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1.
Mikrobiyol Bul ; 54(3): 392-403, 2020 Jul.
Artigo em Turco | MEDLINE | ID: mdl-32755516

RESUMO

Lucilia sericata, a member of the Calliphoridae family, is one of the most common species in the genus Lucilia. Medical importance of L.sericata stems from its use in maggot debridement therapy (MDT). MDT is the name of L.sericata larvae being sterilized and used in the treatment of non-healing wounds. L.sericata maggots used in the treatment of chronic and non-healing wounds (decubitus ulcer, venous leg ulcer, diabetic foot ulcer, etc.) clean the wounds with the help of secreted proteolytic trypsin and lucimycin -like enzymes. The aim of the study was to determine the molecular characterization of lucimycin gene obtained from L.sericata larvae in MDT by using molecular methods and to contribute to the literature. In this study, continuous production of adult colonies of L.sericata species was carried out in insectarium unit where conditions such as light, humidity and temperature were formed. The life cycle of L.sericata was followed and the production of eggs, larvae, pupae, adult flies and fly colonies of the species were formed. In the third stage larvae obtained from adult flies in the insectarium unit, RNA was isolated and subsequently cDNA synthesis was performed by reverse transcription. Polymerase chain reaction (PCR) analysis of the synthesized cDNAs with the specific primers designed for the lucimycin gene of L.sericata was performed and the obtained amplicons were cloned into pJET1.2/blunt vector and the plasmid was purified. The recombinant plasmids were sequenced with vector-specific primers and target gene region sequences were obtained. After the molecular characterization of the isolate with nucleotide sequences was determined, it was registered to GenBank database with the accession number MF964229. The PCR product of 288 bp was obtained from the cDNA obtained from the larvae of L.sericata produced in the insectarium unit by PCR using lucimycin specific primers. The PCR product imaged on the gel was purified by transformation and subsequent colonies were screened to see whether they contained recombinant plasmids. Three of the colonies were identified as recombinant plasmids containing L.sericata lucimycin gene by PCR screening. From three colonies confirmed by PCR screening, recombinant plasmids containing L.sericata lucimycin gene were purified by miniprep. The recombinant plasmid product was confirmed to contain the L.sericata lucimycin gene by PCR from a total of 20 µl of the recombinant plasmid miniprep product. DNA sequencing analysis was performed to confirm the plasmid after cloning. The 288 bp L.sericata lucimycin sequence was confirmed by DNA sequence analysis. The lucimycin gene isolated was confirmed by specific and pJET1.2 forward and reverse primers using Blastn algorithm as a result of species and/or subspecies using the Blastn algorithm and the related isolate was recorded in GenBank database with the MF964229 accessory number. The DNA sequence of the isolated sample was compared with other isolates found in GenBank by Pubmed/Blast program. KJ413251.1 was found to be 99% similar to the GenBank isolate. The 113th nucleotide was C (cytosine) in the sequence of our isolate, while the existence of G (guanine) in the sequence numbered KJ413251.1 GenBank revealed the difference between the two sequences. In this study the molecular characterization of lucimycin gene derived from L.sericata larvae were determined for the first time in Turkey, it is assumed that this molecule which has an antifungal property, can be used in the studies that will be carried out in the future, especially in microorganisms causing cutaneous infections. The study is important since the isolate is registered as a biological asset of Turkey in GenBank and also being the second study in the world.


Assuntos
Dípteros , Genes de Insetos , Lucensomycin , Animais , Dípteros/enzimologia , Dípteros/genética , Genes de Insetos/genética , Larva , Turquia
2.
Proc Biol Sci ; 287(1931): 20201267, 2020 07 29.
Artigo em Inglês | MEDLINE | ID: mdl-32693728

RESUMO

Müllerian mimicry strongly exemplifies the power of natural selection. However, the exact measure of such adaptive phenotypic convergence and the possible causes of its imperfection often remain unidentified. Here, we first quantify wing colour pattern differences in the forewing region of 14 co-mimetic colour pattern morphs of the butterfly species Heliconius erato and Heliconius melpomene and measure the extent to which mimicking colour pattern morphs are not perfectly identical. Next, using gene-editing CRISPR/Cas9 KO experiments of the gene WntA, which has been mapped to colour pattern diversity in these butterflies, we explore the exact areas of the wings in which WntA affects colour pattern formation differently in H. erato and H. melpomene. We find that, while the relative size of the forewing pattern is generally nearly identical between co-mimics, the CRISPR/Cas9 KO results highlight divergent boundaries in the wing that prevent the co-mimics from achieving perfect mimicry. We suggest that this mismatch may be explained by divergence in the gene regulatory network that defines wing colour patterning in both species, thus constraining morphological evolution even between closely related species.


