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The paper presents the results of an experimental and numerical study of some properties of multimode liquid crystal waveguide structures. The nematic 4-cyano-4'-pentylbiphenyl (5CB) was used as a liquid crystal. A description of the experiments performed and some of the techniques used are given. Scattering diagrams are presented that characterize the features of propagation in a multimode liquid-crystal waveguide of one and many modes. It is shown that when an external repetitively pulsed electric field is switched on, the attenuation and size of inhomogeneities decrease. To explain this effect, the classical theory of liquid crystal director fluctuations is used. For the first time some properties of a liquid-crystal waveguide are described with explicit allowance for the two-dimensional Frederiks model. The two-dimensional nature of the liquid crystal director reorientation effects in our case (including under the action of an electric impulse-periodic field) required the involvement of the three-dimensional theory of light scattering in an LC waveguide and, as a consequence, the study of two-dimensional scattering diagrams in experiments, which make it possible to consider the two-dimensional nature of the behavior of the LC director. The relevance and importance of such studies are related both to the practical use and prospects of using liquid crystal materials in various high-speed and low-energy integrated-optical devices, for example, in communication elements, modulators, and sensors.
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Subcritical water has potential as an environmentally friendly solvent for applications including hydrolysis, liquefaction, extraction, and carbonization. Here, we report hydrolysis of sugarcane straw, an abundant byproduct of sugar production, in a semi-continuous reactor at reaction temperatures ranging from 190 to 260°C and at operating pressures of 9 and 16MPa. The target hydrolysis products were total reducing sugars. The main products of sugarcane straw hydrolysis were glucose, xylose, arabinose, and galactose in addition to 5- hydroxymethylfurfural and furfural as minor byproducts. Fourier transform infrared spectroscopy and thermogravimetric analysis provided additional information on the surface and bulk composition of the residual biomass. Char was present on samples treated at temperatures equal to and greater than 190°C. Samples treated at 260°C contained approximately 20wt% char, yet retained substantial hemicellulose and cellulose content. Hydrolysis temperature of 200°C provided the greatest TRS yield while minimizing char formation.
Assuntos
Biomassa , Carboidratos , Saccharum , Hidrólise , ÁguaRESUMO
The ultrasonic propagation in the water-based magnetic fluid with doubled layered surfactant shell was studied. The measurements were carried out both in the presence as well as in the absence of the external magnetic field. The thickness of the surfactant shell was evaluated by comparing the mean size of magnetic grain extracted from magnetization curve with the mean hydrodynamic diameter obtained from differential centrifugal sedimentation method. The thickness of surfactant shell was used to estimate volume fraction of the particle aggregates consisted of magnetite grain and surfactant layer. From the ultrasonic velocity measurements in the absence of the applied magnetic field, the adiabatic compressibility of the particle aggregates was determined. In the external magnetic field, the magnetic fluid studied in this article becomes acoustically anisotropic, i.e., velocity and attenuation of the ultrasonic wave depend on the angle between the wave vector and the direction of the magnetic field. The results of the ultrasonic measurements in the external magnetic field were compared with the hydrodynamic theory of Ovchinnikov and Sokolov (velocity) and with the internal chain dynamics model of Shliomis, Mond and Morozov (attenuation).
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In this paper, the sorption of arsenic onto nanocrystalline magnetite mineral Fe3O4 was studied in a model system. Nanocrystalline magnetite was produced by mechanical activation in a planetary ball mill from natural microcrystalline magnetite. As a consequence of milling, the specific surface area increased from 0.1m(2)/g to 11.9 m(2)/g and the surface site concentration enhanced from 2.2 sites/nm(2) to 8.4 sites/nm(2). These changes in surface properties of magnetite lead to the enhancement of arsenic removal from model system. The best sorption ability was achieved with magnetite sample activated for 90 min. In this case the sample was able to absorb around 4 mg/g. The structural changes of magnetite were also observed and the new hematite phase was detected after 120 min of milling. A good correlation between the decreasing particle size, increasing specific surface area and reduction of saturation magnetization was found. In desorption study, KOH and NaOH were found as the best eluents where more than 70% of arsenic was released back into the solution. The principal novelty of the paper is that mineral magnetite, truly one nature's gift can be used after "smart" milling (mechanical activation) as an effective arsenic sorbent.
