RESUMO
The mammalian Y Chromosome sequence, critical for studying male fertility and dispersal, is enriched in repeats and palindromes, and thus, is the most difficult component of the genome to assemble. Previously, expensive and labor-intensive BAC-based techniques were used to sequence the Y for a handful of mammalian species. Here, we present a much faster and more affordable strategy for sequencing and assembling mammalian Y Chromosomes of sufficient quality for most comparative genomics analyses and for conservation genetics applications. The strategy combines flow sorting, short- and long-read genome and transcriptome sequencing, and droplet digital PCR with novel and existing computational methods. It can be used to reconstruct sex chromosomes in a heterogametic sex of any species. We applied our strategy to produce a draft of the gorilla Y sequence. The resulting assembly allowed us to refine gene content, evaluate copy number of ampliconic gene families, locate species-specific palindromes, examine the repetitive element content, and produce sequence alignments with human and chimpanzee Y Chromosomes. Our results inform the evolution of the hominine (human, chimpanzee, and gorilla) Y Chromosomes. Surprisingly, we found the gorilla Y Chromosome to be similar to the human Y Chromosome, but not to the chimpanzee Y Chromosome. Moreover, we have utilized the assembled gorilla Y Chromosome sequence to design genetic markers for studying the male-specific dispersal of this endangered species.
Assuntos
Biologia Computacional , Sequenciamento de Nucleotídeos em Larga Escala , Mamíferos/genética , Cromossomo Y , Animais , Biologia Computacional/métodos , Rearranjo Gênico , Genoma , Genômica , Gorilla gorilla/genética , Humanos , Sequências Repetidas Invertidas , Masculino , Repetições de Microssatélites , Pan troglodytes/genética , Sequências Repetitivas de Ácido Nucleico , Análise de Sequência de DNARESUMO
MAIN CONCLUSION: Ferulic acid esterases have been identified and partially purified from maize pollen. Results suggest that maize pollen FAEs may play an important role in pollen fertilization. A critical step in maize (Zea mays) seed production involves fertilization of the ovule by pollen, a process that relies on ability of the pollen tube to grow through the highly structured and feruloylated arabinoxylan/cellulose-rich tissue of the silk and stigma. It is known that different cell wall hydrolases are present on the surface of pollen. An important hydrolase reported to date is an endo-xylanase (ZmXYN1). We report presence and characterization of another hydrolase, ferulic acid esterase (FAE), in maize pollen. Using a combination of biochemical approaches, these FAEs were partially purified and characterized with respect to their biochemical properties and putative sequences. Maize pollen FAEs were shown to be expressed early during pollen development, to release significant amounts of both monomeric and dimeric ferulates esterified from maize silks and other grass cell walls, and to synergize with an externally applied fungal endo-1,4-ß-xylanase on the release of cell wall ferulates and diferulates. Preliminary analysis of maize silk cell walls following pollination, showed a significant reduction of esterified ferulates up to 96 h following pollination, compared to unpollinated silks. These results suggest that maize pollen FAEs may play an important biological role in pollen fertilization and possibly in seed production.
Assuntos
Hidrolases de Éster Carboxílico/química , Ácidos Cumáricos/química , Pólen/química , Polinização/fisiologia , Sementes/química , Zea mays/químicaRESUMO
Winged insects underwent an unparalleled evolutionary radiation, but mechanisms underlying the origin and diversification of wings in basal insects are sparsely known compared with more derived holometabolous insects. In the neopteran species Oncopeltus fasciatus, we manipulated wing specification genes and used RNA-seq to obtain both functional and genomic perspectives. Combined with previous studies, our results suggest the following key steps in wing origin and diversification. First, a set of dorsally derived outgrowths evolved along a number of body segments including the first thoracic segment (T1). Homeotic genes were subsequently co-opted to suppress growth of some dorsal flaps in the thorax and abdomen. In T1 this suppression was accomplished by Sex combs reduced, that when experimentally removed, results in an ectopic T1 flap similar to prothoracic winglets present in fossil hemipteroids and other early insects. Global gene-expression differences in ectopic T1 vs. T2/T3 wings suggest that the transition from flaps to wings required ventrally originating cells, homologous with those in ancestral arthropod gill flaps/epipods, to migrate dorsally and fuse with the dorsal flap tissue thereby bringing new functional gene networks; these presumably enabled the T2/T3 wing's increased size and functionality. Third, "fused" wings became both the wing blade and surrounding regions of the dorsal thorax cuticle, providing tissue for subsequent modifications including wing folding and the fit of folded wings. Finally, Ultrabithorax was co-opted to uncouple the morphology of T2 and T3 wings and to act as a general modifier of hindwings, which in turn governed the subsequent diversification of lineage-specific wing forms.
