS) ce qui pourrait expliquer les faibles taux d'OXPHOS observés dans les mitochondries mâles des bivalves à DUI. L'insertion pourrait également modifier le métabolisme mitochondrial mâle en interagissant avec d'autres complexes de la CTE et avec l'ATP synthase. Comme pour les autres modifications de Mcox2 chez les bivalves à DUI, un rôle potentiel dans la détermination du sexe peut être prédit pour MCOX2 chez S. plana., Modifications in the cytochrome c oxidase subunit II gene of male-transmitted genome (Mcox2) have been found in some bivalve species that exhibit a unique mode of mitochondrial transmission named doubly uniparental inheritance (DUI). In DUI, paternal mitochondria (and their male mtDNA) as well as maternal mitochondria (and their female mtDNA) are transmitted to male offspring. Scrobicularia plana, a bivalve specie exhibiting this inheritance model possesses an important in-frame insertion of approximately 4,8 kb in its Mcox2 gene that is translated into a polypeptide of 1 892 amino acids making it the largest metazoan COX2 protein known to date. The aim of this study was to characterize the evolution and possible function of the Mcox2 insertion in S. plana through RT-PCRs, immunoassays, and bioinformatic analysis. The insertion is present amongst individuals from different populations, contains some variations in its sequence length, is rich in intrinsically disordered regions and evolves under purifying selection. The long insertion could modify the 3D structure of complex IV in the electron transport chain (ETC), impacting its function in oxidative phosphorylation (OXPHOS) which could explain low OXPHOS rates that were found in male mitochondria of DUI bivalves. The insertion could also alter male mitochondrial metabolism by interacting with other complexes of the ETC and with ATP synthase. As for other modifications of Mcox2 in DUI bivalves, a role in sex determination can also be predicted for MCOX2 in S.plana.

Poplar and willow species in the Salicaceae are dioecious, yet have been shown to use different sex determination systems located on different chromosomes. Willows in the subgenus Vetrix are interesting for comparative studies of sex determination systems, yet genomic resources for these species are still quite limited. Only a few annotated reference genome assemblies are available, despite many species in use in breeding programs. Here we present de novo assemblies and annotations of 11 shrub willow genomes from six species. Copy number variation of candidate sex determination genes within each genome was characterized and revealed remarkable differences in putative master regulator gene duplication and deletion. We also analyzed copy number and expression of candidate genes involved in floral secondary metabolism, and identified substantial variation across genotypes, which can be used for parental selection in breeding programs. Lastly, we report on a genotype that produces only female descendants and identified gene presence/absence variation in the mitochondrial genome that may be responsible for this unusual inheritance. Published version

Aim : The aim of the present study is to determine sex of the individuals using mandibular ramus width (right side). Introduction: Sex determination as a cornerstone of identification is at the same time an important aspect of forensic anthropology and in forensic examination. Measurements of the mandibular ramus typically reveal more gender differences than those of the mandibular body, and measurements of the mandibular ramus typically reveal more sexual dimorphism Materials and methods: This study was conducted in the department of Forensic Odontology. A retrospective study was conducted using an orthopantomograph of 60 subjects (30 males and 30 females). Their ages ranged from 41 to 50 years. Results and discussion: In the present study comprising 30 males and 30 females; The mean standard deviation for male is 35.9667 + 4.02264 and the mean + standard deviation for female is 36.7567 + 4.90487. The mandible is considered to be the best bone for determining sex because of its distinct characteristics, which include being the largest, strongest, and sexually dimorphic bone of the skull as well as frequently being recovered substantially intact. The condylar area and mandibular ramus undergo the most morphological transformations during growth in terms of size and remodelling. Conclusion: The study concluded that the minimum mandibular ramus width of females is 36.76 and that of male is 35.96 stating that females have larger mandibular width than males. From the results it can be concluded that minimum mandibular ramus width is one of the best parameters in sex determination.

Aim: The aim of the study is to identify gender using the width of the left side of the mandibular condylar process. Introduction: Forensic anthropology is the branch of forensic science which deals with the physical anthropology of the dead remains and helps in the analysis of the skeletal remains with the anatomical knowledge. The mandible is the strongest and the hardest bone of the skull which remains intact. It provides the accurate information to whom it belongs and its has the greater sexual dimorphism among the other skeletal structures. The condyle is the growing interest of the anthropologist because they eventually reported that the condylar width of males is comparatively more than that of females. Sex determination in forensic is foremost because the age and the stature are resting on each other.To do sex determination, the dead remains and the state of dimorphism play a critical role. There is paucity in literature regarding the width of the condylar process in gender determination. Materials and methods: The study consisted of 60 individuals (30 males, 30 females, Age group: 22- 25 years). This study was performed using Orthopantomographs images acquired from patients who were chosen for a variety of dental complaints and needed to be viewed with OPG. The research plan was confirmed by the administration of the Faculty of Dentistry. The study was carried out at the Department of Forensic odontology, Saveetha Dental college, Chennai. Data were analyzed using the Statistical Package for Social Sciences (SPSS), version 20. Regression analysis and chi square tests were performed. Plan maker software was utilized for measuring the condylar process width from OPGs. Results: The mean absolute error of the study came out to be 1.121. The standard deviation for male is 1.63 and for females is 1.69. Chi- square analysis for gender determination was found to be insignificant. One way anova analysis for gender determination is and it was found to be insignificant. Males have higher condylar process width compared to females. Discussion: Ramesh et al in their study suggested that the parameters such as condylar height and coronoid height using orthopantomographs are reliable in gender determination and concluded that the condylar height of the right side is the best parameter in gender determination In the present study of the chi square analysis, one way anova analysis was found to be insignificant. But the condylar process width (left side) of the males were comparatively higher than females. Therefore, this ideology could help in sex determination. Conclusion: Within the limits of the study it can be concluded that condylar process width ( left side) of male were comparatively higher than females. But the study is not significant due to Sample insufficiency and less time duration. Since this study was circumscribed to a particular region and population, with more sample size, extended period of time for analysis and with different ethnicity can make the study significant.

