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1.
Fungal Genet Biol ; 149: 103527, 2021 04.
Artigo em Inglês | MEDLINE | ID: mdl-33524555

RESUMO

Cercospora zeina is a causal pathogen of gray leaf spot (GLS) disease of maize in Africa. This fungal pathogen exhibits a high genetic diversity in South Africa. However, little is known about the pathogen's population structure in the rest of Africa. In this study, we aimed to assess the diversity and gene flow of the pathogen between major maize producing countries in East and Southern Africa (Kenya, Uganda, Zambia, Zimbabwe, and South Africa). A total of 964 single-spore isolates were made from GLS lesions and confirmed as C.zeina using PCR diagnostics. The other causal agent of GLS, Cercospora zeae-maydis, was absent. Genotyping all the C.zeina isolates with 11 microsatellite markers and a mating-type gene diagnostic revealed (i) high genetic diversity with some population structure between the five African countries, (ii) cryptic sexual recombination, (iii) that South Africa and Kenya were the greatest donors of migrants, and (iv) that Zambia had a distinct population. We noted evidence of human-mediated long-distance dispersal, since four haplotypes from one South African site were also present at five sites in Kenya and Uganda. There was no evidence for a single-entry point of the pathogen into Africa. South Africa was the most probable origin of the populations in Kenya, Uganda, and Zimbabwe. Continuous annual maize production in the tropics (Kenya and Uganda) did not result in greater genetic diversity than a single maize season (Southern Africa). Our results will underpin future management of GLS in Africa through effective monitoring of virulent C.zeina strains.


Assuntos
Cercospora/genética , Cercospora/patogenicidade , Zea mays/microbiologia , África Oriental , Ascomicetos/genética , Resistência à Doença/genética , Fluxo Gênico/genética , Variação Genética/genética , Genética Populacional/métodos , Haplótipos/genética , Repetições de Microssatélites/genética , Doenças das Plantas/microbiologia , Locos de Características Quantitativas/genética , África do Sul
2.
Fungal Genet Biol ; 125: 36-44, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-30659907

RESUMO

Gray leaf spot (GLS) is an important foliar disease of maize. This disease, caused by Cercospora zeina, is prevalent in both smallholder and commercial maize farms in South Africa. Notably, smallholder practices are geared towards conservation agriculture, planting diverse maize genotypes within a field and avoiding chemical control. This study examined the population genetic structure of 129 C. zeina isolates from three smallholder farm sites in KwaZulu-Natal in South Africa using 13 microsatellite markers. These were analysed, together with 239 isolates previously analysed from four commercial farms in the same province, to determine whether farming systems influence the genetic diversity of C.zeina. In addition, we wanted to determine whether the smallholder farming system harboured a greater diversity of C.zeina haplotypes due to lack of chemical spraying of these crops. Overall, farming systems exhibited partial, but significant, population differentiation, contributing 10% of the genetic variation observed. A 16% genetic variation conferred between KwaNxamalala (smallholder) and Cedara (commercial) areas that are in close proximity, confirmed this. Private alleles accounted for 29% of the 52 alleles observed in smallholder farms. Smallholder farms harboured a higher gene and genotypic diversity, with a clonal fraction of only 13% compared to 32% in commercial farms. Mating type ratios indicative of sexual recombination and lower linkage disequilibrium in most smallholder populations were consistent with higher levels of diversity. This study suggests that commercial farming practices, such as fungicides and monoculture crop planting, may result in a narrower genetic diversity of the pathogen that is then propagated by asexual reproduction. In contrast, management of GLS disease in smallholder farms should consider the greater diversity of pathogen genotypes, especially if future research shows that this equates to a greater diversity of pathogenicity alleles.


Assuntos
Ascomicetos/genética , Genética Populacional , Doenças das Plantas/genética , Zea mays/microbiologia , Agricultura , Alelos , Ascomicetos/patogenicidade , Produtos Agrícolas , Haplótipos , Humanos , Repetições de Microssatélites/genética , Doenças das Plantas/microbiologia , Locos de Características Quantitativas/genética , Zea mays/genética
3.
Bioorg Med Chem ; 27(7): 1430-1436, 2019 04 01.
Artigo em Inglês | MEDLINE | ID: mdl-30792103

RESUMO

Class C ß-lactamases have previously been shown to be efficiently inactivated by O-aryloxycarbonyl hydroxamates. O-Phenoxycarbonyl-N-benzyloxycarbonylhydroxylamine (1) and O-phenoxycarbonyl-N-(R)-[(4-amino-4-carboxy-1-butyl)oxycarbonyl]hydroxylamine (2), for example, were found to be effective inactivators. The present paper describes a structure-activity study of these molecules to better define the important structural elements for high inhibitory activity. The results show that a well-positioned hydrophobic element (which may interact with the Tyr221 residue of the enzyme) and a negatively charged element, e.g. a carboxylate group (which may interact with Arg204), are required for high reactivity with the enzyme. The new compounds were found to inactivate by forming a carbonyl cross-linked enzyme (probably Ser64OCONHLys 315) as for 1 rather than the inert hydroxamoyl derivative observed with 2.


