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1.
Extremophiles ; 20(1): 19-25, 2016 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-26475627

RESUMO

A Gram-negative, halophilic, heterotrophic, rod-shaped, non-spore-forming bacterium (SV525T) was isolated from the sediment of a hypersaline lake located at 4600 m above sea level (Laguna Vilama, Argentina). Strain SV525T was strictly aerobic and formed pink-to-magenta colonies. Growth occurred at 10­35 °C (optimum 25­30 °C), at pH levels 6.0­8.5 (optimum 7.0) and at NaCl concentrations of 7.5­25 % (w/v) with an optimum at 10­15 % (w/v). The strain required sodium and magnesium but not potassium ions for growth. Grows with tryptone, or Bacto Peptone as sole carbon and energy source and requires yeast extract for growth. It produced catalase and oxidase. The predominant ubiquinone was Q-8 and the major fatty acids comprised C18:1 ω7c, C16:0 and C18:0. The DNA G+C content was 60.4 mol% and its polar lipids consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine and a phosphoglycolipid. Phylogenetic analysis based on 16S rRNA gene indicated that strain SV525T belongs to the family Ectothiorhodospiraceae within the class Gammaproteobacteria. On the basis of phylogenetic and phenotypic data, SV525T represents a novel genus and species, for which the name Halopeptonella vilamensis gen. nov., sp. nov. is proposed. The type strain is SV525T (=DSM 21056T =JCM 16388T =NCIMB 14596T).


Assuntos
Ectothiorhodospiraceae/isolamento & purificação , Filogenia , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Composição de Bases , DNA Bacteriano/química , DNA Bacteriano/genética , Ectothiorhodospiraceae/classificação , Ectothiorhodospiraceae/genética , Ectothiorhodospiraceae/metabolismo , RNA Ribossômico 16S/genética
2.
Proc Natl Acad Sci U S A ; 110(29): 11917-22, 2013 Jul 16.
Artigo em Inglês | MEDLINE | ID: mdl-23798396

RESUMO

Insects are constantly adapting to human-driven landscape changes; however, the roles of their gut microbiota in these processes remain largely unknown. The western corn rootworm (WCR, Diabrotica virgifera virgifera LeConte) (Coleoptera: Chrysomelidae) is a major corn pest that has been controlled via annual rotation between corn (Zea mays) and nonhost soybean (Glycine max) in the United States. This practice selected for a "rotation-resistant" variant (RR-WCR) with reduced ovipositional fidelity to cornfields. When in soybean fields, RR-WCRs also exhibit an elevated tolerance of antiherbivory defenses (i.e., cysteine protease inhibitors) expressed in soybean foliage. Here we show that gut bacterial microbiota is an important factor facilitating this corn specialist's (WCR's) physiological adaptation to brief soybean herbivory. Comparisons of gut microbiota between RR- and wild-type WCR (WT-WCR) revealed concomitant shifts in bacterial community structure with host adaptation to soybean diets. Antibiotic suppression of gut bacteria significantly reduced RR-WCR tolerance of soybean herbivory to the level of WT-WCR, whereas WT-WCR were unaffected. Our findings demonstrate that gut bacteria help to facilitate rapid adaptation of insects in managed ecosystems.


Assuntos
Adaptação Fisiológica/fisiologia , Besouros/microbiologia , Inibidores de Cisteína Proteinase/metabolismo , Glycine max/química , Resistência a Inseticidas/fisiologia , Metagenoma/fisiologia , Zea mays/parasitologia , Acetatos , Animais , Sequência de Bases , Análise por Conglomerados , Besouros/fisiologia , Ciclopentanos , Inibidores de Cisteína Proteinase/análise , DNA Ribossômico/genética , Herbivoria/fisiologia , Interações Hospedeiro-Patógeno/fisiologia , Estimativa de Kaplan-Meier , Metagenoma/genética , Dados de Sequência Molecular , Oxilipinas , Folhas de Planta/química , Análise de Sequência de DNA , Estados Unidos
3.
Pestic Biochem Physiol ; 110: 1-6, 2014 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-24759044

