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1.
FEMS Microbiol Ecol ; 95(8)2019 08 01.
Artigo em Inglês | MEDLINE | ID: mdl-31247638

RESUMO

Acacia spirorbis subsp. spirorbis Labill. is a widespread tree legume endemic to New Caledonia that grows in ultramafic (UF) and volcano-sedimentary (VS) soils. The aim of this study was to assess the symbiotic promiscuity of A. spirorbis with nodulating and nitrogen-fixing rhizobia in harsh edaphic conditions. Forty bacterial strains were isolated from root nodules and characterized through (i) multilocus sequence analyses, (ii) symbiotic efficiency and (iii) tolerance to metals. Notably, 32.5% of the rhizobia belonged to the Paraburkholderia genus and were only found in UF soils. The remaining 67.5%, isolated from both UF and VS soils, belonged to the Bradyrhizobium genus. Strains of the Paraburkholderia genus showed significantly higher nitrogen-fixing capacities than those of Bradyrhizobium genus. Strains of the two genera isolated from UF soils showed high metal tolerance and the respective genes occurred in 50% of strains. This is the first report of both alpha- and beta-rhizobia strains associated to an Acacia species adapted to UF and VS soils. Our findings suggest that A. spirorbis is an adaptive plant that establishes symbioses with whatever rhizobia is present in the soil, thus enabling the colonization of contrasted ecosystems.


Assuntos
Acacia/microbiologia , Bradyrhizobium/metabolismo , Burkholderiaceae/metabolismo , Metais/metabolismo , Poluentes do Solo/metabolismo , Adaptação Fisiológica , Bradyrhizobium/classificação , Bradyrhizobium/isolamento & purificação , Burkholderiaceae/classificação , Burkholderiaceae/isolamento & purificação , Nova Caledônia , Nitrogênio/metabolismo , Filogenia , Microbiologia do Solo , Simbiose
2.
Methods Mol Biol ; 1852: 211-220, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30109633

RESUMO

Rhodopseudomonas palustris is a purple photosynthetic bacterium that accumulates in the inner membrane the photosynthetic pigment spirilloxanthin, formed from lycopene. Here, we describe the procedures used to successfully engineer Rps. palustris strains to reroute the production of lycopene toward the synthesis of ß-carotene or canthaxanthin. The crtCD genes specifically involved in spirilloxanthin were replaced by crtY and crtW genes from Bradyrhizobium ORS278 to synthesize ß-carotene and (or) canthaxanthin, two pigments of industrial interest. Since the synthesis of canthaxanthin depends on the presence of oxygen, the procedure to optimize their production is also proposed. By modulating the light and oxygen during the growth process, a single species of photosynthetic bacteria, with an efficient growth rate, produces various carotenoids of economical interest.


Assuntos
Bioengenharia , Carotenoides/biossíntese , Fotossíntese , Rodopseudomonas/fisiologia , Bioengenharia/métodos , Biotecnologia/métodos , Cantaxantina/biossíntese , Ordem dos Genes , Engenharia Genética , Genoma Bacteriano , Luz , Licopeno/metabolismo , Oxigênio/metabolismo , beta Caroteno/metabolismo
3.
Microb Ecol ; 76(4): 964-975, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-29717331

RESUMO

This study aims to characterize the ectomycorrhizal (ECM) communities associated with Acacia spirorbis, a legume tree widely spread in New Caledonia that spontaneously grows on contrasted edaphic constraints, i.e. calcareous, ferralitic and volcano-sedimentary soils. Soil geochemical parameters and diversity of ECM communities were assessed in 12 sites representative of the three mains categories of soils. The ectomycorrhizal status of Acacia spirorbis was confirmed in all studied soils, with a fungal community dominated at 92% by Basidiomycota, mostly represented by/tomentella-thelephora (27.6%), /boletus (15.8%), /sebacina (10.5%), /russula-lactarius (10.5%) and /pisolithus-scleroderma (7.9%) lineages. The diversity and the proportion of the ECM lineages were similar for the ferralitic and volcano-sedimentary soils but significantly different for the calcareous soils. These differences in the distribution of the ECM communities were statistically correlated with pH, Ca, P and Al in the calcareous soils and with Co in the ferralitic soils. Altogether, these data suggest a high capacity of A. spirorbis to form ECM symbioses with a large spectrum of fungi regardless the soil categories with contrasted edaphic parameters.