Assuntos
Arsênio/análise , Óxido Ferroso-Férrico/química , Nanopartículas/química , Adsorção , Arsênio/química , Monitoramento Ambiental/métodos , Compostos Férricos/química , Concentração de Íons de Hidrogênio , Hidróxidos/química , Cinética , Magnetismo , Microscopia Eletrônica de Transmissão , Minerais/química , Tamanho da Partícula , Compostos de Potássio/química , Hidróxido de Sódio/química , Estresse Mecânico , Propriedades de Superfície , Temperatura , Difração de Raios XRESUMO
Drawing from a series of field measurement activities including the Alternative Aviation Fuels Experiments (AAFEX1 and AAFEX2), we present experimental measurements of particle number, size, and composition-resolved mass that describe the physical and chemical evolution of aircraft exhaust plumes on the time scale of 5 s to 2-3 min. As the plume ages, the particle number emission index initially increases by a factor of 10-50, due to gas-to-particle formation of a nucleation/growth mode, and then begins to fall with increased aging. Increasing the fuel sulfur content causes the initial increase to occur more rapidly. The contribution of the nucleation/growth mode to the overall particle number density is most pronounced at idle power and decreases with increasing engine power. Increasing fuel sulfur content, but not fuel aromatic content causes the nucleation/growth mode to dominate the particle number emissions at higher powers than for a fuel with "normal" sulfur and aromatic content. Particle size measurements indicate that the observed particle number emissions trends are due to continuing gas-to-particle conversion and coagulation growth of the nucleation/growth mode particles, processes which simultaneously increase particle mass and reduce particle number density. Measurements of nucleation/growth mode mass are consistent with the interpretation of particle number and size data and suggest that engine exit plane measurements may underestimate the total particle mass by as much as a factor of between 5 and 10.
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Poluentes Atmosféricos/análise , Aeronaves , Atmosfera/química , Emissões de Veículos/análise , Gasolina/análise , Tamanho da PartículaRESUMO
Striga is a root parasitic weed that attacks many of the staple crops in Africa, India and Southeast Asia, inflicting tremendous losses in yield and for which there are few effective control measures. Studies of parasitic plant virulence and host resistance will be greatly facilitated by the recent emergence of genomic resources that include extensive transcriptome sequence datasets spanning all life stages of S. hermonthica. Functional characterization of Striga genes will require detailed analyses of gene expression patterns. Quantitative real-time PCR is a powerful tool for quantifying gene expression, but correct normalization of expression levels requires identification of control genes that have stable expression across tissues and life stages. Since no S. hermonthica housekeeping genes have been established for this purpose, we evaluated the suitability of six candidate housekeeping genes across key life stages of S. hermonthica from seed conditioning to flower initiation using qRT-PCR and high-throughput cDNA sequencing. Based on gene expression analysis by qRT-PCR and RNA-Seq across heterogeneous Striga life stages, we determined that using the combination of three genes, UBQ1, PP2A and TUB1 provides the best normalization for gene expression throughout the parasitic life cycle. The housekeeping genes characterized here provide robust standards that will facilitate powerful descriptions of parasite gene expression patterns.
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Genes Essenciais , Interações Hospedeiro-Parasita/genética , Plantas Daninhas/genética , Striga/genética , África , Sudeste Asiático , Regulação da Expressão Gênica no Desenvolvimento , Índia , Doenças das Plantas/genética , Doenças das Plantas/parasitologia , Raízes de Plantas/genética , Raízes de Plantas/crescimento & desenvolvimento , Raízes de Plantas/parasitologia , Plantas Daninhas/crescimento & desenvolvimento , RNA/genética , RNA/isolamento & purificação , Reação em Cadeia da Polimerase em Tempo Real , Análise de Sequência de RNA , Striga/crescimento & desenvolvimentoRESUMO
The human lung adenocarcinoma epithelial (A549) cells and the human embryo lung (HEL 12469) cells were used to investigate the uptake and cytotoxicity of magnetite nanoparticles (MNPs) with different chemically modified surfaces. MNPs uptake was an energy-dependent process substantially affected by the serum concentration in the culture medium. Internalized MNPs localized in vesicle-bound aggregates were observed in the cytoplasm, none in the nucleus or in mitochondria. All MNPs induced a dose- and time-dependent increase in cytotoxicity in both human lung cell lines. The cytotoxicity of MNPs increased proportionally with the particle size. Since the cytotoxicity of MNPs was nearly identical when the doses were equalized based on particle surface area, we suppose that the particle surface area rather than the surface modifications per se underlay the cytotoxicity of MNPs. In general, higher internalized amount of MNPs was found in HEL 12469 cells compared with A549 cells. Accordingly, the viability of the human embryo lung cells was reduced more substantially than that of the adenocarcinoma lung cells. The weak MNPs uptake into A549 cells might be of biomedical relevance in cases where MNPs should be used as nanocarriers for targeted drug delivery in tumor tissue derived from alveolar epithelial cells.