RESUMO

RNA interference (RNAi) mediated crop protection against insect pests is a technology that is greatly anticipated by the academic and industrial pest control communities. Prior to commercialization, factors influencing the potential for evolution of insect resistance to RNAi should be evaluated. While mutations in genes encoding the RNAi machinery or the sequences targeted for interference may serve as a prominent mechanism of resistance evolution, differential effects of RNAi on target pests may also facilitate such evolution. However, to date, little is known about how variation of field insect populations could influence the effectiveness of RNAi treatments. To approach this question, we evaluated the effects of RNAi treatments on adults of three western corn rootworm (WCR; Diabrotica virgifera virgifera LeConte) populations exhibiting different levels of gut cysteine protease activity, tolerance of soybean herbivory, and immune gene expression; two populations were collected from crop rotation-resistant (RR) problem areas and one from a location where RR was not observed (wild type; WT). Our results demonstrated that RNAi targeting DvRS5 (a highly expressed cysteine protease gene) reduced gut cysteine protease activity in all three WCR populations. However, the proportion of the cysteine protease activity that was inhibited varied across populations. When WCR adults were treated with double-stranded RNA of an immune gene att1, different changes in survival among WT and RR populations on soybean diets occurred. Notably, for both genes, the sequences targeted for RNAi were the same across all populations examined. These findings indicate that the effectiveness of RNAi treatments could vary among field populations depending on their physiological and genetic backgrounds and that the consistency of an RNAi trait's effectiveness on phenotypically different populations should be considered or tested prior to wide deployment. Also, genes that are potentially subjected to differential selection in the field should be avoided for RNAi-based pest control.


Assuntos
Catepsina L/genética , Besouros/genética , Proteínas de Insetos/genética , Controle Biológico de Vetores , Interferência de RNA , Animais , Sequência de Bases , Trato Gastrointestinal/metabolismo , Genes de Insetos , Variação Genética , Larva/genética , Dados de Sequência Molecular , Glycine max
4.
Proc Natl Acad Sci U S A ; 107(27): 12168-73, 2010 Jul 06.
Artigo em Inglês | MEDLINE | ID: mdl-20566863

RESUMO

As an obligatory parasite of humans, the body louse (Pediculus humanus humanus) is an important vector for human diseases, including epidemic typhus, relapsing fever, and trench fever. Here, we present genome sequences of the body louse and its primary bacterial endosymbiont Candidatus Riesia pediculicola. The body louse has the smallest known insect genome, spanning 108 Mb. Despite its status as an obligate parasite, it retains a remarkably complete basal insect repertoire of 10,773 protein-coding genes and 57 microRNAs. Representing hemimetabolous insects, the genome of the body louse thus provides a reference for studies of holometabolous insects. Compared with other insect genomes, the body louse genome contains significantly fewer genes associated with environmental sensing and response, including odorant and gustatory receptors and detoxifying enzymes. The unique architecture of the 18 minicircular mitochondrial chromosomes of the body louse may be linked to the loss of the gene encoding the mitochondrial single-stranded DNA binding protein. The genome of the obligatory louse endosymbiont Candidatus Riesia pediculicola encodes less than 600 genes on a short, linear chromosome and a circular plasmid. The plasmid harbors a unique arrangement of genes required for the synthesis of pantothenate, an essential vitamin deficient in the louse diet. The human body louse, its primary endosymbiont, and the bacterial pathogens that it vectors all possess genomes reduced in size compared with their free-living close relatives. Thus, the body louse genome project offers unique information and tools to use in advancing understanding of coevolution among vectors, symbionts, and pathogens.


Assuntos
Genoma Bacteriano/genética , Genoma de Inseto/genética , Pediculus/genética , Pediculus/microbiologia , Animais , Enterobacteriaceae/genética , Genes Bacterianos/genética , Genes de Insetos/genética , Genômica/métodos , Humanos , Infestações por Piolhos/parasitologia , Dados de Sequência Molecular , Análise de Sequência de DNA , Simbiose
5.
Int J Syst Evol Microbiol ; 61(Pt 5): 1211-1217, 2011 May.
Artigo em Inglês | MEDLINE | ID: mdl-20584819