Assuntos
Evolução Molecular , Variação Genética , Insetos/genética , Asas de Animais/metabolismo , Animais , Regulação da Expressão Gênica no Desenvolvimento , Genoma de Inseto/genética , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Proteínas de Insetos/genética , Insetos/anatomia & histologia , Insetos/crescimento & desenvolvimento , Interferência de RNA , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Asas de Animais/anatomia & histologia , Asas de Animais/crescimento & desenvolvimentoRESUMO
MAIN CONCLUSION: We show that changing the expression of a putative feruloyl transferase gene belonging to the BAHD acyl-transferase family alters the levels of cell wall esterified ferulates and diferulates in Brachypodium distachyon cell walls. While the potential of grass cell walls for biofuel production has been realized, the technology for lignocellulosic biomass conversion for the production of ethanol is still inefficient because of structural mechanisms that plants have evolved to make the cell wall recalcitrant to enzymatic attack. One of these mechanisms in grasses involves the esterification of arabinoxylans in the cell wall with ferulic acid via an ester linkage to arabinose side chains on xylans. These ferulates undergo oxidative coupling reactions to form ferulate dimers, thus crosslinking polysaccharides. Arabinoxylan feruloylation is an important factor that determines cell wall recalcitrance because it directly cross-links xylans and because ferulates act as nucleating sites for the formation of lignin and for the linkage of lignin to the xylan/cellulose network. Here we report on the effects of changing the expression of Bradi2g43520 (BdAT1), a homologue of the rice feruloyl transferase gene Os01g42880 belonging to the Pfam PF02458 family, in Brachypodium distachyon. Down regulation in several independent RNAi::BdAT1 lines, resulted in up to a 35 % reduction of ferulate levels in both leaves and stems compared to control plants, over 2-3 generations of selfing. In contrast, overexpression of putative BdAT1 resulted in an increase of up to 58 and 47 % of ferulate levels in leaves and stems, respectively, compared to control plants and analyzed over 2-3 generations of selfing. These findings suggest that Bradi2g43520 may be a good candidate for feruloylation of AX in Brachypodium.
Assuntos
Brachypodium/enzimologia , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas/metabolismo , Xilanos/metabolismo , Biocombustíveis , Brachypodium/genética , Parede Celular/metabolismo , Ácidos Cumáricos/metabolismo , Esterificação , Lignina/metabolismo , Oryza/enzimologia , Oryza/genética , Folhas de Planta/enzimologia , Folhas de Planta/genética , Proteínas de Plantas/genética , Caules de Planta/enzimologia , Caules de Planta/genética , Plantas Geneticamente Modificadas , Interferência de RNA , Transferases/genética , Transferases/metabolismoRESUMO
Feruloylation of arabinoxylan in grass cell walls leads to cross-linked xylans. Such cross-linking appears to play a role in plant resistance to pathogens and insect herbivores. In this study, we investigated the effect of ferulate cross-linking on resistance to herbivory by fall armyworm (Spodoptera frugiperda) making use of genetically modified tall fescue [Schedonorus arundinaceus (Festuca arundinacea)] expressing a ferulic acid esterase gene. Mature leaves of these plants have significant reduced levels of cell wall ferulates and diferulates but no change in acid detergent lignin. These reduced levels of esterified cell wall ferulates in transgenic plants had a positive effect on all measures of armyworm larval performance examined. More larvae survived (89 vs. 57 %) and grew faster (pupated 2.1 days sooner) when fed transgenic leaves with reduced levels of cell wall ferulates, than when fed control tall fescue leaves where levels of cell wall ferulates were not altered. Overall, mortality, growth and food utilization were negatively associated with level of esterified cell wall ferulates and diferulates in leaves they were fed. This study is the first to use transgenic plants with modified level of cell wall esterified ferulates to test the role of feruloylation in plant resistance to insects. It is concluded that the accumulation of ferulates and the cross-linking of arabinoxylans via diferulate esters in the leaves of tall fescue underlies the physical barrier to insect herbivory. Reducing ferulate cross-linking in grass cell walls could increase susceptibility of these plants to insect folivores.