Geckos are an excellent group to study the evolution of sex determination, as they possess a remarkable variability ranging from a complete absence of sex chromosomes to highly differentiated sex chromosomes. We explored sex determination in the Madagascar leaf-tail geckos of the genus Uroplatus. The cytogenetic analyses revealed highly heterochromatic W chromosomes in all three examined species (Uroplatus henkeli, U. alluaudi, U. sikorae). The comparative gene coverage analysis between sexes in U. henkeli uncovered an extensive Z-specific region, with a gene content shared with the chicken chromosomes 8, 20, 26 and 28. The genomic region homologous to chicken chromosome 28 has been independently co-opted for the role of sex chromosomes in several vertebrate lineages, including monitors, beaded lizards and monotremes, perhaps because it contains the amh gene, whose homologs were repeatedly recruited as a sex-determining locus. We demonstrate that all tested species of leaf-tail geckos share homologous sex chromosomes despite the differences in shape and size of their W chromosomes, which are not homologous to the sex chromosomes of other closely related genera. The rather old (at least 40 million years), highly differentiated sex chromosomes of Uroplatus geckos can serve as a great system to study the convergence of sex chromosomes evolved from the same genomic region.

Identity verification is a crucial component of forensic sciences, as it necessitates accurately determining an individual's sex, age, and ancestry. The mandible is a commonly studied facial bone for sex determination due to its durability and preservation. This study aims to evaluate various parameters in the mandible for sex determination through morphometric analysis of panoramic radiographs. Fifty-seven panoramic radiographs of patients (22 males and 35 females, 18 – 40 years old) were examined in this study. Ten specific parameters were determined as follows: Bi-condylar Breadth (BCD), Bi-Coronoid Breadth (BCR), Bi-Mental Foramen Breadth (BMF), Bi-Mandibular Notch Breadth (BMN), Bi-Gonial Breadth (BGN), Minimum Ramus Breadth (MRB), Symphysis Height (SYM), Ramus Height (RAH), Mandibular Body Length (MBL), and Mandibular Notch Length (MNL). The morphometric analysis of these parameters was evaluated using Adobe Photoshop®CS6 software. The discriminant function analysis showed a significant difference between males and females in two variables: SYM and RAH, with Wilks’ lambda values of 0.857 and 0.924, respectively, and p

Sex determination (SD) shows huge variation among fish and a high evolutionary rate, as illustrated by the Pleuronectiformes (flatfishes). This order is characterized by its adaptation to demersal life, compact genomes and diversity of SD mechanisms. Here, we assembled the Solea senegalensis genome, a flatfish of great commercial value, into 82 contigs (614 Mb) combining long-and short-read sequencing, which were next scaffolded using a highly dense genetic map (28,838 markers, 21 linkage groups), representing 98.9% of the assembly. Further, we established the correspondence between the assembly and the 21 chromosomes by using BAC-FISH. Whole genome resequencing of six males and six females enabled the identification of 41 single nucleotide polymorphism variants in the follicle stimulating hormone receptor (fshr) consistent with an XX/XY SD system. The observed sex association was validated in a broader independent sample, providing a novel molecular sexing tool. The fshr gene displayed differential expression between male and female gonads from 86 days post-fertilization, when the gonad is still an undifferentiated primordium, concomitant with the activation of amh and cyp19a1a, testis and ovary marker genes, respectively, in males and females. The Y-linked fshr allele, which included 24 nonsynonymous variants and showed a highly divergent 3D protein structure, was overexpressed in males compared to the X-linked allele at all stages of gonadal differentiation. We hypothesize a mechanism hampering the action of the follicle stimulating hormone driving the undifferentiated gonad toward testis., Spanish Government 81792 P20-00938 RTI2018-096847-B-C21, European Commission 81792, Junta de Andalucia-FEDER Grant P20-00938, Spanish Government, European Commission RTI2018-096847-B-C21 RTI2018-096847-B- C22, Ministry of Economy and Competitiveness, Junta de Andalucia

Data de publicació electrònica: 01-01-2023 Includes supplementary materials for the online appendix. Sex determination (SD) shows huge variation among fish and a high evolutionary rate, as illustrated by the Pleuronectiformes (flatfishes). This order is characterized by its adaptation to demersal life, compact genomes and diversity of SD mechanisms. Here, we assembled the Solea senegalensis genome, a flatfish of great commercial value, into 82 contigs (614 Mb) combining long- and short-read sequencing, which were next scaffolded using a highly dense genetic map (28,838 markers, 21 linkage groups), representing 98.9% of the assembly. Further, we established the correspondence between the assembly and the 21 chromosomes by using BAC-FISH. Whole genome resequencing of six males and six females enabled the identification of 41 single nucleotide polymorphism variants in the follicle stimulating hormone receptor (fshr) consistent with an XX/XY SD system. The observed sex association was validated in a broader independent sample, providing a novel molecular sexing tool. The fshr gene displayed differential expression between male and female gonads from 86 days post-fertilization, when the gonad is still an undifferentiated primordium, concomitant with the activation of amh and cyp19a1a, testis and ovary marker genes, respectively, in males and females. The Y-linked fshr allele, which included 24 nonsynonymous variants and showed a highly divergent 3D protein structure, was overexpressed in males compared to the X-linked allele at all stages of gonadal differentiation. We hypothesize a mechanism hampering the action of the follicle stimulating hormone driving the undifferentiated gonad toward testis. This study was supported by the Spanish Ministry of Economy and Competitiveness, FEDER Grants (RTI2018-096847-B-C21, RTI2018-096847-B-C22 and RTI2018-097110-B-C21), Junta de Andalucía-FEDER Grant (P20-00938) and the European Union's Horizon 2020 research and innovation programme under grant agreement No. 817923 (AQUA-FAANG). We thank Geneaqua SL for their participation and financial support of sequencing. We acknowledge the bioinformatic support of the Centro de Supercomputación de Galicia (CESGA).