Assuntos
Mimetismo Biológico , Borboletas/fisiologia , Animais , Borboletas/genética , Borboletas/crescimento & desenvolvimento , Genes de Insetos , Pigmentação/genética , Seleção Genética , Asas de Animais
3.
PLoS Genet ; 16(6): e1008622, 2020 06.
Artigo em Inglês | MEDLINE | ID: mdl-32520935

RESUMO

Insect courtship and mating depend on integration of olfactory, visual, and tactile cues. Compared to other insects, Bombyx mori, the domesticated silkworm, has relatively simple sexual behaviors as it cannot fly. Here by using CRISPR/Cas9 and electrophysiological techniques we found that courtship and mating behaviors are regulated in male silk moths by mutating genes in the sex determination cascade belonging to two conserved pathways. Loss of Bmdsx gene expression significantly reduced the peripheral perception of the major pheromone component bombykol by reducing expression of the product of the BmOR1 gene which completely blocked courtship in adult males. Interestingly, we found that mating behavior was regulated independently by another sexual differentiation gene, Bmfru. Loss of Bmfru completely blocked mating, but males displayed normal courtship behavior. Lack of Bmfru expression significantly reduced the perception of the minor pheromone component bombykal due to the down regulation of BmOR3 expression; further, functional analysis revealed that loss of the product of BmOR3 played a key role in terminating male mating behavior. Our results suggest that Bmdsx and Bmfru are at the base of the two primary pathways that regulate olfactory-based sexual behavior.


Assuntos
Bombyx/genética , Genes de Insetos , Preferência de Acasalamento Animal , Atrativos Sexuais/metabolismo , Processos de Determinação Sexual/genética , Animais , Bombyx/metabolismo , Bombyx/fisiologia , Feminino , Masculino , Receptores de Feromonas/genética , Receptores de Feromonas/metabolismo , Atrativos Sexuais/genética , Olfato
4.
PLoS One ; 15(6): e0234635, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32530959

RESUMO

The entire genome of Helicoverpa armigera single nucleopolyhedrovirus (HearNPV-TR) was sequenced, and compared to genomes of other existing isolates. HearNPV-TR genome is 130.691 base pairs with a 38.9% G+C content and has 137 open reading frames (ORFs) of ≥ 150 nucleotides. Five homologous repeated sequences (hrs) and two baculovirus repeated ORFs (bro-a and bro-b) were identified. Phylogenetic analysis showed that HearNPV-TR is closer to HaSNPV-C1, HaSNPV-G4, HaSNPV-AU and HasNPV. However, there are significant differences in hr3, hr5 regions and in bro-a gene. Pairwise Kimura-2 parameter analysis of 38 core genes sequences of HearNPV-TR and other Helicoverpa NPVs showed that the genetic distances for these sequences were below 0.015 substitutions/site. Genomic differences as revealed by restriction profiles indicated that hr3, hr5 regions and bro-a gene may play a role in the virulence of HearNPV-TR.


Assuntos
Genoma Viral , Mariposas/virologia , Nucleopoliedrovírus/genética , Nucleopoliedrovírus/isolamento & purificação , Análise de Sequência de DNA , Animais , DNA Circular/genética , Genes de Insetos , Fases de Leitura Aberta/genética , Filogenia , Mapeamento por Restrição , Turquia
5.
PLoS One ; 15(6): e0234437, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32511281

RESUMO

Gene flow and genetic variation were examined within and among populations of five of the most common spider species in shrublands of the mountainous Golden Gate Highlands National Park (GGHNP), South Africa. These species included three active hunters, Dendryphantes purcelli Peckham & Peckham, 1903 (Salticidae), Pherecydes tuberculatus O.P.-Cambridge, 1883 (Thomisidae) and Philodromus browningi Lawrence, 1952 (Philodromidae), and two web-builders, Neoscona subfusca (C.L. Koch, 1837) (Araneidae) and a Theridion Walckenaer, 1802 species (Theridiidae). A total of 249 spiders (57 D. purcelli, 69 N. subfusca, 34 P. browningi, 56 P. tuberculatus and 33 Theridion sp.) were collected and analysed from six shrubland localities in the park. Analyses of sequence variation of the mitochondrial cytochrome oxidase c subunit I (COI) gene for each species revealed relatively low nucleotide diversity (π < 0.0420) but high genetic diversity (Hd > 0.6500) within populations for all species, except P. tuberculatus. Genetic differentiation was also noted to differ between species, with only P. tuberculatus indicating very large divergence (Fst > 0.2500). These results were reflected by gene flow, with D. purcelli, N. subfusca and the Theridion sp. estimated as experiencing more than one disperser per generation. Overall, highest gene flow was found in the two web-building species, indicating possible high dispersal ability of these spiders in the GGHNP. Additionally, constructed phylogenies indicated possible cryptic speciation occurring in the majority of the investigated species. Our current results indicate that the five investigated spider species were able to maintain gene flow between shrubland populations within the GGHNP to some degree, despite the mountainous landscape. However, further analyses incorporating additional molecular markers are needed to properly determine the extent of genetic diversity and gene flow of these species within the GGHNP.


