RESUMO
We used the Roche-454 platform to sequence from normalized cDNA libraries from each of two inbred lines of onion (OH1 and 5225). From approximately 1.6 million reads from each inbred, 27,065 and 33,254 cDNA contigs were assembled from OH1 and 5225, respectively. In total, 3,364 well supported single nucleotide polymorphisms (SNPs) on 1,716 cDNA contigs were identified between these two inbreds. One SNP on each of 1,256 contigs was randomly selected for genotyping. OH1 and 5225 were crossed and 182 gynogenic haploids extracted from hybrid plants were used for SNP mapping. A total of 597 SNPs segregated in the OH1 × 5225 haploid family and a genetic map of ten linkage groups (LOD ≥8) was constructed. Three hundred and thirty-nine of the newly identified SNPs were also mapped using a previously developed segregating family from BYG15-23 × AC43, and 223 common SNPs were used to join the two maps. Because these new SNPs are in expressed regions of the genome and commonly occur among onion germplasms, they will be useful for genetic mapping, gene tagging, marker-aided selection, quality control of seed lots, and fingerprinting of cultivars.
Assuntos
Mapeamento Cromossômico/métodos , Cromossomos de Plantas/genética , Genoma de Planta/genética , Cebolas/genética , Polimorfismo de Nucleotídeo Único , Ligação Genética , Variação Genética , Genótipo , Análise de Sequência de DNA , Transcriptoma/genéticaRESUMO
TGA factors play a key role in plant defense by binding to the promoter region of defense genes, inducing expression. Salicylic acid (SA) induces the expression of the gene encoding NIMIN-1, which interacts with NPR1/NIM1, a key regulator of systemic acquired resistance. We investigated whether the TGA2-binding motif TGACG located upstream of the NIMIN-1 gene is necessary for SA induction of NIMIN-1 expression. A mutated version of the NIMIN-1 promoter was created by site-directed mutagenesis. We generated T-DNA constructs in which native NIMIN-1 and mutated promoters were fused to green fluorescent protein and beta-glucuronidase reporters. We produced transgenic Arabidopsis plants and observed NIMIN-1 promoter-driven green fluorescent protein expression in the roots, petiole and leaves. Constructs were agroinfiltrated into the leaves for transient quantitative assays of gene expression. Using quantitative real-time RT-PCR, we characterized the normal gene response to SA and compared it to the response of the mutant version of the NIMIN-1 promoter. Both the native NIMIN-1 construct and an endogenous copy of NIMIN-1 were induced by SA. However, the mutated promoter construct was much less sensitive to SA than the native NIMIN-1 promoter, indicating that this TGA2-binding motif is directly involved in the modulation of SA-induced NIMIN-1 expression in Arabidopsis.
Assuntos
Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Arabidopsis/efeitos dos fármacos , Arabidopsis/genética , Fatores de Transcrição de Zíper de Leucina Básica/metabolismo , Proteínas de Transporte/genética , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Proteínas Nucleares/metabolismo , Regiões Promotoras Genéticas , Ácido Salicílico/farmacologia , Sequência de Bases , Sítios de Ligação , Proteínas de Transporte/metabolismo , Proteínas de Fluorescência Verde/metabolismo , Dados de Sequência Molecular , Mutação/genética , Plantas Geneticamente Modificadas , Ligação Proteica/efeitos dos fármacos , Fatores de TranscriçãoRESUMO
We have used phosphorus-31 nuclear magnetic resonance to determine intracellular pH in the cellular slime mold Dictyostelium discoideum. We devised an air-lift circulator to maintain the dense cell suspensions in a well-oxygenated and well-stirred state while causing minimal perturbation to the sample flowing through the detector coils. Cells continued to develop normally in this set-up. Spectra acquired under these conditions typically show two peaks in the inorganic phosphate region corresponding to pH values of 7.16 +/- 0.03 and 6.48 +/- 0.02. These peaks are believed to represent the mitochondrial and cytosolic compartments respectively, based on a comparison of these values with published data and the collapse of the two compartments upon addition of the mitochondrial uncoupler carbonyl cyanide 4-(trifluoromethoxy)-phenylhydrazone. Dictyostelium cells show a remarkable degree of intracellular pH homeostasis. Both mitochondrial and cytosolic pH remained unchanged as extracellular pH was varied from 4.3 to 8.1. There was also no apparent change in the pH of either compartment after up to 13.5 hours' development in suspension.
