Sexual Phase Research Articles

Integration of GWAS and transcriptomic analyses reveal candidate genes for duck gonadal development during puberty onset

BackgroundPuberty onset signifies the beginning of sexual maturation and reproductive phase in poultry indeed, and plays an essential role in genetics and breeding. Studying gonadal development is one of the important approaches to exploring the genetic mechanism of puberty onset.ResultIn our study, the phenotype data of the testes and ovaries of the 120-day-old Nonghua duck showed a large coefficient of variation, indicating that their gonads were in different developmental states. The CNV-based GWAS results for 358 Nonghua ducks showed two deleted-type CNVRs were associated with testicular weight (TW) and testicular percentage (TP), namely CNVR492 (Chr2: 59473501–59478500 bp) and CNVR494 (Chr2: 59514001–59517000 bp). Additionally, two both-type CNVRs were associated with ovarian weight (OW) and ovarian percentage (OP), namely CNVR557 (Chr2: 99951001–99956500 bp) and CNVR891 (Chr7: 39115001–39122500 bp). RNA-seq analysis showed 6228 and 1070 differentially expressed genes (DEGs) related to the TW and OW. These DEGs were mainly enriched in the MAPK signaling pathway, cytokine-cytokine receptor interaction, and focal adhesion, which were reported to affect gonadal development. Further, by joint analysis of CNV-based GWAS and RNA-seq data, 3 genes, including LOC106019197, CDH19 (LOC101793040), and TYW5 were identified as potential candidate genes for TW and OW. LOC106019197 and CDH19 were down-regulated in the heavier-testes group (> 5 g), while TYW5 was also down-regulated in the heavier-ovaries group (> 3 g). The qRT-PCR revealed that LOC106019197 and CDH19 exhibited higher expression levels in the wild/CN0 and CN0/CN0 genotypes compared to the wild/wild genotype. TYW5 showed the highest expression level in the wild/CN0 genotype and the lowest in the CN2/CN2 genotype. In addition, the expression levels of LOC106019197 and CDH19 were significantly higher at 0w than at 8w and 24w.ConclusionOur results revealed that LOC106019197 and CDH19 may act as inhibitors of duck testicular development. TYW5 may play a role in delaying ovarian development. These findings provide new insights into the mechanism of puberty onset in ducks.

Female Offenders in Human Trafficking: Analyzing Roles in a Spanish Sample

Offenders convicted of trafficking human beings for sexual exploitation involve a high percentage of women compared to other similar offenses. Previous studies have searched for explanations for the dynamics of the illegal sex market and the coercion experienced by victims during the process of exploitation and exit. This article analyses the content of 58 judicial decisions involving a sample of 148 women convicted in Spain of trafficking in human beings between 2017 and 2024. Two main dimensions are explored: the criminal organizations performing the sexual exploitation phase in Spain and the characteristics, positions, and roles of women traffickers within the criminal organizations. Half of the sample occupies leadership roles, while the other half consists of supporters, especially in larger trafficking networks. Regarding the positions and roles of women offenders, supporters and partners in crime are at higher risk of being coerced due to the nature of their roles or partnerships.

Black Crust Complex: Influence of Temperature and Period of Wetness on the Development of Fungi in Hevea brasiliensis.

The objective of this work was to evaluate the development of Davidiella sp. and its asexual form, Cladosporium sp., under different environmental conditions in the rubber tree (Hevea brasiliensis). Rubber tree leaves were inoculated with a spore suspension and kept in a humid chamber under different temperatures and wetness periods. The behavior of the fungi was evaluated using a scanning electron microscope (SEM) and an ultraviolet light microscope (UV). In the images obtained in SEM, four hours after inoculation of the fungus, it was possible to verify the germination and penetration of conidia at temperatures of 10 to 20 °C. The formation of conidiophores was verified from six hours after inoculation, indicating that it is in the reproductive period. In the sexual phase, in SEM, from four hours after inoculation, it was possible to verify the formation of small protuberances at temperatures between 10 and 20 °C. These black dots evolve into circular, protruding black spots, like the symptoms of black crust, with apparent spore formation on them. The data obtained from the UV analyses corroborate those from SEM, showing that the fungus has good development in its two phases between temperatures of 20 and 25 °C and that the period of wetness on the leaf can contribute to the initial development of the pathogen.