Assuntos
Acacia/microbiologia , Basidiomycota/fisiologia , Micorrizas/fisiologia , Microbiologia do Solo , Solo/química , Acacia/crescimento & desenvolvimento , Basidiomycota/classificação , Micorrizas/classificação , Nova Caledônia , Simbiose
4.
Mycorrhiza ; 27(4): 407-413, 2017 May.
Artigo em Inglês | MEDLINE | ID: mdl-28091750

RESUMO

New Caledonian serpentine (ultramafic) soils contain high levels of toxic heavy metals, in particular nickel, (up to 20 g kg-1) and are deficient in essential elements like carbon, nitrogen and phosphorus while having a high magnesium/calcium ratio. Although previous studies showed that ectomycorrhizal symbioses could play an important role in the adaptation of the endemic plants to ultramafic soils (FEMS Microbiol Ecol 72:238-49, 2010), none of them have compared the diversity of microbial communities from ultramafic vs non-ultramafic soils in New Caledonia. We explored the impact of edaphic characteristics on the diversity of ectomycorrhizal (ECM) fungi associated with different endemic species of Tristaniopsis (Myrtaceae) growing under contrasting soil conditions in the natural ecosystems of New Caledonia. ECM root tips were thus sampled from two different ultramafic sites (Koniambo massif and Desmazures forest) vs two volcano-sedimentary ones (Arama and Mont Ninndo). The molecular characterization of the ECM fungi through partial sequencing of the ITS rRNA gene revealed the presence of different dominant fungal genera including, both soil types combined, Cortinarius (36.1%), Pisolithus (18.5%), Russula (13.4%), Heliotales (8.2%) and Boletellus (7.2%). A high diversity of ECM taxa associated with Tristaniopsis species was found in both ultramafic and volcano-sedimentary soils but no significant differences in ECM genera distribution were observed between both soil types. No link could be established between the phylogenetic clustering of ECM taxa and their soil type origin, thus suggesting a possible functional-rather than taxonomical-adaptation of ECM fungal communities to ultramafic soils.


Assuntos
Micorrizas/classificação , Myrtaceae/microbiologia , Microbiologia do Solo , Nova Caledônia , Filogenia , Solo
5.
J Exp Bot ; 65(6): 1551-64, 2014 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-24510940

RESUMO

Nickel is an economically important metal and phytotechnologies are being developed to limit the impact of nickel mining on the environment. More than 300 plant species are known to hyperaccumulate nickel. However, our knowledge of the mechanisms involved in nickel accumulation in plants is very limited because it has not yet been possible to study these hyperaccumulators at the genomic level. Here, we used next-generation sequencing technologies to sequence the transcriptome of the nickel hyperaccumulator Psychotria gabriellae of the Rubiaceae family, and used yeast and Arabidopsis as heterologous systems to study the activity of identified metal transporters. We characterized the activity of three metal transporters from the NRAMP and IREG/FPN families. In particular, we showed that PgIREG1 is able to confer nickel tolerance when expressed in yeast and in transgenic plants, where it localizes in the tonoplast. In addition, PgIREG1 shows higher expression in P. gabriellae than in the related non-accumulator species Psychotria semperflorens. Our results designate PgIREG1 as a candidate gene for nickel tolerance and hyperaccumulation in P. gabriellae. These results also show how next-generation sequencing technologies can be used to access the transcriptome of non-model nickel hyperaccumulators to identify the underlying molecular mechanisms.


Assuntos
Proteínas de Transporte de Cátions/genética , Níquel/metabolismo , Psychotria/genética , Transcriptoma , Sequência de Aminoácidos , Arabidopsis/citologia , Arabidopsis/genética , Arabidopsis/crescimento & desenvolvimento , Arabidopsis/metabolismo , Sequência de Bases , Proteínas de Transporte de Cátions/metabolismo , Genes Reporter , Sequenciamento de Nucleotídeos em Larga Escala , Metais/metabolismo , Anotação de Sequência Molecular , Dados de Sequência Molecular , Mutação , Filogenia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Raízes de Plantas/genética , Raízes de Plantas/crescimento & desenvolvimento , Raízes de Plantas/metabolismo , Plantas Geneticamente Modificadas , Psychotria/metabolismo , Análise de Sequência de DNA , Especificidade por Substrato , Vacúolos/metabolismo
6.
J Plant Physiol ; 171(2): 164-72, 2014 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-24331432

RESUMO

Ectomycorrhizal fungi (ECM) isolates of Pisolithus albus (Cooke and Massee) from nickel-rich ultramafic topsoils in New Caledonia were inoculated onto Acacia spirorbis Labill. (an endemic Fabaceae) and Eucalyptus globulus Labill. (used as a Myrtaceae plant host model). The aim of the study was to analyze the growth of symbiotic ECM plants growing on the ultramafic substrate that is characterized by high and toxic metal concentrations i.e. Co, Cr, Fe, Mn and Ni, deficient concentrations of plant essential nutrients such as N, P, K, and that presents an unbalanced Ca/Mg ratio (1/19). ECM inoculation was successful with a plant level of root mycorrhization up to 6.7%. ECM symbiosis enhanced plant growth as indicated by significant increases in shoot and root biomass. Presence of ECM enhanced uptake of major elements that are deficient in ultramafic substrates; in particular P, K and Ca. On the contrary, the ECM symbioses strongly reduced transfer to plants of element in excess in soils; in particular all metals. ECM-inoculated plants released metal complexing molecules as free thiols and oxalic acid mostly at lower concentrations than in controls. Data showed that ECM symbiosis helped plant growth by supplying uptake of deficient elements while acting as a protective barrier to toxic metals, in particular for plants growing on ultramafic substrate with extreme soil conditions. Isolation of indigenous and stress-adapted beneficial ECM fungi could serve as a potential tool for inoculation of ECM endemic plants for the successful restoration of ultramafic ecosystems degraded by mining activities.


