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1.
Front Plant Sci ; 12: 675859, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34394138

RESUMO

Wheat (Triticum aestivum L.) is the most widely grown cereal crop in the world and is staple food to half the world's population. The current world population is expected to reach 9.8 billion people by 2050, but food production is not expected to keep pace with demand in developing countries. Significant opportunities exist for traditional grain exporters to produce and export greater amounts of wheat to fill the gap. Karnal bunt, however, is a major threat, due to its use as a non-tariff trade barrier by several wheat-importing countries. The cultivation of resistant varieties remains the most cost-effective approach to manage the disease, but in countries that are free of the disease, genetic improvement is difficult due to quarantine restrictions. Here we report a study on pre-emptive breeding designed to identify linked molecular markers, evaluate the prospects of genomic selection as a tool, and prioritise wheat genotypes suitable for use as parents. In a genome-wide association (GWAS) study, we identified six DArTseq markers significantly linked to Karnal bunt resistance, which explained between 7.6 and 29.5% of the observed phenotypic variation. The accuracy of genomic prediction was estimated to vary between 0.53 and 0.56, depending on whether it is based solely on the identified Quantitative trait loci (QTL) markers or the use of genome-wide markers. As genotypes used as parents would be required to possess good yield and phenology, further research was conducted to assess the agronomic value of Karnal bunt resistant germplasm from the International Maize and Wheat Improvement Center (CIMMYT). We identified an ideal genotype, ZVS13_385, which possessed similar agronomic attributes to the highly successful Australian wheat variety, Mace. It is phenotypically resistant to Karnal bunt infection (<1% infection) and carried all the favourable alleles detected for resistance in this study. The identification of a genotype combining Karnal bunt resistance with adaptive agronomic traits overcomes the concerns of breeders regarding yield penalty in the absence of the disease.

2.
Proc Natl Acad Sci U S A ; 118(14)2021 04 06.
Artigo em Inglês | MEDLINE | ID: mdl-33737447

RESUMO

When addressing a genomic question, having a reliable and adequate reference genome is of utmost importance. This drives the necessity to refine and customize reference genomes (RGs). Our laboratory has recently developed a strategy, the Perfect Match Genomic Landscape (PMGL), to detect variation between genomes [K. Palacios-Flores et al.Genetics 208, 1631-1641 (2018)]. The PMGL is precise and sensitive and, in contrast to most currently used algorithms, is nonstatistical in nature. Here we demonstrate the power of PMGL to refine and customize RGs. As a proof-of-concept, we refined different versions of the Saccharomyces cerevisiae RG. We applied the automatic PMGL pipeline to refine the genomes of microorganisms belonging to the three domains of life: the archaea Methanococcus maripaludis and Pyrococcus furiosus; the bacteria Escherichia coli, Staphylococcus aureus, and Bacillus subtilis; and the eukarya Schizosaccharomyces pombe, Aspergillus oryzae, and several strains of Saccharomyces paradoxus. We analyzed the reference genome of the virus SARS-CoV-2 and previously published viral genomes from patients' samples with COVID-19. We performed a mutation-accumulation experiment in E. coli and show that the PMGL strategy can detect specific mutations generated at any desired step of the whole procedure. We propose that PMGL can be used as a final step for the refinement and customization of any haploid genome, independently of the strategies and algorithms used in its assembly.


Assuntos
Variação Genética , Genoma Microbiano , Genômica/métodos , SARS-CoV-2/genética , Algoritmos , Acúmulo de Mutações , Estudo de Prova de Conceito , Saccharomyces cerevisiae/genética
3.
Int J Mol Sci ; 20(13)2019 Jun 26.
Artigo em Inglês | MEDLINE | ID: mdl-31247965