Komunikasi Antarpribadi Orang Tua-Anak Dalam Mengurangi Prilaku Seksual Sebelum Menikah Pada Remaja Hindu

College student who are late teens aged 18-19 years, have sexual development and phases that make them inclined to enter into a heterosexual relationship or relationship such as dating. During this period of dating they have a tendency to engage in sexual behavior, this is a negative thing in Hinduism which prohibits interpersonal communication. Interpersonal communication has the potential to carry out an instrumental function as a tool to influence or persuade others. Parent-adolescent interpersonal communication is a form of parenting process that has an important influence on the formation of adolescents' attitudes regarding sexual relations before marriage. So in this study it is hypothesized that stronger and closer parent-child interpersonal communication can reduce sexual behavior before marriage among Hindu teenagers. This research aims to find the influence of interpersonal communication on sexual behavior before marriage in Hindu teenagers. Quantitative methods were used in this research, collecting data by distributing questionnaires to 53 Hindu teenagers at Bumigora University. The results of this study state that interpersonal communication has a significant negative effect on the attitudes of Hindu teenagers in having sexual relations before marriage. This means that the better the interpersonal communication between Hindu teenagers and their parents, the lower their sexual behavior before marriage.

Floral scents of the protogynous aroid Spathiphyllum cannifolium: Investigating the chemistry and timing of emission at female and male phases

AbstractSpathiphyllum is a Neotropical aroid genus whose inflorescences are composed of protogynous bisexual flowers that emit strong perfumes. These scents serve as both attractants and rewards for male euglossine pollinators. We investigated the inflorescence scent of S. cannifolium, asking whether inflorescence at female and male phases differ in scent traits. For this, we collected scent samples from inflorescence at the two phases using dynamic headspace methods and analyzed them by a gas chromatograph coupled to a mass spectrometer (GC‐MS). The inflorescence scent of S. cannifolium consisted of 26 compounds, with benzyl acetate as major component (ca. 80% of the scent bouquet). We found no quantitative or qualitative differences between scents of inflorescences at female and male phases, nor in the emission time. However, when considering the timing of scent emission among individuals, we observed two general plant groups. Some individuals emitted more scent at 06:00, while others emitted more scent at 09:00, independent of the sexual phase. The similarity of floral scents between the sexes might promote floral constancy by male euglossine pollinators and consequently alternate pollinator flow between inflorescences at female and male phases. The occurrence of the two general patterns of scent emission can be interpreted as a way to ensure the pollination service. Since plants emit scents in the morning, even outside the peak emission period, this would enable these plants to attract pollinators that are active at different times than the peak emission of floral scents. This way, it would ensure the flow of pollen, even if it occurs at a low frequency.

First evidence of late-acting self-incompatibility in the Aristolochiaceae.

Most Aristolochiaceae species studied so far are from temperate regions, bearing self-compatible protogynous trap flowers. Although self-incompatibility has been suggested for tropical species, the causes of self-sterility in this family remain unknown. To fill this gap, we studied the pollination of the tropical Aristolochia esperanzae, including the physical and physiological anti-selfing mechanisms. Floral visitors trapped inside flowers were collected to determine the pollinators. Protogyny was characterized by observing the temporal expression of sexual phases and stigmatic receptivity tests. The breeding system was investigated using hand-pollination treatments. Pollen tube growth was observed using epifluorescence to identify the self-incompatibility mechanism. Flies were the most frequent visitors found inside A. esperanzae trap flowers, with individuals from the family Ulidiidae being potential pollinators since they carried pollen. The characteristic flower odour and presence of larvae indicate that A. esperanzae deceives flies through oviposition-site mimicry. Although this species showed incomplete protogyny, stigmatic receptivity decreased during the male phase, avoiding self-pollination. Fruits developed only after cross- and open pollination, indicating that the population is non-autonomous, non-apomictic, and self-sterile. This occurred through a delay in the growth of geitonogamous pollen tubes to the ovary and lower ovule penetration, indicating a late-acting self-incompatibility mechanism. Our findings expand the number of families in which late-acting self-incompatibility has been reported, demonstrating that it is more widespread than previously thought, especially when considering less-studied tropical species among the basal angiosperms.

