Fly Development Research Articles

A conserved function of Human DLC3 and Drosophila Cv-c in testis development.

The identification of genes affecting gonad development is essential to understand the mechanisms causing Variations/Differences in Sex Development (DSD). Recently, a DLC3 mutation was associated with male gonadal dysgenesis in 46,XY DSD patients. We have studied the requirement of Cv-c, the Drosophila ortholog of DLC3, in Drosophila gonad development, as well as the functional capacity of DLC3 human variants to rescue cv-c gonad defects. We show that Cv-c is required to maintain testis integrity during fly development. We find that Cv-c and human DLC3 can perform the same function in fly embryos, as flies carrying wild type but not patient DLC3 variations can rescue gonadal dysgenesis, suggesting functional conservation. We also demonstrate that the StART domain mediates Cv-c's function in the male gonad independently from the GAP domain's activity. This work demonstrates a role for DLC3/Cv-c in male gonadogenesis and highlights a novel StART domain mediated function required to organize the gonadal mesoderm and maintain its interaction with the germ cells during testis development.

RNAi-mediated knockdown of juvenile hormone acid O-methyltransferase disrupts larval development in the oriental fruit fly, Bactrocera dorsalis (Hendel)

The oriental fruit fly, Bactrocera dorsalis (Hendel), is a notoriously agricultural pest that causes serious economic losses to fruits and vegetables. Widespread insecticide resistance in B. dorsalis is a major obstacle in successful control. Therefore, new pest control strategies, such as those targeting specific genes that can block pest development, are urgently needed. In the current study, the function of JHAMT in B. dorsalis was systematically investigated. A methyltransferase gene in B. dorsalis (BdJHAMT) that is homologous to JHAMT of Drosophila melanogaster was cloned firstly. The subsequently spatiotemporal expression analysis indicated that BdJHAMT mRNA was continuously present in the larval stage, declined sharply immediately before pupation, and then increased in the adult. Subcellular localization showed that BdJHAMT was localized in the adult corpora allata and larval intestinal wall cells. The JH III titer in B. dorsalis was closely related to the transcription level of BdJHAMT in different developmental stages. The dsBdJHAMT feeding-based RNAi resulted in a greatly decreased JH III titer that disrupted fly development. The slow growth caused by BdJHAMT silencing was partially rescued by application of the JH mimic, methoprene. These results demonstrated that BdJHAMT was crucial for JH biosynthesis and thus regulated larval development in B. dorsalis, indicating it may serve as a prospective target for the development of novel control strategies against this pest.

Predict bean production according to bean growth, root rots, fly and weed development under different planting dates and weed control treatments

These two-year experiments determined the best predictors of bean growth and fly, root rots, and weed development in different cultivars, planting dates and weed treatments across 256 plots. Root rot diseases were naturally caused by Fusarium solani and Rhizoctonia solani. Weed management treatments involved: herbicide (Imazethapyr or Trifluralin) application, hand-weeding and control. Parameters estimated by exponential, Gaussian and linear-by-linear models fitted to bean, disease and weed datasets were considered as progress curve elements. Factor analysis detected the most predictive variables to characterize bean growth and production, disease, fly and weed development over season. There were significant correlations between considered plant, disease, pest and weed descriptors. Based on principal component analysis, considered bean-disease-fly-productivity-weed predictors justified 70% of total variance in datasets. Finally, multivariate regression model involving eight selected predictors explained a noticeable part (63%) of yield variations. Such information may improve accuracy of future efforts to monitor bean, disease, fly and weed development, predict bean yield and develop integrative field management programs.