Molecular mechanisms underlying environmental sex determination remained elusive for half a century, until just recently, when new insights into the topic were gained. The thesis summarizes current progress on this issue and focuses, among other things, on the role of oxidative stress and regulation of gene expression in the process of sex determination and sex reversal.

Scincoidea, the reptilian clade that includes girdled lizards, night lizards, plated lizards and skinks, are considered as a lineage with diversity in sex-determining systems. Recently, the hypothesis on the variability in sex determination in skinks and even more the absence of sex chromosomes in some of them has been rivalling. Homologous, evolutionary stable XX/XY sex chromosomes were documented to be widespread across skinks. However, sex determination in the other scincoidean families is highly understudied. ZZ/ZW sex chromosomes have been identified only in night lizards and a single species of plated lizards. It seems that although there are different sex chromosome systems among scincoidean lineages, they share one common trait: they are mostly poorly differentiated and often undetectable by cytogenetic methods. Here, we report one of the exceptions, demonstrating for the first time ZZ/ZW sex chromosomes in the plated lizard Zonosaurus madagascariensis. Its sex chromosomes are morphologically similar, but the W is clearly detectable by comparative genomic hybridization (CGH), suggesting that the Z and W chromosomes highly differ in sequence content. Our findings confirm the presence of female heterogamety in plated lizards and provides novel insights to expand our understanding of sex chromosome evolution in scincoidean lizards.

Paternal factors play an important role in embryonic morphogenesis and contribute to sexual dimorphism in development. To assess the effect of paternal DNA on sexual dimorphism of embryonic genome activation, we compared X and Y sperm and different sexes of embryos before sex determination. Through transcriptome sequencing (RNA-seq) and whole-genome bisulfite sequencing (WGBS) of X and Y sperm, we found a big proportion of upregulated genes in Y sperm, supported by the observation that genome-wide DNA methylation level is slightly lower than in X sperm. Cytokine–cytokine receptor interaction, TGF-beta, and toll-like receptor pathways play important roles in spermatogenesis. Through whole-genome re-sequencing (WGRS) of parental fish and RNA-seq of five early embryonic stages, we found the low-blastocyst time point is a key to maternal transcriptome degradation and zygotic genome activation. Generally, sexual differences emerged from the bud stage. Moreover, through integrated analysis of paternal SNPs and gene expression, we evaluated the influence of paternal inheritance on sexual dimorphism of genome activation. Besides, we screened out gata6 and ddx5 as potential instructors for early sex determination and gonad development in yellow catfish. This work is meaningful for revealing the molecular mechanisms of sex determination and sexual dimorphism of fish species.

Use of morphological characteristics in human identification maybe difficult in conditions where human remains are grossly fragmented, decomposed and/or mutilated, or when only a tooth is available. The study aims to ascertain and compare the quantity of DNA from teeth exposed to different environmental conditions using organic and silica column DNA extraction methods and determine their gender using amelogenin gene primer. Human teeth were used and divided into four major groups based on different exposure types (buried in soil, storage in Lagoon water, normal environmental condition by placing them on the floor at the botanical garden, and exposure to temperature of 210oC for 15mins). The powdered dentin-cementum complex was used for DNA extraction, quantification and purity assessments. Amplification was done using amelogenin gene primers. Mean DNA concentration and purity were 187.59±26.25ng/μl and 0.97±0.07 respectively for silica column method, and, 138.98±35.71ng/μl and 0.91±0.11 respectively for organic extraction. With organic extraction, the least and highest mean DNA concentration are from teeth immersed in Lagoon water (74.86±61.19ng/μl) and teeth exposed to temperature of 210oC for 15mins (385.14±106.39ng/μl). In the Silica column method, teeth heated at 210oC for 15mins showed the lowest mean DNA concentration (165.57±33.82ng/μl) while teeth exposed to "normal environmental conditions", gave the highest DNA concentration (218.46±45.03ng/μl). Positive and negative male identification were 30.8% and 69.2% respectively, while female identification was 100% positive irrespective of the environmental exposure. The dentine-cementum is a valuable DNA source for forensic investigations. Male identification using amelogenin gene, should still be investigated in the Nigerian context.

11 pages, 4 figures, 1 table, supplementary data https://doi.org/10.1016/j.envres.2022.113549, Sex ratio depends on sex determination mechanisms and is a key demographic parameter determining population viability and resilience to natural and anthropogenic stressors. There is increasing evidence that the environment can alter sex ratio even in genetically sex-determined species (GSD), as elevated temperature can cause female-to-male sex reversal (neomales). Alarmingly, neomales are being discovered in natural populations of several fish, amphibian and reptile species worldwide. Understanding the basis of neomale development is important for conservation biology. Among GSD species, it is unknown whether those with chromosomal sex determination (CSD), the most common system, will better resist the influence of high temperature than those with polygenic sex determination (PSD). Here, we compared the effects of elevated temperature in two wild zebrafish strains, Nadia (NA) and Ekkwill (EKW), which have CSD with a ZZ/ZW system, against the AB laboratory strain, which has PSD. First, we uncovered novel sex genotypes and the results showed that, at control temperature, the masculinization rate roughly doubled with the addition of each Z chromosome, while some ZW and WW fish of the wild strains became neomales. Surprisingly, we found that at elevated temperatures WW fish were just as likely as ZW fish to become neomales and that all strains were equally susceptible to masculinization. These results demonstrate that the Z chromosome is not essential for male development and that the dose of W buffers masculinization at the control temperature but not at elevated temperature. Furthermore, at the elevated temperature the testes of neomales, but not of normal males, contained more spermatozoa than at the control temperature. Our results show in an unprecedented way that, in a global warming scenario, CSD species may not necessarily be better protected against the masculinizing effect of elevated temperature than PSD species, and reveal genotype-by-temperature interactions in male sex determination and spermatogenesis, This study was supported by the Spanish Ministry of Science and Innovation grant AGL2016-787107-R “Epimark” to FP and NIH grants R01 GM085318 and 5R35 GM139635 to JHP. With funding from the Spanish government through the ‘Severo Ochoa Centre of Excellence’ accreditation (CEX2019-000928-S). AV was supported by Spanish government scholarships (BES-2014-069051) with two short-term stay grants (EEBB-I-18-12947 and EEBB-I-17-12342) performed in the JHP lab. LR was supported by RYC2018-024017-I. RHN and MSR were supported by São Paulo Research Foundation (FAPESP 14/07620–7 and 17/15793–7)