Phenotypic and genetic variations of endangered Lysimachia maritima at global distribution limit

Species can respond to environmental pressures through genetic and phenotypic variation. Lysimachia maritima (L.) Galasso, Banfi & Soldano, a perennial halophyte, is distributed sporadically along the east coast in south Korea and is designated as an endangered plant species in Korea and ‘Least Concern’ in the IUCN Red List. To understand the phenotypic and genetic variations of L. maritima among sites and the environmental factors contributing to these differences, field surveys and genetic analyses were conducted. The principal component analysis of the environmental characteristics revealed that soil type, temperature, soil moisture content, and organic matter content were major factors determining variations among sites. L. maritima in Goseong (GS) and Yangyang (YY) sites faced significant limitations in its distribution due to competition with Phragmites australis for limited soil water and nutrients. There were significant phenotypic differences of L. maritima among sites. In sand-based sites of GS and YY, studied growth characteristics of L. maritima except stem length in YY and the number of seed-bearing individuals were smaller than those in organic soil substrate sites of US and PH, where L. maritima grew vigorously and were almost exclusively in the sexual reproductive phase. Genetic diversity at species level was high (h= 0.31, I= 0.46) but genetic diversities at site level were low, varying among sites (h= 0.133 ∼ 0.244). Also, genetic variation of L. maritima was higher within sites (58.32%) than among sites (41.68%). This indicates that L. maritima might prefer outcrossing more than selfing. L. maritima were strongly differentiated among sites (Gst= 0.336) because of limited gene flow (Nm= 0.987). Geographical isolation might have led to differentiated genetic structure in L. maritima, contributing to the formation of its spatial genetic structure. This high genetic diversity is crucial for the biodiversity of L. maritima at its global distribution limit, and this study emphasize the necessity of conserving distribution limit in the sense of soil characteristics.

Characterization of a nuclear transport factor 2-like domain-containing protein in Plasmodium berghei

BackgroundPlasmodium lacks an mRNA export receptor ortholog, such as yeast Mex67. Yeast Mex67 contains a nuclear transport factor 2 (NTF2)-like domain, suggesting that NTF2-like domain-containing proteins might be associated with mRNA export in Plasmodium. In this study, the relationship between mRNA export and an NTF2-like domain-containing protein, PBANKA_1019700, was investigated using the ANKA strain of rodent malaria parasite Plasmodium berghei.MethodsThe deletion mutant Δ1019700 was generated by introducing gene-targeting vectors into the P. berghei ANKA genome, and parasite growth and virulence were examined. To investigate whether PBANKA_1019700 is involved in mRNA export, live-cell fluorescence imaging and immunoprecipitation coupled to mass spectrometry (IP-MS) were performed using transgenic parasites expressing fusion proteins (1019700::mCherry).ResultsDeletion of PBANKA_1019700 affected the sexual phase but not the asexual phase of malaria parasites. Live-cell fluorescence imaging showed that PBANKA_1019700 localizes to the cytoplasm. Moreover, IP-MS analysis of 1019700::mCherry indicated that PBANKA_1019700 interacts with ubiquitin-related proteins but not nuclear proteins.ConclusionsPBANKA_1019700 is a noncanonical NTF2-like superfamily protein.

Functional Characterization of the Tmg-4 Magnesium Transporter with the GQN Motif from Neurospora crassa

To unveil the physiological relevance of magnesium and its transport pathways in Neurospora crassa, the vegetative, asexual, and sexual phases of development were investigated. Notably, a regular rate of hyphal growth extension was observed in media without magnesium supplementation. Further, conidia and perithecia formation was completely abolished under the same conditions. By estimating the levels of mycelial cations, magnesium was identified as the 3rd most abundant ion and its transport was found to be mediated by four putative CorA magnesium transporters: Tmg-1, Tmg-2, Tmg-3, and Tmg-4. Among these, the Tmg-4 transporter encoded by the NCU07816.5 (tmg-4) gene possesses a GQN motif instead of the universally conserved GMN motif of CorA magnesium transporters. Phenotypic analysis of the knockout mutant strain, Δtmg-4, revealed stunted vegetative growth, acquired partial cobalt resistance, and reduced levels of mycelial magnesium compared to that of the wild type strain. Further, tmg-4 gene expression remained unchanged during vegetative development but was upregulated by three-fold in the sexual cycle. Collectively, these results validate tmg-4 and its encoded protein as functional novel variant in the CorA superfamily magnesium transporters of fungi.