PS-3 Effects of Feeding Rates on Diflubenzuron Efficacy Against Filth Flies (Diptera: Muscidae) in Manure of Beef Heifers

Abstract The objective of the current study is to determine the effect of feeding rates on the efficacy of diflubenzuron against filth fly development in the manure of beef heifers. Twelve Angus × Hereford crossbred heifers (263 kg ± 16.2 kg BW) were used in a completely randomized design and heifers were assigned to a 2 x 2 factorial arrangement of treatments to evaluate the efficacy of diflubenzuron against horn flies Haematobia irritans (L.), stable flies Stomoxys calcitrans and house fly Musca domestica utilizing fecal bioassays. Treatments consisted of 2 insecticidal treatments [Diflubenzuron (DFB; 0.10 mg per kg BW) and no DFB (CNT)] in a factorial arrangement with 2 dietary fed regimens [1.5% BW alfalfa hay cubes and 3.0% BW alfalfa hay cubes]. All heifers receiving DFB treatments were dosed daily beginning on day 0 and continuing through day 7. Manure samples were collected daily prior to DFB offering and for 11 consecutive days. Efficacy calculations were developed utilizing standard filth fly fecal bioassays accounting for control performance. A treatment × day interaction (P ≤ 0.01) was observed for all species largely associated with the presence of DFB and associated with the on-set of efficacy (day 0–2) and wash out (day 8–10) of the product. Feeding regimen did not (P > 0.05) influence DFB performance against any of the fly species evaluated. Formulations of DFB appear to handle a wide range of animal intake scenarios under controlled situations.

Orsai, the Drosophila homolog of human ETFRF1, links lipid catabolism to growth control

BackgroundLipid homeostasis is an evolutionarily conserved process that is crucial for energy production, storage and consumption. Drosophila larvae feed continuously to achieve the roughly 200-fold increase in size and accumulate sufficient reserves to provide all energy and nutrients necessary for the development of the adult fly. The mechanisms controlling this metabolic program are poorly understood.ResultsHerein we identified a highly conserved gene, orsai (osi), as a key player in lipid metabolism in Drosophila. Lack of osi function in the larval fat body, the regulatory hub of lipid homeostasis, reduces lipid reserves and energy output, evidenced by decreased ATP production and increased ROS levels. Metabolic defects due to reduced Orsai (Osi) in time trigger defective food-seeking behavior and lethality. Further, we demonstrate that downregulation of Lipase 3, a fat body-specific lipase involved in lipid catabolism in response to starvation, rescues the reduced lipid droplet size associated with defective orsai. Finally, we show that osi-related phenotypes are rescued through the expression of its human ortholog ETFRF1/LYRm5, known to modulate the entry of β-oxidation products into the electron transport chain; moreover, knocking down electron transport flavoproteins EtfQ0 and walrus/ETFA rescues osi-related phenotypes, further supporting this mode of action.ConclusionsThese findings suggest that Osi may act in concert with the ETF complex to coordinate lipid homeostasis in the fat body in response to stage-specific demands, supporting cellular functions that in turn result in an adaptive behavioral response.

Identification and Characterization of Small RNA Markers of Age in the Blow Fly Cochliomyia macellaria (Fabricius) (Diptera: Calliphoridae).

Simple SummaryImmature blow fly development information is critical for temporal estimations in death investigations. While there are phenotypic markers of development, they often do not provide adequate precision and resolution. In order to ameliorate these issues, we have investigated microRNAs as putative markers of immature development in the forensically relevant blow fly, Cochliomyia macellaria. Using RNA sequencing, we were able to identify miRNAs present in immature developmental stages, as well as identify miRNA which were significantly differentially expressed. Through these analyses, we found likely markers of development time, some of which were validated with qPCR methods. As a follow-up, we investigated factors which may influence expression of miRNA throughout development, such as selected genotypic variation and sex. It was very clear that genetic and abiotic factors can impact predictions of age, some of which may exhibit interactions. It is important that future work investigate the markers most robust to genetic effects on development.Blow fly development is important in decomposition ecology, agriculture, and forensics. Much of the impact of these species is from immature samples, thus knowledge of their development is important to enhance or ameliorate their effects. One application of this information is the estimation of immature insect age to provide temporal information for death investigations. While traditional markers of age such as stage and size are generally accurate, they lack precision in later developmental stages. We used miRNA sequencing to measure miRNA expression, throughout development, of the secondary screwworm, Cochliomyia macellaria (Fabricius) (Diptera: Calliphoridae) and identified 217 miRNAs present across the samples. Ten were identified to be significantly differentially expressed in larval samples and seventeen were found to be significantly differentially expressed in intrapuparial samples. Twenty-eight miRNAs were identified to be differentially expressed between sexes. Expression patterns of two miRNAs, miR-92b and bantam, were qPCR-validated in intrapuparial samples; these and likely food-derived miRNAs appear to be stable markers of age in C. macellaria. Our results support the use of miRNAs for developmental markers of age and suggest further investigations across species and under a range of abiotic and biotic conditions.