RESUMO

Karnal bunt disease of wheat, caused by the fungus Neovossia indica, is one of the most important challenges to the grain industry as it affects the grain quality and also restricts the international movement of infected grain. It is a seed-, soil- and airborne disease with limited effect of chemical control. Currently, this disease is contained through the deployment of host resistance but further improvement is limited as only a few genotypes have been found to carry partial resistance. To identify genomic regions responsible for resistance in a set of 339 wheat accessions, genome-wide association study (GWAS) was undertaken using the DArTSeq® technology, in which 18 genomic regions for Karnal bunt resistance were identified, explaining 5-20% of the phenotypic variation. The identified quantitative trait loci (QTL) on chromosome 2BL showed consistently significant effects across all four experiments, whereas another QTL on 5BL was significant in three experiments. Additional QTLs were mapped on chromosomes 1DL, 2DL, 4AL, 5AS, 6BL, 6BS, 7BS and 7DL that have not been mapped previously, and on chromosomes 4B, 5AL, 5BL and 6BS, which have been reported in previous studies. Germplasm with less than 1% Karnal bunt infection have been identified and can be used for resistance breeding. The SNP markers linked to the genomic regions conferring resistance to Karnal bunt could be used to improve Karnal bunt resistance through marker-assisted selection.


Assuntos
Basidiomycota , Resistência à Doença/genética , Estudo de Associação Genômica Ampla , Interações Hospedeiro-Patógeno/genética , Doenças das Plantas/genética , Doenças das Plantas/microbiologia , Triticum/genética , Marcadores Genéticos , Variação Genética , Genoma Fúngico , Melhoramento Vegetal , Polimorfismo de Nucleotídeo Único , Locos de Características Quantitativas , Sementes
4.
Proc Natl Acad Sci U S A ; 116(17): 8445-8450, 2019 04 23.
Artigo em Inglês | MEDLINE | ID: mdl-30962378

RESUMO

Genomes are dynamic structures. Different mechanisms participate in the generation of genomic rearrangements. One of them is nonallelic homologous recombination (NAHR). This rearrangement is generated by recombination between pairs of repeated sequences with high identity. We analyzed rearrangements mediated by repeated sequences located in different chromosomes. Such rearrangements generate chimeric chromosomes. Potential rearrangements were predicted by localizing interchromosomal identical repeated sequences along the nuclear genome of the Saccharomyces cerevisiae S288C strain. Rearrangements were identified by a PCR-based experimental strategy. PCR primers are located in the unique regions bordering each repeated region of interest. When the PCR is performed using forward primers from one chromosome and reverse primers from another chromosome, the break point of the chimeric chromosome structure is revealed. In all cases analyzed, the corresponding chimeric structures were found. Furthermore, the nucleotide sequence of chimeric structures was obtained, and the origin of the unique regions bordering the repeated sequence was located in the expected chromosomes, using the perfect-match genomic landscape strategy (PMGL). Several chimeric structures were searched in colonies derived from single cells. All of the structures were found in DNA isolated from each of the colonies. Our findings indicate that interchromosomal rearrangements that generate chimeric chromosomes are recurrent and occur, at a relatively high frequency, in cell populations of S. cerevisiae.


Assuntos
Cromossomos Fúngicos/genética , Rearranjo Gênico/genética , Genoma Fúngico/genética , Saccharomyces cerevisiae/genética , Genômica , Modelos Genéticos , Reação em Cadeia da Polimerase , Sequências Repetitivas de Ácido Nucleico/genética , Análise de Sequência de DNA
5.
G3 (Bethesda) ; 9(5): 1437-1447, 2019 05 07.
Artigo em Inglês | MEDLINE | ID: mdl-30824480

RESUMO

Karnal bunt caused by Tilletia indica Mitra [syn. Neovossia indica (Mitra) Mundkur] is a significant biosecurity concern for wheat-exporting countries that are free of the disease. It is a seed-, soil-and air-borne disease with no effective chemical control measures. The current study used data from multi-year field experiments of two bi-parental populations and a genome-wide association (GWA) mapping panel to unravel the genetic basis for resistance in common wheat. Broad-sense heritability for Karnal bunt resistance in the populations varied from 0.52 in the WH542×HD29 population, to 0.61 in the WH542×W485 cross and 0.71 in a GWAS panel. Quantitative trait locus (QTL) analysis with seven years of phenotypic data identified a major locus on chromosome 3B (R2 = 27.8%) and a minor locus on chromosome 1A (R2 = 12.2%), in the WH542×HD29 population, with both parents contributing the high-value alleles. A major locus (R2 = 27.8%) and seven minor loci (R2 = 4.4-15.8%) were detected in the WH542×W485 population. GWA mapping validated QTL regions in the bi-parent populations, but also identified novel loci not previously associated with Karnal bunt resistance. Meta-QTL analysis aligned the results from this study with those reported in wheat over the last two decades. Two major clusters were detected, the first on chromosome 4B, which clustered with Qkb.ksu-4B, QKb.cimmyt-4BL, Qkb.cim-4BL, and the second on chromosome 3B, which clustered with Qkb.cnl-3B, QKb.cimmyt-3BS and Qkb.cim-3BS1 The results provide definitive chromosomal assignments for QTL/genes controlling Karnal bunt resistance in common wheat, and will be useful in pre-emptive breeding against the pathogen in wheat-producing areas that are free of the disease.