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Adenocarcinoma/tratamento farmacológico , Proliferação de Células/efeitos dos fármacos , Sistemas de Liberação de Medicamentos , Compostos Férricos/farmacologia , Neoplasias Pulmonares/tratamento farmacológico , Pulmão/efeitos dos fármacos , Nanopartículas de Magnetita , Adenocarcinoma/patologia , Células Cultivadas , Diploide , Embrião de Mamíferos/citologia , Embrião de Mamíferos/efeitos dos fármacos , Humanos , Pulmão/citologia , Neoplasias Pulmonares/patologia , Microscopia Eletrônica de Transmissão , Tamanho da Partícula , Espectroscopia de Infravermelho com Transformada de Fourier , Propriedades de SuperfícieRESUMO
Genetic diversity and phylogenetic relationships among 22 local cowpea (Vigna unguiculata) varieties and inbred lines collected throughout Senegal were evaluated using simple sequence repeat molecular markers. A set of 49 primer combinations were developed from cowpea genomic/expressed sequence tags and evaluated for their ability to detect polymorphisms among the various cowpea genotypes. Forty-four primer combinations detected polymorphisms, with the remaining five primer sets failing to yield PCR amplification products. From one to 16 alleles were found among the informative primer combinations; their frequencies ranged from 0.60 to 0.95 (mean = 0.79). The genetic diversity of the sample varied from 0.08 to 0.42 (mean = 0.28). The polymorphic information content ranged from 0.08 to 0.33 (mean = 0.23). The local varieties clustered in the same group, except 53-3, 58-53, and 58-57; while Ndoute yellow pods, Ndoute violet pods and Baye Ngagne were in the second group. The photosensitive varieties (Ndoute yellow pods and Ndoute violet pods) were closely clustered in the second group and so were inbred line Mouride and local cultivar 58-57, which is also one of the parents for inbred line Mouride. These molecular markers could be used for selection and identification of elite varieties for cowpea improvement and germplasm management in Senegal.
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Fabaceae/classificação , Fabaceae/genética , Repetições de Microssatélites/genética , Alelos , Primers do DNA/genética , DNA de Plantas/genética , Variação Genética , Filogenia , Polimorfismo Genético , Técnica de Amplificação ao Acaso de DNA Polimórfico , SenegalRESUMO
The effect of magnetic field on the structure formation in an oil-based magnetic fluid with various concentrations of magnetite particles was studied. The evaluation of the experimental data obtained from small-angle X-ray scattering and ultrasonic attenuation indicates the formation of chain-like aggregates composed of magnetite particles. The experimental data obtained from ultrasonic spectroscopy fit well with the recent theoretical model by Shliomis, Mond and Morozov but only for a diluted magnetic fluid. In this model it is assumed that a dimer is the main building block of a B -field-induced chain-like structure, thus the estimation of the nematic order parameter does not depend on the actual length of the structure. The scattering method used reveals information about the aggregated structure size and relative changes in the degree of anisotropy in qualitative terms. The coupling constant [Formula: see text] , concentrations [Formula: see text] , average particle size d and its polydispersity [Formula: see text] were initially obtained using the vibrating sample magnetometry and these results were further confirmed by rheometry and scattering methods. Both the particles' orientational distribution and the nematic order parameter S were inferred from the ultrasonic measurements. The investigation of SAXS patterns reveals the orientation and sizes of aggregated structures under application of different magnetic-field strengths. In addition, the magnetic-field-dependent yield stress was measured, and a relationship between the yield stress and magnetic-field strength up to 0.5 T was established.