Filth Fly Parasitoid (Hymenoptera: Pteromalidae) Monitoring Techniques and Species Composition in Poultry Layer Facilities.

Muscid flies, especially house flies (Musca domestica L.) (Diptera: Muscidae), are a major pest of poultry layer facilities. Augmentative biological control of muscid flies with pteromalid wasps has gained increased attention in recent years. Knowing which pteromalid species are present in a specific area could produce more effective filth fly control. The purpose of this project was to survey parasitoid populations in poultry layer facilities in central and southeastern Pennsylvania from June through September. Two genera of parasitoids, Spalangia and Trichomalopsis, were collected over the course of the survey. Overall, out of 3,724 parasitized pupae the species collected in order of most to least common were Spalangia cameroni Perkins, Spalangia nigroaenea Curtis, Trichomalopsis spp., and Spalangia endius Walker. House fly parasitism overall and by each parasitoid species varied by location and over the four study months. A second objective was to evaluate a new parasitoid trap for surveying parasitoid wasp populations. This device uses a combination of house fly third instars and development media. This was compared to a more traditional method, the sentinel bag, which uses only fly pupae. A higher proportion of Spalangia spp. emerged from the new trap design and more Trichomalopsis spp. emerged from the sentinel bag. This suggests that using this new device alongside the traditional collection method may result in more accurate sampling of pteromalid populations.

Metabolic reserves of diapausing western cherry fruit fly (Diptera: Tephritidae) pupae in relation to chill duration and post-chill rearing conditions.

How different macronutrients are utilized at various stages of pupal diapause and the effects of winter length on nutrient reserves remain poorly studied for most insects. Western cherry fruit fly, Rhagoletis indifferens (Diptera: Tephritidae), is a specialist on cherries in higher latitudes or elevations in western North America that exhibits a obligate pupal diapause requiring chilling before adult development can occur. We determined the relationship between metabolic reserves and diapause status in R. indifferens pupae, testing the hypotheses that lipids are the primary reserves utilized during diapause and that long periods of warmth deplete these reserves more than periods of cold. Effects of 0- to 20-week durations at 3°C and subsequent exposure to 23°C and 16:8 L:D (warm rearing conditions) for 0 to 7 weeks on lipid, protein, soluble carbohydrates, and glycogen reserves of R. indifferens pupae were determined. During diapause, lipid reserves were the primary source of energy utilized by R. indifferens, while protein and soluble carbohydrates levels were stable throughout diapause and thus less utilized. At post-diapause, glycogen levels fluctuated the most, indicating that lipid reserves were utilized to produce glycogen to support metabolism for adult fly development. Unchilled pupae did not deplete lipid reserves, unlike chilled pupae, likely because unchilled pupae remained in diapause. Rhagoletis indifferens may have evolved a nutrient utilization strategy typical of rigid diapausing insects in higher latitude environments.

Optimization of the production of black soldier flies Hermetia Illucens by controlling biological parameters in Côte d'Ivoire

This study was conducted at the ENS vivarium located at the UFHB of Cocody in the city of Abidjan in the south of Ivory Coast, with a temperature of about 28° C and a humidity of about 70%. It aimed to study some biological parameters of the black soldier fly Hermetia illucens. For achieve this goal, it was a question of studying the different stages of development of the black soldier fly and also studying the evolution of its growth parameters. The flies were captured, kept in an aviary and observed from the fly stage to the end of the pupal stage (emergence of the first flies). Data on the duration of the different stages of development and the evolution of larval growth parameters (average weight, length, width, Fulton index, length/width ratio, survival and mortality rate of larvae) have been taken. This study revealed that the life cycle of Hermetia illucens is between 31 and 43 days, with an incubation time of eggs that lasts 2 days, a larval phase of 14 days, a pupation of 10 days and a duration of adult life of 8 days on average. In addition, this study reports a low mortality rate of larvae (3.45%), larvae much longer (19.46 mm), wider (6 mm) and plumper.