RESUMO

A Gram-stain-negative, aerobic, rod-shaped, non-spore-forming bacterium (SV325(T)) was isolated from the sediment of a hypersaline lake located 4600 m above sea level (Laguna Vilama, Argentina). Strain SV325(T) formed cream to pink colonies, was motile and moderately halophilic, and tolerated NaCl concentrations of 1-25 % (w/v) with an optimum of 5-10 % (w/v). Growth occurred at 5-40 °C (optimum around 30 °C) and at pH 5.0-10.0 (optimum 7.0-8.0). The bacterium did not produce exopolysaccharides and stained positively for intracellular polyphosphate granules but not for poly-ß-hydroxyalkanoates. It produced catalase and oxidase, reduced nitrate to nitrite, hydrolysed gelatin, did not produce acids from sugars and utilized a limited range of substrates as carbon and energy sources: acetate, caproate, fumarate, dl-ß-hydroxybutyrate, malate, maleate, malonate and succinate. The predominant ubiquinones were Q-9 (92.5 %) and Q-8 (7.5 %), the major fatty acids were C(19 : 0) cyclo ω8c, C(16 : 0), C(17 : 0) cyclo and C(16 : 1)ω7c/iso-C(15:0) 2-OH, and the DNA G+C content was 55.0 mol%. Phylogenetic analyses based on the 16S rRNA gene indicated that strain SV325(T) belongs to the genus Halomonas in the class Gammaproteobacteria. Physiological and biochemical tests allowed phenotypic differentiation of strain SV325(T) from closely related species with validly published names. We therefore propose a novel species, Halomonas vilamensis sp. nov., with type strain SV325(T) ( = DSM 21020(T)  = LMG 24332(T)).


Assuntos
Água Doce/microbiologia , Sedimentos Geológicos/microbiologia , Halomonas/classificação , Halomonas/isolamento & purificação , Altitude , DNA Bacteriano/genética , Ácidos Graxos/metabolismo , Halomonas/genética , Halomonas/metabolismo , Dados de Sequência Molecular , Filogenia , RNA Ribossômico 16S/genética , Cloreto de Sódio/metabolismo
6.
BMC Struct Biol ; 10: 43, 2010 Dec 29.
Artigo em Inglês | MEDLINE | ID: mdl-21190582

RESUMO

BACKGROUND: The completion of maize genome sequencing has resulted in the identification of a large number of uncharacterized genes. Gene annotation and functional characterization of gene products are important to uncover novel protein functionality. RESULTS: In this paper, we identify, and annotate members of all the maize aldehyde dehydrogenase (ALDH) gene superfamily according to the revised nomenclature criteria developed by ALDH Gene Nomenclature Committee (AGNC). The maize genome contains 24 unique ALDH sequences encoding members of ten ALDH protein families including the previously identified male fertility restoration RF2A gene, which encodes a member of mitochondrial class 2 ALDHs. Using computational modeling analysis we report here the identification, the physico-chemical properties, and the amino acid residue analysis of a novel tunnel like cavity exclusively found in the maize sterility restorer protein, RF2A/ALDH2B2 by which this protein is suggested to bind variably long chain molecular ligands and/or potentially harmful molecules. CONCLUSIONS: Our finding indicates that maize ALDH superfamily is the most expanded of plant ALDHs ever characterized, and the mitochondrial maize RF2A/ALDH2B2 is the only plant ALDH that harbors a newly defined pocket/cavity with suggested functional specificity.


Assuntos
Aldeído Desidrogenase/química , Aldeído Desidrogenase/metabolismo , Proteínas Mitocondriais/química , Proteínas Mitocondriais/metabolismo , Zea mays/enzimologia , Zea mays/genética , Aldeído Desidrogenase/genética , Sequência de Aminoácidos , Sequência de Bases , Sítios de Ligação , Fenômenos Químicos , Mitocôndrias/genética , Mitocôndrias/metabolismo , Proteínas Mitocondriais/genética , Proteínas Mitocondriais/isolamento & purificação , Modelos Moleculares , Anotação de Sequência Molecular , Família Multigênica , Filogenia , Ligação Proteica , Estrutura Quaternária de Proteína , Alinhamento de Sequência , Eletricidade Estática , Zea mays/metabolismo
7.
Mol Biol Rep ; 37(7): 3583-91, 2010 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-20204524

RESUMO

Previous studies have shown the role of melanized appressoria in the pathogenicity of various fungi. Diplocarpon rosae is a worldwide outdoor fungal pathogen of rose plants causing black spot disease of rose leaves. To fully understand how this fungus colonizes its host, which is critical for the development of an efficient and sustainable disease management program, we studied the fungal (especially the appressoria) structures of D. rosae in detail at an early stage of infection. Using both microscopic and biochemical analyses, we observed strong melanized appressoria formation localized at the point of D. rosae penetration, which forms the pathogen-plant interface. Treatment of infected plants with melanin biosynthesis inhibitors (MBIs) prevented melanization of D. rosae appressoria and positively correlated with significant reductions in black spot disease symptoms, suggesting that melanization of appressoria might be a critical factor for the pathogenicity of D. rosae. Our findings were confirmed and validated by the lack of melanized appressorial ring formation on an artificial surface and on a D. rosae-non host plant system, Arabidopsis thaliana. Our findings suggest that localized melanization of appressoria is a crucial factor for the pathogenicity of D. rosae and treatment of the fungus with MBIs seems to be a promising disease management alternative for black spot disease of roses.