Spolni kromosomi definiraju se kao kromosomi koji sadrže spolno-određujuće gene koji preusmjeravaju razvoj jedinke na ženski ili muški spol. Teorije o njihovom nastanku i evoluciji pojavile su se još početkom 20. stoljeća i neke pretpostavke su prihvaćene i danas. Osnovna teorija o nastanku XX/XY heteromorfnih spolnih kromosoma je o primarnoj pojavi gena koji je glavni regulator spola na jednom od autosoma te on i njegov homologni kromosom postaju spolni kromosomi. Predak na kojem se pojavljuju takvi spolni kromosomi smatra se ili hermafrodit ili jedinka odvojenih spolova koje determiniraju okolišni faktori. Zatim kroz niz genskih promjena, od kojih je najistaknutija inverzija, dolazi do supresije njihove rekombinacije. Zatim dolazi do degradacije Y kromosoma i pojave kompenzacijske doze. Analogno se objašnjava i ZZ/ZW sustav ženske heterogametnosti. Međutim novija istraživanja pokazuju kako evolucija spolnih kromosoma nije toliko linearna te se dovodi u pitanje mehanizam svake od pretpostavljenih stadija evolucije. Također kroz razvoj teorija o evoluciji spolnih kromosoma kroz prošlost bio je zanemaren i ekološki utjecaj koji se u novije vrijeme više istražuje. Ovaj rad predstavlja pregled i usporedbu nekih od glavnih točaka klasičnih i novijih teorija evolucije spolnih kromosoma. Sex chromosomes are defined as chromosomes which contain sex-defining genes which direct the development of an individual to either the female or the male sex. Theories of their emergence and evolution were made at the beginning of the 20th century and some of them are considered valid, even today. The classical theory of the creation of XX/XY heteromorphic sex chromosomes is based on the emergence of a master sex-regulator on one of the autosomes by which it and its homolog became sex chromosomes. The ancestor which obtained such sex chromosomes is considered to be either a hermaphrodite or a species which has environmental sex determination. The next step is suppression of recombination through an assortment of different genetic changes, the most important one being inversion. Afterwards would be the degradation of the Y chromosome and the emergence of dosage compensation. The ZZ/ZW system of female heterogamety is explained accordingly. Despite classical theories having abundance of evidence, newer research shows that the evolution of sex chromosomes is not that linear, and the mechanisms of each stage are being questioned. Also, in forming classical theories of sex chromosomes evolution in the past the influence of ecology was often overlooked, which will be briefly touched upon here. This paper introduces a review and comparison of some key points in classical and alternative theories of sex chromosome evolution.

Background: One of the main issues of general personality identification is the ascertainment of the sex of impersonated or fragmented bodies. Sporadic studies in the forensic medical literature have reported larynx cartilage sexual dimorphism, among which arytenoid cartilages are absent. The publications have morphometric focus and do not consider the practical aspects of sex diagnostics, which have been determined in this study. Aim: To develop a method for determining sex using the features of sexually dimorphic anatomical and morphological arytenoid cartilage of an adult. Materials and methods: The anatomical and morphological features of arytenoid cartilage from 80 males and 80 females aged 2078 years were studied. The visual examination revealed differences related to sex based on nine characteristics. The reliability of the sexual dimorphism of traits was verified using the Chi-square test and proportionality coefficients. Results: The informational significance of the features was clarified, and the individual observations were digitized using the formula. The possibility of sex determination by arytenoid cartilage was established in 93.75% of cases. In the remaining 6.25% cases, it was deemed impossible to solve this problem using the given set of features. Conclusion: The method developed for determining sex is highly accurate and can be used in sex diagnostics during a forensic medical larynx examination of a decayed or a fragmented corpse.

International audience; A vast diversity of types of life cycles exists in nature, and several theories have been advanced to explain how this diversity has evolved and how each type of life cycle is retained over evolutionary time. Here, we exploited the diversity of life cycles and reproductive traits of the brown algae (Phaeophyceae) to test several hypotheses on the evolution of life cycles. We investigated the evolutionary dynamics of four life-history traits: life cycle, sexual system, level of gamete dimorphism and gamete parthenogenetic capacity. We assigned states to up to 77 representative species of the taxonomic diversity of the brown algal group, in a multi-gene phylogeny. We used maximum likelihood and Bayesian analyses of correlated evolution, while taking the phylogeny into account, to test for correlations between traits, and to investigate the chronological sequence of trait acquisition. Our analyses are consistent with the prediction that diploid growth evolves when sexual reproduction is preferred over asexual reproduction, possibly because it allows the complementation of deleterious mutations. We also found that haploid sex determination is ancestral in relation to diploid sex determination. However, our results could not address whether increased zygotic and diploid growth are associated with increased sexual dimorphism. Our analyses suggest that in the brown algae, isogamous species evolved from anisogamous ancestors, contrary to the commonly reported pattern where evolution proceeds from isogamy to anisogamy.