Pilot study of a new methodology to study the development of the blue bottle fly (Calliphora vomitoria) under exposure to radio-frequency electromagnetic fields at 5.4 GHz

Purpose Exposure of insects to radio-frequency electromagnetic fields (RF-EMFs) can have developmental effects. However, there is currently no clear understanding of the exposure level that can lead to such effects. Therefore, the goal of this study was to, for the first time, study the development of the Blue Bottle Fly (Calliphora vomitoria, CV) under exposure to RF-EMFs at 5.4 GHz, using both numerical RF-EMF dosimetry with anatomically accurate 3 D models of insects and an RF-EMF exposure experiment. Materials and Methods CV was chosen as a model organism in this study because CV’s development can be influenced thermally and CV’s pupal stage presents a window of several days in which immobile pupae can be exposed to RF-EMFs. The 5.4 GHz frequency was used because it allowed us the license-free operation of the exposure setup. Numerical, EM simulations with 3 D anatomically accurate models of CV, obtained using micro-CT scanning, were used in this study. These simulations enable the estimation of the absorbed power and the whole-body averaged specific absorption rate in CV during RF exposure experiments. An experiment with three exposure conditions was designed and executed in which 400 pupae were split into an exposed group that was placed inside the TEM cell for 48 h and concurrent control. Two exposure conditions used RF-EMF input power into the TEM cell at 5.4 GHz on two different levels. One exposure condition was sham exposure. Electric field strength measurements were used to validate the proper functioning of the exposure setups and to quantify the RF-EMF exposure of the control groups. Results and conclusions All studied groups of pupae – exposed to RF-EMFs, sham, and control groups- showed similar (evolutions of) masses, lengths and diameters during their development. The total rate of pupal emergence was reduced in one of the studied RF-EMF exposures in comparison to its concurrent control, while the other RF-EMF exposure and the sham exposure did not alter the total rate of pupal emergence. The sham exposure and the lowest of the two studied RF-EMF exposure conditions (19.4 V/m) caused a median delay in pupal emergence of 4 and 8 hours, respectively, in comparison to concurrent control groups. The higher studied exposure of 55 V/m caused a median relative acceleration in the development of 8 h.

Penggunaan Probiotik Dan Magot Untuk Mereduksi Lalat di Peternakan Ayam Pedaging Muara Selayar Desa Pijot

Chicken manure waste is one of the main problems for large-scale broiler cultivation, especially in areas close to residential areas. The accumulation of manure that accumulates under the cage attracts the arrival and development of the flies in the cultivation site. The development of these flies will spread to the houses of residents who live around the cage. As a result, many farmers stop cultivating the broilers because of public complaints. To overcome this problem, the use of probiotics from acetic acid-producing bacteria can be used to reduce the development of flies because of the ability of the bacteria to reduce the pH of the media and create an uncomfortable environment for egg growth. Likewise, the development of maggot can be a competing medium that beats the flay. In community service activities regarding "The use of probiotics and maggot production to reduce flies in the Muara Selayar broiler farm, Pijot Village, Keruak District, East Lombok Regency". Through this activity, the development of flies can be emphasized by the use of probiotics and the development of magot which can be used as chicken feed.

Argonaute1 and Gawky Are Required for the Development and Reproduction of Melon fly, Zeugodacus cucurbitae.