Assuntos
Genes de Insetos/genética , Especiação Genética , Variação Genética , Aranhas/genética , Animais , DNA Mitocondrial/genética , Complexo IV da Cadeia de Transporte de Elétrons/genética , Comportamento Alimentar , Fluxo Gênico , Pradaria , Proteínas de Insetos/genética , Filogenia , África do Sul
6.
PLoS One ; 15(5): e0233231, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32437379

RESUMO

Environmental changes cause stress, Reactive Oxygen Species and unfolded protein accumulation which hamper synaptic activity and trigger cell death. Heat shock proteins (HSPs) assist protein refolding to maintain proteostasis and cellular integrity. Mechanisms regulating the activity of HSPs include transcription factors and posttranslational modifications that ensure a rapid response. HSPs preserve synaptic function in the nervous system upon environmental insults or pathological factors and contribute to the coupling between environmental cues and neuron control of development. We have performed a biased screening in Drosophila melanogaster searching for synaptogenic modulators among HSPs during development. We explore the role of two small-HSPs (sHSPs), sHSP23 and sHSP26 in synaptogenesis and neuronal activity. Both sHSPs immunoprecipitate together and the equilibrium between both chaperones is required for neuronal development and activity. The molecular mechanism controlling HSP23 and HSP26 accumulation in neurons relies on a novel gene (CG1561), which we name Pinkman (pkm). We propose that sHSPs and Pkm are targets to modulate the impact of stress in neurons and to prevent synapse loss.


Assuntos
Proteínas de Drosophila/metabolismo , Drosophila melanogaster/crescimento & desenvolvimento , Drosophila melanogaster/metabolismo , Proteínas de Choque Térmico/metabolismo , Animais , Animais Geneticamente Modificados , Sistema Nervoso Central/crescimento & desenvolvimento , Sistema Nervoso Central/metabolismo , Proteínas de Drosophila/genética , Drosophila melanogaster/genética , Genes de Insetos , Proteínas de Choque Térmico/genética , Proteínas de Choque Térmico Pequenas/genética , Proteínas de Choque Térmico Pequenas/metabolismo , Modelos Neurológicos , Neurogênese/genética , Neurônios/metabolismo , Sinapses/metabolismo
7.
Nat Commun ; 11(1): 2631, 2020 05 26.
Artigo em Inglês | MEDLINE | ID: mdl-32457347

RESUMO

The evolution of winged insects revolutionized terrestrial ecosystems and led to the largest animal radiation on Earth. However, we still have an incomplete picture of the genomic changes that underlay this diversification. Mayflies, as one of the sister groups of all other winged insects, are key to understanding this radiation. Here, we describe the genome of the mayfly Cloeon dipterum and its gene expression throughout its aquatic and aerial life cycle and specific organs. We discover an expansion of odorant-binding-protein genes, some expressed specifically in breathing gills of aquatic nymphs, suggesting a novel sensory role for this organ. In contrast, flying adults use an enlarged opsin set in a sexually dimorphic manner, with some expressed only in males. Finally, we identify a set of wing-associated genes deeply conserved in the pterygote insects and find transcriptomic similarities between gills and wings, suggesting a common genetic program. Globally, this comprehensive genomic and transcriptomic study uncovers the genetic basis of key evolutionary adaptations in mayflies and winged insects.


Assuntos
Adaptação Fisiológica/genética , Ephemeroptera/genética , Evolução Molecular , Asas de Animais , Animais , Ephemeroptera/classificação , Ephemeroptera/crescimento & desenvolvimento , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Genes de Insetos/genética , Genoma de Inseto/genética , Brânquias , Insetos/classificação , Insetos/genética , Estágios do Ciclo de Vida/genética , Masculino , Filogenia
8.
PLoS One ; 15(5): e0232991, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32407359