Assuntos
Hidrolases de Éster Carboxílico/metabolismo , Parede Celular/química , Ácidos Cumáricos/metabolismo , Festuca/metabolismo , Spodoptera/fisiologia , Animais , Hidrolases de Éster Carboxílico/genética , Ácidos Cumáricos/análise , Resistência à Doença , Festuca/química , Festuca/genética , Festuca/parasitologia , Herbivoria , Larva , Lignina/análise , Fenóis/análise , Fenóis/metabolismo , Doenças das Plantas/parasitologia , Folhas de Planta/química , Folhas de Planta/genética , Folhas de Planta/metabolismo , Folhas de Planta/parasitologia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas , Spodoptera/crescimento & desenvolvimento , Xilanos/análise , Xilanos/metabolismoRESUMO
In fragmented landscapes, small populations frequently go extinct and new ones are established with poorly understood consequences for genetic diversity and evolution of life history traits. Here, we apply functional genomic tools to an ecological model system, the well-studied metapopulation of the Glanville fritillary butterfly. We investigate how dispersal and colonization select upon existing genetic variation affecting life history traits by comparing common-garden reared 2-day adult females from new populations with those from established older populations. New-population females had higher expression of abdomen genes involved in egg provisioning and thorax genes involved in the maintenance of flight muscle proteins. Physiological studies confirmed that new-population butterflies have accelerated egg maturation, apparently regulated by higher juvenile hormone titer and angiotensin converting enzyme mRNA, as well as enhanced flight metabolism. Gene expression varied between allelic forms of two metabolic genes (Pgi and Sdhd), which themselves were associated with differences in flight metabolic rate, population age and population growth rate. These results identify likely molecular mechanisms underpinning life history variation that is maintained by extinction-colonization dynamics in metapopulations.
Assuntos
Borboletas/genética , Borboletas/metabolismo , Metabolismo Energético , Animais , Borboletas/fisiologia , Sistema Enzimático do Citocromo P-450/genética , Ecossistema , Feminino , Voo Animal , Expressão Gênica , Variação Genética , Genômica , Glucose-6-Fosfato Isomerase/genética , Glucose-6-Fosfato Isomerase/metabolismo , Oxirredutases Intramoleculares/genética , Hormônios Juvenis/genética , Hormônios Juvenis/fisiologia , Peptidil Dipeptidase A/genética , Dinâmica Populacional , Succinato Desidrogenase/genética , Succinato Desidrogenase/metabolismoRESUMO
Genes with opposing effects on fitness at different life stages are the mechanistic basis for evolutionary theories of aging and life history. Examples come from studies of mutations in model organisms, but there is little knowledge of genetic bases of life history tradeoffs in natural populations. Here, we test the hypothesis that alleles affecting oxygen sensing in Glanville fritillary butterflies have opposing effects on larval versus adult fitness-related traits. Intermediate-frequency alleles in Succinate dehydrogenase d, and to a lesser extent Hypoxia inducible factor 1α, are associated in larvae with variation in metabolic rate and activation of the hypoxia inducible factor (HIF) pathway, which affects tracheal development and delivery of oxygen to adult flight muscles. A dominant Sdhd allele is likely to cause antagonistic pleiotropy for fitness through its opposing effects on larval metabolic and growth rate versus adult flight and dispersal, and may have additional effects arising from sensitivity to low-iron host plants. Prior results in Glanville fritillaries indicate that fitness of alleles in Sdhd and another antagonistically pleiotropic metabolic gene, Phosphoglucose isomerase, depend strongly on the size and distribution of host plant patches. Hence, these intermediate-frequency alleles are involved in ecoevolutionary dynamics involving life history tradeoffs.