Argonaute family genes encode a highly conserved group of proteins that have been associated with RNA silencing in both animals and plants. This study investigates the importance of microRNA biogenesis key regulators Argonaute1 (Ago1) and Gawky genes in the post-embryonic and ovarian development of the melon fly, Zeugodacus cucurbitae. The expression levels of these genes were mapped in all developmental stages and different adult tissues. Their roles in development were investigated using RNA interference (RNAi) via two different dsRNA delivery techniques. Embryo microinjection and oral feeding of third instar larvae successfully knocked down and greatly reduced the expression level of the target genes. Additionally, ex vivo essays revealed the stability of dsRNA in food was sufficient for gene silencing, although its integrity was affected in midgut. A wide range of phenotypes were observed on pupation, segmentation, pigmentation, and ovarian development. RNAi-mediated silencing of Gawky caused high mortality and loss of body segmentation, while Ago1 knockdown affected ovarian development and pigmentation. Developmental abnormalities and ovarian malformations caused by silencing these genes suggest that these genes are crucial for viability and reproductive capacity of Z. cucurbitae, and may be used as potential target genes in pest management.

The Role of GCN2 Kinase in Mediating the Effects of Amino Acids on Longevity and Feeding Behaviour in Drosophila.

Restriction of amino acids in the diet can extend lifespan in diverse species ranging from flies to mammals. However, the role of individual amino acids and the underlying molecular mechanisms are only partially understood. The evolutionarily conserved serine/threonine kinase General Control Nonderepressible 2 (GCN2) is a key sensor of amino acid deficiency and has been implicated in the response of lifespan to dietary restriction (DR). Here, we generated a novel Drosophila GCN2 null mutant and analyzed its response to individual amino acid deficiency. We show that GCN2 function is essential for fly development, longevity and feeding behaviour under long-term, but not short-term, deprivation of all individual essential amino acids (EAAs) except for methionine. GCN2 mutants were longer-lived than control flies and showed normal feeding behaviour under methionine restriction. Thus, in flies at least two systems regulate these responses to amino acid deprivation. Methionine deprivation acts via a GCN2-independent mechanism, while all other EAA are sensed by GCN2. Combined deficiency of methionine and a second EAA blocked the response of GCN2 mutants to methionine, suggesting that these two pathways are interconnected. Wild type flies showed a short-term rejection of food lacking individual EAA, followed by a long-term compensatory increase in food uptake. GCN2 mutants also showed a short-term rejection of food deprived of individual EAA, but were unable to mount the compensatory long-term increase in food uptake. Over-expression of the downstream transcription factor ATF4 partially rescued the response of feeding behaviour in GCN2 mutants to amino acid deficiency. Phenotypes of GCN2 mutants induced by leucine and tryptophan, but not isoleucine, deficiency were partially rescued by ATF4 over-expression. The exact function of GCN2 as an amino acid sensor in vivo and the downstream action of its transcription factor effector ATF4 are thus context-specific with respect to the EAA involved.

A prominent gene activation role for C-terminal binding protein in mediating PcG/trxG proteins through Hox gene regulation.

The evolutionarily conserved C-terminal binding protein (CtBP) has been well characterized as a transcriptional co-repressor. Herein, we report a previously unreported function for CtBP, showing that lowering CtBP dosage genetically suppresses Polycomb group (PcG) loss-of-function phenotypes while enhancing that of trithorax group (trxG) in Drosophila, suggesting that the role of CtBP in gene activation is more pronounced in fly development than previously thought. In fly cells, we show that CtBP is required for the derepression of the most direct PcG target genes, which are highly enriched by homeobox transcription factors, including Hox genes. Using ChIP and co-IP assays, we demonstrate that CtBP is directly required for the molecular switch between H3K27me3 and H3K27ac in the derepressed Hox loci. In addition, CtBP physically interacts with many proteins, such as UTX, CBP, Fs(1)h and RNA Pol II, that have activation roles, potentially assisting in their recruitment to promoters and Polycomb response elements that control Hox gene expression. Therefore, we reveal a prominent activation function for CtBP that confers a major role for the epigenetic program of fly segmentation and development.

Assessing the Tsetse Fly Microbiome Composition and the Potential Association of Some Bacteria Taxa with Trypanosome Establishment.