Assuntos
Ascomicetos/patogenicidade , Melaninas/metabolismo , Esporos Fúngicos/metabolismo , Acetatos/farmacologia , Ascomicetos/citologia , Ascomicetos/efeitos dos fármacos , Ascomicetos/crescimento & desenvolvimento , Meios de Cultura/farmacologia , Germinação/efeitos dos fármacos , Iminas/farmacologia , Metacrilatos/farmacologia , Doenças das Plantas/microbiologia , Folhas de Planta/citologia , Folhas de Planta/efeitos dos fármacos , Folhas de Planta/microbiologia , Rosa/microbiologia , Esporos Fúngicos/citologia , Esporos Fúngicos/efeitos dos fármacos , Esporos Fúngicos/crescimento & desenvolvimento , Estrobilurinas
8.
Mol Biol Rep ; 37(7): 3631-5, 2010 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-20204529

RESUMO

Genomic DNA extraction protocols generally require the use of expensive and hazardous reagents necessary for decontamination of phenolic compounds from the extracts. In addition, they are lengthy, hindering large-scale sample extractions necessary for high-throughput analyses. Here we describe a simple, time and cost-efficient method for genomic DNA extraction from insects. The extracted DNA was successfully used in a Polymerase Chain Reaction (PCR), making it suitable for automation for large-scale genetic analysis and barcoding studies. The protocol employs a single purification step to remove polysaccharides and other contaminating compounds using a non-hazardous reagent buffer. In addition, we conducted a bioinformatics database analysis as proof of concept for the efficiency of the DNA extraction protocol by using universal barcoding primers specific for cytochrome c oxidase I gene to identify different arthropod specimens through Barcode of Life Database (BOLD) database search. The usefulness of this protocol in various molecular biology and biodiversity studies is further discussed.


Assuntos
Artrópodes/classificação , Artrópodes/genética , Código de Barras de DNA Taxonômico/métodos , DNA/genética , DNA/isolamento & purificação , Genoma/genética , Reação em Cadeia da Polimerase/métodos , Animais
9.
PLoS One ; 10(3): e0118566, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25768110

RESUMO

Gut bacterial communities of bumble bees are correlated with defense against pathogens. Further understanding this host-microbe association is vitally important as bumble bees are currently experiencing global population declines, potentially due in part to emergent diseases. In this study, we used pyrosequencing and community fingerprinting (ARISA) to characterize the gut microbial communities of nine bumble species from across the Bombus phylogeny. Overall, we delimited 74 bacterial taxa (operational taxonomic units or OTUs) belonging to Betaproteobacteria, Gammaproteobacteria, Bacilli, Actinobacteria, Flavobacteria and Alphaproteobacteria. Each bacterial community was taxonomically simple, containing an average of 1.9 common (relative abundance per sample > 5%) bacterial OTUs. The most abundant and prevalent (occurring in 92% of the samples) bacterial OTU, based on 16S rRNA sequences, closely matched that of the previously described Betaproteobacteria species Snodgrassella alvi. Bacteria that were first described in bee-related external environments dominated a number of gut bacterial communities, suggesting that they are not strictly dependent on the internal gut environment. The ARISA data showed a correlation between bacterial community structures and the geographic locations where the bees were sampled, suggesting that at least a subset of the bacterial species may be transmitted environmentally. Using light and fluorescent microscopy, we demonstrated that the gut bacteria form a biofilm on the internal epithelial surface of the ileum, corroborating results obtained from Apis mellifera.


Assuntos
Bactérias/genética , Abelhas/microbiologia , Trato Gastrointestinal/microbiologia , Animais , Biofilmes/crescimento & desenvolvimento , Ecologia , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência/métodos , Simbiose/genética
10.
Evol Appl ; 8(7): 692-704, 2015 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-26240606

RESUMO

The western corn rootworm (WCR,Diabrotica virgifera virgifera LeConte) is an important pest of corn. Annual crop rotation between corn and soybean disrupts the corn-dependent WCR life cycle and is widely adopted to manage this pest. This strategy selected for rotation-resistant (RR) WCR with reduced ovipositional fidelity to corn. Previous studies revealed that RR-WCR adults exhibit greater tolerance of soybean diets, different gut physiology, and host-microbe interactions compared to rotation-susceptible wild types (WT). To identify the genetic mechanisms underlying these phenotypic changes, a de novo assembly of the WCR adult gut transcriptome was constructed and used for RNA-sequencing analyses of RNA libraries from different WCR phenotypes fed with corn or soybean diets. Global gene expression profiles of WT- and RR-WCR were similar when feeding on corn diets, but different when feeding on soybean. Using network-based methods, we identified gene modules transcriptionally correlated with the RR phenotype. Gene ontology enrichment analyses indicated that the functions of these modules were related to metabolic processes, immune responses, biological adhesion, and other functions/processes that appear to correlate to documented traits in RR populations. These results suggest that gut transcriptomic divergence correlated with brief soybean feeding and other physiological traits may exist between RR- and WT-WCR adults.