Assuntos
Ácidos Hidroxâmicos/farmacologia , Inibidores de beta-Lactamases/farmacologia , beta-Lactamases/metabolismo , Relação Dose-Resposta a Droga , Enterobacter cloacae/enzimologia , Ácidos Hidroxâmicos/síntese química , Ácidos Hidroxâmicos/química , Hidroxilaminas/síntese química , Hidroxilaminas/química , Hidroxilaminas/farmacologia , Cinética , Estrutura Molecular , Relação Estrutura-Atividade , Inibidores de beta-Lactamases/síntese química , Inibidores de beta-Lactamases/química
4.
Plant J ; 89(4): 746-763, 2017 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-27862526

RESUMO

We used a systems genetics approach to elucidate the molecular mechanisms of the responses of maize to grey leaf spot (GLS) disease caused by Cercospora zeina, a threat to maize production globally. Expression analysis of earleaf samples in a subtropical maize recombinant inbred line population (CML444 × SC Malawi) subjected in the field to C. zeina infection allowed detection of 20 206 expression quantitative trait loci (eQTLs). Four trans-eQTL hotspots coincided with GLS disease QTLs mapped in the same field experiment. Co-expression network analysis identified three expression modules correlated with GLS disease scores. The module (GY-s) most highly correlated with susceptibility (r = 0.71; 179 genes) was enriched for the glyoxylate pathway, lipid metabolism, diterpenoid biosynthesis and responses to pathogen molecules such as chitin. The GY-s module was enriched for genes with trans-eQTLs in hotspots on chromosomes 9 and 10, which also coincided with phenotypic QTLs for susceptibility to GLS. This transcriptional network has significant overlap with the GLS susceptibility response of maize line B73, and may reflect pathogen manipulation for nutrient acquisition and/or unsuccessful defence responses, such as kauralexin production by the diterpenoid biosynthesis pathway. The co-expression module that correlated best with resistance (TQ-r; 1498 genes) was enriched for genes with trans-eQTLs in hotspots coinciding with GLS resistance QTLs on chromosome 9. Jasmonate responses were implicated in resistance to GLS through co-expression of COI1 and enrichment of genes with the Gene Ontology term 'cullin-RING ubiquitin ligase complex' in the TQ-r module. Consistent with this, JAZ repressor expression was highly correlated with the severity of GLS disease in the GY-s susceptibility network.


Assuntos
Folhas de Planta/genética , Folhas de Planta/microbiologia , Zea mays/genética , Zea mays/microbiologia , Ascomicetos/patogenicidade , Cromossomos de Plantas/genética , Redes Reguladoras de Genes/genética , Redes Reguladoras de Genes/fisiologia , Doenças das Plantas/genética , Doenças das Plantas/microbiologia , Locos de Características Quantitativas/genética
5.
J Mol Evol ; 86(3-4): 216-239, 2018 04.
Artigo em Inglês | MEDLINE | ID: mdl-29556741

RESUMO

Amaranthus species are an emerging and promising nutritious traditional vegetable food source. Morphological plasticity and poorly resolved dendrograms have led to the need for well resolved species phylogenies. We hypothesized that whole chloroplast phylogenomics would result in more reliable differentiation between closely related amaranth species. The aims of the study were therefore: to construct a fully assembled, annotated chloroplast genome sequence of Amaranthus tricolor; to characterize Amaranthus accessions phylogenetically by comparing barcoding genes (matK, rbcL, ITS) with whole chloroplast sequencing; and to use whole chloroplast phylogenomics to resolve deeper phylogenetic relationships. We generated a complete A. tricolor chloroplast sequence of 150,027 bp. The three barcoding genes revealed poor inter- and intra-species resolution with low bootstrap support. Whole chloroplast phylogenomics of 59 Amaranthus accessions increased the number of parsimoniously informative sites from 92 to 481 compared to the barcoding genes, allowing improved separation of amaranth species. Our results support previous findings that two geographically independent domestication events of Amaranthus hybridus likely gave rise to several species within the Hybridus complex, namely Amaranthus dubius, Amaranthus quitensis, Amaranthus caudatus, Amaranthus cruentus and Amaranthus hypochondriacus. Poor resolution of species within the Hybridus complex supports the recent and ongoing domestication within the complex, and highlights the limitation of chloroplast data for resolving recent evolution. The weedy Amaranthus retroflexus and Amaranthus powellii was found to share a common ancestor with the Hybridus complex. Leafy amaranth, Amaranthus tricolor, Amaranthus blitum, Amaranthus viridis and Amaranthus graecizans formed a stable sister lineage to the aforementioned species across the phylogenetic trees. This study demonstrates the power of next-generation sequencing data and reference-based assemblies to resolve phylogenies, and also facilitated the identification of unknown Amaranthus accessions from a local genebank. The informative phylogeny of the Amaranthus genus will aid in selecting accessions for breeding advanced genotypes to satisfy global food demand.