The tsetse flies, biological vectors of African trypanosomes, harbour a variety of bacteria involved in their vector competence that may help in developing novel vector control tools. This study provides an inventory of tsetse bacterial communities in Cameroon and explores their possible associations with trypanosome establishment in Glossina palpalis palpalis. High throughput sequencing of the V3-V4 hypervariable region of the bacterial 16S rRNA gene, with subsequent metagenomic, multivariate, and association analyses, were used to investigate the levels and patterns of microbial diversity in four tsetse species. Overall, 31 bacterial genera and four phyla were identified. The primary symbiont Wigglesworthia dominated almost all the samples, with an overall relative abundance of 47.29%, and seemed to be replaced by Serratia or Burkholderia in some G. tachinoides flies. Globally, significant differences were observed in the microbiome diversity and composition among tsetse species and between teneral and non-teneral flies, or between flies displaying or not displaying mature trypanosome infections. In addition, differential abundance testing showed some OTUs, or some bacteria taxa, associated with trypanosome maturation in tsetse flies. These bacteria could be further investigated for an understanding of their mechanism of action and alternatively, transformed and used to block trypanosome development in tsetse flies.

Reduced body length and morphological disorders in Chrysomya albiceps (Diptera: Calliphoridae) larvae reared on aluminum phosphide-treated rabbits

Assessing the time of death based on the growth and development of insects is a critical task in forensic entomology. The rate of larvae development can be affected by a variety of toxins, including pesticides. Aluminum phosphide (AlP) is a low-cost insecticide that has yet to be tested for entomotoxicological significance, despite the fact that it is frequently the cause of fatal poisoning. In this study, we measured the body length of Chrysomya albiceps larvae reared on the carcasses of rabbits poisoned with AlP and analyzed the morphological changes of the larvae reared on the carcasses of rabbits poisoned with AlP. The concentration of AlP in the body of the larvae was significantly lower than in rabbit tissues. Insects from the AlP group had a significantly lower gain in body length. Furthermore, deformities in the larvae were found. Smaller respiratory spiracles were found, as well as a deformed small posterior end with hypogenesis of the posterior respiratory spiracles. Thus, disturbed growth and development of carrion flies found at a crime scene could indicate pesticide poisoning, such as aluminum phosphide.

The Drosophila melanogaster enzyme glycerol-3-phosphate dehydrogenase 1 is required for oogenesis, embryonic development, and amino acid homeostasis

As the fruit fly, Drosophila melanogaster, progresses from one life stage to the next, many of the enzymes that compose intermediary metabolism undergo substantial changes in both expression and activity. These predictable shifts in metabolic flux allow the fly meet stage-specific requirements for energy production and biosynthesis. In this regard, the enzyme glycerol-3-phosphate dehydrogenase 1 (GPDH1) has been the focus of biochemical genetics studies for several decades and, as a result, is one of the most well-characterized Drosophila enzymes. Among the findings of these earlier studies is that GPDH1 acts throughout the fly lifecycle to promote mitochondrial energy production and triglyceride accumulation while also serving a key role in maintaining redox balance. Here, we expand upon the known roles of GPDH1 during fly development by examining how depletion of both the maternal and zygotic pools of this enzyme influences development, metabolism, and viability. Our findings not only confirm previous observations that Gpdh1 mutants exhibit defects in larval development, lifespan, and fat storage but also reveal that GPDH1 serves essential roles in oogenesis and embryogenesis. Moreover, metabolomics analysis reveals that a Gpdh1 mutant stock maintained in a homozygous state exhibits larval metabolic defects that significantly differ from those observed in the F1 mutant generation. Overall, our findings highlight unappreciated roles for GPDH1 in early development and uncover previously undescribed metabolic adaptations that could allow flies to survive the loss of this key enzyme.