12.
J Mol Microbiol Biotechnol ; 23(1-2): 152-77, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23615203

RESUMO

The origin and evolution of modern biochemistry and cellular structure is a complex problem that has puzzled scientists for almost a century. While comparative, functional and structural genomics has unraveled considerable complexity at the molecular level, there is very little understanding of the origin, evolution and structure of the molecules responsible for cellular or viral features in life. Recent efforts, however, have dissected the emergence of the very early molecules that populated primordial cells. Deep historical signal was retrieved from a census of molecular structures and functions in thousands of nucleic acid and protein structures and hundreds of genomes using powerful phylogenomic methods. Together with structural, chemical and cell biology considerations, this information reveals that modern biochemistry is the result of the gradual evolutionary appearance and accretion of molecular parts and molecules. These patterns comply with the principle of continuity and lead to molecular and cellular complexity. Here, we review findings and report possible origins of molecular and cellular structure, the early rise of lipid biosynthetic pathways and components of cytoskeletal microstructures, the piecemeal accumulation of domains in ATP synthase complexes and the origin and evolution of the ribosome. Phylogenomic studies suggest the last universal common ancestor of life, the 'urancestor', had already developed complex cellular structure and bioenergetics. Remarkably, our findings falsify the existence of an ancient RNA world. Instead they are compatible with gradually coevolving nucleic acids and proteins in interaction with increasingly complex cofactors, lipid membrane structures and other cellular components. This changes the perception we have of the rise of modern biochemistry and prompts further analysis of the emergence of biological complexity in an ever-expanding coevolving world of macromolecules.


Assuntos
Evolução Química , Evolução Molecular , Membranas/ultraestrutura , Proteínas/química , RNA/química , Fenômenos Bioquímicos , Evolução Biológica , Código Genético , Lipídeos/biossíntese , Membranas/química , ATPases Mitocondriais Próton-Translocadoras/química , Organelas/ultraestrutura , Filogenia , Proteínas/genética , Ribonucleoproteínas/química , Ribossomos/genética , Ribossomos/metabolismo
13.
J Mol Microbiol Biotechnol ; 23(1-2): 178-91, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23615204

RESUMO

Cells and viruses are structured and harbor complex organization. This manifests in intra- and extracellular compartments such as reticuli and periplasmic spaces, storage and energy-harvesting organelles such as acidocalcisomes and mitochondria, and specialized structures that hold genomic repositories such as nuclei and capsids. Structural phylogenomic reconstruction of the protein repertoire of the common ancestor of life, the urancestor, suggests these entities that existed 2.9 billion years ago were not only complex from a structural and functional point of view, but were also cellularly structured. We also provide support to the existence of urancestral storage organelles that were analogous to acidocalcisomes. These cellular structures probably accumulated compounds that stored energy in their phosphoanhydride bonds, such as polyphosphates. These energy-rich compounds necessary for thioester and pyrophosphate intermediates would have channeled the abundant redox energy of early Earth to the early metabolic needs of the primordial cells. Our findings are compatible with a relatively complex urancestral cell and with abundant microfossil evidence supporting the existence of primordial microbial communities as far back as 3.4 billion years ago. Results highlight the centrality of the cellular compartment and bioenergetics in the early evolutionary stages of life.


Assuntos
Evolução Biológica , Compartimento Celular , Metabolismo Energético , Evolução Molecular , Origem da Vida , Archaea/citologia , Archaea/genética , Archaea/metabolismo , Bactérias/citologia , Bactérias/genética , Bactérias/metabolismo , Membrana Celular , Eucariotos/citologia , Eucariotos/genética , Eucariotos/metabolismo , Genômica , Organelas/metabolismo , Organelas/ultraestrutura , Filogenia
14.
Ecol Evol ; 2(8): 2005-17, 2012 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-22957201