RESUMO

Following nerve stimulation, there are two distinct phases of Ca2+-dependent neurotransmitter release: a fast, synchronous release phase, and a prolonged, asynchronous release phase. Each of these phases is tightly regulated and mediated by distinct mechanisms. Synaptotagmin 1 is the major Ca2+ sensor that triggers fast, synchronous neurotransmitter release upon Ca2+ binding by its C2A and C2B domains. It has also been implicated in the inhibition of asynchronous neurotransmitter release, as blocking Ca2+ binding by the C2A domain of synaptotagmin 1 results in increased asynchronous release. However, the mutation used to block Ca2+ binding in the previous experiments (aspartate to asparagine mutations, sytD-N) had the unintended side effect of mimicking Ca2+ binding, raising the possibility that the increase in asynchronous release was directly caused by ostensibly constitutive Ca2+ binding. Thus, rather than modulating an asynchronous sensor, sytD-N may be mimicking one. To directly test the C2A inhibition hypothesis, we utilized an alternate C2A mutation that we designed to block Ca2+ binding without mimicking it (an aspartate to glutamate mutation, sytD-E). Analysis of both the original sytD-N mutation and our alternate sytD-E mutation at the Drosophila neuromuscular junction showed differential effects on asynchronous release, as well as on synchronous release and the frequency of spontaneous release. Importantly, we found that asynchronous release is not increased in the sytD-E mutant. Thus, our work provides new mechanistic insight into synaptotagmin 1 function during Ca2+-evoked synaptic transmission and demonstrates that Ca2+ binding by the C2A domain of synaptotagmin 1 does not inhibit asynchronous neurotransmitter release in vivo.


Assuntos
Proteínas de Drosophila/metabolismo , Neurotransmissores/metabolismo , Sinaptotagmina I/metabolismo , Substituição de Aminoácidos , Animais , Animais Geneticamente Modificados , Sítios de Ligação/genética , Cálcio/metabolismo , Proteínas de Drosophila/química , Proteínas de Drosophila/genética , Drosophila melanogaster/genética , Drosophila melanogaster/metabolismo , Genes de Insetos , Mutagênese Sítio-Dirigida , Domínios Proteicos , Transmissão Sináptica , Vesículas Sinápticas/metabolismo , Sinaptotagmina I/química , Sinaptotagmina I/genética
9.
Parasitol Res ; 119(7): 2067-2073, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32468188

RESUMO

Fleas are ectoparasites of mammals and birds. In livestock such as sheep and goat, flea bites cause many clinical signs. Several types of insecticides including pyrethroids are used to struggle against fleas. The widespread use of these insecticides causes an increase in the number of resistant individuals in flea populations. T929V and L1014F mutations corresponding to pyrethroid resistance have been found in the para gene of cat fleas. We aimed to investigate T929V and L1014F mutations in flea samples (n:162) collected from goats in seven different farms where cypermethrin, a synthetic pyrethroid, had been used intensively. To achieve this aim, collected flea samples were morphologically identified under a stereo microscope and DNA isolation was conducted by HotSHOT method. Later, a bi-PASA targeting the para gene was applied to identify both mutations in corresponding samples. According to the results obtained, all fleas were Ctenocephalides felis. Frequencies of T929V and L1014F mutations in fleas were 92.6% (150/162) and 95.7% (155/162), respectively. In conclusion, the frequency of mutations related to pyrethroid resistance was very high in the fleas collected from all the farms and it was thought that the high frequency of these mutations can be attributed to intensive use of pyrethroids.


Assuntos
Ctenocephalides/genética , Infestações por Pulgas/veterinária , Genes de Insetos/genética , Doenças das Cabras/parasitologia , Resistência a Inseticidas/genética , Piretrinas , Animais , Infestações por Pulgas/parasitologia , Cabras , Inseticidas , Mutação
10.
Gene ; 749: 144712, 2020 Jul 30.
Artigo em Inglês | MEDLINE | ID: mdl-32360412

RESUMO

The quantitative real-time polymerase chain reaction (qRT-PCR) has rapidly become the most sensitive and accurate method for the quantitative analysis of gene expression. Normalization of gene expression to that of relatively stably expressed housekeeping genes is required to facilitate the study of gene expression and to obtain more accurate RT-PCR data. However, no studies of the stability of expression of housekeeping genes in Lymantria dispar have been reported. In the present study, BestKeeper, GeNorm and NormFinder statistical software was used to evaluate the expression of thirteen candidate reference genes in L. dispar under different conditions. The expression levels of candidate reference genes were determined for two biological factors (developmental stages and tissues) and four abiotic treatments (temperature, insecticide, CO2 and starvation). The results showed that the best candidate reference genes in L. dispar were TUB, AK, RPS15 for developmental stages, RPL32 and GAPDH for tissues, ACTB and EF1-α for CO2 stress, GAPDH and RPL32 for temperature stress, RPS3 and GAPDH for insecticide stress, and GAPDH and RPS3 for starvation stress. In summary, EF1-α and TUB are preferential housekeeping genes in L. dispar under various conditions. These results provide a basis for the further study of functional genes of L. dispar.