Assuntos
Acacia/microbiologia , Basidiomycota/fisiologia , Eucalyptus/microbiologia , Metais Pesados/metabolismo , Micorrizas/fisiologia , Acacia/crescimento & desenvolvimento , Acacia/metabolismo , Adaptação Biológica , Eucalyptus/crescimento & desenvolvimento , Eucalyptus/metabolismo , Ácido Oxálico/análise , Exsudatos de Plantas/química , Raízes de Plantas/microbiologia , Raízes de Plantas/fisiologia , Solo/química , Compostos de Sulfidrila/análise
7.
Environ Microbiol Rep ; 6(5): 510-8, 2014 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-25646544

RESUMO

Nickel (Ni)-tolerant ectomycorrhizal Pisolithus albus was isolated from extreme ultramafic soils that are naturally rich in heavy metals. This study aimed to identify the specific molecular mechanisms associated with the response of P. albus to nickel. In presence of high concentration of nickel, P. albus Ni-tolerant isolate showed a low basal accumulation of nickel in its fungal tissues and was able to perform a metal efflux mechanism. Three genes putatively involved in metal efflux were identified from the P. albus transcriptome, and their overexpression was confirmed in the mycelium that was cultivated in vitro in the presence of nickel and in fungal tissues that were sampled in situ. Cloning these genes in yeast provided significant advantages in terms of nickel tolerance (+ 31% Ni EC50) and growth (+ 83% µ) compared with controls. Furthermore, nickel efflux was also detected in the transformed yeast cells. Protein sequence analysis indicated that the genes encoded a P-type-ATPase, an ABC transporter and a major facilitator superfamily permease (MFS). This study sheds light on a global mechanism of metal efflux by P. albus cells that supports nickel tolerance. These specific responses to nickel might contribute to the fungal adaptation in ultramafic soil.


Assuntos
Basidiomycota/metabolismo , Micorrizas/isolamento & purificação , Micorrizas/metabolismo , Níquel/metabolismo , Microbiologia do Solo , Basidiomycota/genética , Basidiomycota/isolamento & purificação , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Micélio/classificação , Micélio/genética , Micélio/metabolismo , Micorrizas/classificação , Micorrizas/genética
8.
Mol Ecol ; 21(9): 2208-23, 2012 May.
Artigo em Inglês | MEDLINE | ID: mdl-22429322

RESUMO

The fungus Pisolithus albus forms ectomycorrhizal (ECM) associations with plants growing on extreme ultramafic soils, which are naturally rich in heavy metals such as nickel. Both nickel-tolerant and nickel-sensitive isolates of P. albus are found in ultramafic soils in New Caledonia, a biodiversity hotspot in the Southwest Pacific. The aim of this work was to monitor the expression of genes involved in the specific molecular response to nickel in a nickel-tolerant P. albus isolate. We used pyrosequencing and quantitative polymerase chain reaction (qPCR) approaches to investigate and compare the transcriptomes of the nickel-tolerant isolate MD06-337 in the presence and absence of nickel. A total of 1,071,375 sequencing reads were assembled to infer expression patterns of 19,518 putative genes. Comparison of expression levels revealed that 30% of the identified genes were modulated by nickel treatment. The genes, for which expression was induced most markedly by nickel, encoded products that were putatively involved in a variety of biological functions, such as the modification of cellular components (53%), regulation of biological processes (27%) and molecular functions (20%). The 10 genes that pyrosequencing analysis indicated were induced the most by nickel were characterized further by qPCR analysis of both nickel-tolerant and nickel-sensitive P. albus isolates. Five of these genes were expressed exclusively in nickel-tolerant isolates as well as in ECM samples in situ, which identified them as potential biomarkers for nickel tolerance in this species. These results clearly suggest a positive transcriptomic response of the fungus to nickel-rich environments. The presence of both nickel-tolerant and nickel-sensitive fungal phenotypes in ultramafic soils might reflect environment-dependent phenotypic responses to variations in the effective concentrations of nickel in heterogeneous ultramafic habitats.