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The dielectric properties (permittivity, loss factor, dielectric breakdown strength) of magnetic liquids were investigated. The magnetic liquids were composed of magnetite particles coated with oleic acid as surfactant and dispersed in transformer oil. To determine their dielectric properties they were subjected to a uniform magnetic field at high alternating electric fields up to 14 MV m(-1). Nearly constant permittivity of magnetic liquid with particle volume concentration Φ = 0.0019 as a function of electric field was observed. Magnetic liquids with concentrations Φ = 0.019 and 0.032 showed significant changes of permittivity and loss factor dependent on electric and magnetic fields. The best concentration of magnetic fluid was found at which partial current impulse magnitudes were the lowest. The breakdown strength distribution of the magnetic liquid with Φ = 0.0025 was fitted with the Duxbury-Leath, Weibull and Gauss distribution functions.
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In this work we describe the observations of structural transitions in ferronematics based on the thermotropic nematics 6CHBT (4-trans-4'-n-hexyl-cyclohexyl-isothiocyanato-benzene). The ferronematic droplets were observed in solutions of nematogenic 6CHBT dissolved in phenyl isocyanate and doped with fine magnetic particles. The phase diagram of the transitions from the isotropic phase to the nematic phase via a droplet state was found. Magneto-dielectric measurements of various structural transitions in this new system enabled us to estimate the type of anchoring of the nematic molecules on the magnetic particle surfaces in the droplets.
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Control of the size and spatial distribution of materials at multiple length scales is one of the most compelling issues in nanotechnology research. We report a multiple-length-scale patterning of pure magnetic particles as well as biocompatible magnetic particles based on a printing technique named micro-injection molding in capillaries. The magnetic particles were prepared by a technique of co-precipitation of ferric and ferrous salts in an alkali medium. We demonstrate that the morphology and the size of the patterning nanoparticles can be controlled by simply controlling the concentration of the solution. Our method exploits the self-organization of the nanoparticles in a solution confined between a stamp and the surfaces of a substrate, exploiting confinement and competing interactions between the adsorbate and the substrate. Our approach represents a remarkable example of an integrated top-down/bottom-up process.
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In this study, we have prepared PLGA (poly-D,L-lactide-co-glycolide) nanospheres loaded with biocompatible magnetic fluid and anticancer drug taxol by a modified nanoprecipitation technique and investigated their magnetic properties. A magnetic fluid, MF-PEG, with a biocompatible layer of polyethylene glycol (PEG), was chosen as a magnetic carrier. The PLGA, whose copolymer ratio of D,L-lactide to glycolide is 85:15, was utilized as a capsulation material. Taxol, as an important anticancer drug, was chosen for its significant role against a wide range of tumours. The morphology and particle size distributions of the prepared nanospheres were investigated by transmission electron microscopy (TEM) and scanning electron microscopy (SEM) and showed a spherical shape of prepared nanospheres with size 250 nm. Infrared spectroscopy (FTIR), differential scanning calorimetry (DSC) and thermogravimetry (TGA) analysis confirmed incorporation of magnetic particles and taxol into the PLGA polymer. The results showed good encapsulation with magnetite content 21.5 wt% and taxol 0.5 wt%. Magnetic properties of magnetic fluids and taxol within the PLGA polymer matrix were investigated by SQUID magnetometry from 4.2 to 300 K. The SQUID measurements showed superparamagnetism of prepared nanospheres with a blocking temperature of 160 K and saturation magnetization 1.4 mT.
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An improved genetic linkage map has been constructed for cowpea (Vigna unguiculata L. Walp.) based on the segregation of various molecular markers and biological resistance traits in a population of 94 recombinant inbred lines (RILs) derived from the cross between 'IT84S-2049' and '524B'. A set of 242 molecular markers, mostly amplified fragment length polymorphism (AFLP), linked to 17 biological resistance traits, resistance genes, and resistance gene analogs (RGAs) were scored for segregation within the parental and recombinant inbred lines. These data were used in conjunction with the 181 random amplified polymorphic DNA (RAPD), restriction fragment length polymorphism (RFLP), AFLP, and biochemical markers previously mapped to construct an integrated linkage map for cowpea. The new genetic map of cowpea consists of 11 linkage groups (LGs) spanning a total of 2670 cM, with an average distance of 6.43 cM between markers. Astonishingly, a large, contiguous portion of LG1 that had been undetected in previous mapping work was discovered. This region, spanning about 580 cM, is composed entirely of AFLP markers (54 in total). In addition to the construction of a new map, molecular markers associated with various biological resistance and (or) tolerance traits, resistance genes, and RGAs were also placed on the map, including markers for resistance to Striga gesnerioides races 1 and 3, CPMV, CPSMV, B1CMV, SBMV, Fusarium wilt, and root-knot nematodes. These markers will be useful for the development of tools for marker-assisted selection in cowpea breeding, as well as for subsequent map-based cloning of the various resistance genes.