Assuntos
Genes de Insetos , Mariposas/genética , Animais , Dióxido de Carbono , Primers do DNA , Privação de Alimentos , Expressão Gênica , Genes Essenciais , Inseticidas , Mariposas/crescimento & desenvolvimento , Mariposas/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Estresse Fisiológico/genética , Temperatura
11.
PLoS One ; 15(5): e0222256, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32374761

RESUMO

kakusei is a non-coding RNA that is overexpressed in foraging bee brain. This study describes a possible role of the IEG kakusei during the daily foraging of honey bees. kakusei was found to be transiently upregulated within two hours during rewarded foraging. Interestingly, during unrewarded foraging the gene was also found to be up-regulated, but immediately lowered when food was not rewarded. Moreover, the kakusei overexpression was diminished within a very short time when the time schedule of feeding was changed. This indicates the potential role of kakusei on the motivation of learned reward foraging. These results provide evidence for a dynamic role of kakusei during for aging of bees, and eventually its possible involvement in learning and memory. Thus the kakusei gene could be used as search tool in finding distinct molecular pathways that mediate diverse behavioral components of foraging.


Assuntos
Abelhas/genética , Abelhas/fisiologia , Comportamento Alimentar , Genes Precoces/fisiologia , Genes de Insetos/fisiologia , Envelhecimento/genética , Animais , Aprendizagem , RNA não Traduzido/genética
12.
PLoS Negl Trop Dis ; 14(4): e0008219, 2020 04.
Artigo em Inglês | MEDLINE | ID: mdl-32298261

RESUMO

Aedes aegypti is the primary vector of dengue, chikungunya, Zika, and urban yellow fever. Insecticides are often the most effective tools to rapidly decrease the density of vector populations, especially during arbovirus disease outbreaks. However, the intense use of insecticides, particularly pyrethroids, has selected for resistant mosquito populations worldwide. Mutations in the voltage gated sodium channel (NaV) are among the principal mechanisms of resistance to pyrethroids and DDT, also known as "knockdown resistance," kdr. Here we report studies on the origin and dispersion of kdr haplotypes in samples of Ae. aegypti from its worldwide distribution. We amplified the IIS6 and IIIS6 NaV segments from pools of Ae. aegypti populations from 15 countries, in South and North America, Africa, Asia, Pacific, and Australia. The amplicons were barcoded and sequenced using NGS Ion Torrent. Output data were filtered and analyzed using the bioinformatic pipeline Seekdeep to determine frequencies of the IIS6 and IIIS6 haplotypes per population. Phylogenetic relationships among the haplotypes were used to infer whether the kdr mutations have a single or multiple origin. We found 26 and 18 haplotypes, respectively for the IIS6 and IIIS6 segments, among which were the known kdr mutations 989P, 1011M, 1016I and 1016G (IIS6), 1520I, and 1534C (IIIS6). The highest diversity of haplotypes was found in African samples. Kdr mutations 1011M and 1016I were found only in American and African populations, 989P + 1016G and 1520I + 1534C in Asia, while 1534C was present in samples from all continents, except Australia. Based primarily on the intron sequence, IIS6 haplotypes were subdivided into two well-defined clades (A and B). Subsequent phasing of the IIS6 + IIIS6 haplotypes indicates two distinct origins for the 1534C kdr mutation. These results provide evidence of kdr mutations arising de novo at specific locations within the Ae. aegypti geographic distribution. In addition, our results suggest that the 1534C kdr mutation had at least two independent origins. We can thus conclude that insecticide selection pressure with DDT and more recently with pyrethroids is selecting for independent convergent mutations in NaV.


Assuntos
Aedes/efeitos dos fármacos , Aedes/genética , Genes de Insetos , Resistência a Inseticidas/genética , Mutação , Aedes/virologia , África , Alelos , Animais , Ásia , Austrália , Febre de Chikungunya/prevenção & controle , Febre de Chikungunya/transmissão , DNA/genética , Dengue/prevenção & controle , Dengue/transmissão , Frequência do Gene , Genótipo , Inseticidas , Mosquitos Vetores/genética , Mosquitos Vetores/virologia , América do Norte , Piretrinas , Febre Amarela/prevenção & controle , Febre Amarela/transmissão , Infecção por Zika virus/prevenção & controle , Infecção por Zika virus/transmissão
13.
Proc Biol Sci ; 287(1926): 20200443, 2020 05 13.
Artigo em Inglês | MEDLINE | ID: mdl-32345166

RESUMO

Polymorphic Batesian mimics exhibit multiple protective morphs that each mimic a different noxious model. Here, we study the genomic transitions leading to the evolution of different mimetic wing patterns in the polymorphic Mocker Swallowtail Papilio dardanus. We generated a draft genome (231 Mb over 30 chromosomes) and re-sequenced individuals of three morphs. Genome-wide single nucleotide polymorphism (SNP) analysis revealed elevated linkage disequilibrium and divergence between morphs in the regulatory region of engrailed, a developmental gene previously implicated in the mimicry switch. The diverged region exhibits a discrete chromosomal inversion (of 40 kb) relative to the ancestral orientation that is associated with the cenea morph, but not with the bottom-recessive hippocoonides morph or with non-mimetic allopatric populations. The functional role of this inversion in the expression of the novel phenotype is currently unknown, but by preventing recombination, it allows the stable inheritance of divergent alleles enabling geographic spread and local coexistence of multiple adaptive morphs.