Assuntos
Basidiomycota , Resistência à Doença/genética , Ligação Genética , Estudo de Associação Genômica Ampla , Doenças das Plantas/genética , Doenças das Plantas/microbiologia , Triticum/genética , Triticum/microbiologia , Mapeamento Cromossômico , Genética Populacional , Estudo de Associação Genômica Ampla/métodos , Genótipo , Endogamia , Fenótipo , Melhoramento Vegetal , Polimorfismo de Nucleotídeo Único , Locos de Características Quantitativas
6.
Front Plant Sci ; 9: 1497, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30386358

RESUMO

Karnal bunt (KB) of wheat, caused by Tilletia indica, is one of the greatest challenges to grain industry, not because of yield loss, but quarantine regulations that restrict international movement and trade of affected stocks. Genetic resistance is the best way to manage this disease. Although several different sources of resistance have been identified to date, very few of those have been subjected to genetic analyses. Understanding the genetics of resistance, characterization and mapping of new resistance loci can help in development of improved germplasm. The objective of this study was to identify and characterize resistance loci (QTL) in two independent recombinant inbred lines (RILs) populations utilizing different wheat lines as resistance donors. Elite CIMMYT wheat lines Blouk#1 and Huirivis#1 were used as susceptible female parents and WHEAR/KUKUNA/3/C80.1/3∗BATAVIA//2∗WBLL1 (WKCBW) and Mutus as moderately resistant male parents in Pop1 and Pop2 populations, respectively. Populations were evaluated for KB resistance in 2015-16 and 2016-17 cropping seasons at two seeding dates (total four environments) in Cd. Obregon, Mexico. Two stable QTL from each population were identified in each environment: QKb.cim-2B and QKb.cim-3D (Pop1), QKb.cim-3B1 and QKb.cim-5B2 (Pop2). Other than those four QTL, other QTL were detected in each population which were specific to environments: QKb.cim-5B1, QKb.cim-6A, and QKb.cim-7A (Pop1), QKb.cim-3B2, QKb.cim-4A1, QKb.cim-4A2, QKb.cim-4B, QKb.cim-5A1, QKb.cim-5A2, and QKb.cim-7A2 (Pop2). Among the four stable QTL, all but QKb.cim-3B1 were derived from the resistant parent. QKb.cim-2B and QKb.cim-3D in Pop1 and QKb.cim-3B1 and QKb.cim-5B2 in Pop2 explained 5.0-11.4% and 3.3-7.1% phenotypic variance, respectively. A combination of two stable QTL in each population reduced KB infection by 24-33%, respectively. Transgressive resistant segregants lines derived with resistance alleles from both parents in each population were identified. Single nucleotide polymorphism (SNP) markers flanking these QTL regions may be amenable to marker-assisted selection. The best lines from both populations (in agronomy, end-use quality and KB resistance) carrying resistance alleles at all identified loci, may be used for inter-crossing and selection of improved germplasm in future. Markers flanking these QTL may assist in selection of such lines.