Background: This study aimed to assess the effectiveness of mandibular ramus in gender identification in Haryana population. Materials and Methods: One hundred patients were assessed. Panoramic radiological examination was done. Following parameters were recorded: MAX. RM. WDTH (Maximum ramus breadth), MIN. RM. WDTH (Minimum ramus breadth), CND. HGT. RM (Condylar height), PRJ. HGT. RM (Projective height of the ramus), and CRND. HGT. RM (Coronoid height). All the results were recorded in Microsoft excel sheet and were analyzed by SPSS software version 16.0. Results: There is significant difference found in males and females in maximum ramus width, minimum ramus width, condylar height, and coronoid height and between male and females in condylar height and coronoid height. Furthermore, there was statistical significant difference between male and females in coronoid height. Conclusion: Mandibular ramus can be used for gender assessment as a part of forensic investigation.

While the process of meiosis is highly conserved across eukaryotes, the sexual systems that govern life cycle phase transitions are surprisingly labile. Switches between sexual systems have profound evolutionary and ecological consequences, in particular for plants, but our understanding of the fundamental mechanisms and ultimate causes underlying these transitions is still surprisingly incomplete. We explore here the idea that brown and green algae may be interesting comparative models that can increase our understanding of relevant processes in plant reproductive biology, from evolution of gamete dimorphism, gametogenesis, sex determination and transitions in sex-determining systems.

Nowadays, illegal trafficking and smuggling of animals are among the greatest threats to many avian species. Most commonly smuggled birds belong to parrots, song birds, raptor species and owls. All of these species are protected by national and international legislations. In order to prevent and reduce wildlife trafficking, DNA methods have become an important forensic tool in species and sex identification. In this study, CHD1R/CHD1F primer pair was used to amplify a part of the CHD1 gene from 65 birds that belong to 43 species. For 36 species this is the first time that the length of CHD1 amplicons was measured for the purpose of species determination. The results were visualized using capillary electrophoresis and enabled simultaneous determination of sex and species. Based on the number of amplicons (two in females, one in males) sex was successfully determined in all species, even in cases where gel electrophoresis failed to give results. Moreover, the species was successfully determined in most bird species based on the species-specific sizes of CHD1 amplicon. The method used in this study is of great importance for veterinary forensic medicine and the prevention of wildlife smuggling. Still, further work is necessary to confirm the effectiveness of the method in all bird species. Ilegalna trgovina i krijumčarenje divljim vrstama danas predstavlja jednu od najvećih pretnji koje ugrožavaju populacije velikog broja ptica. Najčešće vrste kojima se ilegalno trguje su papagaji, ptice pevačice, grabljivice i sove. Sve ove vrste ptica su zaštićene nacionalnim ili međunarodnim zakonskim regulativama. U cilju prevencije i smanjenja ilegalne trgovine, molekularne metode su postale bitno forenzičko oruđe u identifi kaciji vrste i pola zaštićenih vrsta ptica. U ovom istraživanju ispitano je ukupno 65 uzoraka ptica (ukupno 43 vrste), a u cilju umnožavanja dela CHD1 gena korišćen je CHD1R/CHD1F par prajmera. Za 36 vrsta ovo je prvi put da je dužina CHD1 gena određivana u cilju identifi kacije vrste. Dobijeni produkti su vizuelizovani koristeći kapilarnu elektroforezu i omogućili su simultano određivanje pola i vrste. Na osnovu analize amplikona (dva kod ženke, jedan kod mužjaka), pol je uspešno identifi kovan kod svih vrsta, čak i kod onih kod kojih gel elektroforeza nije bila uspešna.Vrsta je uspešno određena kod većine ptica na osnovu veličine CHD1 amplikona specifične za vrstu. Metoda korišćena u ovoj studiji je od velikog značaja u veterinarskoj forenzici i prevenciji ilegalne trgovine životinjama. Ipak, dalji rad na ovoj metodi je neophodan da bi se potvrdila efi kasnost metode na svim vrstama ptica.

Alternative splicing is an important way of regulating gene functions in eukaryotes. Several key genes involved in sex determination and gonadal differentiation, such as nr5a1 and ddx4, have sex-biased transcripts between males and females, suggesting a potential regulatory role of alternative splicing in gonads. Currently, the sex-specific alternative splicing events and genes have not been comprehensively studied at the genome-wide level in zebrafish. In this study, through global splicing analysis on three independent sets of RNA-seq data from matched zebrafish testes and ovaries, we identified 120 differentially spliced genes shared by the three datasets, most of which haven’t been reported before. Functional enrichment analysis showed that the GO terms of mRNA processing, mRNA metabolism and microtubule-based process were strongly enriched. The testis- and ovary-biased alternative splicing genes were identified, and part of them (tp53bp1, tpx2, mapre1a, kif2c, and ncoa5) were further validated by RT-PCR. Sequence characteristics analysis suggested that the lengths, GC contents, and splice site strengths of the alternative exons or introns may have different influences in different types of alternative splicing events. Interestingly, we identified an unexpected high proportion (over 70%) of non-frameshift exon-skipping events, suggesting that in these cases the two protein isoforms derived from alternative splicing may both have functions. Furthermore, as a representative example, we found that the alternative splicing of ncoa5 causes the loss of a conserved RRM domain in the short transcript predominantly produced in testes. Our study discovers novel sex-specific alternative splicing events and genes with high reliabilities in zebrafish testes and ovaries, which would provide attractive targets for follow-up studies to reveal the biological significances of alternative splicing events and genes in sex determination and gonadal differentiation.