Assuntos
Mimetismo Biológico/fisiologia , Borboletas/parasitologia , Inversão Cromossômica , Animais , Genes de Insetos , Genômica , Desequilíbrio de Ligação , Fenótipo , Sequências Reguladoras de Ácido Nucleico , Asas de Animais
14.
PLoS Genet ; 16(4): e1008762, 2020 04.
Artigo em Inglês | MEDLINE | ID: mdl-32348297

RESUMO

Animals often exhibit dramatically behavioral plasticity depending on their internal physiological state, yet little is known about the underlying molecular mechanisms. The migratory locust, Locusta migratoria, provides an excellent model for addressing these questions because of their famous phase polyphenism involving remarkably behavioral plasticity between gregarious and solitarious phases. Here, we report that a major insect hormone, juvenile hormone, is involved in the regulation of this behavioral plasticity related to phase change by influencing the expression levels of olfactory-related genes in the migratory locust. We found that the treatment of juvenile hormone analog, methoprene, can significantly shift the olfactory responses of gregarious nymphs from attraction to repulsion to the volatiles released by gregarious nymphs. In contrast, the repulsion behavior of solitarious nymphs significantly decreased when they were treated with precocene or injected with double-stranded RNA of JHAMT, a juvenile hormone acid O-methyltransferase. Further, JH receptor Met or JH-response gene Kr-h1 knockdown phenocopied the JH-deprivation effects on olfactory behavior. RNA-seq analysis identified 122 differentially expressed genes in antennae after methoprene application on gregarious nymphs. Interestingly, several olfactory-related genes were especially enriched, including takeout (TO) and chemosensory protein (CSP) which have key roles in behavioral phase change of locusts. Furthermore, methoprene application and Met or Kr-h1 knockdown resulted in simultaneous changes of both TO1 and CSP3 expression to reverse pattern, which mediated the transition between repulsion and attraction responses to gregarious volatiles. Our results suggest the regulatory roles of a pleiotropic hormone in locust behavioral plasticity through modulating gene expression in the peripheral olfactory system.


Assuntos
Antenas de Artrópodes/metabolismo , Comportamento Animal/efeitos dos fármacos , Hormônios Juvenis/farmacologia , Comportamento Social , Transcriptoma/efeitos dos fármacos , Animais , Antenas de Artrópodes/efeitos dos fármacos , Genes de Insetos , Gafanhotos , Proteínas de Insetos/genética , Proteínas de Insetos/metabolismo , Metoprene/farmacologia , Metiltransferases/genética , Metiltransferases/metabolismo
15.
PLoS One ; 15(4): e0232192, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32343725

RESUMO

The introduction of exotic disease vectors into a new habitat can drastically change the local epidemiological situation. During 2012-2015, larvae and an adult of the yellow-fever mosquito, Aedes aegypti, were captured alive at two international airports serving the Greater Tokyo Area, Japan. Because this species does not naturally distribute in this country, those mosquitoes were considered to be introduced from overseas via air-transportation. To infer the places of origin of those mosquitoes, we genotyped the 12 microsatellite loci for which the most comprehensive population genetic reference is currently available. Although clustering by Bayesian and multivariate methods both suggested that all those mosquitoes captured at the airports in Japan belonged to the Asia/Pacific populations, they were not clustered into a single cluster. Moreover, there was variation in mitochondrial cytochrome oxidase I gene (CoxI) haplotypes among mosquitoes collected in different incidents of discovery which indicated the existence of multiple maternal origins. We conclude there is little evidence to support the overwintering of Ae. aegypti at the airports; nevertheless, special attention is still needed to prevent the invasion of this prominent arbovirus vector.


Assuntos
Aedes/genética , Aeroportos , Mosquitos Vetores/genética , Aedes/classificação , Aedes/virologia , Animais , Infecções por Arbovirus/transmissão , Infecções por Arbovirus/virologia , Arbovirus/isolamento & purificação , Arbovirus/patogenicidade , Teorema de Bayes , Ecossistema , Complexo IV da Cadeia de Transporte de Elétrons/genética , Genes de Insetos , Genes Mitocondriais , Variação Genética , Genética Populacional , Genótipo , Haplótipos , Humanos , Proteínas de Insetos/genética , Repetições de Microssatélites , Mosquitos Vetores/classificação , Mosquitos Vetores/virologia , Tóquio
16.
Vet Parasitol ; 281: 109102, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32289653