Assuntos
Citoplasma/fisiologia , Dictyostelium/fisiologia , Concentração de Íons de Hidrogênio , 3',5'-AMP Cíclico Fosfodiesterases/metabolismo , Nucleotídeos de Adenina/metabolismo , Fatores Etários , Carbonil Cianeto p-Trifluormetoxifenil Hidrazona/farmacologia , Fosfato de Di-Hidroxiacetona/metabolismo , Galactosiltransferases/metabolismo , Espectroscopia de Ressonância Magnética/métodos , Mitocôndrias/fisiologia , Fosforilases/metabolismoRESUMO
A much higher yield of DNA single-strand breaks was obtained in the DNA polymerase-deficient mutant Escherichia coli K-12 pol A1 after a given dose of x-rays than had been found before in Escherichia coli. The increased yield of single-strand breaks was due to the absence of a rapid repair system, which had not been described in Escherichia coli K-12. This absence probably accounts for the x-ray sensitivity of the pol A1 mutant. The rapid repair system can be reversibly inhibited in pol+ cells.
Assuntos
DNA Nucleotidiltransferases , DNA Bacteriano/efeitos da radiação , Efeitos da Radiação , Centrifugação com Gradiente de Concentração , Cromossomos Bacterianos , Escherichia coli , Genética Microbiana , MutaçãoRESUMO
Although the nuclear genome of banana (Musa spp.) is relatively small (1C approximately 610 Mbp for M. acuminata), the results obtained from other sequenced genomes suggest that more than half of the banana genome may be composed of repetitive and non-coding DNA sequences. Knowledge of repetitive DNA can facilitate mapping of important traits, phylogenetic studies, BAC-based physical mapping, and genome sequencing/annotation. However, only a few repetitive DNA sequences have been characterized in banana. In this work, we used DNA reassociation kinetics to isolate the highly repeated fraction of the banana genome (M. acuminata 'Calcutta 4'). Two libraries, one prepared from Cot =0.05 DNA (2,688 clones) and one from Cot =0.1 sequences (4,608 clones), were constructed, and 614 DNA clones were chosen randomly for sequencing and further characterization. Dot-plot analysis revealed that 14% of the sequenced clones contained various semi-tandem and palindromic repeated sequences. 'BLAST' homology searches showed that, in addition to tandem repeats, the Cot libraries were composed mainly of different types of retrotransposons, the most frequent being the Ty3/gypsy type monkey retrotransposon. Selected sequences displaying tandem organization properties were mapped by PRimed IN Situ DNA labeling (PRINS) to the secondary constriction on metaphase chromosomes of M. acuminata 'Calcutta 4'. Southern hybridization with selected BAC clones carrying 45S rDNA confirmed the presence of the tandem repeats in the 45S rDNA unit. This work significantly expands the knowledge of the repetitive fraction of the Musa genome and organization of its chromosomes.
Assuntos
DNA de Plantas/genética , DNA de Plantas/isolamento & purificação , Genoma de Planta/genética , Musa/genética , Sequências Repetitivas de Ácido Nucleico/genética , Sequência de Bases , Cromossomos Artificiais Bacterianos , Células Clonais , DNA Ribossômico/genética , Dosagem de Genes , Biblioteca Gênica , Marcação in Situ com Primers , Homologia de Sequência do Ácido Nucleico , Sequências de Repetição em TandemRESUMO
Arabidopsis thaliana, a small annual plant belonging to the mustard family, is the subject of study by an estimated 7000 researchers around the world. In addition to the large body of genetic, physiological and biochemical data gathered for this plant, it will be the first higher plant genome to be completely sequenced, with completion expected at the end of the year 2000. The sequencing effort has been coordinated by an international collaboration, the Arabidopsis Genome Initiative (AGI). The rationale for intensive investigation of Arabidopsis is that it is an excellent model for higher plants. In order to maximize use of the knowledge gained about this plant, there is a need for a comprehensive database and information retrieval and analysis system that will provide user-friendly access to Arabidopsis information. This paper describes the initial steps we have taken toward realizing these goals in a project called The Arabidopsis Information Resource (TAIR) (www.arabidopsis.org).