Assuntos
Amaranthus/classificação , Genoma de Cloroplastos , Genoma de Planta , Filogenia , Código de Barras de DNA Taxonômico , Genômica
6.
J Neurosci ; 36(38): 9896-907, 2016 09 21.
Artigo em Inglês | MEDLINE | ID: mdl-27656027

RESUMO

UNLABELLED: Vascular cognitive impairment and dementia (VCID) is the second most common form of dementia behind Alzheimer's disease (AD). It is estimated that 40% of AD patients also have some form of VCID. One promising therapeutic for AD is anti-Aß immunotherapy, which uses antibodies against Aß to clear it from the brain. While successful in clearing Aß and improving cognition in mice, anti-Aß immunotherapy failed to reach primary cognitive outcomes in several different clinical trials. We hypothesized that one potential reason the anti-Aß immunotherapy clinical trials were unsuccessful was due to this high percentage of VCID comorbidity in the AD population. We used our unique model of VCID-amyloid comorbidity to test this hypothesis. We placed 9-month-old wild-type and APP/PS1 mice on either a control diet or a diet that induces hyperhomocysteinemia (HHcy). After being placed on the diet for 3 months, the mice then received intraperotineal injections of either IgG2a control or 3D6 for another 3 months. While we found that treatment of our comorbidity model with 3D6 resulted in decreased total Aß levels, there was no cognitive benefit of the anti-Aß immunotherapy in our AD/VCID mice. Further, microhemorrhages were increased by 3D6 in the APP/PS1/control but further increased in an additive fashion when 3D6 was administered to the APP/PS1/HHcy mice. This suggests that the use of anti-Aß immunotherapy in patients with both AD and VCID would be ineffective on cognitive outcomes. SIGNIFICANCE STATEMENT: Despite significant mouse model data demonstrating both pathological and cognitive efficacy of anti-Aß immunotherapy for the treatment of Alzheimer's disease, clinical trial outcomes have been underwhelming, failing to meet any primary endpoints. We show here that vascular cognitive impairment and dementia (VCID) comorbidity eliminates cognitive efficacy of anti-Aß immunotherapy, despite amyloid clearance. Further, cerebrovascular adverse events of the anti-Aß immunotherapy are significantly exacerbated by the VCID comorbidity. These data suggest that VCID comorbidity with Alzheimer's disease may mute the response to anti-Aß immunotherapy.


Assuntos
Peptídeos beta-Amiloides/imunologia , Demência Vascular , Imunoterapia/métodos , Precursor de Proteína beta-Amiloide/genética , Precursor de Proteína beta-Amiloide/metabolismo , Animais , Antígeno CD11b/metabolismo , Demência Vascular/complicações , Demência Vascular/diagnóstico por imagem , Demência Vascular/genética , Demência Vascular/terapia , Modelos Animais de Doenças , Feminino , Humanos , Hiper-Homocisteinemia/complicações , Hiper-Homocisteinemia/etiologia , Hiper-Homocisteinemia/genética , Hiper-Homocisteinemia/terapia , Imageamento por Ressonância Magnética , Masculino , Aprendizagem em Labirinto , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Presenilina-1/genética , Presenilina-1/metabolismo , RNA Mensageiro/metabolismo
7.
Mol Plant Microbe Interact ; 30(9): 710-724, 2017 09.
Artigo em Inglês | MEDLINE | ID: mdl-28535078

RESUMO

Gray leaf spot (GLS), caused by the sibling species Cercospora zeina or Cercospora zeae-maydis, is cited as one of the most important diseases threatening global maize production. C. zeina fails to produce cercosporin in vitro and, in most cases, causes large coalescing lesions during maize infection, a symptom generally absent from cercosporin-deficient mutants in other Cercospora spp. Here, we describe the C. zeina cercosporin toxin biosynthetic (CTB) gene cluster. The oxidoreductase gene CTB7 contained several insertions and deletions as compared with the C. zeae-maydis ortholog. We set out to determine whether complementing the defective CTB7 gene with the full-length gene from C. zeae-maydis could confer in vitro cercosporin production. C. zeina transformants containing C. zeae-maydis CTB7 were generated by Agrobacterium tumefaciens-mediated transformation and were evaluated for in vitro cercosporin production. When grown on nitrogen-limited medium in the light-conditions conducive to cercosporin production in other Cercospora spp.-one transformant accumulated a red pigment that was confirmed to be cercosporin by the KOH assay, thin-layer chromatography, and ultra performance liquid chromatography-quadrupole-time-of-flight mass spectrometry. Our results indicated that C. zeina has a defective CTB7, but all other necessary machinery required for synthesizing cercosporin-like molecules and, thus, C. zeina may produce a structural variant of cercosporin during maize infection.