Assuntos
Borboletas/genética , Pleiotropia Genética , Glucose-6-Fosfato Isomerase/genética , Características de História de Vida , Succinato Desidrogenase/genética , Alelos , Animais , Borboletas/enzimologia , Feminino , Fator 1 Induzível por Hipóxia/genética , Larva/metabolismoRESUMO
Migrant populations of Helicoverpa zea (Boddie) captured during 2002, 2005, 2016, and 2018 from Landisville and Rock Springs in Pennsylvania, USA were genotyped using 85 single nucleotide polymorphism (SNP) markers. Samples (n = 702) genotyped were divided into 16 putative populations based on collection time and site. Fixation indices (F-statistics), analysis of molecular variance, and discriminant analysis of principal components were used to examine within and among population genetic variation. The observed and expected heterozygosity in putative populations ranged from 0.317-0.418 and 0.320-0.359, respectively. Broad range of FST (0.0-0.2742) and FIS (0.0-0.2330) values indicated different genotype frequencies between and within the populations, respectively. High genetic diversity within and low genetic differentiation between populations was found in 2002 and 2005. Interestingly, high genetic differentiation between populations from two collection sites observed in 2018 populations was not evident in within-site comparisons of putative populations collected on different dates during the season. The shift of H. zea population genetic makeup in 2018 may be influenced by multiple biotic and abiotic factors including tropical storms. Continued assessment of these peripheral populations of H. zea will be needed to assess the impacts of genetic changes on pest control and resistance management tactics.
RESUMO
Glycans are multi-branched sugars that are displayed from lipids and proteins. Through their diverse polysaccharide structures they can potentiate a myriad of cellular signaling pathways involved in development, growth, immuno-communication and survival. Not surprisingly, disruption of glycan synthesis is fundamental to various human diseases; including cancer, where aberrant glycosylation drives malignancy. Here, we report the discovery of a novel mannose-binding lectin, ML6, which selectively recognizes and binds to these irregular tumor-specific glycans to elicit potent and rapid cancer cell death. This lectin was engineered from gene models identified in a tropical rainforest tree root transcriptome and is unusual in its six canonical mannose binding domains (QxDxNxVxY), each with a unique amino acid sequence. Remarkably, ML6 displays antitumor activity that is >105 times more potent than standard chemotherapeutics, while being almost completely inactive towards non-transformed, healthy cells. This activity, in combination with results from glycan binding studies, suggests ML6 differentiates healthy and malignant cells by exploiting divergent glycosylation pathways that yield naïve and incomplete cell surface glycans in tumors. Thus, ML6 and other high-valence lectins may serve as novel biochemical tools to elucidate the glycomic signature of different human tumors and aid in the rational design of carbohydrate-directed therapies. Further, understanding how nature evolves proteins, like ML6, to combat the changing defenses of competing microorganisms may allow for fundamental advances in the way we approach combinatorial therapies to fight therapeutic resistance in cancer.
Assuntos
Antineoplásicos/farmacologia , Lectinas de Ligação a Manose/farmacologia , Neoplasias/tratamento farmacológico , Extratos Vegetais/farmacologia , Raízes de Plantas/química , Transcriptoma , Árvores/química , Sequência de Aminoácidos , Antineoplásicos/química , Apoptose , Proliferação de Células , Descoberta de Drogas , Glicosilação , Humanos , Lectinas de Ligação a Manose/química , Modelos Moleculares , Neoplasias/genética , Neoplasias/patologia , Polissacarídeos/metabolismo , Conformação Proteica , Floresta Úmida , Células Tumorais CultivadasRESUMO
We present a de novo assembly of a eukaryote transcriptome using 454 pyrosequencing data. The Glanville fritillary butterfly (Melitaea cinxia; Lepidoptera: Nymphalidae) is a prominent species in population biology but had no previous genomic data. Sequencing runs using two normalized complementary DNA collections from a genetically diverse pool of larvae, pupae, and adults yielded 608,053 expressed sequence tags (mean length = 110 nucleotides), which assembled into 48,354 contigs (sets of overlapping DNA segments) and 59,943 singletons. BLAST comparisons confirmed the accuracy of the sequencing and assembly, and indicated the presence of c. 9000 unique genes, along with > 6000 additional microarray-confirmed unannotated contigs. Average depth of coverage was 6.5-fold for the longest 4800 contigs (348-2849 bp in length), sufficient for detecting large numbers of single nucleotide polymorphisms. Oligonucleotide microarray probes designed from the assembled sequences showed highly repeatable hybridization intensity and revealed biological differences among individuals. We conclude that 454 sequencing, when performed to provide sufficient coverage depth, allows de novo transcriptome assembly and a fast, cost-effective, and reliable method for development of functional genomic tools for nonmodel species. This development narrows the gap between approaches based on model organisms with rich genetic resources vs. species that are most tractable for ecological and evolutionary studies.