Assuntos
Arabidopsis/genética , Bases de Dados Factuais , Mapeamento Cromossômico , Genoma de Planta , Serviços de Informação , Armazenamento e Recuperação da Informação , InternetRESUMO
We have isolated a genomic clone containing Arabidopsis thaliana 5S ribosomal RNA (rRNA)-encoding genes (rDNA) by screening an A. thaliana library with a 5S rDNA probe from flax. The clone isolated contains seven repeat units of 497 bp, plus 11 kb of flanking genomic sequence at one border. Sequencing of individual subcloned repeat units shows that the sequence of the 5S rRNA coding region is very similar to that reported for other flowering plants. Four A. thaliana ecotypes were found to contain approx. 1000 copies of 5S rDNA per haploid genome. Southern-blot analysis of genomic DNA indicates that 5S rDNA occurs in long tandem arrays, and shows the presence of numerous restriction-site polymorphisms among the six ecotypes studied.
Assuntos
DNA Ribossômico/genética , Família Multigênica/genética , Plantas/genética , Polimorfismo de Fragmento de Restrição , RNA Ribossômico 5S/genética , Sequência de Bases , Southern Blotting , Clonagem Molecular , Sondas de DNA/genética , Dados de Sequência Molecular , Sequências Repetitivas de Ácido Nucleico/genéticaRESUMO
In most microarray assays, labeled cDNA molecules derived from reference and query RNA samples are co-hybridized to probes arrayed on a glass surface. Gene expression profiles are then calculated for each gene based on the relative hybridization intensities measured between the two samples. The most commonly used reference samples are typically isolates from a single representative RNA source (RNA-0) or pooled mixtures of RNA derived from a plurality of sources (RNA-p). Genomic DNA offers an alternative reference nucleic acid with a number of potential advantages, including stability, reproducibility, and a potentially uniform representation of all genes, as each unique gene should have equal representation in a haploid genome. Using hydrogen peroxide-treated Arabidopsis thaliana plants as a model, we evaluated genomic DNA and RNA-p as reference samples and compared expression levels inferred through the reference relative to unexposed plants with expression levels measured directly using an RNA-0 reference. Our analysis demonstrates that while genomic DNA can serve as a reasonable reference source for microarray assays, a much greater correlation with direct measurements can be achieved using an RNA-based reference sample.
Assuntos
Perfilação da Expressão Gênica/métodos , Análise de Sequência com Séries de Oligonucleotídeos , Padrões de Referência , Arabidopsis/efeitos dos fármacos , Arabidopsis/genética , DNA Complementar/genética , DNA de Plantas/genética , Genoma de Planta , Proteínas de Choque Térmico HSP70/genética , Peróxido de Hidrogênio/farmacologia , Hibridização in Situ Fluorescente , Hibridização de Ácido Nucleico , Análise de Sequência com Séries de Oligonucleotídeos/normas , RNA/genética , RNA de Plantas/isolamento & purificaçãoRESUMO
Pigeonpea (Cajanus cajan), an important food legume crop in the semi-arid regions of the world and the second most important pulse crop in India, has an average crop productivity of 780 kg/ha. The relatively low crop yields may be attributed to non-availability of improved cultivars, poor crop husbandry and exposure to a number of biotic and abiotic stresses in pigeonpea growing regions. Narrow genetic diversity in cultivated germplasm has further hampered the effective utilization of conventional breeding as well as development and utilization of genomic tools, resulting in pigeonpea being often referred to as an 'orphan crop legume'. To enable genomics-assisted breeding in this crop, the pigeonpea genomics initiative (PGI) was initiated in late 2006 with funding from Indian Council of Agricultural Research under the umbrella of Indo-US agricultural knowledge initiative, which was further expanded with financial support from the US National Science Foundation's Plant Genome Research Program and the Generation Challenge Program. As a result of the PGI, the last 3 years have witnessed significant progress in development of both genetic as well as genomic resources in this crop through effective collaborations and coordination of genomics activities across several institutes and countries. For instance, 25 mapping populations segregating for a number of biotic and abiotic stresses have been developed or are under development. An 