Assuntos
Basidiomycota/genética , Regulação Fúngica da Expressão Gênica , Micorrizas/genética , Níquel/farmacologia , Microbiologia do Solo , Adaptação Fisiológica/genética , Basidiomycota/efeitos dos fármacos , Basidiomycota/isolamento & purificação , Basidiomycota/metabolismo , Biodiversidade , Biomarcadores , Ecossistema , Variação Genética , Anotação de Sequência Molecular , Micorrizas/efeitos dos fármacos , Micorrizas/isolamento & purificação , Micorrizas/metabolismo , Nova Caledônia , Níquel/metabolismo , Filogenia , Plantas/microbiologia , Análise de Sequência de DNA , Transcriptoma
9.
Mol Plant Microbe Interact ; 23(6): 760-70, 2010 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-20459315

RESUMO

Photosynthetic Bradyrhizobium strains possess the unusual ability to form nitrogen-fixing nodules on a specific group of legumes in the absence of Nod factors. To obtain insight into the bacterial genes involved in this Nod-independent symbiosis, we screened 15,648 Tn5 mutants of Bradyrhizobium sp. strain ORS278 for clones affected in root symbiosis with Aeschynomene indica. From the 268 isolated mutants, 120 mutants were altered in nodule development (Ndv(-)) and 148 mutants were found to be deficient in nitrogen fixation (Fix(-)). More than 50% of the Ndv(-) mutants were found to be altered in purine biosynthesis, strengthening the previous hypothesis of a symbiotic role of a bacterial purine derivative during the Nod-independent symbiosis. The other Ndv(-) mutants were auxotrophic for pyrimidines and amino acids (leucine, glutamate, and lysine) or impaired in genes encoding proteins of unknown function. The Fix(-) mutants were found to be affected in a wide variety of cellular processes, including both novel (n = 56) and previously identified (n = 31) genes important in symbiosis. Among the novel genes identified, several were involved in the Calvin cycle, suggesting that CO(2) fixation could play an important role during this symbiosis.


Assuntos
Bradyrhizobium/genética , Bradyrhizobium/metabolismo , Fabaceae/microbiologia , Genes Bacterianos , Fotossíntese/fisiologia , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Metabolismo dos Carboidratos , Metabolismo Energético , Fabaceae/fisiologia , Perfilação da Expressão Gênica , Regulação Bacteriana da Expressão Gênica , Mutagênese Insercional , Nucleotídeos/biossíntese , Oxigenases/genética , Oxigenases/metabolismo , Fotossíntese/genética , Simbiose
10.
FEMS Microbiol Ecol ; 72(2): 238-49, 2010 May.
Artigo em Inglês | MEDLINE | ID: mdl-20199570

RESUMO

Isolates of ectomycorrhizal Pisolithus albus were sampled from both ultramafic and volcano-sedimentary soils in New Caledonia, a tropical hotspot of biodiversity, to investigate the relationships between genetic diversity and edaphic constraint through tolerance to nickel (Ni). Carpophore description, spore morphology and phylogenetic analysis based on internal transcribed spacer (ITS) rDNA sequences confirmed that all isolates belong to P. albus and are closely related to other Australasian specimens. Using molecular tools, ITS-restriction fragment length polymorphism and amplified fragment length polymorphism markers, we showed the existence of two distinct genetic clusters within P. albus: ultramafic and volcano-sedimentary. Mycelia response to Ni toxicity supports such a population structure. Pisolithus albus from ultramafic soils included isolates with a high diversity of in vitro Ni tolerance, with both Ni-tolerant isolates (average Ni EC(50) at 575 microM) and Ni-sensitive isolates (average Ni EC(50) at 37 microM). In contrast, all isolates from volcano-sedimentary soils were found to be Ni sensitive (average Ni EC(50) at 32 microM). We highlight that (1) P. albus population from ultramafic soils of New Caledonia are genetically structured in ecotype, and that (2) Ni tolerance among ultramafic isolates suggests an adaptive physiological response to Ni toxicity.


Assuntos
Micorrizas/genética , Microbiologia do Solo , Solo/análise , Análise do Polimorfismo de Comprimento de Fragmentos Amplificados , DNA Fúngico/genética , DNA Espaçador Ribossômico/genética , Variação Genética , Micélio/metabolismo , Micorrizas/isolamento & purificação , Nova Caledônia , Níquel/metabolismo , Fenótipo , Filogenia , Polimorfismo de Fragmento de Restrição , Análise de Sequência de DNA
11.
Photosynth Res ; 100(2): 97-105, 2009 May.
Artigo em Inglês | MEDLINE | ID: mdl-19452262

RESUMO

In aerobic anoxygenic phototrophs, oxygen is required for both the formation of the photosynthetic apparatus and an efficient cyclic electron transfer. Mutants of Bradyrhizobium sp. ORS278 affected in photosystem synthesis were selected by a bacteriochlorophyll fluorescence-based screening. Out of the 9,600 mutants of a random Tn5 insertion library, 50 clones, corresponding to insertions in 28 different genes, present a difference in fluorescence intensity compared to the WT. Besides enzymes and regulators known to be involved in photosystem synthesis, 14 novel components of the photosynthesis control are identified. Among them, two genes, hsIU and hsIV, encode components of a protein degradation complex, probably linked to the renewal of photosystem, an important issue in Bradyrhizobia which have to deal with harmful reactive oxygen species. The presence of homologs in non-photosynthetic bacteria for most of the regulatory genes identified during study suggests that they could be global regulators, as the RegA-RegB system.