Assuntos
Borboletas/genética , Etiquetas de Sequências Expressas , Perfilação da Expressão Gênica , Análise de Sequência de DNA/métodos , Processamento Alternativo , Animais , Sequência de Bases , DNA Complementar/genética , Análise de Sequência com Séries de Oligonucleotídeos , Polimorfismo de Nucleotídeo Único , Alinhamento de SequênciaRESUMO
Emergence of polyphagous herbivorous insects entails significant adaptation to recognize, detoxify and digest a variety of host-plants. Despite of its biological and practical importance - since insects eat 20% of crops - no exhaustive analysis of gene repertoires required for adaptations in generalist insect herbivores has previously been performed. The noctuid moth Spodoptera frugiperda ranks as one of the world's worst agricultural pests. This insect is polyphagous while the majority of other lepidopteran herbivores are specialist. It consists of two morphologically indistinguishable strains ("C" and "R") that have different host plant ranges. To describe the evolutionary mechanisms that both enable the emergence of polyphagous herbivory and lead to the shift in the host preference, we analyzed whole genome sequences from laboratory and natural populations of both strains. We observed huge expansions of genes associated with chemosensation and detoxification compared with specialist Lepidoptera. These expansions are largely due to tandem duplication, a possible adaptation mechanism enabling polyphagy. Individuals from natural C and R populations show significant genomic differentiation. We found signatures of positive selection in genes involved in chemoreception, detoxification and digestion, and copy number variation in the two latter gene families, suggesting an adaptive role for structural variation.
Assuntos
Adaptação Fisiológica/genética , Genoma de Inseto , Herbivoria , Spodoptera/genética , Animais , Produtos Agrícolas , Larva/genética , Especificidade da EspécieRESUMO
Seasonal climatic shifts create peripheral habitats that alternate between habitable and uninhabitable for migratory species. Such dynamic peripheral habitats are potential sites where migratory species could evolve high genetic diversity resulting from convergence of immigrants from multiple regionally distant areas. Migrant populations of Helicoverpa zea (Boddie) captured during two different seasons were assessed for genetic structure using microsatellite markers and for host plant type using stable carbon isotope analysis. Individuals (N = 568) were genotyped and divided into 13 putative populations based on collection site and time. Fixation indices (F-statistics), analysis of molecular variance (AMOVA), and discriminant analysis of principal components (DAPC) were used to examine within and among population genetic variation. Mean number of alleles per locus was 10.25 (± 3.2 SD), and allelic richness ranged from 2.38 to 5.13 (± 3.2 SD). The observed and expected heterozygosity ranged from 0.07 to 0.48 and 0.08 to 0.62, respectively. Low F ST (0.01 to 0.02) and high F IS (0.08 to 0.33) values suggest captured migrants originated from breeding populations with different allele frequencies. We postulate that high genetic diversity within migrant populations and low genetic differentiation among migrant populations of H. zea are the result of asymmetrical immigration due to the high dispersal and reproductive behavior of H. zea, which may hinder the adaptation and establishment of H. zea to peripheral habitat. These findings highlight the importance of assessing peripheral population structure in relation to ecological and evolutionary dynamics of this and other highly reproductive and dispersive species.
RESUMO
Understanding the molecular mechanisms underlying insect compensatory responses to plant defenses could lead to improved plant resistance to herbivores. The Mp708 inbred line of maize produces the maize insect resistant 1-cysteine protease (Mir1-CP) toxin. Reduced feeding and growth of fall armyworm larvae fed on Mp708 was previously linked to impairment of nutrient utilization and degradation of the midgut (MG) peritrophic matrix (PM) by Mir1-CP. Here we examine the biochemical and transcriptional responses of fall armyworm larvae to Mir1-CP. Insect Intestinal Mucin (IIM) was severely depleted from pure PMs treated in vitro with recombinant Mir1-CP. Larvae fed on Mp708 midwhorls excrete frass largely depleted of IIM. Cracks, fissures and increased porosity previously observed in the PM of larvae fed on Mp708 midwhorls could ensue when Mir1-CP degrades the IIM that cross-links chitin fibrils in the PM. Both targeted and global transcriptome analyses were performed to determine how complete dissolution of the structure and function of the PM is prevented, enabling larvae to continue growing in the presence of Mir1-CP. The MGs from fall armyworm fed on Mp708 upregulate expression of genes encoding proteins involved in PM production as an apparent compensation to replace the disrupted PM structure and restore appropriate counter-current MG gradients. Also, several families of digestive enzymes (endopeptidases, aminopeptidases, lipases, amylase) were more highly expressed in MGs from larvae fed on Mp708 than MGs from larvae fed on diets lacking Mir1-CP (artificial diet, midwhorls from Tx601 or B73 maize). Impaired growth of larvae fed on Mp708 probably results from metabolic costs associated with higher production of PM constituents and digestive enzymes in a compensatory attempt to maintain MG function.