Lineage-specific genes are clustered with HET-domain genes and respond to environmental and genetic manipulations regulating reproduction in Neurospora.

Lineage-specific genes (LSGs) have long been postulated to play roles in the establishment of genetic barriers to intercrossing and speciation. In the genome of Neurospora crassa, most of the 670 Neurospora LSGs that are aggregated adjacent to the telomeres are clustered with 61% of the HET-domain genes, some of which regulate self-recognition and define vegetative incompatibility groups. In contrast, the LSG-encoding proteins possess few to no domains that would help to identify potential functional roles. Possible functional roles of LSGs were further assessed by performing transcriptomic profiling in genetic mutants and in response to environmental alterations, as well as examining gene knockouts for phenotypes. Among the 342 LSGs that are dynamically expressed during both asexual and sexual phases, 64% were detectable on unusual carbon sources such as furfural, a wildfire-produced chemical that is a strong inducer of sexual development, and the structurally-related furan 5-hydroxymethyl furfural (HMF). Expression of a significant portion of the LSGs was sensitive to light and temperature, factors that also regulate the switch from asexual to sexual reproduction. Furthermore, expression of the LSGs was significantly affected in the knockouts of adv-1 and pp-1 that regulate hyphal communication, and expression of more than one quarter of the LSGs was affected by perturbation of the mating locus. These observations encouraged further investigation of the roles of clustered lineage-specific and HET-domain genes in ecology and reproduction regulation in Neurospora, especially the regulation of the switch from the asexual growth to sexual reproduction, in response to dramatic environmental conditions changes.

Mbari and uncle Nicodemus: Male representations in the heterosexual discourse among female undergraduates in Nigeria

This paper examines the various labels ascribed to males in the heterosexual discourse among female undergraduates in Nigeria. Insights from Community of Practice and Sexual Script Theory were used to demonstrate the use of situated linguistic choices to establish the differentials in male sexual behaviours. Data for the study were generated through participant observation and unstructured interviews with the aid of audiotape recordings and field notes. The study involves a representative sample of 22 female undergraduates who served as participants and research assistants. The findings reveal that situated language practices stimulate heterosexual discourses where female undergraduates creatively ascribe labels to the different males’ sexual scripts. The labels include those related to male sexual performance, body size and size of sex organs, financial and marital status, familial and gatekeeping roles, and other routine sexual behaviours shown by men in sexual phases such as advances, arousal and intercourse. The scripts are also combined to re-enact the peculiar sexual narratives by the female students that show emerging sexual orientations. It is believed that the categorisation of such males’ sexual behaviours in the University social contexts, would provide a basis for comparison with the sexual practices of other young adults’ social groups across the globe.

Plasmodium microtubule-binding protein EB1 is critical for partitioning of nuclei in male gametogenesis.