RESUMO

The present study investigated the genetic profile of the cosmopolitan cat flea, Ctenocephalides felis (Siphonaptera: Pulicidae) from Malaysia and the reference data available in the National Center for Biotechnology Information (NCBI) GenBank. A set of sequences of 100 Malaysian samples aligned as 550 characters of the cytochrome c oxidase subunit I (cox1) and 706 characters of the II (cox2) genes revealed ten haplotypes (A1-A10) and eight haplotypes (B1-B8), respectively. The concatenated sequences of cox1 and cox2 genes with a total of 1256 characters revealed 15 haplotypes (AB1-AB15). Analyses indicated that haplotype AB1 was the most frequent and the most widespread haplotype in Malaysia. Overall haplotype and nucleotide diversities of the concatenated sequences were 0.52909 and 0.00424, respectively, with moderate genetic differentiation (FST = 0.17522) and high gene flow (Nm = 1.18). The western population presented the highest genetic diversity (Hd = 0.78333, Pi = 0.01269, Nh = 9), whereas the southern population demonstrated the lowest diversity (Hd = 0.15667, Pi = 0.00019, Nh = 3). The concatenated sequences showed genetic distances ranged from 0.08 % to 4.39 %. There were three aberrant haplotypes in cox2 sequences that highly divergent, suggesting the presence of cryptic species or occurrence of introgression. In the global point of view, the aligned sequences of C. felis revealed 65 haplotypes (AA1-AA65) by the cox1 gene (n = 586), and 27 haplotypes (BB1-BB27) by the cox2 gene (n = 204). Mapping of the haplotype network showed that Malaysian C. felis possesses seven unique haplotypes in both genes with the common haplotypes demonstrated genetic affinity with C. felis from Southeast Asia for cox1 and South America for cox2. The topologies of cox1 and cox2 phylogenetic trees were concordant with relevant grouping pattern of haplotypes in the network but revealed two major lineages by which Malaysian haplotypes were closely related with haplotypes from the tropical region.


Assuntos
Ctenocephalides/genética , Genes de Insetos/genética , Variação Genética , Animais , Bases de Dados Genéticas , Haplótipos/genética , Malásia
17.
Nat Commun ; 11(1): 1388, 2020 03 13.
Artigo em Inglês | MEDLINE | ID: mdl-32170121

RESUMO

Transcription factors (TFs) control cell fates by precisely orchestrating gene expression. However, how individual TFs promote transcriptional diversity remains unclear. Here, we use the Hox TF Ultrabithorax (Ubx) as a model to explore how a single TF specifies multiple cell types. Using proximity-dependent Biotin IDentification in Drosophila, we identify Ubx interactomes in three embryonic tissues. We find that Ubx interacts with largely non-overlapping sets of proteins with few having tissue-specific RNA expression. Instead most interactors are active in many cell types, controlling gene expression from chromatin regulation to the initiation of translation. Genetic interaction assays in vivo confirm that they act strictly lineage- and process-specific. Thus, functional specificity of Ubx seems to play out at several regulatory levels and to result from the controlled restriction of the interaction potential by the cellular environment. Thereby, it challenges long-standing assumptions such as differential RNA expression as determinant for protein complexes.


Assuntos
Linhagem da Célula/fisiologia , Proteínas de Drosophila/metabolismo , Drosophila/embriologia , Drosophila/metabolismo , Proteínas de Homeodomínio/metabolismo , Fatores de Transcrição/metabolismo , Animais , Cromatina/metabolismo , Drosophila/genética , Proteínas de Drosophila/genética , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Genes de Insetos , Proteínas de Homeodomínio/genética , Masculino , Mesoderma/citologia , Mesoderma/metabolismo , Domínios e Motivos de Interação entre Proteínas , Mapas de Interação de Proteínas , RNA/metabolismo , Fatores de Transcrição/genética
18.
PLoS One ; 15(3): e0230435, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32191747

RESUMO

Harmonia axyridis is a major bio-control agent of pests in agriculture and forest ecosystems. It is also a globally important invasive insect species. To test whether dark elytra colour is associated with greater cold hardiness, we compared the survival rate of prolonged cold exposure in both yellow and black colour morphs of female and male H. axyridis. We determined the trehalose and glycogen content, trehalase activity, and the dynamics of genes associated with the trehalose metabolic pathway. Yellow forms predominated before winter began, however black forms increased from 11.15 to 30.46% after overwintering. There was no significant difference in trehalose content between the females and males during overwintering. Glycogen content in over-wintering yellow females and black males increased significantly, while it decreased in black females. Soluble trehalase activity increased significantly in all the insects except black females. Membrane-bound trehalase activity increased in black males, and decreased in black females. Trehalose and glycogen content and trehalase activity were regulated by differential expression of TRE and TPS genes. Female beetles weighed more than males and survived in low temperatures for longer periods of time, regardless of elytra colour, suggesting that mass is a stronger predictor of overwintering survival rather than colour morph. Our results provide a guide for comparing cold resistance in insects and a theoretical basis for cold storage of H. axyridis for use as natural enemies of pests in biological control programs.