Assuntos
Ascomicetos/genética , Proteínas Fúngicas/genética , Teste de Complementação Genética , Perileno/análogos & derivados , Zea mays/microbiologia , Processamento Alternativo/genética , Sequência de Aminoácidos , Ascomicetos/isolamento & purificação , Sequência de Bases , Vias Biossintéticas/genética , Simulação por Computador , Sequência Conservada/genética , DNA Fúngico/genética , Proteínas Fúngicas/química , Proteínas Fúngicas/metabolismo , Deleção de Genes , Regulação Fúngica da Expressão Gênica , Genes Fúngicos , Íntrons/genética , Espectrometria de Massas , Família Multigênica , Oxirredutases/metabolismo , Perileno/metabolismo , Transcrição Gênica , Transformação Genética
8.
Neurobiol Dis ; 103: 174-183, 2017 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-28433741

RESUMO

The zinc-finger SWIM domain-containing protein 6 (ZSWIM6) is a protein of unknown function that has been associated with schizophrenia and limited educational attainment by three independent genome-wide association studies. Additionally, a putatively causal point mutation in ZSWIM6 has been identified in several cases of acromelic frontonasal dysostosis with severe intellectual disability. Despite the growing number of studies implicating ZSWIM6 as an important regulator of brain development, its role in this process has never been examined. Here, we report the generation of Zswim6 knockout mice and provide a detailed anatomical and behavioral characterization of the resulting phenotype. We show that Zswim6 is initially expressed widely during embryonic brain development but becomes restricted to the striatum postnatally. Loss of Zswim6 causes a reduction in striatal volume and changes in medium spiny neuron morphology. These changes are associated with alterations in motor control, including hyperactivity, impaired rotarod performance, repetitive movements, and behavioral hyperresponsiveness to amphetamine. Together, our results show that Zswim6 is indispensable to normal brain function and support the notion that Zswim6 might serve as an important contributor to the pathogenesis of schizophrenia and other neurodevelopmental disorders.


Assuntos
Corpo Estriado/metabolismo , Corpo Estriado/patologia , Proteínas de Ligação a DNA/deficiência , Hipercinese/metabolismo , Hipercinese/patologia , Animais , Corpo Estriado/crescimento & desenvolvimento , Proteínas de Ligação a DNA/genética , Hipercinese/genética , Locomoção/fisiologia , Camundongos , Camundongos da Linhagem 129 , Camundongos Endogâmicos C57BL , Camundongos Knockout , Transtornos do Neurodesenvolvimento/genética , Transtornos do Neurodesenvolvimento/metabolismo , Transtornos do Neurodesenvolvimento/patologia
9.
BMC Plant Biol ; 17(1): 197, 2017 Nov 13.
Artigo em Inglês | MEDLINE | ID: mdl-29132306

RESUMO

BACKGROUND: Cercospora zeina is a foliar pathogen responsible for maize grey leaf spot in southern Africa that negatively impacts maize production. Plants use a variety of chemical and structural mechanisms to defend themselves against invading pathogens such as C. zeina, including the production of secondary metabolites with antimicrobial properties. In maize, a variety of biotic and abiotic stressors induce the accumulation of the terpenoid phytoalexins, zealexins and kauralexins. RESULTS: C. zeina-susceptible line displayed pervasive rectangular grey leaf spot lesions, running parallel with the leaf veins in contrast to C. zeina-resistant line that had restricted disease symptoms. Analysis of the transcriptome of both lines indicated that genes involved in primary and secondary metabolism were up-regualted, and although different pathways were prioritized in each line, production of terpenoid compounds were common to both. Targeted phytoalexin analysis revealed that C. zeina-inoculated leaves accumulated zealexins and kauralexins. The resistant line shows a propensity toward accumulation of the kauralexin B series metabolites in response to infection, which contrasts with the susceptible line that preferentially accumulates the kauralexin A series. Kauralexin accumulation was correlated to expression of the kauralexin biosynthetic gene, ZmAn2 and a candidate biosynthetic gene, ZmKSL2. We report the expression of a putative copalyl diphosphate synthase gene that is induced by C. zeina in the resistant line exclusively. DISCUSSION: This study shows that zealexins and kauralexins, and expression of their biosynthetic genes, are induced by C. zeina in both resistant and susceptible germplasm adapted to the southern African climate. The data presented here indicates that different forms of kauralexins accumulate in the resistant and susceptible maize lines in response to C. zeina, with the accumulation of kauralexin B compounds in a resistant maize line and kauralexin A compounds accumulating in the susceptible line.


Assuntos
Ascomicetos/patogenicidade , Resistência à Doença/genética , Doenças das Plantas/genética , Terpenos/metabolismo , Zea mays/genética , Ontologia Genética , Doenças das Plantas/microbiologia , Folhas de Planta/microbiologia , Análise de Sequência de RNA , Zea mays/metabolismo , Zea mays/microbiologia
10.
Environ Sci Technol ; 50(3): 1536-43, 2016 Feb 02.
Artigo em Inglês | MEDLINE | ID: mdl-26726952