Cannabis (Cannabis sativa L.) is a dioecious plant that produces both male and female inflorescences. In nature, male and female plants can be found with nearly equal frequency, which determines species out-crossing. In cannabis farming, only female plants are preferred due to their high yield of cannabinoids. In addition to unfavorable male plants, commercial production of cannabis faces the appearance of hermaphroditic inflorescences, species displaying both pistillate flowers and anthers. Such plants can out-cross female plants, simultaneously producing undesired seeds. The problem of hermaphroditic cannabis triggered a search for analytical tools that can be used for their rapid detection and identification. In this study, we investigate the potential of Raman spectroscopy (RS), an emerging sensing technique that can be used to probe plant biochemistry. Our results show that the biochemistry of male, female and hermaphroditic cannabis plants is drastically different which allows for their confirmatory identification using a hand-held Raman spectrometer. Furthermore, the coupling of machine learning approaches enables the identification of hermaphrodites with 98.7% accuracy, whereas both male and female plants can be identified with 100% accuracy. Considering the label-free, non-invasive and non-destructive nature of RS, the developed optical sensing approach can transform cannabis farming in the U.S. and overseas.

Ethylene biosynthesis and signal transduction play critical roles in plant sex differentiation. ACS (1-aminocyclopropane-1-carboxylic acid synthase) is a rate-limiting enzyme in ethylene biosynthesis. However, the understanding of the ACS gene family in Cucurbita maxima is limited. Here, we identified and characterized 13 ACS genes in the C. maxima genome. All ACS genes could be divided into three groups according to a conserved serine residue at the C-terminus. Thirteen CmaACS genes were found to be randomly distributed on 10 of the 20 chromosomes of C. maxima. The ACS gene exhibits different tissue-specific expression patterns in pumpkin, and four ACS genes (CmaACS1, CmaACS4, CmaACS7, and CmaACS9) were expressed specifically in both the female and male flowers of C. maxima. In addition, the expression levels of CmaACS4 and CmaACS7 were upregulated after ethephon and IAA treatments, which ultimately increased the number of female flowers, decreased the position of the first female flower and decreased the number of bisexual flowers per plant. These results provide relevant information for determining the function of the ACS genes in C. maxima, especially for regulating the function of ethylene in sex determination.

The background of this study is related to the tendencies in caviar production and prices in the world. The objective of the study is to evaluate and compare the profitability of different sturgeon sex determination methods for the conditions of aquaculture farms in northern Israel. We present the economic valuation of the technological innovation of early sex determination in this aquaculture industry. The production of sturgeon biomass exceeded the fishery harvest during the 1970–1980s by more than four times. There was a significant decline in world caviar prices the last time. The aquafarming business has a high potential economic value in the region of northern Israel. Based on the empirical data of the Caviar Galilee farm in this region, we examine the economic benefits of the innovation in sturgeon sex determination, including the case of a possible decrease in caviar price. Using the bio-economic model for a sturgeon caviar farm in two versions (for each of the considered sex determination methods, the endoscopy method and the early sex determination method), we show the economic advantages of this innovation of 9–34% for different financial highlights compared to the traditional method of sturgeon sex determination. The study empirically illustrates the economic advantages of the cooperation between aquaculture farming and research institutes. Our study suggests that additional directions in research should be sought in order to increase the profitability of sturgeon farms, especially when the world caviar prices decline. Further research can be conducted for additional regions and data.

Temperature-dependent sex determination (TSD) decides the sex fate of an individual based on incubation temperature. However, other environmental factors, such as pollutants, could derail TSD sexual development. Cadmium is one such contaminant of soils and water bodies known to affect DNA methylation, an epigenetic DNA modification with a key role in sexual development of TSD vertebrate embryos. Yet, whether cadmium alters DNA methylation of genes underlying gonadal formation in turtles remains unknown. Here, we investigated the effects of cadmium on the expression of two gene regulators of TSD in the painted turtle, Chrysemys picta, incubated at male-producing and female-producing temperatures using qPCR. Results revealed that cadmium alters transcription of Dmrt1 and aromatase, overriding the normal thermal effects during embryogenesis, which could potentially disrupt the sexual development of TSD turtles. Results from a preliminary DNA methylation-sensitive PCR assay implicate changes in DNA methylation of Dmrt1 as a potential cause that requires further testing (aromatase methylation assays were precluded).

Haplodiploidy and paternal genome elimination (HD/PGE) are common in invertebrates, having evolved at least two dozen times, all from male heterogamety (i.e., systems with X chromosomes). However, why X chromosomes are important for the evolution of HD/PGE remains debated. The Haploid Viability Hypothesis posits that X-linked genes promote the evolution of male haploidy by facilitating purging recessive deleterious mutations. The Intragenomic Conflict Hypothesis holds that conflict between genes drives genetic system turnover; under this model, X-linked genes could promote the evolution of male haploidy due to conflicts with autosomes over sex ratios and genetic transmission. We studied lineages where we can distinguish these hypotheses: species with germline PGE that retain an XX/X0 sex determination system (gPGE+X). Because evolving PGE in these cases involves changes in transmission without increases in male hemizygosity, a high degree of X linkage in these systems is predicted by the Intragenomic Conflict Hypothesis but not the Haploid Viability Hypothesis. To quantify the degree of X linkage, we sequenced and compared 7 gPGE+X species’ genomes with 11 related species with typical XX/XY or XX/X0 genetic systems, representing three transitions to gPGE. We find highly increased X linkage in both modern and ancestral genomes of gPGE+X species compared to non-gPGE relatives, and recover a significant positive correlation between percent X linkage and the evolution of gPGE. These are among the first empirical results suggesting a role for intragenomic conflict in the evolution of novel genetic systems like HD/PGE.Significance StatementSex determination systems such as haplodiploidy, in which males’ gene transmission is haploid, are surprisingly common, however, the evolutionary paths to these systems are poorly understood. X chromosomes may play a particularly important role, either by increasing survival of males with only maternal genomes, or due to conflicts between X-chromosomal and autosomal genes. We studied X-chromosome gene richness in three arthropod lineages in which males are diploid as adults but only transmit their maternally-inherited haploid genome. We find that species with such atypical systems have far more X chromosomal genes than related diploid species. These results suggest that conflict between genetic elements within the genome drives the evolution of unusual sex determination systems.