Assuntos
Bradyrhizobium/genética , Bradyrhizobium/metabolismo , Genes Bacterianos , Mutação/genética , Complexo de Proteínas do Centro de Reação Fotossintética/biossíntese , Bacterioclorofilas/metabolismo , Bradyrhizobium/efeitos da radiação , Células Clonais , Luz , Fenótipo , Fotossíntese/genética , Fotossíntese/efeitos da radiação , Espectrometria de Fluorescência
12.
J Bacteriol ; 190(17): 5824-31, 2008 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-18606738

RESUMO

The recent sequence analysis of the photosynthetic and plant-symbiotic Bradyrhizobium sp. strain BTAi1 revealed the unexpected presence of a pucBA operon encoding the apoproteins of peripheral light-harvesting (LH) complexes. This pucBA operon is found close to a bacteriophytochrome gene (BphP3(B BTAi1)) and a two-component transcriptional regulator gene (TF(BTAi1) gene). In this study, we show that BphP3(B BTAi1) acts as a bona fide bacteriophytochrome and controls, according to light conditions, the expression of the pucBA operon found in its vicinity. This light regulatory pathway is very similar to the one previously described for chromo-BphP4(Rp) in Rhodopseudomonas palustris and conducts the synthesis of a peripheral LH complex. This LH complex presents a single absorption band at low temperature, centered at 803 nm. Fluorescence emission analysis of intact cells indicates that this peripheral LH complex does not act as an efficient light antenna. One putative function of this LH complex could be to evacuate excess light energy in order to protect Bradyrhizobium strain BTAi1, an aerobic anoxygenic photosynthetic bacterium, against photooxidative damage during photosynthesis.


Assuntos
Proteínas de Bactérias/metabolismo , Bradyrhizobium/metabolismo , Complexos de Proteínas Captadores de Luz/metabolismo , Fotossíntese/fisiologia , Aerobiose , Proteínas de Bactérias/classificação , Proteínas de Bactérias/genética , Bradyrhizobium/genética , Histidina Quinase , Complexos de Proteínas Captadores de Luz/genética , Óperon/genética , Fenótipo , Fotossíntese/genética , Filogenia , Proteínas Quinases/metabolismo , Transdução de Sinais/genética , Transdução de Sinais/fisiologia
13.
Biochim Biophys Acta ; 1777(2): 163-72, 2008 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-17988648

RESUMO

In the purple photosynthetic bacterium Rhodopseudomonas palustris, far-red illumination induces photosystem synthesis via the action of the bacteriophytochrome RpBphP1. This bacteriophytochrome antagonizes the repressive effect of the transcriptional regulator PpsR2 under aerobic condition. We show here that, in addition to photosystem synthesis, far-red light induces a significant growth rate limitation, compared to cells grown in the dark, linked to a decrease in the respiratory activity. The phenotypes of mutants inactivated in RpBphP1 and PpsR2 show their involvement in this regulation. Based on enzymatic and transcriptional studies, a 30% decrease in the expression of the alpha-ketoglutarate dehydrogenase complex, a central enzyme of the Krebs cycle, is observed under far-red light. We propose that this decrease is responsible for the down-regulation of respiration in this condition. This regulation mechanism at the Krebs cycle level still allows the formation of the photosynthetic apparatus via the synthesis of key biosynthesis precursors but lowers the production of NADH, i.e. the respiratory activity. Overall, the dual action of RpBphP1 on the regulation of both the photosynthesis genes and the Krebs cycle allows a fine adaptation of bacteria to environmental conditions by enhancement of the most favorable bioenergetic process in the light, photosynthesis versus respiration.


Assuntos
Proteínas de Bactérias/fisiologia , Complexo Cetoglutarato Desidrogenase/metabolismo , Consumo de Oxigênio/fisiologia , Fotossíntese/fisiologia , Complexo de Proteína do Fotossistema I/biossíntese , Pigmentos Biológicos/fisiologia , Rodopseudomonas/fisiologia , Rodopseudomonas/genética , Rodopseudomonas/crescimento & desenvolvimento , Rodopseudomonas/efeitos da radiação
14.
Science ; 316(5829): 1307-12, 2007 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-17540897

RESUMO

Leguminous plants (such as peas and soybeans) and rhizobial soil bacteria are symbiotic partners that communicate through molecular signaling pathways, resulting in the formation of nodules on legume roots and occasionally stems that house nitrogen-fixing bacteria. Nodule formation has been assumed to be exclusively initiated by the binding of bacterial, host-specific lipochito-oligosaccharidic Nod factors, encoded by the nodABC genes, to kinase-like receptors of the plant. Here we show by complete genome sequencing of two symbiotic, photosynthetic, Bradyrhizobium strains, BTAi1 and ORS278, that canonical nodABC genes and typical lipochito-oligosaccharidic Nod factors are not required for symbiosis in some legumes. Mutational analyses indicated that these unique rhizobia use an alternative pathway to initiate symbioses, where a purine derivative may play a key role in triggering nodule formation.


