Steinernema Feltiae- Xenorhabdus Bovienii: More Information about this Bactohelminthic Complex from Iran

Background: Entomopathogenic nematodes (EPNs) of the families Steinernematidae and Heterorhabditidae that are symbiotically associated with Xenorhabdus and Photorhabdus bacteria are one of the effective biological control agents of insect pests. Native isolates can probably be more efficacious to control insect pests than exotic ones due to their adaptability to indigenous environmental conditions. Results: In this study, Steinernema feltiae isolate FUM221 was recovered from soil samples collected from the Fandoghloo pasture, Ardabil province, Iran. Morphological investigations of the first and second-generation adults, infective juveniles, and molecular characterizations were given based on ITS and 18S rDNA genes. Besides, molecular analysis based on the 16S rRNA region and phenetic data recognized Xenorhabdus bovienii as its symbiont bacterium. The scanning electron microscopy (SEM) images verified the identification of this isolate.Conclusion: The molecular characterization using two loci and phylogenetic analyses provided more evidence for the classification of this steinernematid and its difference of the same species from other countries. Moreover, molecular and phenetic characterizations of its symbiotic bacterium were provided with low variations compared to other isolates. Herein, the comprehensive taxonomic data of this steinernematid from Iran is presented.

Description of Some Aquatic Insect Genera in Greater Zab River Branches, North of Iraq

Aquatic insects samples were collected from 6 sites along the Greater Zab River in the northern Iraq from Duhok and Erbil governorates over 12 months during September 2016 to August2017, which belong to seven orders (Ephemeroptera, Plecoptera, Trichoptera, Odonatan, Diptera, Coleoptera, and Megaloptera). Clustering mitochondrial DNA cytochrome c oxidase and 16S rDNA genes, morphological keys, and matches in the Barcode of Life Database, we identified 24 species return to 7 orders and 12 families, as indicated in the results. The reported species were: Ephemeroptera 5 members of the family Heptageniidae (Maccaffertium meririvulanum, Raptoheptagenia cruentata, Ecdyonurus dispar, Anepeorus rusticus, Stenonema femoratum), 1 Ephemerellidae (Seratella ignita), 1 Arthropleidae (Arthroplea bipunctata), 6 Baetidae (Baetis alpinus, Baetis braaschi, Baetis noa, Baetis harrisoni, Iswaeon anoka, Heterocloeon amplum), 1 member for each of Diptera, Coleoptera, Megaloptera and Odonatan orders, while Plecoptera 2 members Leuctridae (Leuctra hippopoides, Leuctra inermis) and Tricoptera 4 members 3 Hydropsychidae (Leptonema albovirens, Hydropsyche simulans, Arctopsyche irrorate), 1 Hydroptilidae (Ochrotrichia tenuata). Most of these recorded species and genera were mentioned for the first time and represent new records in Iraq. Presence and distribution of identified species varied between studied sites, as a result of differences in biogeographical and physical conditions.

Distribution of GC-rich heterochromatin and ribosomal genes in three fungus-farming ants (Myrmicinae, Attini, Attina): insights on chromosomal evolution