RESUMO

Western corn rootworm (Diabrotica virgifera) (WCR) depends on the continuous availability of corn. Broad adoption of annual crop rotation between corn (Zea mays) and nonhost soybean (Glycine max) exploited WCR biology to provide excellent WCR control, but this practice dramatically reduced landscape heterogeneity in East-central Illinois and imposed intense selection pressure. This selection resulted in behavioral changes and "rotation-resistant" (RR) WCR adults. Although soybeans are well defended against Coleopteran insects by cysteine protease inhibitors, RR-WCR feed on soybean foliage and remain long enough to deposit eggs that will hatch the following spring and larvae will feed on roots of planted corn. Other than documenting changes in insect mobility and egg laying behavior, 15 years of research have failed to identify any diagnostic differences between wild-type (WT)- and RR-WCR or a mechanism that allows for prolonged RR-WCR feeding and survival in soybean fields. We documented differences in behavior, physiology, digestive protease activity (threefold to fourfold increases), and protease gene expression in the gut of RR-WCR adults. Our data suggest that higher constitutive activity levels of cathepsin L are part of the mechanism that enables populations of WCR to circumvent soybean defenses, and thus, crop rotation. These new insights into the mechanism of WCR tolerance of soybean herbivory transcend the issue of RR-WCR diagnostics and management to link changes in insect gut proteolytic activity and behavior with landscape heterogeneity. The RR-WCR illustrates how agro-ecological factors can affect the evolution of insects in human-altered ecosystems.

15.
Biol Direct ; 6: 50, 2011 Oct 05.
Artigo em Inglês | MEDLINE | ID: mdl-21974828

RESUMO

BACKGROUND: Volutin granules appear to be universally distributed and are morphologically and chemically identical to acidocalcisomes, which are electron-dense granular organelles rich in calcium and phosphate, whose functions include storage of phosphorus and various metal ions, metabolism of polyphosphate, maintenance of intracellular pH, osmoregulation and calcium homeostasis. Prokaryotes are thought to differ from eukaryotes in that they lack membrane-bounded organelles. However, it has been demonstrated that as in acidocalcisomes, the calcium and polyphosphate-rich intracellular "volutin granules (polyphosphate bodies)" in two bacterial species, Agrobacterium tumefaciens, and Rhodospirillum rubrum, are membrane bound and that the vacuolar proton-translocating pyrophosphatases (V-H+PPases) are present in their surrounding membranes. Volutin granules and acidocalcisomes have been found in organisms as diverse as bacteria and humans. RESULTS: Here, we show volutin granules also occur in Archaea and are, therefore, present in the three superkingdoms of life (Archaea, Bacteria and Eukarya). Molecular analyses of V-H+PPase pumps, which acidify the acidocalcisome lumen and are diagnostic proteins of the organelle, also reveal the presence of this enzyme in all three superkingdoms suggesting it is ancient and universal. Since V-H+PPase sequences contained limited phylogenetic signal to fully resolve the ancestral nodes of the tree, we investigated the divergence of protein domains in the V-H+PPase molecules. Using Protein family (Pfam) database, we found a domain in the protein, PF03030. The domain is shared by 31 species in Eukarya, 231 in Bacteria, and 17 in Archaea. The universal distribution of the V-H+PPase PF03030 domain, which is associated with the V-H+PPase function, suggests the domain and the enzyme were already present in the Last Universal Common Ancestor (LUCA). CONCLUSION: The importance of the V-H+PPase function and the evolutionary dynamics of these domains support the early origin of the acidocalcisome organelle. In particular, the universality of volutin granules and presence of a functional V-H+PPase domain in the three superkingdoms of life reveals that the acidocalcisomes may have appeared earlier than the divergence of the superkingdoms. This result is remarkable and highlights the possibility that a high degree of cellular compartmentalization could already have been present in the LUCA.


Assuntos
Bactérias/enzimologia , Grânulos Citoplasmáticos/metabolismo , Evolução Molecular , Pirofosfatase Inorgânica/metabolismo , Vacúolos/enzimologia , Ácidos/metabolismo , Archaea/classificação , Archaea/enzimologia , Bactérias/classificação , Bactérias/ultraestrutura , Teorema de Bayes , Cálcio/metabolismo , Bases de Dados de Proteínas , Eucariotos/classificação , Eucariotos/enzimologia , Microscopia Eletrônica , Filogenia , Polifosfatos/metabolismo , Estrutura Terciária de Proteína , Alinhamento de Sequência , Vacúolos/metabolismo
16.
Commun Integr Biol ; 4(2): 188-91, 2011 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-21655436