11X-genome coverage bacterial artificial chromosome (BAC) library comprising of 69,120 clones have been developed of which 50,000 clones were end sequenced to generate 87,590 BAC-end sequences (BESs). About 10,000 expressed sequence tags (ESTs) from Sanger sequencing and ca. 2 million short ESTs by 454/FLX sequencing have been generated. A variety of molecular markers have been developed from BESs, microsatellite or simple sequence repeat (SSR)-enriched libraries and mining of ESTs and genomic amplicon sequencing. Of about 21,000 SSRs identified, 6,698 SSRs are under analysis along with 670 orthologous genes using a GoldenGate SNP (single nucleotide polymorphism) genotyping platform, with large scale SNP discovery using Solexa, a next generation sequencing technology, is in progress. Similarly a diversity array technology array comprising of ca. 15,000 features has been developed. In addition, >600 unique nucleotide binding site (NBS) domain containing members of the NBS-leucine rich repeat disease resistance homologs were cloned in pigeonpea; 960 BACs containing these sequences were identified by filter hybridization, BES physical maps developed using high information content fingerprinting. To enrich the genomic resources further, sequenced soybean genome is being analyzed to establish the anchor points between pigeonpea and soybean genomes. In addition, Solexa sequencing is being used to explore the feasibility of generating whole genome sequence. In summary, the collaborative efforts of several research groups under the umbrella of PGI are making significant progress in improving molecular tools in pigeonpea and should significantly benefit pigeonpea genetics and breeding. As these efforts come to fruition, and expanded (depending on funding), pigeonpea would move from an 'orphan legume crop' to one where genomics-assisted breeding approaches for a sustainable crop improvement are routine.
Assuntos
DNA Nucleotidiltransferases , DNA Bacteriano/efeitos da radiação , Escherichia coli/efeitos da radiação , Oxigênio , Efeitos da Radiação , Células Cultivadas , Centrifugação com Gradiente de Concentração , Cianetos/farmacologia , Reparo do DNA/efeitos dos fármacos , Reparo do DNA/efeitos da radiação , DNA Bacteriano/farmacologia , DNA de Cadeia Simples/efeitos da radiação , Etilmaleimida/farmacologia , Hidroxiureia/farmacologia , Iodoacetatos/farmacologia , Quinacrina/farmacologiaAssuntos
DNA Bacteriano/efeitos da radiação , Escherichia coli/efeitos da radiação , Oxigênio/farmacologia , Efeitos da Radiação , Sobrevivência Celular/efeitos da radiação , Colífagos/efeitos da radiação , Escherichia coli/ultraestrutura , Peso Molecular , Doses de Radiação , Timidina/metabolismo , Raios XRESUMO
The rhg1 gene or genes lie at a recessive or co-dominant locus, necessary for resistance to all Hg types of the soybean (Glycine max (L.) Merr.) cyst nematode (Heterodera glycines I.). The aim here was to identify nucleotide changes within a candidate gene found at the rhg1 locus that were capable of altering resistance to Hg types 0 (race 3). A 1.5 +/- 0.25 cM region of chromosome 18 (linkage group G) was shown to encompass rhg1 using recombination events from four near isogenic line populations and nine DNA markers. The DNA markers anchored two bacterial artificial chromosome (BAC) clones 21d9 and 73p6. A single receptor like kinase (RLK; leucine rich repeat-transmembrane-protein kinase) candidate resistance gene was amplified from both BACs using redundant primers. The DNA sequence showed nine alleles of the RLK at Rhg1 in the soybean germplasm. Markers designed to detect alleles showed perfect association between allele 1 and resistance to soybean cyst nematode Hg types 0 in three segregating populations, fifteen additional selected recombination events and twenty-two Plant Introductions. A quantitative trait nucleotide (QTN) [corrected] in the RLK at rhg1 was inferred that alters A87 to V87 in the context of H274 rather than N274. [corrected] Contiguous DNA sequence of 315 kbp of chromosome 18 (about 2 cM) contained additional gene candidates that may modulate resistance to other Hg-types including a variant laccase, a hydrogen-sodium ion antiport and two proteins of unknown function. A molecular basis for recessive and co-dominant resistance that involves interactions among paralagous disease-resistance genes was inferred that would improve methods for developing new nematode-resistant soybean cultivars.