7.
Sci Rep ; 8(1): 12527, 2018 08 21.
Artigo em Inglês | MEDLINE | ID: mdl-30131572

RESUMO

The value of exotic wheat genetic resources for accelerating grain yield gains is largely unproven and unrealized. We used next-generation sequencing, together with multi-environment phenotyping, to study the contribution of exotic genomes to 984 three-way-cross-derived (exotic/elite1//elite2) pre-breeding lines (PBLs). Genomic characterization of these lines with haplotype map-based and SNP marker approaches revealed exotic specific imprints of 16.1 to 25.1%, which compares to theoretical expectation of 25%. A rare and favorable haplotype (GT) with 0.4% frequency in gene bank identified on chromosome 6D minimized grain yield (GY) loss under heat stress without GY penalty under irrigated conditions. More specifically, the 'T' allele of the haplotype GT originated in Aegilops tauschii and was absent in all elite lines used in study. In silico analysis of the SNP showed hits with a candidate gene coding for isoflavone reductase IRL-like protein in Ae. tauschii. Rare haplotypes were also identified on chromosomes 1A, 6A and 2B effective against abiotic/biotic stresses. Results demonstrate positive contributions of exotic germplasm to PBLs derived from crosses of exotics with CIMMYT's best elite lines. This is a major impact-oriented pre-breeding effort at CIMMYT, resulting in large-scale development of PBLs for deployment in breeding programs addressing food security under climate change scenarios.


Assuntos
Sequenciamento de Nucleotídeos em Larga Escala/métodos , Polimorfismo de Nucleotídeo Único , Locos de Características Quantitativas , Triticum/genética , Mapeamento Cromossômico , Grão Comestível/genética , Abastecimento de Alimentos , Frequência do Gene , Haplótipos , Temperatura Alta , Melhoramento Vegetal , Banco de Sementes , Análise de Sequência de DNA , Estresse Fisiológico , Triticum/classificação , Triticum/crescimento & desenvolvimento
8.
Proc Natl Acad Sci U S A ; 115(21): 5516-5521, 2018 05 22.
Artigo em Inglês | MEDLINE | ID: mdl-29735690

RESUMO

The precise determination of de novo genetic variants has enormous implications across different fields of biology and medicine, particularly personalized medicine. Currently, de novo variations are identified by mapping sample reads from a parent-offspring trio to a reference genome, allowing for a certain degree of differences. While widely used, this approach often introduces false-positive (FP) results due to misaligned reads and mischaracterized sequencing errors. In a previous study, we developed an alternative approach to accurately identify single nucleotide variants (SNVs) using only perfect matches. However, this approach could be applied only to haploid regions of the genome and was computationally intensive. In this study, we present a unique approach, coverage-based single nucleotide variant identification (COBASI), which allows the exploration of the entire genome using second-generation short sequence reads without extensive computing requirements. COBASI identifies SNVs using changes in coverage of exactly matching unique substrings, and is particularly suited for pinpointing de novo SNVs. Unlike other approaches that require population frequencies across hundreds of samples to filter out any methodological biases, COBASI can be applied to detect de novo SNVs within isolated families. We demonstrate this capability through extensive simulation studies and by studying a parent-offspring trio we sequenced using short reads. Experimental validation of all 58 candidate de novo SNVs and a selection of non-de novo SNVs found in the trio confirmed zero FP calls. COBASI is available as open source at https://github.com/Laura-Gomez/COBASI for any researcher to use.


Assuntos
Variações do Número de Cópias de DNA , Genoma Humano , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Pais , Polimorfismo de Nucleotídeo Único , Análise de Sequência de DNA/métodos , Software , Algoritmos , Criança , Humanos
9.
Genetics ; 208(4): 1631-1641, 2018 04.
Artigo em Inglês | MEDLINE | ID: mdl-29367403

RESUMO

We present a conceptually simple, sensitive, precise, and essentially nonstatistical solution for the analysis of genome variation in haploid organisms. The generation of a Perfect Match Genomic Landscape (PMGL), which computes intergenome identity with single nucleotide resolution, reveals signatures of variation wherever a query genome differs from a reference genome. Such signatures encode the precise location of different types of variants, including single nucleotide variants, deletions, insertions, and amplifications, effectively introducing the concept of a general signature of variation. The precise nature of variants is then resolved through the generation of targeted alignments between specific sets of sequence reads and known regions of the reference genome. Thus, the perfect match logic decouples the identification of the location of variants from the characterization of their nature, providing a unified framework for the detection of genome variation. We assessed the performance of the PMGL strategy via simulation experiments. We determined the variation profiles of natural genomes and of a synthetic chromosome, both in the context of haploid yeast strains. Our approach uncovered variants that have previously escaped detection. Moreover, our strategy is ideally suited for further refining high-quality reference genomes. The source codes for the automated PMGL pipeline have been deposited in a public repository.