Cytogenetic studies on fungus-farming ants have shown remarkable karyotype diversity, suggesting different chromosomal rearrangements involved in karyotype evolution in some genera. A notable cytogenetic characteristic in this ant group is the presence of GC-rich heterochromatin in the karyotypes of some ancient and derivative species. It was hypothesized that this GC-rich heterochromatin may have a common origin in fungus-farming ants, and the increase in species studied is important for understanding this question. In addition, many genera within the subtribe Attina have few or no cytogenetically studied species; therefore, the processes that shaped their chromosomal evolution remain obscure. Thus, in this study, we karyotyped, through classical and molecular cytogenetic techniques, the fungus-farming ants Cyphomyrmex transversus Emery, 1894, Sericomyrmex maravalhas Ješovnik et Schultz, 2017, and Mycetomoellerius relictus (Borgmeier, 1934), to provide insights into the chromosomal evolution in these genera and to investigate the presence the GC-rich heterochromatin in these species. Cyphomyrmex transversus (2n = 18, 10m + 2sm + 6a) and S. maravalhas (2n = 48, 28m + 20sm) showed karyotypes distinct from other species from their genera. Mycetomoellerius relictus (2n = 20, 20m) presented the same karyotype as the colonies previously studied. Notably, C. transversus presented the lowest chromosomal number for the genus and a distinct karyotype from the other two previously observed for this species, showing the existence of a possible species complex and the need for its taxonomic revision. Chromosomal banding data revealed GC-rich heterochromatin in all three species, which increased the number of genera with this characteristic, supporting the hypothesis of a common origin of GC-rich heterochromatin in Attina. Although a single chromosomal pair carries rDNA genes in all studied species, the positions of these rDNA clusters varied. The rDNA genes were located in the intrachromosomal region in C. transversus and M. relictus, and in the terminal region of S. maravalhas. The combination of our molecular cytogenetic data and observations from previous studies corroborates that a single rDNA site located in the intrachromosomal region is a plesiomorphic condition in Attina. In addition, cytogenetic data obtained suggest centric fission events in Sericomyrmex Mayr, 1865, and the occurrence of inversions as the origin of the location of the ribosomal genes in M. relictus and S. maravalhas. This study provides new insights into the chromosomal evolution of fungus-farming ants.

Chd1 protects genome integrity at promoters to sustain hypertranscription in embryonic stem cells

Stem and progenitor cells undergo a global elevation of nascent transcription, or hypertranscription, during key developmental transitions involving rapid cell proliferation. The chromatin remodeler Chd1 mediates hypertranscription in pluripotent cells but its mechanism of action remains poorly understood. Here we report a novel role for Chd1 in protecting genome integrity at promoter regions by preventing DNA double-stranded break (DSB) accumulation in ES cells. Chd1 interacts with several DNA repair factors including Atm, Parp1, Kap1 and Topoisomerase 2β and its absence leads to an accumulation of DSBs at Chd1-bound Pol II-transcribed genes and rDNA. Genes prone to DNA breaks in Chd1 KO ES cells are longer genes with GC-rich promoters, a more labile nucleosomal structure and roles in chromatin regulation, transcription and signaling. These results reveal a vulnerability of hypertranscribing stem cells to accumulation of endogenous DNA breaks, with important implications for developmental and cancer biology.

Non-Coding, RNAPII-Dependent Transcription at the Promoters of rRNA Genes Regulates Their Chromatin State in S. cerevisiae

Pervasive transcription is widespread in eukaryotes, generating large families of non-coding RNAs. Such pervasive transcription is a key player in the regulatory pathways controlling chromatin state and gene expression. Here, we describe long non-coding RNAs generated from the ribosomal RNA gene promoter called UPStream-initiating transcripts (UPS). In yeast, rDNA genes are organized in tandem repeats in at least two different chromatin states, either transcribed and largely depleted of nucleosomes (open) or assembled in regular arrays of nucleosomes (closed). The production of UPS transcripts by RNA Polymerase II from endogenous rDNA genes was initially documented in mutants defective for rRNA production by RNA polymerase I. We show here that UPS are produced in wild-type cells from closed rDNA genes but are hidden within the enormous production of rRNA. UPS levels are increased when rDNA chromatin states are modified at high temperatures or entering/leaving quiescence. We discuss their role in the regulation of rDNA chromatin states and rRNA production.

Microorganisms that participate in biochemical cycles in wetlands

Several biochemical cycles are performed in natural wetlands (NWs) and constructed wetlands (CWs). The knowledge of the microorganisms could be used to monitor the restoration of wetlands or the performance of the wastewater treatment. Regarding bacteria, Proteobacteria phylum is the most abundant in NWs and CWs, which possesses a role in N, P, and S cycles, and in the degradation of organic matter. Other phyla are present in lower abundance. Archaea participate in methanogenesis, methane oxidation, and the methanogenic N2 fixation. Sulfur and phosphorus cycles are also performed by other microorganisms, such as Chloroflexi or Nitrospirae phyla. In general, there is more information about the N cycle, especially nitrification and denitrification. Processes where archaea participate (e.g. methane oxidation, methanogenic N2 fixation) are still unclear their metabolic role and several of these microorganisms have not been isolated so far. The study can use 16S rDNA genes or functional genes. The use of functional genes gives information to monitor specific microbial populations and 16S rDNA is more suitable to perform the taxonomic classification. Also, there are several Candidatus microorganisms, which have not been isolated so far. However, it has been described their metabolic role in the biochemical cycles in wetlands.