Assuntos
Mapeamento Cromossômico , Genes de Plantas/genética , Glycine max/genética , Imunidade Inata/genética , Doenças das Plantas/parasitologia , Tylenchoidea , Animais , Sequência de Bases , Southern Blotting , Cromossomos Artificiais Bacterianos , Cruzamentos Genéticos , Genômica , Repetições de Microssatélites/genética , Dados de Sequência Molecular , Doenças das Plantas/genética , Análise de Sequência de DNARESUMO
We have compared the pattern of enzyme expression in cyclic AMP-induced monolayer cultures of Dictyostelium discoideum with that found during normal development. We find that both the temporal and quantitative pattern of enzyme expression are initially similar in the two situations, although the developmental sequence is more protracted and terminal cell differentiation is delayed in the monolayer situation. We describe differentiation conditions that permit the expression of only one terminal phenotype, which may be useful for further biochemical studies. Enzyme accumulation patterns under these conditions indicate that UDP gal transferase is not required for stalk cell differentiation (i.e., it is a prespore enzyme). We have shown that, when cell monolayers are incubated with cAMP, the presence of a weak acid at low extracellular pH favors stalk-cell differentiation, while a weak base at high extracellular pH favors spore differentiation. Finally, we show that variations in the monovalent cation content of the buffer, or the addition of an ion transport inhibitor (scillaren), or an ionophore (valinomycin) all affect the ratio of stalk cells to spores. Taken together, these results suggest that intracellular H+ and/or other cations may play an important role in regulating differentiation of specific cell types in D. discoideum.
Assuntos
Dictyostelium/citologia , 3',5'-AMP Cíclico Fosfodiesterases/metabolismo , Fosfatase Alcalina/metabolismo , Amônia/farmacologia , Cátions Monovalentes , Adesão Celular , Diferenciação Celular , Células Cultivadas , AMP Cíclico/farmacologia , Dictyostelium/enzimologia , Concentração de Íons de Hidrogênio , Fosforilases/metabolismo , Pronase/metabolismo , UDPglucose 4-EpimeraseRESUMO
gamma-Radiation-induced tumors of Arabidopsis thaliana L. have been produced as a novel approach to isolation of genes that regulate plant development. Tumors excised from irradiated plants are hormone autonomous in culture and have been maintained on hormone-free medium for up to 4 years. Five tumor tissue lines having different morphologies and growth rates were analyzed for auxin, cytokinin, and 1-aminocyclopropane-1-carboxylic acid (ACC) content, ethylene production, and response to exogenous growth regulators. Normal tissues and two crown gall tissue lines were analyzed for comparison. Rosettes and whole seedlings each contained approximately 30 nanograms. (gram fresh weight)(-1) free indoleacetic acid (IAA), 150 nanograms. (gram fresh weight)(-1) ester-conjugated IAA, and 10 to 20 micrograms. (gram fresh weight)(-1) amide-conjugated IAA. The crown gall lines contained similar amounts of free and ester-conjugated IAA but less amide conjugates. Whereas three of the radiation-induced tumor lines had IAA profiles similar to normal tissues, one line had 10- to 100-fold more free IAA and three- to 10-fold less amide-conjugated IAA. The fifth line had normal free IAA levels but more conjugated IAA than control tissues. Whole seedlings contained approximately 2 nanograms. (gram fresh weight)(-1) of both zeatin riboside and isopentenyladenosine. The crown gall lines had 100- to 1000-fold higher levels of each cytokinin. In contrast, the three radiation-induced tumor lines analyzed contained cytokinin levels similar to the control tissue. The radiation-induced tumor tissues produced very little ethylene, although each contained relatively high levels of ACC. Normal callus contained similar amounts of ACC but produced several times more ethylene than the radiation-induced tumor lines. Each of the radiation-induced tumor tissues displayed a unique set of responses to exogenously supplied growth regulators. Only one tumor line showed the same response as normal callus to both auxin and cytokinin feeding. In some cases, one or more tumor lines showed increased sensitivity to certain growth substances. In other cases, growth regulator feeding had no significant effect on tumor tissue growth. Morphology of the radiation-induced tumor tissues generally did not correlate with auxin to cytokinin ratio in the expected manner. The results suggest that a different primary genetic event led to the formation of each tumor and that growth and differentiation in the tumor tissue lines are uncoupled from the normal hormonal controls.