RESUMO

The body louse, with its recently sequenced genome, is now primed to serve as a powerful model organism for addressing fundamental questions relating to how insects interact with their environment. One characteristic of the body louse that facilitates this research is the size of its genome-the smallest insect genome sequenced to date. This diminutive genome must nonetheless control an organism that senses and responds to its environment, reacting to threats of corporal and genomic integrity. Additionally, the body louse transmits several important human diseases compared to its very close relative, the head louse, which does not. Therefore, these two organisms comprise an excellent model system for studying molecular mechanisms associated with vector competence. To understand more fully the development of vector/pathogen interactions, we have developed an in vitro bioassay system and determined that the body louse genome appears to contain the genes necessary for RNAi. The body louse will therefore be useful for determining the set of conditions permissive to the evolution of vector competence.

17.
PLoS One ; 6(2): e16444, 2011 Feb 02.
Artigo em Inglês | MEDLINE | ID: mdl-21311752

RESUMO

We report the assembly of the 14,054 bp near complete sequencing of the mitochondrial genome of the legume pod borer (LPB), Maruca vitrata (Lepidoptera: Crambidae), which we subsequently used to estimate divergence and relationships within the lepidopteran lineage. The arrangement and orientation of the 13 protein-coding, 2 rRNA, and 19 tRNA genes sequenced was typical of insect mitochondrial DNA sequences described to date. The sequence contained a high A+T content of 80.1% and a bias for the use of codons with A or T nucleotides in the 3rd position. Transcript mapping with midgut and salivary gland ESTs for mitochondrial genome annotation showed that translation from protein-coding genes initiates and terminates at standard mitochondrial codons, except for the coxI gene, which may start from an arginine CGA codon. The genomic copy of coxII terminates at a T nucleotide, and a proposed polyadenylation mechanism for completion of the TAA stop codon was confirmed by comparisons to EST data. EST contig data further showed that mature M. vitrata mitochondrial transcripts are monocistronic, except for bicistronic transcripts for overlapping genes nd4/nd4L and nd6/cytb, and a tricistronic transcript for atp8/atp6/coxIII. This processing of polycistronic mitochondrial transcripts adheres to the tRNA punctuated cleavage mechanism, whereby mature transcripts are cleaved only at intervening tRNA gene sequences. In contrast, the tricistronic atp8/atp6/coxIII in Drosophila is present as separate atp8/atp6 and coxIII transcripts despite the lack of an intervening tRNA. Our results indicate that mitochondrial processing mechanisms vary between arthropod species, and that it is crucial to use transcriptional information to obtain full annotation of mitochondrial genomes.


Assuntos
Perfilação da Expressão Gênica , Genoma Mitocondrial/genética , Lepidópteros/genética , Análise de Sequência de DNA , Animais , Mapeamento Cromossômico , Cromossomos de Insetos , Códon/análise , Códon/genética , Etiquetas de Sequências Expressas , Frutas/parasitologia , Genoma de Inseto/genética , Lepidópteros/fisiologia , Anotação de Sequência Molecular , Dados de Sequência Molecular , Filogenia
18.
PLoS One ; 6(7): e21388, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-21754987

RESUMO

The legume pod borer, Maruca vitrata (Lepidoptera: Crambidae), is an insect pest species of crops grown by subsistence farmers in tropical regions of Africa. We present the de novo assembly of 3729 contigs from 454- and Sanger-derived sequencing reads for midgut, salivary, and whole adult tissues of this non-model species. Functional annotation predicted that 1320 M. vitrata protein coding genes are present, of which 631 have orthologs within the Bombyx mori gene model. A homology-based analysis assigned M. vitrata genes into a group of paralogs, but these were subsequently partitioned into putative orthologs following phylogenetic analyses. Following sequence quality filtering, a total of 1542 putative single nucleotide polymorphisms (SNPs) were predicted within M. vitrata contig assemblies. Seventy one of 1078 designed molecular genetic markers were used to screen M. vitrata samples from five collection sites in West Africa. Population substructure may be present with significant implications in the insect resistance management recommendations pertaining to the release of biological control agents or transgenic cowpea that express Bacillus thuringiensis crystal toxins. Mutation data derived from transcriptome sequencing is an expeditious and economical source for genetic markers that allow evaluation of ecological differentiation.


