Variation and Variability in Drosophila Grooming Behavior

Behavioral differences can be observed between species or populations (variation) or between individuals in a genetically similar population (variability). Here, we investigate genetic differences as a possible source of variation and variability in Drosophila grooming. Grooming confers survival and social benefits. Grooming features of five Drosophila species exposed to a dust irritant were analyzed. Aspects of grooming behavior, such as anterior to posterior progression, were conserved between and within species. However, significant differences in activity levels, proportion of time spent in different cleaning movements, and grooming syntax were identified between species. All species tested showed individual variability in the order and duration of action sequences. Genetic diversity was not found to correlate with grooming variability within a species: melanogaster flies bred to increase or decrease genetic heterogeneity exhibited similar variability in grooming syntax. Individual flies observed on consecutive days also showed grooming sequence variability. Standardization of sensory input using optogenetics reduced but did not eliminate this variability. In aggregate, these data suggest that sequence variability may be a conserved feature of grooming behavior itself. These results also demonstrate that large genetic differences result in distinguishable grooming phenotypes (variation), but that genetic heterogeneity within a population does not necessarily correspond to an increase in the range of grooming behavior (variability).

Genetic Variability of Acetolactate Synthase (ALS) Sequence in Centaurea cyanus Plants Resistant and Susceptible to Tribenuron-Methyl

Centaurea cyanus, belonging to the Asteraceae family, is an arable weed species encountered mainly in fields with cereals, sugar beet, and maize. The high genetic variability of C. cyanus has been recently reported; however, little is known about its sequence variability in the context of its herbicide resistance. C. cyanus resistance was found mainly against acetolactate synthase (ALS) inhibitors, but no ALS sequence information concerning the herbicide resistance mechanism has been published yet. The aim of this study was to determine the ALS sequences for biotypes susceptible and resistant to tribenuron-methyl in order to identify mutations that may be associated with the resistance emergence. DNA isolation from susceptible and resistant plants was followed by PCR amplification and ALS sequencing. As a result, different lengths of DNA products were obtained. Moreover, both nucleotide and amino acid sequence analysis revealed high sequence variability within one plant as well as between plants from the same biotype. In a few resistant plants, four changes in the amino acid sequence were identified in comparison to those in the susceptible ones. However, these preliminary studies require further investigation toward confirming the significance of these mutations in herbicide resistance development. This study provides preliminary information contributing to the research on the C. cyanus target-site resistance mechanism.

Software choice and depth of sequence coverage can impact plastid genome assembly - A case study in the narrow endemic Calligonum bakuense

Most plastid genome sequences are assembled from short-read whole-genome sequencing data, yet the impact that sequence coverage and the choice of assembly software can have on the accuracy of the resulting assemblies is poorly understood. In this study, we test the impact of both factors on plastid genome assembly in the threatened and rare endemic shrub Calligonum bakuense, which forms a distinct lineage in the genus Calligonum. We aim to characterize the differences across plastid genome assemblies generated by different assembly software tools and levels of sequence coverage and to determine if these differences are large enough to affect the phylogenetic position inferred for C. bakuense. Four assembly software tools (FastPlast, GetOrganelle, IOGA, and NOVOPlasty) and three levels of sequence coverage (original depth, 2,000x, and 500x) are compared in our analyses. The resulting assemblies are evaluated with regard to reproducibility, contig number, gene complement, inverted repeat length, and computation time; the impact of sequence differences on phylogenetic tree inference is also assessed. Our results show that software choice can have a considerable impact on the accuracy and reproducibility of plastid genome assembly and that GetOrganelle produced the most consistent assemblies for C. bakuense. Moreover, we found that a cap in sequence coverage can reduce both the sequence variability across assembly contigs and computation time. While no evidence was found that the sequence variability across assemblies was large enough to affect the phylogenetic position inferred for C. bakuense, differences among the assemblies may influence genotype recognition at the population level.

Sequence Variability of Hepatitis B Virus (HBV) Surface Antigen Gene (S Gene) Among the Non-responders of Antiviral Therapy

BackgroundHepatitis B Virus (HBV), has been among the wide spread lethal causative agent of mortality in the population of Pakistan. Prolonged administration of antiviral therapy for chronic hepatitis B may result in the development of hepatitis B viral mutants. ObjectivesTo gain insight into the mechanism involved in the sequence variability of Hepatitis B Virus (HBV) surface antigen gene (S gene) among responders and non-responders to antiviral therapy, baseline characteristics of the patients and sequences within the S region were investigated in pre-treatment serum samples of responders and post-treatment serum samples of non-responders. Data collection and methodologyThe data was collected from 15 individuals with chronic hepatitis B from Khyber Pakhtunkhwa (KPK) province, Pakistan. The antiviral response was independent of viral genotypes, and nonresponse to antiviral therapy was associated with a complex variability of the viral mutants as determined by PCR. ResultsThe sequence analysis of the S gene among responders and non-responders patients of pre and post-treatment with antiviral therapy showed variability in DNA sequence marked as Pakistani isolates make a distinct cluster in the phylogenetic tree. The S gene of HBV isolates from KPK province shows some similarities with isolates of other countries. No significant variations of nucleotides in the S gene of HBV was found among the responders and non-responders receiving antiviral therapy indicating that S gene may not be important with respect to treatment outcome. ConclusionIt illustrates that antigenicity of other various HBV proteins can be targeted in order to design more effective vaccines against the local strains.