Assuntos
Bradyrhizobium/genética , Bradyrhizobium/fisiologia , Fabaceae/microbiologia , Caules de Planta/microbiologia , Nódulos Radiculares de Plantas/fisiologia , Simbiose , Aciltransferases/genética , Aciltransferases/metabolismo , Amidoidrolases/genética , Amidoidrolases/metabolismo , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Bradyrhizobium/crescimento & desenvolvimento , Citocininas/metabolismo , Genes Bacterianos , Genoma Bacteriano , Genômica , Lipopolissacarídeos/metabolismo , Dados de Sequência Molecular , Mutação , N-Acetilglucosaminiltransferases/genética , N-Acetilglucosaminiltransferases/metabolismo , Fotossíntese , Raízes de Plantas/microbiologia , Purinas/biossíntese , Nódulos Radiculares de Plantas/microbiologia , Transdução de Sinais
15.
EMBO J ; 26(14): 3322-31, 2007 Jul 25.
Artigo em Inglês | MEDLINE | ID: mdl-17581629

RESUMO

Bacteriophytochromes are red/far-red photoreceptors that bacteria use to mediate sensory responses to their light environment. Here, we show that the photosynthetic bacterium Rhodopseudomonas palustris has two distinct types of bacteriophytochrome-related protein (RpBphP4) depending upon the strain considered. The first type binds the chromophore biliverdin and acts as a light-sensitive kinase, thus behaving as a bona fide bacteriophytochrome. However, in most strains, RpBphP4 does not to bind this chromophore. This loss of light sensing is replaced by a redox-sensing ability coupled to kinase activity. Phylogenetic analysis is consistent with an evolutionary scenario, where a bacteriophytochrome ancestor has adapted from light to redox sensing. Both types of RpBphP4 regulate the synthesis of light harvesting (LH2) complexes according to the light or redox conditions, respectively. They modulate the affinity of a transcription factor binding to the promoter regions of LH2 complex genes by controlling its phosphorylation status. This is the first complete description of a bacteriophytochrome signal transduction pathway involving a two-component system.


Assuntos
Proteínas de Bactérias/metabolismo , Evolução Molecular , Luz , Rodopseudomonas/metabolismo , Sequência de Aminoácidos , Proteínas de Bactérias/química , Proteínas de Bactérias/genética , Proteínas de Bactérias/isolamento & purificação , Complexos de Proteínas Captadores de Luz/biossíntese , Complexos de Proteínas Captadores de Luz/efeitos dos fármacos , Complexos de Proteínas Captadores de Luz/efeitos da radiação , Modelos Biológicos , Dados de Sequência Molecular , Oxirredução/efeitos da radiação , Oxigênio/farmacologia , Fotossíntese/efeitos dos fármacos , Fotossíntese/efeitos da radiação , Filogenia , Fitocromo/química , Fitocromo/genética , Fitocromo/isolamento & purificação , Fitocromo/metabolismo , Regiões Promotoras Genéticas/genética , Proteínas Recombinantes/genética , Proteínas Recombinantes/isolamento & purificação , Rodopseudomonas/efeitos dos fármacos , Rodopseudomonas/genética , Rodopseudomonas/efeitos da radiação , Transdução de Sinais/efeitos dos fármacos , Fatores de Transcrição/metabolismo
16.
J Biol Chem ; 282(10): 7320-8, 2007 Mar 09.
Artigo em Inglês | MEDLINE | ID: mdl-17218312

RESUMO

Bacteriophytochromes are phytochrome-like proteins that mediate photosensory responses in various bacteria according to their light environment. The genome of the photosynthetic and plant-symbiotic Bradyrhizobium sp. strain ORS278 revealed the presence of a genomic island acquired by lateral transfer harboring a bacteriophytochrome gene, BrBphP3.ORS278, and genes involved in the synthesis of phycocyanobilin and gas vesicles. The corresponding protein BrBphP3.ORS278 is phylogenetically distant from the other (bacterio)phytochromes described thus far and displays a series of unusual properties. It binds phycocyanobilin as a chromophore, a unique feature for a bacteriophytochrome. Moreover, its C-terminal region is short and displays no homology with any known functional domain. Its dark-adapted state absorbs maximally around 610 nm, an unusually short wavelength for (bacterio)phytochromes. This form is designated as Po for orange-absorbing form. Upon illumination, a photo-reversible switch occurs between the Po form and a red (670 nm)-absorbing form (Pr), which rapidly backreacts in the dark. Because of this instability, illumination results in a mixture of the Po and Pr states in proportions that depend on the intensity. These uncommon features suggest that BrBphP3.ORS278 could be fitted to measure light intensity rather than color.