Assuntos
Temperatura Baixa , Besouros/anatomia & histologia , Besouros/metabolismo , Estações do Ano , Trealose/metabolismo , Animais , Peso Corporal , Besouros/genética , Feminino , Regulação da Expressão Gênica , Genes de Insetos , Glicogênio/metabolismo , Masculino , Fenótipo , Análise de Sobrevida , Fatores de Tempo , Trealase/metabolismo
19.
Gene ; 743: 144605, 2020 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-32199950

RESUMO

Atrijuglans hetaohei Yang (Lepidoptera: Gelechioidea), is one of the major pests that can seriously damage the walnut fruits. Neuropeptides and their receptors regulate most physiological functions in insects and represent new targets for the development of control agents. To identify the neuropeptides and their receptors from A. hetaohei, we sequenced and analyzed its head transcriptomic data, identified 32 neuropeptides and 39 neuropeptide receptor genes. Sequence comparisons and phylogenetic analyses suggest that A. hetaohei neuropeptides and receptor genes have high homology with those in Bombyx mori, Chilo suppressalis, Plutella xylostella and Helicoverpa armigera. Moreover, gene expression patterns revealed that neuropeptide genes such as AKH1, CP, MS and PTTH were expressed specifically in male head, while CAP3, DH, NPLP1, PBAN and SIF showed higher expression in the female head. Bur showed abdomen biased expression in both male and female. Neuropeptide receptor genes such as A8, A11, A15 and LGR were highly expressed in male head, whereas A24 and LGR2 were preferentially expressed in female head. This is the first sequencing, identification and expression analyses of neuropeptides and neuropeptide receptor genes from A. hetaohei. Our results could provide a powerful background that will facilitate the further investigations using transcriptomics to determine neuropeptides and their receptors presence, functions, and indicates potential targets in A. hetaohei for a novel pest management strategy.


Assuntos
Proteínas de Insetos/genética , Mariposas/genética , Neuropeptídeos/genética , Receptores de Neuropeptídeos/genética , Transcriptoma/genética , Animais , Clonagem Molecular , Feminino , Perfilação da Expressão Gênica , Genes de Insetos , Juglans/parasitologia , Masculino , Controle de Pragas/métodos , Filogenia , Fatores Sexuais
20.
BMC Evol Biol ; 20(1): 17, 2020 01 31.
Artigo em Inglês | MEDLINE | ID: mdl-32005133

RESUMO

BACKGROUND: Drosophila subobscura exhibits a rich inversion polymorphism, with some adaptive inversions showing repeatable spatiotemporal patterns in frequencies related to temperature. Previous studies reported increased basal HSP70 protein levels in homokaryotypic strains for a warm-climate arrangement compared to a cold-climate one. These findings do not match the similar hsp70 genomic organization between arrangements, where gene expression levels are expected to be similar. In order to test this hypothesis and understand the molecular basis for hsp70 expression, we compared basal hsp70 mRNA levels in males and females, and analysed the 5' and 3' regulatory regions of hsp70 genes in warm- and cold-climate isochromosomal O3 + 4 + 7 and OST lines of D. subobscura. RESULTS: We observed comparable mRNA levels between the two arrangements and a sex-biased hsp70 gene expression. The number of heat-shock elements (HSEs) and GAGA sites on the promoters were identical amongst the OST and O3 + 4 + 7 lines analysed. This is also true for 3' AU-rich elements where most A and B copies of hsp70 have, respectively, two and one element in both arrangements. Beyond the regulatory elements, the only notable difference between both arrangements is the presence in 3' UTR of a 14 bp additional fragment after the stop codon in the hsp70A copy in five O3 + 4 + 7 lines, which was not found in any of the six OST lines. CONCLUSIONS: The equivalent hsp70 mRNA amounts in OST and O3 + 4 + 7 arrangements provide the first evidence of a parallelism between gene expression and genetic organization in D. subobscura lines having these arrangements. This is reinforced by the lack of important differential features in the number and structure of regulatory elements between both arrangements, despite the genetic differentiation observed when the complete 5' and 3' regulatory regions were considered. Therefore, the basal levels of hsp70 mRNA cannot account, in principle, for the adaptive variation of the two arrangements studied. Consequently, further studies are necessary to understand the intricate molecular mechanisms of hsp70 gene regulation in D. subobscura.


Assuntos
Adaptação Fisiológica/genética , Clima , Drosophila/genética , Regulação da Expressão Gênica , Rearranjo Gênico/genética , Genes de Insetos , Variação Genética , Proteínas de Choque Térmico HSP70/genética , Regiões 3' não Traduzidas/genética , Análise de Variância , Animais , Sequência de Bases , Sequência Conservada , Drosophila/fisiologia , Feminino , Proteínas de Choque Térmico HSP70/metabolismo , Masculino , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Sequências Reguladoras de Ácido Nucleico/genética
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