Sexual reproduction of the malaria parasites is critical for their transmission to a mosquito vector. Several signaling molecules, such as kinases and phosphatases, are known to regulate this process. We previously demonstrated that Plasmodium falciparum (Pf) Ca2+-dependent protein kinase 4 (CDPK4) and serine/arginine-rich protein kinase 1 (SRPK1) are critical for axoneme formation during male gametogenesis, with genetic deletion of either gene causing a complete block in parasite transmission to the mosquito. A comparative phospho-proteome analysis of Pfcdpk4- and RNA-seq analysis of Pfsrpk1- gametocytes showed that these kinases regulate similar biological processes linked to both microtubule (MT) dynamics and cell motility. One of these proteins was a nuclear MT-associated End Binding protein 1 (EB1), which was hypophosphorylated in Pfcdpk4- gametocytes. To study the functional relevance of EB1, we created gene deletion parasites for EB1. We further demonstrate that Pfeb1- parasites like WT NF54 parasites proliferate normally as asexuals and undergo gametocytogenesis and gametogenesis. Strikingly, these parasites suffer a severe defect in nuclear segregation and partitioning of nuclei into emerging microgametes. Further genetic crosses utilizing male- and female-sterile parasites revealed that Pfeb1- parasites only suffer a male fertility defect. Overall, our study reveals an essential function for PfEB1 in male gamete nuclear segregation and suggests a potential therapeutic avenue in the design of transmission-blocking drugs to prevent malaria transmission from humans to mosquito. IMPORTANCE Gametogenesis and subsequent gamete fusion are central to successful transmission of the malaria parasites to a female Anopheles mosquito vector and completion of the sexual phase of the parasite life cycle. Male gametogenesis involves the formation of axonemes inside male gametes from male gametocytes via active cytoskeleton remodeling. The tubulin and tubulin-binding proteins are, thus, attractive anti-malarial drug targets. In the present study, we demonstrate that a microtubule-binding protein PfEB1 is essential for male gamete fertility, specifically for the inheritance of nuclei from activated male gametocytes. Targeting PfEB1 function may provide new avenues into designing interventions to prevent malaria transmission and disease spread.

Theoretical assessment of persistence and adaptation in weeds with complex life cycles

Herbicide-resistant weeds pose a substantial threat to global food security. Perennial weed species are particularly troublesome. Such perennials as Sorghum halepense spread quickly and are difficult to manage due to their ability to reproduce sexually via seeds and asexually through rhizomes. Our theoretical study of S. halepense incorporates this complex life cycle with control measures of herbicide application and tillage. Rooted in the biology and experimental data of S. halepense, our population-based model predicts population dynamics and target-site resistance evolution in this perennial weed. We found that the resistance cost determines the standing genetic variation for herbicide resistance. The sexual phase of the life cycle, including self-pollination and seed bank dynamics, contributes substantially to the persistence and rapid adaptation of S. halepense. While self-pollination accelerates target-site resistance evolution, seed banks considerably increase the probability of escape from control strategies and maintain genetic variation. Combining tillage and herbicide application effectively reduces weed densities and the risk of control failure without delaying resistance adaptation. We also show how mixtures of different herbicide classes are superior to rotations and mono-treatment in controlling perennial weeds and resistance evolution. Thus, by integrating experimental data and agronomic views, our theoretical study synergistically contributes to understanding and tackling the global threat to food security from resistant weeds.

Weight-Length and Length-Length Relationships of the Endangered Zebra Pleco Hypancistrus zebra (Siluriformes, Loricariidae) from the Xingu River, Amazon, Brazil

Hypancistrus zebra is a Loricariid endemic to a very limited stretch of the Xingu River and has a high value in the ornamental fish trade. The natural populations have experienced high fishing pressure since the late 80s and were recently impacted by the construction of the third largest hydroelectric dam complex in the world, the Belo Monte Project. Since 2004, it has been considered critically endangered and had its capture and trade forbidden. Even with its capture and trade prohibited since 2004, the species continues to be one of the most trafficked fish in the world. The biometric knowledge of the species is essential for monitoring its natural populations and management in ex situ environments. This work aims to establish the weight-length relationship (WLR) and length-length relationship (LLR) of the species and to evaluate the condition factor (K) of H. zebra rescued from trafficking. The study was conducted at the Laboratório de Aquicultura de Peixes Ornamentais do Xingu from February 2022 to June 2022, estimating the parameters of WLR and LLR of 1165 specimens divided into four simple linear regression (SLR) models: all specimens, only juveniles, only females, and only males. The K of 190 specimens rescued from trafficking by the Brazilian Federal Police was evaluated. The WLRs and LLRs were estimated using SLR. The difference between the allometry coefficients (b) and between the residuals of the models (juveniles, females, and males) was evaluated using the ANOVA significance test. The total length of the species ranged from 1.6 to 9.3 cm. The coefficient of allometry (b) of the WLR of the species is 3.22 and of the LLR is 1.04, demonstrating that H. zebra presents positive allometric growth. The evaluation of the residuals of the models showed differences between ontogenetic stages and between sexes. The development of juveniles is isometric until the first sexual maturation phase and differs in negative allometric for females and positive allometric for males, showing sexual dimorphism in the species. Among the specimens rescued from trafficking, 54.74% were underweight, and 13.16% were under 60% of the expected K value, demonstrating the poor conditions in which these animals are transported by trafficking.