RESUMO

Smoke from wood-fueled fires is one of the most common hazards encountered by firefighters worldwide. Wood smoke is complex in nature and contains numerous compounds, including methoxyphenols (MPs) and polycyclic aromatic hydrocarbons (PAHs), some of which are carcinogenic. Chronic exposure to wood smoke can lead to adverse health outcomes, including respiratory infections, impaired lung function, cardiac infarctions, and cancers. At training exercises held in burn houses at four fire departments across Ontario, air samples, skin wipes, and urine specimens from a cohort of firefighters (n = 28) were collected prior to and after exposure. Wood was the primary fuel used in these training exercises. Air samples showed that MP concentrations were on average 5-fold greater than those of PAHs. Skin wipe samples acquired from multiple body sites of firefighters indicated whole-body smoke exposure. A suite of MPs (methyl-, ethyl-, and propylsyringol) and deconjugated PAH metabolites (hydroxynaphthalene, hydroxyfluorene, hydroxyphenanthrene, and their isomers) were found to be sensitive markers of smoke exposure in urine. Creatinine-normalized levels of these markers were significantly elevated (p < 0.05) in 24 h postexposure urine despite large between-subject variations that were dependent on the specific operational roles of firefighters while using personal protective equipment. This work offers deeper insight into potential health risk from smoke exposure that is needed for translation of better mitigation policies, including improved equipment to reduce direct skin absorption and standardized hygiene practices implemented at different regional fire services.


Assuntos
Bombeiros/estatística & dados numéricos , Exposição Ocupacional/análise , Fenóis/urina , Hidrocarbonetos Policíclicos Aromáticos/urina , Fumaça/análise , Poluentes Atmosféricos/urina , Carcinógenos/análise , Feminino , Incêndios , Humanos , Masculino , Ontário , Medição de Risco , Absorção Cutânea , Madeira
11.
Phytopathology ; 106(10): 1194-1205, 2016 10.
Artigo em Inglês | MEDLINE | ID: mdl-27392176

RESUMO

South Africa is one of the leading maize-producing countries in sub-Saharan Africa. Since the 1980s, Cercospora zeina, a causal agent of gray leaf spot of maize, has become endemic in South Africa, and is responsible for substantial yield reductions. To assess genetic diversity and population structure of C. zeina in South Africa, 369 isolates were collected from commercial maize farms in three provinces (KwaZulu-Natal, Mpumalanga, and North West). These isolates were evaluated with 14 microsatellite markers and species-specific mating type markers that were designed from draft genome sequences of C. zeina isolates from Africa (CMW 25467) and the United States (USPA-4). Sixty alleles were identified across 14 loci, and gene diversity values within each province ranged from 0.18 to 0.35. High levels of gene flow were observed (Nm = 5.51), and in a few cases, identical multilocus haplotypes were found in different provinces. Overall, 242 unique multilocus haplotypes were identified with a low clonal fraction of 34%. No distinct population clusters were identified using STRUCTURE, principal coordinate analysis, or Weir's theta θ statistic. The lack of population differentiation was supported by analysis of molecular variance tests, which indicated that only 2% of the variation was attributed to variability between populations from each province. Mating type ratios of MAT1-1 and MAT1-2 idiomorphs from 335 isolates were not significantly different from a 1:1 ratio in all provinces, which provided evidence for sexual reproduction. The draft genome of C. zeina CMW 25467 exhibited a complete genomic copy of the MAT1-1 idiomorph as well as exonic fragments of MAT genes from both idiomorphs. The high level of gene diversity, shared haplotypes at different geographical locations within South Africa, and presence of both MAT idiomorphs at all sites indicates widespread dispersal of C. zeina between maize fields in the country as well as evidence for sexual recombination. The outcomes of this genome-enabled study are important for disease management since the high diversity has implications for dispersal of fungicide resistance should it emerge and the need for diversified resistance breeding.


Assuntos
Ascomicetos/genética , Variação Genética , Genética Populacional , Genoma Fúngico/genética , Doenças das Plantas/microbiologia , Zea mays/microbiologia , Ascomicetos/isolamento & purificação , Fluxo Gênico , Geografia , Repetições de Microssatélites/genética , Análise de Sequência de DNA , África do Sul
12.
BMC Plant Biol ; 15: 117, 2015 May 10.
Artigo em Inglês | MEDLINE | ID: mdl-25957781