Assuntos
Proteínas de Bactérias/genética , Bradyrhizobium/genética , Transferência Genética Horizontal , Fitocromo/genética , Sequência de Aminoácidos , Proteínas de Bactérias/química , Fluorescência , Dados de Sequência Molecular , Ficobilinas/metabolismo , Ficocianina/metabolismo , Fitocromo/química , Temperatura
17.
J Biol Chem ; 280(37): 32389-97, 2005 Sep 16.
Artigo em Inglês | MEDLINE | ID: mdl-16009707

RESUMO

Phytochromes are chromoproteins found in plants and bacteria that switch between two photointerconvertible forms via the photoisomerization of their chromophore. These two forms, Pr and Pfr, absorb red and far-red light, respectively. We have characterized the biophysical and biochemical properties of two bacteriophytochromes, RpBphP2 and RpBphP3, from the photosynthetic bacterium Rhodopseudomonas palustris. Their genes are contiguous and localized near the pucBAd genes encoding the polypeptides of the light harvesting complexes LH4, whose synthesis depends on the light intensity. At variance with all (bacterio)phytochromes studied so far, the light-induced isomerization of the chromophore of RpBphP3 converts the Pr form to a form absorbing at shorter wavelength around 645 nm, designated as Pnr for near red. The quantum yield for the transformation of Pr into Pnr is about 6-fold smaller than for the reverse reaction. Both RpBphP2 and RpBphP3 autophosphorylate in their dark-adapted Pr forms and transfer their phosphate to a common response regulator Rpa3017. Under semiaerobic conditions, LH4 complexes replace specifically the LH2 complexes in wild-type cells illuminated by wavelengths comprised between 680 and 730 nm. In contrast, mutants deleted in each of these two bacteriophytochromes display no variation in the composition of their light harvesting complexes whatever the light intensity. From both the peculiar properties of these bacteriophytochromes and the phenotypes of their deletion mutants, we propose that they operate in tandem to control the synthesis of LH4 complexes by measuring the relative intensities of 645 and 710 nm lights.


Assuntos
Fitocromo/química , Rodopseudomonas/fisiologia , Sequência de Aminoácidos , Fenômenos Bioquímicos , Bioquímica , Fenômenos Biofísicos , Biofísica , Cisteína/química , Eletroforese em Gel de Poliacrilamida , Luz , Modelos Químicos , Dados de Sequência Molecular , Mutação , Fenótipo , Fosforilação , Complexo de Proteínas do Centro de Reação Fotossintética/química , Fitocromo/classificação , Estrutura Terciária de Proteína , Proteínas Recombinantes/química , Homologia de Sequência de Aminoácidos , Espectrometria de Fluorescência , Espectrofotometria , Espectroscopia de Infravermelho com Transformada de Fourier , Fatores de Tempo
18.
J Biol Chem ; 279(43): 44407-16, 2004 Oct 22.
Artigo em Inglês | MEDLINE | ID: mdl-15304477

RESUMO

The two closely related bacteria Bradyrhizobium and Rhodopseudomonas palustris show an unusual mechanism of regulation of photosystem formation by light thanks to a bacteriophytochrome that antirepresses the regulator PpsR. In these two bacteria, we found out, unexpectedly, that two ppsR genes are present. We show that the two Bradyrhizobium PpsR proteins exert antagonistic effects in the regulation of photosystem formation with a classical repressor role for PpsR2 and an unexpected activator role for PpsR1. DNase I footprint analysis show that both PpsR bind to the same DNA TGTN12ACA motif that is present in tandem in the bchC promoter and the crtED intergenic region. Interestingly, the cycA and aerR promoter regions that contain only one conserved palindrome are recognized by PpsR2, but not PpsR1. Further biochemical analyses indicate that PpsR1 only is redox sensitive through the formation of an intermolecular disulfide bond, which changes its oligomerization state from a tetramer to an octamer under oxidizing conditions. Moreover, PpsR1 presents a higher DNA affinity under its reduced form in contrast to what has been previously found for PpsR or its homolog CrtJ from the Rhodobacter species. These results suggest that regulation of photosystem synthesis in Bradyrhizobium involves two PpsR competing for the binding to the same photosynthesis genes and this competition might be modulated by two factors: light via the antagonistic action of a bacteriophytochrome on PpsR2 and redox potential via the switch of PpsR1 oligomerization state.