Assuntos
Variação Genética , Genoma , Genômica , Haploidia , Cromossomos , Biologia Computacional , Simulação por Computador , Testes Genéticos , Genoma Fúngico , Genoma Humano , Estudo de Associação Genômica Ampla , Genômica/métodos , Humanos , Polimorfismo de Nucleotídeo Único , Sequenciamento Completo do Genoma , Leveduras/genética
10.
Genome Announc ; 5(30)2017 Jul 27.
Artigo em Inglês | MEDLINE | ID: mdl-28751391

RESUMO

We present here the high-quality complete genome sequences of eight strains of Rhizobium-nodulating Phaseolus vulgaris Comparative analyses showed that some of them belonged to different genomic and evolutionary lineages with common symbiotic properties. Two novel symbiotic plasmids (pSyms) with P. vulgaris specificity are reported here.

11.
J Vis Exp ; (124)2017 06 25.
Artigo em Inglês | MEDLINE | ID: mdl-28671650

RESUMO

Here, we present chimera assembly by plasmid recovery and restriction enzyme site insertion (CAPRRESI). CAPRRESI benefits from many strengths of the original plasmid recovery method and introduces restriction enzyme digestion to ease DNA ligation reactions (required for chimera assembly). For this protocol, users clone wildtype genes into the same plasmid (pUC18 or pUC19). After the in silico selection of amino acid sequence regions where chimeras should be assembled, users obtain all the synonym DNA sequences that encode them. Ad hoc Perl scripts enable users to determine all synonym DNA sequences. After this step, another Perl script searches for restriction enzyme sites on all synonym DNA sequences. This in silico analysis is also performed using the ampicillin resistance gene (ampR) found on pUC18/19 plasmids. Users design oligonucleotides inside synonym regions to disrupt wildtype and ampR genes by PCR. After obtaining and purifying complementary DNA fragments, restriction enzyme digestion is accomplished. Chimera assembly is achieved by ligating appropriate complementary DNA fragments. pUC18/19 vectors are selected for CAPRRESI because they offer technical advantages, such as small size (2,686 base pairs), high copy number, advantageous sequencing reaction features, and commercial availability. The usage of restriction enzymes for chimera assembly eliminates the need for DNA polymerases yielding blunt-ended products. CAPRRESI is a fast and low-cost method for fusing protein-coding genes.


Assuntos
Enzimas de Restrição do DNA , Vetores Genéticos , Plasmídeos , Sequência de Aminoácidos , Enzimas de Restrição do DNA/genética , Vetores Genéticos/genética , Plasmídeos/química
12.
Genome Announc ; 5(11)2017 Mar 16.
Artigo em Inglês | MEDLINE | ID: mdl-28302777

RESUMO

The whole-genome sequences of three strains of Rhizobium gallicum reported here support the concept that the distinct nodulation host ranges displayed by the symbiovars gallicum and phaseoli can be largely explained by different symbiotic plasmids.

13.
Int Urogynecol J ; 28(7): 1101-1102, 2017 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-28032188

RESUMO

INTRODUCTION AND HYPOTHESIS: We present a simple approach to the marsupialization of a Skene's gland cyst. METHODS: Our technique facilitates suture placement to exteriorize the cyst wall to efficiently treat a distal Skene's gland cyst and reduce the risk of recurrence. CONCLUSION: Marsupialization is an accepted option for the surgical management of Skene's gland cyst. This technique is an effective and streamlined approach to Skene's gland marsupialization.