RESUMO

BACKGROUND: Histone modifications play an integral role in plant development, but have been poorly studied in woody plants. Investigating chromatin organization in wood-forming tissue and its role in regulating gene expression allows us to understand the mechanisms underlying cellular differentiation during xylogenesis (wood formation) and identify novel functional regions in plant genomes. However, woody tissue poses unique challenges for using high-throughput chromatin immunoprecipitation (ChIP) techniques for studying genome-wide histone modifications in vivo. We investigated the role of the modified histone H3K4me3 (trimethylated lysine 4 of histone H3) in gene expression during the early stages of wood formation using ChIP-seq in Eucalyptus grandis, a woody biomass model. RESULTS: Plant chromatin fixation and isolation protocols were optimized for developing xylem tissue collected from field-grown E. grandis trees. A "nano-ChIP-seq" procedure was employed for ChIP DNA amplification. Over 9 million H3K4me3 ChIP-seq and 18 million control paired-end reads were mapped to the E. grandis reference genome for peak-calling using Model-based Analysis of ChIP-Seq. The 12,177 significant H3K4me3 peaks identified covered ~1.5% of the genome and overlapped some 9,623 protein-coding genes and 38 noncoding RNAs. H3K4me3 library coverage, peaking ~600 - 700 bp downstream of the transcription start site, was highly correlated with gene expression levels measured with RNA-seq. Overall, H3K4me3-enriched genes tended to be less tissue-specific than unenriched genes and were overrepresented for general cellular metabolism and development gene ontology terms. Relative expression of H3K4me3-enriched genes in developing secondary xylem was higher than unenriched genes, however, and highly expressed secondary cell wall-related genes were enriched for H3K4me3 as validated using ChIP-qPCR. CONCLUSIONS: In this first genome-wide analysis of a modified histone in a woody tissue, we optimized a ChIP-seq procedure suitable for field-collected samples. In developing E. grandis xylem, H3K4me3 enrichment is an indicator of active transcription, consistent with its known role in sustaining pre-initiation complex formation in yeast. The H3K4me3 ChIP-seq data from this study paves the way to understanding the chromatin landscape and epigenomic architecture of xylogenesis in plants, and complements RNA-seq evidence of gene expression for the future improvement of the E. grandis genome annotation.


Assuntos
Mapeamento Cromossômico/métodos , Eucalyptus/genética , Genoma de Planta , Histonas/metabolismo , Lisina/metabolismo , Xilema/genética , Parede Celular/genética , Imunoprecipitação da Cromatina , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Estudos de Associação Genética , Metilação , Regiões Promotoras Genéticas/genética , Reação em Cadeia da Polimerase em Tempo Real , Reprodutibilidade dos Testes , Análise de Sequência de DNA , Madeira/genética
13.
New Phytol ; 206(4): 1351-63, 2015 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-25388807

RESUMO

Tension wood has distinct physical and chemical properties, including altered fibre properties, cell wall composition and ultrastructure. It serves as a good system for investigating the genetic regulation of secondary cell wall biosynthesis and wood formation. The reference genome sequence for Eucalyptus grandis allows investigation of the global transcriptional reprogramming that accompanies tension wood formation in this global wood fibre crop. We report the first comprehensive analysis of physicochemical wood property changes in tension wood of Eucalyptus measured in a hybrid (E. grandis × Eucalyptus urophylla) clone, as well as genome-wide gene expression changes in xylem tissues 3 wk post-induction using RNA sequencing. We found that Eucalyptus tension wood in field-grown trees is characterized by an increase in cellulose, a reduction in lignin, xylose and mannose, and a marked increase in galactose. Gene expression profiling in tension wood-forming tissue showed corresponding down-regulation of monolignol biosynthetic genes, and differential expression of several carbohydrate active enzymes. We conclude that alterations of cell wall traits induced by tension wood formation in Eucalyptus are a consequence of a combination of down-regulation of lignin biosynthesis and hemicellulose remodelling, rather than the often proposed up-regulation of the cellulose biosynthetic pathway.


Assuntos
Carbono/metabolismo , Eucalyptus/anatomia & histologia , Eucalyptus/crescimento & desenvolvimento , Madeira/anatomia & histologia , Madeira/crescimento & desenvolvimento , Vias Biossintéticas , Carboidratos/genética , Parede Celular/metabolismo , Celulose/metabolismo , Cruzamentos Genéticos , Regulação para Baixo , Eucalyptus/genética , Eucalyptus/metabolismo , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Lignina/metabolismo , Madeira/genética , Madeira/metabolismo , Xilema/metabolismo
14.
J Occup Environ Hyg ; 12(12): 855-65, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26252188

RESUMO

This article outlines a hierarchy of data required for retrospective exposure assessment for occupational disease of an individual worker. It then outlines in a step-wise manner how trend analysis using a relatively large exposure database can be used to estimate such exposure. The process of how a large database containing exposure measurements can be prepared for estimating historic occupational exposures of individual workers in relation to their illnesses is described. The asbestos subset from a large government collected air monitoring database called Medical Surveillance (MESU) was selected to illustrate the cleaning and analysis processes. After unidentifiable values were removed, the cleaned dataset was examined for possible sources of variability such as changes to sampling protocol. Limit of detection (LOD) values were substituted for all non-detectable values prior to the calculation of descriptive statistic using left censored analysis methods (i.e., maximum likelihood estimation (MLE), Kaplan Meier (KM), and simple substitution). The JoinPoint Regression Program was used to perform trend analysis and calculate an annual percentage change (APC) value for the available sampling period. An asbestos case study is presented to illustrate how the APC can then be combined with more recent job and/or process specific exposure data to estimate historic levels. The MESU asbestos dataset contained 1,610 samples from 1984-1995. An average of 17% of this data was left censored. The asbestos air sampling methods in Ontario changed around 1990. LOD values of 0.06 f/cc and 0.02 f/cc were substituted for LOD values pre- and post-1990, respectively. The annual mean fiber levels for the MLE method were an average of 44% lower than KM and substitution methods. The corresponding APC for MLE method was -6.5% and -7.7% for KM and simple substitution. The findings of this paper illustrate how the temporal trend of an exposure databases can be used to efficiently estimate historic contaminant levels in the presence of limited historical information.