Assuntos
Proteínas de Bactérias/química , Bradyrhizobium/metabolismo , Proteínas de Ligação a DNA/química , Regulação da Expressão Gênica , Oxirredução , Proteínas Repressoras/química , Motivos de Aminoácidos , Proteínas de Bactérias/fisiologia , Sequência de Bases , Cromatografia em Gel , Clonagem Molecular , DNA/química , DNA/metabolismo , Proteínas de Ligação a DNA/fisiologia , Desoxirribonuclease I/química , Desoxirribonuclease I/metabolismo , Dissulfetos , Ditiotreitol/farmacologia , Relação Dose-Resposta a Droga , Eletroforese em Gel de Poliacrilamida , Ligantes , Luz , Modelos Genéticos , Dados de Sequência Molecular , Mutagênese Sítio-Dirigida , Mutação , Oxigênio/metabolismo , Fotossíntese , Filogenia , Regiões Promotoras Genéticas , Ligação Proteica , Proteínas Repressoras/fisiologia , Rhodobacter/metabolismo , beta-Galactosidase/metabolismo
19.
Photochem Photobiol Sci ; 3(6): 587-91, 2004 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-15170489

RESUMO

The synthesis of the photosynthetic apparatus of different strains of Rhodopseudomonas palustris has been studied as a function of the oxygen concentration and far-red light. For strain CEA001, only a small amount of photosynthetic apparatus is synthesized in the dark for oxygen concentration higher than 8% whereas synthesis is strongly enhanced by far-red light illumination. This enhancement is due to the action of a bacteriophytochrome (ORF2127/ORF2128), which antagonizes the repressor PpsR. On the contrary, a large fraction of photosystem is synthesized in the dark and far-red illumination induces no enhancement in strain CGA009. This difference in phenotype of strain CGA009 is explained by a single point-mutation R428C in the helix-turn-helix DNA binding motif of PpsR, rendering it inactive. In addition, a frame-shift mutation had occurred in the gene encoding bacteriophytochrome (ORF2127/ORF2128), conducting to a truncated inactive sensor. We propose that these mutations occurred in culture. Bacteria have developed a sophisticated regulatory process to synthesize their photosynthetic apparatus when light is available. This process is a critical advantage for the bacteria under natural conditions since they optimize their development depending on the available energy resources. On the contrary, under laboratory growth conditions where there is no substrate limitation, there is no crucial need for such a regulation and deleterious mutations affecting this process are of no importance.


Assuntos
Proteínas de Bactérias/metabolismo , Fotossíntese/fisiologia , Fitocromo/fisiologia , Rodopseudomonas/metabolismo , Sequência de Aminoácidos , Proteínas de Bactérias/genética , Sequência de Bases , Bradyrhizobium/genética , Clonagem Molecular , Primers do DNA , DNA Bacteriano/genética , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Mapeamento por Restrição , Rodopseudomonas/classificação , Rodopseudomonas/genética , Rodopseudomonas/crescimento & desenvolvimento , Alinhamento de Sequência , Homologia de Sequência de Aminoácidos , Homologia de Sequência do Ácido Nucleico , Espectrofotometria
20.
J Biol Chem ; 279(15): 15076-83, 2004 Apr 09.
Artigo em Inglês | MEDLINE | ID: mdl-14734565

RESUMO

Aerobic photosynthetic bacteria possess the unusual characteristic of producing different classes of carotenoids. In this study, we demonstrate the presence of two distinct crt gene clusters involved in the synthesis of spirilloxanthin and canthaxanthin in a Bradyrhizobium strain. Each cluster contains the genes crtE, crtB, and crtI leading to the common precursor lycopene. We show that spirilloxanthin is associated with the photosynthetic complexes, while canthaxanthin protects the bacteria from oxidative stress. Only the spirilloxanthin crt genes are regulated by light via the control of a bacteriophytochrome. Despite this difference in regulation, the biosyntheses of both carotenoids are strongly interconnected at the level of the common precursors. Phylogenetic analysis suggests that the canthaxanthin crt gene cluster has been acquired by a lateral gene transfer. This acquisition may constitute a major selective advantage for this class of bacteria, which photosynthesize only under conditions where harmful reactive oxygen species are generated.


Assuntos
Bradyrhizobium/genética , Cantaxantina/biossíntese , Carotenoides/genética , Família Multigênica , Xantofilas/análogos & derivados , Xantofilas/biossíntese , Cantaxantina/química , Carotenoides/química , Carotenoides/metabolismo , Membrana Celular/metabolismo , Relação Dose-Resposta a Droga , Luz , Licopeno , Modelos Químicos , Dados de Sequência Molecular , Mutação , Estresse Oxidativo , Paraquat/farmacologia , Filogenia , Pigmentos Biológicos , Espécies Reativas de Oxigênio , Espectrometria de Fluorescência
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