Assuntos
Cistos/cirurgia , Procedimentos Cirúrgicos em Ginecologia/métodos , Doenças da Vulva/cirurgia , Feminino , Humanos
14.
Front Microbiol ; 7: 1078, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27468278

RESUMO

Sigma factors are RNA polymerase subunits engaged in promoter recognition and DNA strand separation during transcription initiation in bacteria. Primary sigma factors are responsible for the expression of housekeeping genes and are essential for survival. RpoD, the primary sigma factor of Escherichia coli, a γ-proteobacteria, recognizes consensus promoter sequences highly similar to those of some α-proteobacteria species. Despite this resemblance, RpoD is unable to sustain transcription from most of the α-proteobacterial promoters tested so far. In contrast, we have found that SigA, the primary sigma factor of Rhizobium etli, an α-proteobacteria, is able to transcribe E. coli promoters, although it exhibits only 48% identity (98% coverage) to RpoD. We have called this the transcriptional laxity phenomenon. Here, we show that SigA partially complements the thermo-sensitive deficiency of RpoD285 from E. coli strain UQ285 and that the SigA region σ4 is responsible for this phenotype. Sixteen out of 74 residues (21.6%) within region σ4 are variable between RpoD and SigA. Mutating these residues significantly improves SigA ability to complement E. coli UQ285. Only six of these residues fall into positions already known to interact with promoter DNA and to comprise a helix-turn-helix motif. The remaining variable positions are located on previously unexplored sites inside region σ4, specifically into the first two α-helices of the region. Neither of the variable positions confined to these helices seem to interact directly with promoter sequence; instead, we adduce that these residues participate allosterically by contributing to correct region folding and/or positioning of the HTH motif. We propose that transcriptional laxity is a mechanism for ensuring transcription in spite of naturally occurring mutations from endogenous promoters and/or horizontally transferred DNA sequences, allowing survival and fast environmental adaptation of α-proteobacteria.

16.
Sci Rep ; 6: 23092, 2016 03 15.
Artigo em Inglês | MEDLINE | ID: mdl-26976656

RESUMO

Climate change and slow yield gains pose a major threat to global wheat production. Underutilized genetic resources including landraces and wild relatives are key elements for developing high-yielding and climate-resilient wheat varieties. Landraces introduced into Mexico from Europe, also known as Creole wheats, are adapted to a wide range of climatic regimes and represent a unique genetic resource. Eight thousand four hundred and sixteen wheat landraces representing all dimensions of Mexico were characterized through genotyping-by-sequencing technology. Results revealed sub-groups adapted to specific environments of Mexico. Broadly, accessions from north and south of Mexico showed considerable genetic differentiation. However, a large percentage of landrace accessions were genetically very close, although belonged to different regions most likely due to the recent (nearly five centuries before) introduction of wheat in Mexico. Some of the groups adapted to extreme environments and accumulated high number of rare alleles. Core reference sets were assembled simultaneously using multiple variables, capturing 89% of the rare alleles present in the complete set. Genetic information about Mexican wheat landraces and core reference set can be effectively utilized in next generation wheat varietal improvement.


Assuntos
Cromossomos de Plantas/genética , Variação Genética , Genoma de Planta/genética , Triticum/genética , Algoritmos , Alelos , Fluxo Gênico , Frequência do Gene , Genótipo , Geografia , México , Modelos Genéticos , Fenótipo , Filogenia , Melhoramento Vegetal , Polimorfismo de Nucleotídeo Único , Poliploidia , Análise de Componente Principal , Especificidade da Espécie , Triticum/classificação
17.
BMC Genomics ; 15: 770, 2014 Sep 08.
Artigo em Inglês | MEDLINE | ID: mdl-25201548

RESUMO

BACKGROUND: Regulation of transcription is essential for any organism and Rhizobium etli (a multi-replicon, nitrogen-fixing symbiotic bacterium) is no exception. This bacterium is commonly found in the rhizosphere (free-living) or inside of root-nodules of the common bean (Phaseolus vulgaris) in a symbiotic relationship. Abiotic stresses, such as high soil temperatures and salinity, compromise the genetic stability of R. etli and therefore its symbiotic interaction with P. vulgaris. However, it is still unclear which genes are up- or down-regulated to cope with these stress conditions. The aim of this study was to identify the genes and non-coding RNAs (ncRNAs) that are differentially expressed under heat and saline shock, as well as the promoter regions of the up-regulated loci. RESULTS: Analysing the heat and saline shock responses of R. etli CE3 through RNA-Seq, we identified 756 and 392 differentially expressed genes, respectively, and 106 were up-regulated under both conditions. Notably, the set of genes over-expressed under either condition was preferentially encoded on plasmids, although this observation was more significant for the heat shock response. In contrast, during either saline shock or heat shock, the down-regulated genes were principally chromosomally encoded. Our functional analysis shows that genes encoding chaperone proteins were up-regulated during the heat shock response, whereas genes involved in the metabolism of compatible solutes were up-regulated following saline shock. Furthermore, we identified thirteen and nine ncRNAs that were differentially expressed under heat and saline shock, respectively, as well as eleven ncRNAs that had not been previously identified. Finally, using an in silico analysis, we studied the promoter motifs in all of the non-coding regions associated with the genes and ncRNAs up-regulated under both conditions. CONCLUSIONS: Our data suggest that the replicon contribution is different for different stress responses and that the heat shock response is more complex than the saline shock response. In general, this work exemplifies how strategies that not only consider differentially regulated genes but also regulatory elements of the stress response provide a more comprehensive view of bacterial gene regulation.