Assuntos
Amianto/análise , Doenças Profissionais/epidemiologia , Exposição Ocupacional/história , Exposição Ocupacional/estatística & dados numéricos , Poluentes Ocupacionais do Ar/análise , Bases de Dados Factuais , História do Século XX , Humanos , Funções Verossimilhança , Limite de Detecção , Ontário , Crescimento Demográfico , Estudos Retrospectivos
15.
Anal Biochem ; 458: 66-8, 2014 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-24792153

RESUMO

Metal affinity chromatography using polyhistidine tags is a standard laboratory technique for the general purification of proteins from cellular systems, but there have been no attempts to explore whether the surface character of a protein may be engineered to similar affinity. We present the Arg160His mutation of Haemophilus influenzae carbonic anhydrase (HICA), which mimics the endogenous metal affinity of Escherichia coli carbonic anhydrase (ECCA). The purity and activity of the mutant are reported, and the purification is discussed. This is the first step toward developing a general method to engineer surface metal affinity for use in purification and metal labeling techniques.


Assuntos
Anidrases Carbônicas/isolamento & purificação , Cromatografia de Afinidade , Histidina/metabolismo , Substituição de Aminoácidos , Arginina/metabolismo , Anidrases Carbônicas/genética , Anidrases Carbônicas/metabolismo , Escherichia coli/enzimologia , Haemophilus influenzae/enzimologia , Cinética , Propriedades de Superfície
16.
BMC Genet ; 15: 60, 2014 May 22.
Artigo em Inglês | MEDLINE | ID: mdl-24885661

RESUMO

BACKGROUND: Gray leaf spot (GLS) is a globally important foliar disease of maize. Cercospora zeina, one of the two fungal species that cause the disease, is prevalent in southern Africa, China, Brazil and the eastern corn belt of the USA. Identification of QTL for GLS resistance in subtropical germplasm is important to support breeding programmes in developing countries where C. zeina limits production of this staple food crop. RESULTS: A maize RIL population (F7:S6) from a cross between CML444 and SC Malawi was field-tested under GLS disease pressure at five field sites over three seasons in KwaZulu-Natal, South Africa. Thirty QTL identified from eleven field trials (environments) were consolidated to seven QTL for GLS resistance based on their expression in at least two environments and location in the same core maize bins. Four GLS resistance alleles were derived from the more resistant parent CML444 (bin 1.10, 4.08, 9.04/9.05, 10.06/10.07), whereas the remainder were from SC Malawi (bin 6.06/6.07, 7.02/7.03, 9.06). QTLs in bin 4.08 and bin 6.06/6.07 were also detected as joint QTLs, each explained more than 11% of the phenotypic variation, and were identified in four and seven environments, respectively. Common markers were used to allocate GLS QTL from eleven previous studies to bins on the IBM2005 map, and GLS QTL "hotspots" were noted. Bin 4.08 and 7.02/7.03 GLS QTL from this study overlapped with hotspots, whereas the bin 6.06/6.07 and bin 9.06 QTLs appeared to be unique. QTL for flowering time (bin 1.07, 4.09) in this population did not correspond to QTL for GLS resistance. CONCLUSIONS: QTL mapping of a RIL population from the subtropical maize parents CML444 and SC Malawi identified seven QTL for resistance to gray leaf spot disease caused by C. zeina. These QTL together with QTL from eleven studies were allocated to bins on the IBM2005 map to provide a basis for comparison. Hotspots of GLS QTL were identified on chromosomes one, two, four, five and seven, with QTL in the current study overlapping with two of these. Two QTL from this study did not overlap with previously reported QTL.


Assuntos
Resistência à Doença/genética , Doenças das Plantas/genética , Locos de Características Quantitativas , Zea mays/genética , Alelos , Ascomicetos , Mapeamento Cromossômico , Fenótipo , África do Sul , Zea mays/microbiologia
17.
Am J Ind Med ; 57(12): 1319-30, 2014 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-25242446

RESUMO

BACKGROUND: The Strain Index (SI) has been developed to evaluate the risk for developing a distal upper extremity disorder. The objective of this study is to determine if the SI predicts incidence cases of work-related lateral, medial, or any epicondylities (LEPI, MEPI, and EPI). METHODS: Six hundred seven workers were followed for up to 3.5 years, 70 developed EPI on the dominant side (44 LEPI, 13 MEPI, and 13 both). Survival analyses were conducted adjusting for demographic, psychosocial, and work organizational factors, with the SI as time-dependent variable. RESULTS: High exposure (SI > 5), older age, and self-perceived poor general health were associated with incidence of LEPI and EPI, but not MEPI. There was a significant relationship between higher scores of SI and LEPI, hazard ratio (HR) 2.00 (95% CI 1.04-3.87) for SI 5.1-12, HR 2.12 (95% CI 1.11-4.05) for SI > 12. CONCLUSIONS: The SI can effectively identify jobs with increased risk of developing incidence of LEPI.