Funder: The Branco Weiss Fellowship – Society in Science; doi: http://dx.doi.org/10.13039/501100001710, UNLABELLED: Sex determination systems are highly conserved among most vertebrates with genetic sex determination, but can be variable and evolve rapidly in some. Here, we study sex determination in a clade with exceptionally high sex chromosome turnover rates. We identify the sex determining chromosomes in three interspecific crosses of haplochromine cichlid fishes from Lakes Victoria and Malawi. We find evidence for different sex determiners in each cross. In the Malawi cross and one Victoria cross the same chromosome is sex-linked but while females are the heterogametic sex in the Malawi species, males are the heterogametic sex in the Victoria species. This chromosome has not previously been reported to be sex determining in cichlids, increasing the number of different chromosomes shown to be sex determining in cichlids to 12. All Lake Victoria species of our crosses are less than 15,000 years divergent, and we identified different sex determiners among them. Our study provides further evidence for the diversity and evolutionary flexibility of sex determination in cichlids, factors which might contribute to their rapid adaptive radiations. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s10750-021-04560-7.

Pre-harvest male maturation is problematic for Atlantic salmon (Salmo salar) farmers and is regulated by the environment and genetics (e.g., vgll3). Five families of all-male salmon parr (produced using YY males crossed with XX females) with different vgll3 genotypes were split between three environmental regimes in January 2018. The “advanced maturation” regime used elevated temperature (16 °C) and continuous light from January 2018 with post-smolt maturation assessed in March 2018. The “extended freshwater” regime used ambient freshwater (1–16 °C) and simulated natural photoperiod (SNP) with post-smolt maturation assessed in November 2018. The “sea transfer” regime used ambient temperatures (1–14 °C) and SNP in freshwater until May 2018 when they were transferred to 9 °C seawater with natural photoperiod for 2.5 years (final mean weight of circa. 14 kg) and assessed for post-smolt maturation, 1 sea-winter (1 SW) maturation, and 2 sea-winter (2 SW) maturation in the autumn (November/December) of 2018, 2019, and 2020, respectively. Post-smolt maturation was highest in the advanced maturation and extended freshwater regimes (39–99% depending on family) and lowest in the sea transfer regime (0–95% depending on family). In the sea transfer regime, maturity incidence increased over time (0–95% post-smolt maturation, 1–100% 1 SW, and 50–90% 2 SW maturation, depending on family). In all regimes, those homozygous for the pre-designated vgll3 “early” maturing allele had the highest incidences of maturation whilst those homozygous for the “late” allele had the lowest. A low percentage of 2 SW phenotypic and genetic females were found (0–5% depending on family), one of which was successfully crossed with an XY male resulting in progeny with an approx. 50/50 sex ratio. These results show (i) post-smolt maturation varies dramatically depending on environment although genetic regulation by vgll3 was as expected, and (ii) crossing YY sperm with XX eggs can result in XX progeny which can themselves produce viable progeny with an equal sex ratio when crossed with an XY male.

Primordial germ cells (PGCs) are germline-restricted embryonic cells that form the functional gametes of the adult animal. The use of avian PGCs in biobanking and producing genetically modified birds has driven research on the in vitro propagation and manipulation of these embryonic cells. In avian species, PGCs are hypothesized to be sexually undetermined at an early embryonic stage and undergo differentiation into an oocyte or spermatogonial fate dictated by extrinsic factors present in the gonad. However, chicken male and female PGCs require different culture conditions, suggesting that there are sex-specific differences, even at early stages. To understand potential differences between male and female chicken PGCs during migratory stages, we studied the transcriptomes of circulatory stage male and female PGCs propagated in a serum-free medium. We found that in vitro cultured PGCs were transcriptionally similar to their in ovo counterparts, with differences in cell proliferation pathways. Our analysis also revealed sex-specific transcriptome differences between male and female cultured PGCs, with notable differences in Smad7 and NCAM2 expression. A comparison of chicken PGCs with pluripotent and somatic cell types identified a set of genes that are exclusive to germ cells, enriched in the germplasm, and associated with germ cell development.

The challenge in extracting high-quality RNA impedes the investigation of the transcriptome of developing salmonid embryos. Furthermore, the mRNA expression pattern of important PGC and SD genes during the initial embryonic development of Salmo salar is yet to be studied. So, in the present study, we aimed to isolate high-quality RNA from eggs and developing embryos to check vasa, dnd1, nanos3a, sdf1, gsdf, amh, cyp19a, dmrt1 and foxl2 expression by qPCR. Additionally, four HKGs (GAPDH, UB2L3, eEf1a and β-actin) were validated to select the best internal control for qPCR. High-quality RNA was extracted, which was confirmed by spectrophotometer, agarose gel electrophoresis and Agilent TapeStation analysis. UB2L3 was chosen as a reference gene because it exhibited lower intra- and inter-sample variation. vasa transcripts were expressed in all the developmental stages, while dnd1 was expressed only up to 40 d°C. Nanos3a was expressed in later stages and remained at its peak for a shorter period, while sdf1 showed an irregular pattern of mRNA expression. The mRNA expression levels of SD genes were observed to be upregulated during the later stages of development, prior to hatching. This study presents a straightforward methodology for isolating high-quality RNA from salmon eggs, and the resulting transcript profiles of significant PGC and SD genes in S. salar could aid in improving our comprehension of reproductive development in this commercially important species.