Assuntos
Genoma Bacteriano , Temperatura Alta , Replicon , Rhizobium etli/genética , Salinidade , Estresse Fisiológico/genética , Sítios de Ligação , Expressão Gênica , Perfilação da Expressão Gênica , Regulação Bacteriana da Expressão Gênica , Resposta ao Choque Térmico/genética , Proteínas de Membrana Transportadoras/genética , Proteínas de Membrana Transportadoras/metabolismo , Motivos de Nucleotídeos , Plasmídeos/genética , Matrizes de Pontuação de Posição Específica , Regiões Promotoras Genéticas , Ligação Proteica , RNA não Traduzido/genética , Rhizobium etli/metabolismo , Metabolismo Secundário/genética , Análise de Sequência de RNA , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo
18.
Appl Environ Microbiol ; 80(2): 446-54, 2014 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-24185856

RESUMO

In this work, we isolated and characterized 14 bacteriophages that infect Rhizobium etli. They were obtained from rhizosphere soil of bean plants from agricultural lands in Mexico using an enrichment method. The host range of these phages was narrow but variable within a collection of 48 R. etli strains. We obtained the complete genome sequence of nine phages. Four phages were resistant to several restriction enzymes and in vivo cloning, probably due to nucleotide modifications. The genome size of the sequenced phages varied from 43 kb to 115 kb, with a median size of ≈ 45 to 50 kb. A large proportion of open reading frames of these phage genomes (65 to 70%) consisted of hypothetical and orphan genes. The remainder encoded proteins needed for phage morphogenesis and DNA synthesis and processing, among other functions, and a minor percentage represented genes of bacterial origin. We classified these phages into four genomic types on the basis of their genomic similarity, gene content, and host range. Since there are no reports of similar sequences, we propose that these bacteriophages correspond to novel species.


Assuntos
Bacteriófagos/genética , Bacteriófagos/isolamento & purificação , Genoma Viral , Especificidade de Hospedeiro , Rhizobium etli/virologia , México , Dados de Sequência Molecular , Fases de Leitura Aberta , Filogenia , Rizosfera , Microbiologia do Solo
19.
Rev Bras Ginecol Obstet ; 35(7): 290-4, 2013 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-24080840

RESUMO

PURPOSE: To identify the predictive factors for voiding dysfunction after transobturator slings. METHODS: We retrospectively reviewed the records of all patients who underwent a transobturator sling between March 2003 and December 2008. A total of 514 women had available data with at least a six-week follow-up. Patients' demographics, preoperative symptoms, urodynamic testing including multichannel voiding studies and surgical variables were tabulated. Voiding dysfunction was defined by a catheterized or ultrasonographic postvoid residual greater than 100 cc (≥six weeks after the procedure) associated with any complaints of abnormal voiding. Univariate logistic regression analysis was performed with respect to postoperative voiding dysfunction. RESULTS: The patient population had a mean age of 58.5 ± 12.9 years. Thirty-three out of 514 patients (6.4%) had postoperative voiding dysfunction according to our definition, and 4 (0.78%) required sling transection. No differences were observed between normal and dysfunctional voiders in age, associated prolapse surgery, preoperative postvoid residual, preoperative urinary flow rate, prior pelvic surgery, and menopausal status. Valsalva efforts during the preoperative pressure flow study was the only predictive factor for postoperative voiding dysfunction, 72.4% dysfunctional versus 27.6% normal (p<0.001). CONCLUSION: Preoperative Valsalva maneuver during the micturition could identify those at risk for voiding dysfunction after transobturator sling, and it should be noted during preoperative counseling.