Assuntos
Cisteína Proteases/metabolismo , Proteínas de Insetos/metabolismo , Proteínas de Plantas/metabolismo , Spodoptera/metabolismo , Zea mays/enzimologia , Animais , Trato Gastrointestinal/metabolismo , Herbivoria , Larva/metabolismo , Mucinas/metabolismo , Toxinas Biológicas/metabolismo , TranscriptomaRESUMO
Oxygen conductance to the tissues determines aerobic metabolic performance in most eukaryotes but has cost/benefit tradeoffs. Here we examine in lowland populations of a butterfly a genetic polymorphism affecting oxygen conductance via the hypoxia-inducible factor (HIF) pathway, which senses intracellular oxygen and controls the development of oxygen delivery networks. Genetically distinct clades of Glanville fritillary (Melitaea cinxia) across a continental scale maintain, at intermediate frequencies, alleles in a metabolic enzyme (succinate dehydrogenase, SDH) that regulates HIF-1α. One Sdhd allele was associated with reduced SDH activity rate, twofold greater cross-sectional area of tracheoles in flight muscle, and better flight performance. Butterflies with less tracheal development had greater post-flight hypoxia signaling, swollen & disrupted mitochondria, and accelerated aging of flight metabolic performance. Allelic associations with metabolic and aging phenotypes were replicated in samples from different clades. Experimentally elevated succinate in pupae increased the abundance of HIF-1α and expression of genes responsive to HIF activation, including tracheal morphogenesis genes. These results indicate that the hypoxia inducible pathway, even in lowland populations, can be an important axis for genetic variation underlying intraspecific differences in oxygen delivery, physiological performance, and life history.
Assuntos
Borboletas/genética , Variação Genética , Fator 1 Induzível por Hipóxia/genética , Transdução de Sinais/genética , Envelhecimento , Alelos , Altitude , Animais , Borboletas/metabolismo , Ecossistema , Voo Animal , Genes de Insetos , Fator 1 Induzível por Hipóxia/metabolismo , Proteínas de Insetos/genética , Proteínas de Insetos/metabolismo , Mitocôndrias Musculares/metabolismo , Músculo Esquelético/metabolismo , Músculo Esquelético/fisiologia , População/genética , Succinato Desidrogenase/genética , Succinato Desidrogenase/metabolismo , Ácido Succínico/metabolismo , Traqueia/fisiologia , Transcrição GênicaRESUMO
Adipokinetic neuropeptides from the corpora cardiaca of 17 species of Odonata encompassing mainly the families Corduliidae and Libellulidae were isolated and structurally elucidated using liquid chromatography coupled with ion trap electrospray ionization mass spectrometry. It became evident that all species of the family Corduliidae studied express the peptide code-named Libau-AKH (pGlu-Val-Asn-Phe-Thr-Pro-Ser-Trp amide), which is also present in all but one libellulid species, Erythemis simplicicollis which expresses Erysi-AKH (pGlu-Leu-Asn-Phe-Thr-Pro-Ser-Trp amide). This divergence from all other Libellulids is due to a nonsynonymous missense single nucleotide polymorphism (SNP) in the nucleotide coding sequence (CDS) of prepro-AKH CDS and supports the polyphyletic nature of Sympetrinae and other subfamilies of libellulids. Despite this exception, these findings then support the hypothesis that Corduliidae and Libellulidae are closely related as stated in most phylogenies. The presence of Anaim-AKH (pGlu-Val-Asn-Phe-Ser-Pro-Ser-Trp amide) in Macromiidae likely distinguishes species in this family from Corduliidae. Current molecular genetic phylogenies and our AKH findings suggest that Syncordulia gracilis, which expresses Anaim-AKH, does not belong in Corduliidae. Evolution of AKHs in anisopteran Odonata are likely due to nucleotide substitution involving nonsynonymous missense SNPs in the CDS of prepro-AKH.