A meiotic switch in lysosome activity supports spermatocyte development in young flies but collapses with age

SummaryGamete development ultimately influences animal fertility. Identifying mechanisms that direct gametogenesis, and how they deteriorate with age, may inform ways to combat infertility. Recently, we found that lysosomes acidify during oocyte maturation in Caenorhabditis elegans, suggesting that a meiotic switch in lysosome activity promotes female germ-cell health. Using Drosophila melanogaster, we report that lysosomes likewise acidify in male germ cells during meiosis. Inhibiting lysosomes in young-male testes causes E-cadherin accumulation and loss of germ-cell partitioning membranes. Notably, analogous changes occur naturally during aging; in older testes, a reduction in lysosome acidity precedes E-cadherin accumulation and membrane dissolution, suggesting one potential cause of age-related spermatocyte abnormalities. Consistent with lysosomes governing the production of mature sperm, germ cells with homozygous-null mutations in lysosome-acidifying machinery fail to survive through meiosis. Thus, lysosome activation is entrained to meiotic progression in developing sperm, as in oocytes, and lysosomal dysfunction may instigate male reproductive aging.

The DNA Binding Diversity of the SIX Homeodomain Transcription Factor Family

Transcription factor (TF) DNA‐binding specificity is determined by how their DNA‐binding domain (DBD) interacts with DNA. TFs are identified by the sequence homology shared with described DBDs, which allows them to be classified into families. It is highly widely accepted that similar DBDs recognize similar DNA motifs. However, changes in a TF can lead to changes in its DNA recognition. TFs are critical for development and have been found since the beginning of animals, how their DNA binding changes through time has not been fully studied. TFs members of the sine oculis homeobox (SIX) homeodomain family are found from sponges to humans and are considered atypical members of the homeodomain family. They regulate numerous developmental processes, with phenotypic features spanning from eye development in flies, red color patterning in Heliconius butterfly wings to brain development in humans. How evolutionary related TFs diversify their function has not been fully understood, especially changes to their DNA binding specificity. Using full length SIX proteins from Drosophila melanogaster, Heliconius erato, and Homo sapiens, we performed in vitro Systematic Evolution of Ligands by Exponential Enrichment (SELEX‐seq) to identify the DNA binding specificity. Our preliminary data shows the majority bind to their canonical binding motif (TGATAC), except for six4 members which prefer TGACAC. Interestingly, the way they bind to these motifs differs. Both sine oculis and six4 homologs require a 5’‐GA flanking the core motif. While optix related members prefer a shorter flaking region with less dependence on 5’‐GA. This is interesting since optix is more evolutionarily related to six4 than to sine oculis. In addition, we found that Heliconius erato optix can bind both as a homodimer with a preferred spacing of 2‐bp between binding sites or as a monomer. Our results provide new insights in the binding diversity found in the SIX family of TFs and predict genomic targets of these understudied TF family.

Blow Flies (Diptera: Calliphoridae) and the American Diet: Effects of Fat Content on Blow Fly Development.

Forensic entomology is an important field of forensic science that utilizes insect evidence in criminal investigations. Blow flies (Diptera: Calliphoridae) are among the first colonizers of remains and are frequently used in determining the minimum postmortem interval (mPMI). Blow fly development is influenced by a variety of factors including temperature and feeding substrate. Unfortunately, dietary fat content remains an understudied factor in the development process, which is problematic given the high rates of obesity in the United States. To study the effects of fat content on blow flies we investigated survivorship, adult weight, and development time (overall and by sex) of Lucilia sericata (Meigen) and Phormia regina (Meigen) on ground beef with 10%, 20%, or 27% fat. As fat content increased, survivorship decreased across both species with significant impact to P. regina. While P. regina adults were generally larger than L. sericata across all fat levels, only L. sericata demonstrated a significant (P < 0.05) difference in weight by sex. Overall development times did not vary by fat content, excluding 27% for P. regina. Additionally, development times did not vary by sex for P. regina but did differ (P < 0.05) for L. sericata with females taking longer to develop. These findings provide insight into the effect of fat content on blow fly development, a factor that should be considered when estimating an mPMI. By understanding how fat levels affect the survivorship and development of the species studied here, we can begin improving the practice of insect evidence analysis in casework.