Conulariid soft parts replicated in silica from the Scotch Grove Formation (lower Middle Silurian) of east-central Iowa

AbstractTwo specimens of Metaconularia manni (Roy, 1935) from the lower Middle Silurian Scotch Grove Formation (eastern Iowa) exhibit well-defined, relict soft parts replicated in silica. One of these specimens bears phosphatic periderm, whereas the other specimen is a mold. Present within the erect, undistorted apical region of the specimen preserving periderm, on opposite sides of the peridermal cavity, are two small, elongate masses of silica located near the midlines of two of the four faces. Present in the central portion of the other specimen, at a somewhat greater distance from the apex, are five pairs of hollow, elongate, keeled pouch-like bodies (hereafter pouches), the long axes of which converge on the center of the fossil. Each pair of pouches is associated with a short, narrow, gently curved or broadly U-shaped tube, also composed of silica. Additionally, two of the pouch/tube combinations are associated with a pair of rectilinear furrows that correspond to the paired internal carinae that straddled the conulariid's facial midlines. We interpret the paired pouches and short tubes in the moldic specimen as relic conulariid soft parts homologous, respectively, to the interradial gonads and retractor muscles of extant, stauromedusan and polypoid scyphozoan cnidarians. Unlike most conulariids, which exhibit four faces, this individual had five faces, an aberrant morphology known in one other conulariid. The two small masses in the other specimen are more difficult to interpret, but they, too, could be relic gonads or longitudinal muscles. These interpretations suggest that, as in certain extant scyphozoans, at least one conulariid lost the free-living, sexual medusoid life phase.

Self-compatibility in yeast is selected for reproductive assurance not population-level compatibility.

In haploid species, sexual reproduction by selfing lacks the common benefits from recombination and is indistinguishable from asexual reproduction at the genetic level. Nevertheless, the evolution of self-compatibility, known as homothallism in organisms with mating types, has occurred hundreds of times in fungi. Two main hypotheses have been proposed for the evolution of homothallism. First, that homothallism offers reproductive assurance, which is especially important when species have an obligatory sexual phase in their lifecycle. Second, that homothallism is associated with population-level compatibility, increasing the chance of outbreeding. Here, we test these hypotheses using the fission yeast Schizosaccharomyces pombe, which is homothallic by mating-type switching, leveraging natural variation for switching efficiency in this species. Combining empirical tests with cellular automaton simulations, we show that homothallism by switching increases mating success of switching genotypes, but does not affect population-level compatibility. Experiments show that outcrossing is actually reduced under homothallism. This reduction in outcrossing is explained by our simulations, which show that due to local mating, gametes that mated through intraclonal selfing are no longer available for outcrossing. Our results suggest that the recurrent evolution of haploid self-compatibility is likely driven by selection for mating assurance, not to increase the potential for outcrossing.

A DEAD-box RNA helicase mediates meiotic silencing by unpaired DNA.

During the sexual phase of Neurospora crassa, unpaired genes are subject to a silencing mechanism known as meiotic silencing by unpaired DNA (MSUD). MSUD targets the transcripts of an unpaired gene and utilizes typical RNA interference factors for its process. Using a reverse genetic screen, we have identified a meiotic silencing gene called sad-9, which encodes a DEAD-box RNA helicase. While not essential for vegetative growth, SAD-9 plays a crucial role in both sexual development and MSUD. Our results suggest that SAD-9, with the help of the SAD-2 scaffold protein, recruits the SMS-2 Argonaute to the perinuclear region, the center of MSUD activity.