Assuntos
Slings Suburetrais/efeitos adversos , Transtornos Urinários/etiologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Pessoa de Meia-Idade , Prognóstico , Estudos Retrospectivos , Transtornos Urinários/epidemiologia , Adulto Jovem
20.
Rev. bras. ginecol. obstet ; 35(7): 290-294, July 2013. tab
Artigo em Inglês | LILACS | ID: lil-687370

RESUMO

PURPOSE: To identify the predictive factors for voiding dysfunction after transobturator slings. METHODS: We retrospectively reviewed the records of all patients who underwent a transobturator sling between March 2003 and December 2008. A total of 514 women had available data with at least a six-week follow-up. Patients' demographics, preoperative symptoms, urodynamic testing including multichannel voiding studies and surgical variables were tabulated. Voiding dysfunction was defined by a catheterized or ultrasonographic postvoid residual greater than 100 cc (≥six weeks after the procedure) associated with any complaints of abnormal voiding. Univariate logistic regression analysis was performed with respect to postoperative voiding dysfunction. RESULTS: The patient population had a mean age of 58.5±12.9 years. Thirty-three out of 514 patients (6.4%) had postoperative voiding dysfunction according to our definition, and 4 (0.78%) required sling transection. No differences were observed between normal and dysfunctional voiders in age, associated prolapse surgery, preoperative postvoid residual, preoperative urinary flow rate, prior pelvic surgery, and menopausal status. Valsalva efforts during the preoperative pressure flow study was the only predictive factor for postoperative voiding dysfunction, 72.4% dysfunctional versus 27.6% normal (p<0.001). CONCLUSION: Preoperative Valsalva maneuver during the micturition could identify those at risk for voiding dysfunction after transobturator sling, and it should be noted during preoperative counseling.


OBJETIVO: Identificar fatores preditivos para disfunção miccional após a cirurgia de sling transobturador. MÉTODOS: Foram revisados, retrospectivamente, os protocolos de todas as pacientes que foram submetidas à cirurgia de sling transobturador. Entre março de 2003 e dezembro de 2008, 514 mulheres apresentavam dados disponíveis com ao menos seis semanas de seguimento. Foram avaliados os dados demográficos, os sintomas pré-operatórios, o estudo urodinâmico e as variáveis cirúrgicas. A disfunção miccional foi definida como o resíduo pós-miccional (verificado por sondagem vesical ou ecografia) superior a 100 mL (≥seis semanas após procedimento cirúrgico), que foi associado à queixa de micção anormal. Realizou-se análise por regressão logística univariada com relação à disfunção miccional pós-operatória. RESULTADOS: A população de pacientes tinha uma média de idade de 58,5±12,9 anos. Trinta e três das 514 (6,4%) participantes apresentavam disfunção miccional pós-operatória de acordo com a nossa definição e 4 (0,78%) necessitaram secção do sling. Não houve diferenças com relação à cirurgia para prolapso associada, ao resíduo pós-miccional pré-operatório, à urofluxometria pré-operatória, à cirurgia pélvica prévia e ao estado menopausal entre aquelas que apresentaram disfunção miccional quando comparadas às outras. A identificação da manobra de Valsalva durante o estudo miccional pré-operatório foi o único fator preditivo para disfunção miccional pós-operatória, 72,4% no grupo com disfunção versus 27,6% nas normais (p<0,001). CONCLUSÃO: A manobra de Valsalva pré-operatória durante a micção pôde identificar as mulheres que apresentavam maior risco para disfunção miccional após cirurgia de sling transobturador, e deve ser levada em consideração no aconselhamento pré-operatório das mesmas.


Assuntos
Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Pessoa de Meia-Idade , Adulto Jovem , Slings Suburetrais/efeitos adversos , Transtornos Urinários/etiologia , Prognóstico , Estudos Retrospectivos , Transtornos Urinários/epidemiologia
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