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Mapeamento Cromossômico , Magnoliopsida/genética , Biomarcadores , Polimorfismo Genético , Polimorfismo de Fragmento de Restrição , Técnica de Amplificação ao Acaso de DNA PolimórficoRESUMO
Fluorescence spectroscopic and kinetic analysis of photochemical activity, cofactor and substrate binding, and enzyme denaturation studies were performed with highly purified, recombinant pea NADPH:protochlorophyllide oxidoreductase (POR) heterologously expressed in Escherichia coli. The results obtained with an individual stereoisomer of the substrate [C8-ethyl-C13(2)-(R)-protochlorophyllide] demonstrate that the enzyme photoactive state possesses a characteristic fluorescence maximum at 646 nm that is due to the presence of specific charged amino acids in the enzyme catalytic site. The photoactive state is converted directly into an intermediate having fluorescence at 685 nm in a reaction involving direct hydrogen transfer from the cofactor (NADPH). Site-directed mutagenesis of the highly conserved Tyr275 (Y275F) and Lys279 (K279I and K279R) residues in the enzyme catalytic pocket demonstrated that the presence of these two amino acids in the wild-type POR considerably increases the probability of photoactive state formation following cofactor and substrate binding by the enzyme. At the same time, the presence of these two amino acids destabilizes POR and increases the rate of enzyme denaturation. Neither Tyr275 nor Lys279 plays a crucial role in the binding of the substrate or cofactor by the enzyme. In addition, the presence of Tyr275 is absolutely necessary for the second step of the protochlorophyllide reduction reaction, "dark" conversion of the 685 nm fluorescence intermediate and the formation of the final product, chlorophyllide. We propose that Tyr275 and Lys279 participate in the proper coordination of NADPH and PChlide in the enzyme catalytic site and thereby control the efficiency of the formation of the POR photoactive state.
Assuntos
Domínio Catalítico , Lisina/metabolismo , NADP/metabolismo , Oxirredutases atuantes sobre Doadores de Grupo CH-CH , Oxirredutases/metabolismo , Tirosina/metabolismo , Substituição de Aminoácidos/genética , Sítios de Ligação/genética , Domínio Catalítico/genética , Ativação Enzimática/genética , Estabilidade Enzimática/genética , Congelamento , Cinética , Lisina/genética , Mutagênese Sítio-Dirigida , NADP/genética , Oxirredutases/antagonistas & inibidores , Oxirredutases/genética , Pisum sativum/enzimologia , Pisum sativum/genética , Fotoquímica , Desnaturação Proteica , Protoclorifilida/análise , Protoclorifilida/genética , Protoclorifilida/isolamento & purificação , Rhodobacter capsulatus/genética , Espectrometria de Fluorescência , Especificidade por Substrato/genética , Temperatura , Tirosina/genéticaRESUMO
Membrane association of NADPH:protochlorophyllide oxidoreductase (POR, EC: 1.6.99.1) with isolated prolamellar bodies (PLBs) and prothylakoids (PTs) from wheat etioplasts was investigated. In vitro-expressed radiolabelled POR, with or without transit peptide, was used to characterize membrane association conditions. Proper association of POR with PLBs and PTs did not require the presequence, whereas NADPH and hydrolysable ATP were vital for the process. After treating the membranes with thermolysin, sodium hydroxide or carbonate, a firm attachment of the POR protein to the membrane was found. Although the PLBs and PTs differ significantly in their relative amount of POR in vivo, no major differences in POR association capacity could be observed between the two membrane systems when exogenous NADPH was added. Experiments run with only an endogenous NADPH source almost abolished association of POR with both PLBs and PTs. In addition, POR protein carrying a mutation in the putative nucleotide-binding site (ALA06) was unable to bind to the inner membranes in the presence of NADPH, which further demonstrates that the co-factor is essential for proper membrane association. POR protein carrying a mutation in the substrate-binding site (ALA24) showed less binding to the membranes as compared to the wild type. The results presented here introduce studies of a novel area of protein-membrane interaction, namely the association of proteins with a paracrystalline membrane structure, the PLB.