Assuntos
Doenças Profissionais/epidemiologia , Cotovelo de Tenista/epidemiologia , Humanos , Incidência , Satisfação no Emprego , Doenças Profissionais/diagnóstico , Estudos Prospectivos , Apoio Social , Estresse Mecânico , Cotovelo de Tenista/diagnóstico
18.
J Occup Environ Hyg ; 11(2): 111-6, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24369933

RESUMO

A comprehensive survey of respirable dust and respirable silica in Ontario gold mines was conducted by the Ontario Ministry of Labor during 1978-1979. The aim was to assess the feasibility of introducing gravimetric sampling to replace the assessment method which used konimeters, a device which gave results in terms of number of particles per cubic centimeter (ppcc) of air. The study involved both laboratory and field assessments. The field assessment involved measurement of airborne respirable dust and respirable silica at all eight operating gold mines of the time. This article describes the details of the field assessment. A total of 288 long-term (7-8 hr) personal respirable dust air samples were collected from seven occupational categories in eight gold mines. The respirable silica (α-quartz) was determined by x-ray diffraction method. The results show that during 1978-1979, the industry wide mean respirable dust was about 1 mg/m(3), and the mean respirable silica was 0.08 mg/m(3.)The mean% silica in respirable dust was 7.5%. The data set would be useful in future epidemiological and health studies, as well as in assessment of workers' compensation claims for occupational diseases such as silicosis, chronic obstructive pulmonary disease (COPD), and autoimmune diseases such as renal disease and rheumatoid arthritis.


Assuntos
Poeira/análise , Monitoramento Ambiental/história , Ouro , Exposição por Inalação/análise , Mineração/história , Exposição Ocupacional/história , Dióxido de Silício/análise , Monitoramento Ambiental/instrumentação , Monitoramento Ambiental/métodos , História do Século XX , Ontário , Estados Unidos , United States Occupational Safety and Health Administration
19.
Front Plant Sci ; 15: 1404483, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-39148617

RESUMO

Maize is the most widely cultivated and major security crop in sub-Saharan Africa. Three foliar diseases threaten maize production on the continent, namely northern leaf blight, gray leaf spot, and southern corn leaf blight. These are caused by the fungi Exserohilum turcicum, Cercospora zeina, and Bipolaris maydis, respectively. Yield losses of more than 10% can occur if these pathogens are diagnosed inaccurately or managed ineffectively. Here, we review recent advances in understanding the population biology and management of the three pathogens, which are present in Africa and thrive under similar environmental conditions during a single growing season. To effectively manage these pathogens, there is an increasing adoption of breeding for resistance at the small-scale level combined with cultural practices. Fungicide usage in African cropping systems is limited due to high costs and avoidance of chemical control. Currently, there is limited knowledge available on the population biology and genetics of these pathogens in Africa. The evolutionary potential of these pathogens to overcome host resistance has not been fully established. There is a need to conduct large-scale sampling of isolates to study their diversity and trace their migration patterns across the continent.

20.
Mol Plant Microbe Interact ; 26(4): 398-406, 2013 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-23234403

RESUMO

Bacterial wilt caused by Ralstonia solanacearum is a disease of widespread economic importance that affects numerous plant species, including Arabidopsis thaliana. We describe a pathosystem between A. thaliana and biovar 3 phylotype I strain BCCF402 of R. solanacearum isolated from Eucalyptus trees. A. thaliana accession Be-0 was susceptible and accession Kil-0 was tolerant. Kil-0 exhibited no wilting symptoms and no significant reduction in fitness (biomass, seed yield, and germination efficiency) after inoculation with R. solanacearum BCCF402, despite high bacterial numbers in planta. This was in contrast to the well-characterized resistance response in the accession Nd-1, which limits bacterial multiplication at early stages of infection and does not wilt. R. solanacearum BCCF402 was highly virulent because the susceptible accession Be-0 was completely wilted after inoculation. Genetic analyses, allelism studies with Nd-1, and RRS1 cleaved amplified polymorphic sequence marker analysis showed that the tolerance phenotype in Kil-0 was dependent upon the resistance gene RRS1. Knockout and complementation studies of the R. solanacearum BCCF402 effector PopP2 confirmed that the tolerance response in Kil-0 was dependent upon the RRS1-PopP2 interaction. Our data indicate that the gene-for-gene interaction between RRS1 and PopP2 can contribute to tolerance, as well as resistance, which makes it a useful model system for evolutionary studies of the arms race between plants and bacterial pathogens. In addition, the results alert biotechnologists to the risk that deployment of RRS1 in transgenic crops may result in persistence of the pathogen in the field.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Arabidopsis/microbiologia , Proteínas de Bactérias/metabolismo , Ralstonia solanacearum/metabolismo , Ralstonia solanacearum/patogenicidade , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Proteínas de Bactérias/genética , Doenças das Plantas/microbiologia , Ligação Proteica/genética , Ligação Proteica/fisiologia
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