DNA methylation is a prevalent epigenetic modification in vertebrates, and it has been shown to be involved the regulation of gene expression and embryo development. However, it remains unclear how DNA methylation regulates sexual development, especially in species without sex chromosomes. To determine this, we utilized zebrafish to investigate DNA methylation reprogramming during juvenile germ cell development and adult female-to-male sex transition. We reveal that primordial germ cells (PGCs) undergo significant DNA methylation reprogramming during germ cell development, and the methylome of PGCs is reset to an oocyte/ovary-like pattern at 9 days post fertilization (9 dpf). When DNA methyltransferase (DNMT) activity in juveniles was blocked after 9 dpf, the zebrafish developed into females. We also show that Tet3 is involved in PGC development. Notably, we find that DNA methylome reprogramming during adult zebrafish sex transition is similar to the reprogramming during the sex differentiation from 9 dpf PGCs to sperm. Furthermore, inhibiting DNMT activity can prevent the female-to-male sex transition, suggesting that methylation reprogramming is required for zebrafish sex transition. In summary, DNA methylation plays important roles in zebrafish germ cell development and sexual plasticity.

Background: Sex ratio is an important sociodemographic characteristic of the country reflecting the status of its women and underlying socioeconomic and cultural patterns. PC-PNDT law is in place since so many years still sex determination is prevalent. Here not only doctors but the parents are also equally responsible for propagating this social evil as most of them are unaware about the Act and its legal implications and consequences of the declining sex ratio. Methods: Study was conducted over a period of 24 months from December 2018 to November 2018 in Immunisation OPD of a tertiary health care centre and a primary health centre with a sample size of 117 in both areas. Simple Random sampling method was used and analysis was done using SPSS software. Results: Out of 234 mothers approximately 70% were aware that pre-natal sex determination can be done. About 40% were aware that it is illegal, 30% said that ultrasonography is used for doing it, 20% said that both doctor and parents are punished under this act. Only 5% were aware about the correct punishment under this Act. Awareness regarding the place, method, legality and punishment was more among mothers of urban area as compared to rural area. Education and socioeconomic status was associated with awareness. Conclusion: Creating awareness in the general public and specifically mothers regarding the Act and consequences of its violation. Comprehensive behavioural change communication activities can be done through mass media for educating the people regarding the effect of declining sex ratio.

Biological control is widely successful at controlling pests, but effective biocontrol agents are now more difficult to import from countries of origin due to more restrictive international trade laws (the Nagoya Protocol). Coupled with increasing demand, the efficacy of existing and new biocontrol agents needs to be improved with genetic and genomic approaches. Although they have been underutilised in the past, application of genetic and genomic techniques is becoming more feasible from both technological and economic perspectives. We review current methods and provide a framework for using them. First, it is necessary to identify which biocontrol trait to select and in what direction. Next, the genes or markers linked to these traits need be determined, including how to implement this information into a selective breeding program. Choosing a trait can be assisted by modelling to account for the proper agro-ecological context, and by knowing which traits have sufficiently high heritability values. We provide guidelines for designing genomic strategies in biocontrol programs, which depend on the organism, budget, and desired objective. Genomic approaches start with genome sequencing and assembly. We provide a guide for deciding the most successful sequencing strategy for biocontrol agents. Gene discovery involves quantitative trait loci analyses, transcriptomic and proteomic studies, and gene editing. Improving biocontrol practices includes marker-assisted selection, genomic selection and microbiome manipulation of biocontrol agents, and monitoring for genetic variation during rearing and post-release. We conclude by identifying the most promising applications of genetic and genomic methods to improve biological control efficacy

Fishes provide a unique and intriguing model system for studying the genomic origin and evolutionary mechanisms underlying sex determination and high sex-chromosome turnover. In this study, the mode of sex determination was investigated in Siamese fighting fish, a species of commercial importance. Genome-wide SNP analyses were performed on 75 individuals (40 males and 35 females) across commercial populations to determine candidate sex-specific/sex-linked loci. In total, 73 male-specific loci were identified and mapped to a 5.6 kb region on chromosome 9, suggesting a putative male-determining region (pMDR) containing localized dmrt1 and znrf3 functional sex developmental genes. Repeat annotations of the pMDR revealed an abundance of transposable elements, particularly Ty3/Gypsy and novel repeats. Remarkably, two out of the 73 male-specific loci were located on chromosomes 7 and 19, implying the existence of polygenic sex determination. Besides male-specific loci, five female-specific loci on chromosome 9 were also observed in certain populations, indicating the possibility of a female-determining region and the polygenic nature of sex determination. An alternative explanation is that male-specific loci derived from other chromosomes or female-specific loci in Siamese fighting fish recently emerged as new sex-determining loci during domestication and repeated hybridization.

Sex determination in birds, due to the very common lack of sexual dimorphism, is challenging. Therefore, molecular sexing is often the only reliable way to differentiate between the sexes. However, for many bird species, very few genetic markers are available to accurately, quickly, and cost-effectively type sex. Therefore, in our study, using 14 species belonging to the order Musophagiformes, we tested the usefulness of seven PCR markers (three of which have never been used to determine the sex of turacos), developed based on the CHD1, NIPBL, and SPIN genes, to validate existing and develop new strategies/methods of sex determination. After in silico analysis, for which we used the three turaco nuclear genomes available in GenBank, the suitability of the seven selected markers for sexing turacos was tested in the laboratory. It turned out that the best of the markers tested was the 17th intron in the NIPBL gene (not previously tested in turacos), allowing reliable sex determination in 13 of the 14 species tested. For the one species not sexed by this marker, the 9th intron in the CHD1 gene proved to be effective. The remaining markers were of little (4 markers developed based on the CHD1 gene) or no use (marker developed based on the SPIN gene).

Regulated or programmed cell death (RCD or PCD) is a fundamental biological principle integral to a considerable variety of functions in multicellular organisms. In plants, different PCD processes are part of biotic and abiotic stress responses, but also occur as an essential aspect of unperturbed plant development. PCD is particularly abundant during plant reproduction, eliminating unwanted or no longer needed cells, tissues, or organs in a precisely controlled manner. Failure in reproductive PCD can have detrimental consequences for plant reproduction. Here we shed a light on the latest research into PCD mechanisms in plant reproduction from sex determination over sporogenesis to pollination and fertilization.