Assuntos
Hormônios de Inseto/genética , Insetos/classificação , Insetos/genética , Oligopeptídeos/genética , Filogenia , Ácido Pirrolidonocarboxílico/análogos & derivados , Sequência de Aminoácidos , Animais , Sequência de Bases , Evolução Molecular , Hormônios de Inseto/química , Hormônios de Inseto/metabolismo , Insetos/química , Insetos/metabolismo , Dados de Sequência Molecular , Oligopeptídeos/química , Oligopeptídeos/metabolismo , Ácido Pirrolidonocarboxílico/química , Ácido Pirrolidonocarboxílico/metabolismo , Alinhamento de SequênciaRESUMO
A fundamental feature of gene expression in multicellular organisms is the production of distinct transcripts from single genes by alternative splicing (AS), which amplifies protein and functional diversity. In spite of the likely consequences for organismal biology, little is known about how AS varies among individuals or responds to body condition, environmental variation or extracellular signals in general. Here we show that evolutionarily conserved AS of troponin-t in flight muscle of adult moths responds in a quantitative fashion to experimental manipulation of larval nutrition and adult body weight. Troponin-t (Tnt) isoform composition is known to affect muscle force and power output in other animals, and is shown here to be associated with the thorax mass-specific rate of energy consumption during flight. Loading of adults with external weights for 5 days caused an AS response nearly identical to equal increases in actual body weight. In addition, there were effects of larval feeding history on adult Tnt isoform composition that were independent of body weight, with moths from poorer larval feeding regimes producing isoform profiles associated with reduced muscle performance and energy consumption rate. Thus, Tnt isoform composition in striated muscle is responsive to both weight-sensing and nutrition-sensing mechanisms, with consequent effects on function. In free-living butterflies, Tnt isoform composition was also associated with activity level and very strongly with the rate of egg production. Overall, these results show that AS of a muscle gene responds in a quantitative fashion to whole-organism variables, which apparently serves to coordinate muscle strength and energy expenditure with body condition and life history.
Assuntos
Processamento Alternativo , Mariposas/genética , Precursores de RNA/metabolismo , RNA Mensageiro/metabolismo , Troponina T/genética , Sequência de Aminoácidos , Fenômenos Fisiológicos da Nutrição Animal , Animais , Peso Corporal , Dieta , Voo Animal , Larva/crescimento & desenvolvimento , Larva/fisiologia , Dados de Sequência Molecular , Mariposas/anatomia & histologia , Mariposas/crescimento & desenvolvimento , Músculos/metabolismo , Isoformas de Proteínas/química , Isoformas de Proteínas/metabolismo , Precursores de RNA/química , RNA Mensageiro/química , Alinhamento de Sequência , Troponina T/químicaRESUMO
The genomes of three closely related bordetellae are currently being sequenced, thus providing an opportunity for comparative genomic approaches driven by an understanding of the comparative biology of these three bacteria. Although the other strains being sequenced are well studied, the strain of Bordetella parapertussis chosen for sequencing is a recent human clinical isolate (strain 12822) that has yet to be characterized in detail. This investigation reports the first phenotypic characterization of this strain, which will likely become the prototype for this species in comparison with the prototype strains of B. pertussis (Tohama I), B. bronchiseptica (RB50), and other isolates of B. parapertussis. Multiple in vitro and in vivo assays distinguished each species. B. parapertussis was more similar to B. bronchiseptica than to B. pertussis in many assays, including in BvgS signaling characteristics, presence of urease activity, regulation of urease expression by BvgAS, virulence in the respiratory tracts of immunocompromised mice, induction of anti-Bordetella antibodies, and serum antimicrobial resistance. In other assays, B. parapertussis was distinct from all other species (in pigment production) or more similar to B. pertussis (by lack of motility and cytotoxicity to a macrophage-like cell line). These results begin to provide phenotypes that can be related to genetic differences identified in the genomic sequences of bordetellae.