Chromosomal Analysis of Ctenolucius hujeta Valenciennes, 1850 (Characiformes): A New Piece in the Chromosomal Evolution of the Ctenoluciidae

Ctenoluciidae (Characiformes), a family of freshwater fishes, comprises 2 genera, <i>Ctenolucius</i> and <i>Boulengerella</i>, with 7 recognized species. Up to now, only species of the genus <i>Boulengerella</i> have been subjected to cytogenetic studies. Here, we investigated the karyotype and other cytogenetic features of pike characin, <i>Ctenolucius hujeta</i>, using conventional (Giemsa staining, C-banding, Ag-NOR staining) and molecular (rDNA, telomeric sequences, and fiber-FISH mapping) procedures. This species has a diploid chromosome number of 2n = 36, and a karyotype composed of 12m + 20sm + 4a and FN = 68, similar to that found in <i>Boulengerella</i> species. However, differences regarding the number and distribution of several chromosomal markers support a distinct generic status. Colocalization of the 18S and 5S rDNA genes is an exclusive characteristic of the <i>C. hujeta</i> genome, with an interspersed distribution in the chromosomal fiber, an unusual phenomenon among eukaryotes. Additionally, our results support the view that Ctenoluciidae and Lebiasinidae families are closely related.

Large-scale comparative analysis of cytogenetic markers across Lepidoptera

Fluorescence in situ hybridization (FISH) allows identification of particular chromosomes and their rearrangements. Using FISH with signal enhancement via antibody amplification and enzymatically catalysed reporter deposition, we evaluated applicability of universal cytogenetic markers, namely 18S and 5S rDNA genes, U1 and U2 snRNA genes, and histone H3 genes, in the study of the karyotype evolution in moths and butterflies. Major rDNA underwent rather erratic evolution, which does not always reflect chromosomal changes. In contrast, the hybridization pattern of histone H3 genes was well conserved, reflecting the stable organisation of lepidopteran genomes. Unlike 5S rDNA and U1 and U2 snRNA genes which we failed to detect, except for 5S rDNA in a few representatives of early diverging lepidopteran lineages. To explain the negative FISH results, we used quantitative PCR and Southern hybridization to estimate the copy number and organization of the studied genes in selected species. The results suggested that their detection was hampered by long spacers between the genes and/or their scattered distribution. Our results question homology of 5S rDNA and U1 and U2 snRNA loci in comparative studies. We recommend the use of histone H3 in studies of karyotype evolution.

Cytogenetic Analysis, Heterochromatin Characterization and Location of the rDNA Genes of Hycleus scutellatus (Coleoptera, Meloidae); A Species with an Unexpected High Number of rDNA Clusters

Meloidae are commonly known as blister beetles, so called for the secretion of cantharidin, a toxic substance that causes irritation and blistering. There has been a recent increase in the interest of the cantharidin anticancer potential of this insect group. Cytogenetic and molecular data in this group are scarce. In this study, we performed a karyotype analysis of Hycleus scutellatus, an endemic species of the Iberian Peninsula. We determined its chromosome number, 2n = 20, as well as the presence of the X and Y sex chromosomes. In addition to a karyotype analysis, we carried out DAPI staining. By fluorescence in situ hybridization we mapped the rDNA clusters on 12 different chromosomes. Compared to others, this species shows an unusually high number of chromosomes carrying rDNA. This is one of the highest numbers of rDNA sites found in the Polyphaga suborder (Coleoptera). Additionally, we isolated a satellite DNA family (Hyscu-H), which was located within the pericentromeric regions of all chromosomes, including the sex chromosomes. The results suggest that Hyscu-H is likely to be one of the most abundant satellite DNA repeats in H. scutellatus.