Analysis of Sequence Variability and Transcriptional Profile of Cannabinoid synthase Genes in Cannabis sativa L. Chemotypes with a Focus on Cannabichromenic acid synthase

Cannabis sativa L. has been long cultivated for its narcotic potential due to the accumulation of tetrahydrocannabinolic acid (THCA) in female inflorescences, but nowadays its production for fiber, seeds, edible oil and bioactive compounds has spread throughout the world. However, some hemp varieties still accumulate traces of residual THCA close to the 0.20% limit set by European Union, despite the functional gene encoding for THCA synthase (THCAS) is lacking. Even if some hypotheses have been produced, studies are often in disagreement especially on the role of the cannabichromenic acid synthase (CBCAS). In this work a set of European Cannabis genotypes, representative of all chemotypes, were investigated from a chemical and molecular point of view. Highly specific primer pairs were developed to allow an accurate distinction of different cannabinoid synthases genes. In addition to their use as markers to detect the presence of CBCAS at genomic level, they allowed the analysis of transcriptional profiles in hemp or marijuana plants. While the high level of transcription of THCAS and cannabidiolic acid synthase (CBDAS) clearly reflects the chemical phenotype of the plants, the low but stable transcriptional level of CBCAS in all genotypes suggests that these genes are active and might contribute to the final amount of cannabinoids.

Genetic Variability of Acetolactate Synthase (ALS) Sequence in Centaurea Cyanus Plants Resistant and Susceptible to Tribenuron-methyl

Centaurea cyanus, belonging to the Asteraceae family, is an arable weed species being encountered mainly in the fields with cereals, sugar beet, and corn. C. cyanus high genetic variability has recently been reported, however, little is known about sequence variability in the context of herbicide resistance. C. cyanus resistance was found mainly against acetolactate inhibitors (ALS) inhibitors, but no ALS sequence information concerning herbicide resistance mechanism has been published yet. Therefore, the aim of this study was to determine ALS sequences for biotypes susceptible and resistant to tribenuron-methyl in order to identify possible mutations conferring the resistance. DNA isolation from susceptible and resistant plants was followed by PCR amplification and sequencing of ALS sequence. As a result different lengths of DNA products were obtained. Moreover, both nucleotide and amino acid sequence analysis revealed high sequence variability within one plant as well as between plants from the same biotype. In a few resistant plants, six changes in amino acid sequence were identified in comparison to susceptible ones. However, these preliminary studies require further investigation toward confirming the significance of these mutations in herbicide resistance development. This study provides the first attempt in the research on C. cyanus target-site resistance mechanism.

Multivalency transforms SARS-CoV-2 antibodies into ultrapotent neutralizers

SARS-CoV-2, the virus responsible for COVID-19, has caused a global pandemic. Antibodies can be powerful biotherapeutics to fight viral infections. Here, we use the human apoferritin protomer as a modular subunit to drive oligomerization of antibody fragments and transform antibodies targeting SARS-CoV-2 into exceptionally potent neutralizers. Using this platform, half-maximal inhibitory concentration (IC50) values as low as 9 × 10−14 M are achieved as a result of up to 10,000-fold potency enhancements compared to corresponding IgGs. Combination of three different antibody specificities and the fragment crystallizable (Fc) domain on a single multivalent molecule conferred the ability to overcome viral sequence variability together with outstanding potency and IgG-like bioavailability. The MULTi-specific, multi-Affinity antiBODY (Multabody or MB) platform thus uniquely leverages binding avidity together with multi-specificity to deliver ultrapotent and broad neutralizers against SARS-CoV-2. The modularity of the platform also makes it relevant for rapid evaluation against other infectious diseases of global health importance. Neutralizing antibodies are a promising therapeutic for SARS-CoV-2.

Genetic Variation in the Glycoprotein B Sequence of Equid Herpesvirus 5 among Horses of Various Breeds at Polish National Studs

Equid herpesvirus 5 (EHV-5) is one of two γ-herpesviruses that commonly infect horses worldwide. The objective of the study was to estimate the genetic variability within EHV-5 viruses circulating among horses in Poland. Partial glycoprotein B (gB) sequences from 92 Polish horses from 13 studs throughout Poland were compared to each other and to three EHV-5 sequences from other countries. Despite the overall high level of conservation, considerable variability was observed around the putative furin cleavage site. Based on phylogenetic analysis, the viruses clustered within two major lineages (A and B), with further sub-clustering within group A. The clustering of EHV-5 sequences was independent of age or geographical origin of the sampled horses. Recombination was identified as one of the factors contributing to the genomic heterogeneity. Viruses from unweaned foals were more similar to viruses from other foals at the same stud than to viruses form their dams, suggesting the horizontal transfer and/or evolution of EHV-5 within individual hosts. Our data indicate that the gB sequence is not suitable for tracking the source of EHV-5 infection. Further research is needed to elucidate the importance of the sequence variability around the EHV-5 gB furin cleavage site on the biology of the virus.

Genome Sequences and Phylogeny of Two Duck Hepatitis B Viruses

ABSTRACT Duck hepatitis B virus (DHBV) infection in Pekin ducks is a model for human hepatitis B. Sequence variations may contribute to host therapy responses against the virus. We provide full genome sequences of two DHBVs from France, their phylogenetic classification, and their sequence variability.