Study of Estrus Induction in the Female Dog by PMSG and HCG

Our study, spread over four (4) months (February to June), aims at heat induction and monitoring within a population of 10 female dogs of different breeds (German shepherd, Rottweiler and Pit-bull). Aged from 2.5 to 4.5. Two of them were used as controls. To do this, we carried out the following investigations: Monitoring the sexual cycle of the female dogs, prior to heat induction, by gynecological and cytological examination (vaginal smears) to confirm the sexual rest phase (anestrus); hormonal induction of heat by PMCG and hCG; monitoring the induced heat by the vaginal smear method (Giemsa stain); and a breeding of female dogs in estrus confirmation of the pregnancy of the mated female dogs (palpation and ultrasound) by comparing the vaginal smears of the treated female dogs with those of the control female dogs. At the end of our study, the results obtained were encouraging, since we succeeded in obtaining the appearance of heat in 60% of the treated female dogs.

Follicular Atresia, Cell Proliferation, and Anti-Mullerian Hormone in Two Neotropical Primates (Aotus nancymae and Sapajus macrocephalus).

This study evaluated the follicular atresia, cell proliferation, and anti-Mullerian hormone action in Aotus nancymae and Sapajus macrocephalus during three sexual phases (follicular, luteal, and gestational). Follicular quantification and immunolocalization of Caspase-3 protein, B-cell lymphoma 2 (BCL-2), proliferating cell nuclear antigen (PCNA), and anti-Mullerian hormone (AMH) were performed. A significant difference in the quantification between preantral and antral follicles, with a progressive decrease in the antrals, was identified. Protein and hormonal markers varied significantly between follicle cell types (A. nancymae p = 0.001; S. macrocephalus, p = 0.002). Immunostaining in the preantral and antral follicles was present in all sexual phases; for Caspase-3, in granulosa cells, oocytes, and stroma; for BCL-2, in granulosa cells, oocytes, and theca; and for PCNA and AMH, in oocytes and granulosa cells. The immunostaining for Caspase-3 was more expressive in the preantral follicles (follicular phase, p < 0.05), while that for BCL-2 and PCNA was more expressive in the antral follicles of the follicular phase. The AMH was more expressive in the primary and antral follicles of nonpregnant females, in both the follicular and luteal phases. Our results contribute to understanding the ovarian follicular selection, recruitment, and degeneration of these species.

Genetic Diversity of Trichoderma and Hypocrea lixii Iraqi Strains by Using Random Amplified Polymorphic DNA (RAPD) Technique

As the diversity and characteristics of Trichoderma species are difficult to determine using morphological methods, henceforth molecular tools are crucial. This study utilized random amplified polymorphic DNA (RAPD) technique to investigate the genetic diversity of Trichoderma with sexual phase Hypocrea and to identify similarities and differences in the phylogenetic tree. Nine Iraqi Trichoderma strains (four strains of T. atroviride, one strain of Hypocrea lixii, two strains of T. gamsii and two strains of T. longibriantium) were examined in this research. The genomic DNA of each species was extracted and amplified with each of the five primers. 197 bands were obtained by using five oligodeoxynucleotide primers of which 98.47% were polymorphic and about 1.52% were monomorphic. When primers OPA4, OPA8, and OPA10 were used, the genetic variability was about 100%. Whereas, after using primers OPA7 and OPA5, the obtained genetic variability was 95.7% and 92.6% respectively. Gel images of the RAPD were processed with photo-cap program which detected the bands of isolates based on the ladder. The detected bands were then clustered based on the Jaccard method in Past software that showed T. atroviride, T. gamsii, Hypocrea lixii, and T. longibrachiatum isolates were grouped as clades and lineages. Although all strains belonged to the same species and group in one clade, they differed in size and number of bands. The Jaccard cluster analysis showed that the three isolates of T. atroviride were closely related to each other, while the four isolates of T. atroviride in one cluster were same as Hypocrea lixii, the isolates of T. gamsii and two strains of T. longibrachiatum formed one cluster. Thus, the high reliability of RAPD markers could be applied to identify Trichoderma species and create genetic maps instead of other DNA-based methods which are not only costly but time-consuming too.