RESUMO
A central feature of the processes of aggregation and differentiation in the cellular slime mould Dictyostelium discoideum is the periodic excitatory cycle. Originally thought to involve primarily fluctuations in cyclic AMP levels, this excitatory cycle has since been shown to involve changes in several other second messengers including cyclic GMP, calcium and inositol trisphosphate. Previous work from this laboratory using specific inhibitors strongly suggested a role for eicosanoids in this stimulus-response process. Production of eicosanoids from fatty acid precursors is an oxygen-consuming process. In this paper, we report on oxygen consumption measurements in intact D. discoideum cells and in cell extracts. We demonstrate the existence of an azide-insensitive component of oxygen consumption which can be stimulated by the addition of arachidonate and other polyunsaturated fatty acids, and at least partially inhibited by meclofenamate and eicosatetraynoic acid, both of which block eicosanoid biosynthesis in higher organisms. These observations provide further evidence for the existence of an eicosanoid-metabolizing system in D. discoideum.
Assuntos
Ácidos Araquidônicos/metabolismo , Dictyostelium/metabolismo , Consumo de Oxigênio/efeitos dos fármacos , Ácido 5,8,11,14-Eicosatetrainoico/farmacologia , Ácido Araquidônico , Dictyostelium/efeitos dos fármacos , Ácidos Eicosanoicos/metabolismo , Indometacina/farmacologia , Ácido Meclofenâmico/farmacologiaRESUMO
The terminal differentiation of Dictyostelium discoideum cells plated as monolayers with cyclic AMP is dramatically affected by developmental buffer conditions. High pH and addition of weak bases induces spore differentiation while low pH and weak acids favour stalk cell formation. In order to analyse the timing and nature of this regulation we have raised and characterized an anti-stalk serum which we have used together with an anti-spore serum to monitor developmental progression in the monolayer system and to detect the phenotypic effects of pH at earlier stages of development. The stalk serum detects both polysaccharide and protein antigens expressed during the terminal stages of normal development. In monolayer culture, the stalk-specific protein antigen appears precociously, while the timing of prespore vacuole appearance is unaffected. Expression of stalk polysaccharide antigens in monolayer cultures occurs as early as 12 h and is localized in a single subset of cells or region of extracellular space within the small cell clumps that form. The effects of pH (and acid/base) on these phenotype-specific antigens can be detected early in development, shortly after their first appearance. In monolayers of wild-type V12 M2 cells, the low pH regimes appear to act more by suppressing the spore than enhancing the stalk pathway, while the high pH regimes both suppress stalk and enhance spore antigen expression. In monolayers of the sporogenous mutant HM29, low pH regimes both enhance stalk antigen and suppress spore antigen expression. These results show that extracellular pH regulates phenotypic expression during a large part of the differentiation process and is not simply restricted to terminal cytodifferentiation.
Assuntos
Antígenos de Fungos/análise , Dictyostelium/citologia , Diferenciação Celular , Células Cultivadas , Dictyostelium/imunologia , Imunofluorescência , Concentração de Íons de Hidrogênio , Fenótipo , Esporos FúngicosRESUMO
To investigate the molecular mechanisms of hormonal control of growth, we constructed a subtracted cDNA library enriched for sequences expressed more in a hormone-autonomous, radiation-induced tumor tissue line of Arabidopsis thaliana than in normal, hormone-dependent callus. Ten cDNA clones, which are expressed 1.3- to 10-fold more in the tumor line, were isolated and partially characterized. The clones differ greatly in their level of expression in tumor tissue and in their pattern of expression in plant organs. Southern blot hybridization and sequence analysis showed that this group contains three pairs of closely related clones. Northern blot analysis indicates that one pair of clones represents two members of a gene family that are expressed in different plant organs. One of the isolated sequences shows strong sequence similarity to a cDNA encoding a lipid transfer protein. Two sequences are highly similar to those of previously described membrane channel proteins but have different organ specificities. Two other cDNAs have significant sequence similarity to glycine-rich proteins and hydroxy-proline-rich glycoproteins. When used to probe Southern blots, none of the cDNAs identified polymorphisms between tumor and callus DNA, which might be expected if their overexpression were due to local genome rearrangements induced by radiation. The diversity observed among these 10 clones suggests that some are likely to be involved in tumorous growth and not simply specific to a certain cell or tissue type present in the tumor.