Assuntos
Frutas/parasitologia , Perfilação da Expressão Gênica/métodos , Lepidópteros/genética , Polimorfismo de Nucleotídeo Único/genética , Análise de Sequência de DNA/métodos , África , Animais , DNA Complementar/genética , Bases de Dados Genéticas , Sistema Digestório/metabolismo , Etiquetas de Sequências Expressas , Genes de Insetos/genética , Marcadores Genéticos , Genética Populacional , Genótipo , Geografia , Larva/genética , Anotação de Sequência Molecular , Dados de Sequência Molecular , Filogenia , Análise de Regressão , Glândulas Salivares/metabolismo , Homologia de Sequência do Ácido Nucleico , Software
19.
PLoS One ; 6(4): e18215, 2011 Apr 20.
Artigo em Inglês | MEDLINE | ID: mdl-21533132

RESUMO

BACKGROUND: Methamphetamine (METH), an abused illicit drug, disrupts many cellular processes, including energy metabolism, spermatogenesis, and maintenance of oxidative status. However, many components of the molecular underpinnings of METH toxicity have yet to be established. Network analyses of integrated proteomic, transcriptomic and metabolomic data are particularly well suited for identifying cellular responses to toxins, such as METH, which might otherwise be obscured by the numerous and dynamic changes that are induced. METHODOLOGY/RESULTS: We used network analyses of proteomic and transcriptomic data to evaluate pathways in Drosophila melanogaster that are affected by acute METH toxicity. METH exposure caused changes in the expression of genes involved with energy metabolism, suggesting a Warburg-like effect (aerobic glycolysis), which is normally associated with cancerous cells. Therefore, we tested the hypothesis that carbohydrate metabolism plays an important role in METH toxicity. In agreement with our hypothesis, we observed that increased dietary sugars partially alleviated the toxic effects of METH. Our systems analysis also showed that METH impacted genes and proteins known to be associated with muscular homeostasis/contraction, maintenance of oxidative status, oxidative phosphorylation, spermatogenesis, iron and calcium homeostasis. Our results also provide numerous candidate genes for the METH-induced dysfunction of spermatogenesis, which have not been previously characterized at the molecular level. CONCLUSION: Our results support our overall hypothesis that METH causes a toxic syndrome that is characterized by the altered carbohydrate metabolism, dysregulation of calcium and iron homeostasis, increased oxidative stress, and disruption of mitochondrial functions.


Assuntos
Drosophila melanogaster/efeitos dos fármacos , Metanfetamina/farmacologia , Biologia de Sistemas , Animais , Cromatografia Líquida de Alta Pressão , Carboidratos da Dieta/administração & dosagem , Drosophila melanogaster/citologia , Drosophila melanogaster/metabolismo , Transporte de Elétrons , Metabolismo Energético/genética , Perfilação da Expressão Gênica , Homeostase , Masculino , Metabolômica , Análise de Sequência com Séries de Oligonucleotídeos , Estresse Oxidativo , Proteômica , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Espectrometria de Massas em Tandem , Trealose/administração & dosagem
20.
PLoS One ; 5(12): e15906, 2010 Dec 28.
Artigo em Inglês | MEDLINE | ID: mdl-21203394

RESUMO

The male fertility restorer (RF) proteins belong to extended protein families associated with the cytoplasmic male sterility in higher plants. Up till now, there is no devised nomenclature for naming the RF proteins. The systematic sequencing of new plant species in recent years has uncovered the existence of several novel RF genes and their encoded proteins. Their naming has been simply arbitrary and could not be adequately handled in the context of comparative functional genomics. We propose in this study a unified nomenclature for the RF extended protein families across all plant species. This new and unified nomenclature relies upon previously developed nomenclature for the first ever characterized RF gene, RF2A/ALDH2B2, a member of ALDH gene superfamily, and adheres to the guidelines issued by the ALDH Genome Nomenclature Committees. The proposed nomenclature reveals that RF gene superfamily encodes currently members of 51 families. This unified nomenclature accommodates functional RF genes and pseudogenes, and offers the flexibility needed to incorporate additional RFs as they become available in future. In addition, we provide a phylogenetic relationship between the RF extended families and use computational protein modeling to demonstrate the high divergence of RF functional specializations through specific structural features of selected members of RF superfamily.


Assuntos
Isoenzimas/genética , Retinal Desidrogenase/genética , Família Aldeído Desidrogenase 1 , Biologia Computacional/métodos , Bases de Dados de Proteínas , Genes de Plantas , Genômica , Isoenzimas/metabolismo , Conformação Molecular , Fases de Leitura Aberta , Filogenia , Infertilidade das Plantas , Proteínas de Plantas/química , Proteínas de Plantas/genética , Plantas/metabolismo , Conformação Proteica , Pseudogenes , Retinal Desidrogenase/metabolismo , Software , Zea mays/genética
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