Assuntos
Membranas Intracelulares/enzimologia , Organelas/enzimologia , Oxirredutases atuantes sobre Doadores de Grupo CH-CH , Oxirredutases/metabolismo , Triticum/metabolismo , Trifosfato de Adenosina/farmacologia , Membranas Intracelulares/efeitos dos fármacos , Mutação , NADP/farmacologia , Organelas/efeitos dos fármacos , Plastídeos , Especificidade por Substrato , Triticum/genéticaRESUMO
The distribution of protochlorophyllide (Pchlide) and NADPH-Pchlide oxidoreductase (POR) was characterized in the epicotyls and roots of wild-type pea (Pisum sativum L. cv. Alaska) and lip1, a mutant with light-independent photomorphogenesis caused by a mutation in the COP1 locus. The upper part of the dark-grown lip1 mutant epicotyls had a high Pchlide content that decreased downward the organ. The elevated Pchlide level in lip1 seedlings was a result of the differentiation of more proplastids into Pchlide-containing plastids. The cortex cells in the lip1 epicotyl were filled with such plastids in contrast to the cortex cells of wild-type seedlings. The mutant also developed Pchlide-containing plastids in the roots, indicating the suppressing effect of the COP1 locus on development of plastids in the corresponding tissues in dark-grown wild-type plants. The distribution of Pchlide-containing plastids in dark-grown lip1 mutant stem and root was similar to the distribution of chloroplasts in irradiated wild-type plants. Both wild-type and lip1 epicotyls contained mostly short wavelength Pchlide fluorescing at 631 nm with only a small shoulder at 654 nm, which was transformed to a minute amount of chlorophyllide (Chlide) by flash irradiation. In contrast, with continuous irradiation a considerable amount of Chlide was formed especially in the lip1 epicotyls. Immunoblots indicated the presence of POR, as a 36 kDa band, in epicotyls of both dark-grown wild-type and lip1 mutant seedlings. However, lip1 stem tissue had a higher content of POR than the wild-type pea. The high content of POR was unexpected as lip1 lacked both the 654 nm fluorescing Pchlide form and the regular PLBs. In light, a significant amount of chlorophyll was formed also in the roots of the lip1 seedlings.
Assuntos
Proteínas de Arabidopsis , Clorofila/metabolismo , Oxirredutases atuantes sobre Doadores de Grupo CH-CH , Pisum sativum/metabolismo , Protoclorifilida/metabolismo , Ubiquitina-Proteína Ligases , Proteínas de Transporte/metabolismo , Clorofilídeos/metabolismo , Escuridão , Mutação , Oxirredutases/biossíntese , Pisum sativum/enzimologia , Pisum sativum/genética , Proteínas de Plantas/metabolismo , Raízes de Plantas/enzimologia , Caules de Planta/enzimologia , Plastídeos/fisiologia , Sementes , Análise EspectralRESUMO
Amplified fragment length polymorphism (AFLP) analysis was used to determine the degree of intra- and inter-specific genetic variation in the genus Nicotiana. Forty-six lines of cultivated tobacco (Nicotiana tabacum L.) and seven wild Nicotiana species, including N. sylvestris, N. tomentosiformis, N. otophora, N. glutinosa, N. suaveolens, N. rustica, and N. longiflora, were analyzed, using at least eight different oligonucleotide primer combinations capable of detecting a minimum of 50 polymorphic bands per primer pair. The amount of genetic polymorphism present among cultivated tobacco lines (N. tabacum) was limited, as evidenced by the high degree of similarity in the AFLP profiles of cultivars collected worldwide. Six major clusters were found within cultivated tobacco that were primarily based upon geographic origin and manufacturing quality traits. A greater amount of genetic polymorphism exists among wild species of Nicotiana than among cultivated forms. Pairwise comparisons of the AFLP profiles of wild and cultivated Nicotiana species show that polymorphic bands present in N. tabacum can be found in at least one of three proposed wild progenitor species (i.e., N. sylvestris, N. tomentosiformis, and N. otophora). This observation provides additional support for these species contributing to the origin of N. tabacum.
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Evolução Molecular , Genes de Plantas , Nicotiana/genética , Polimorfismo Genético , DNA/genética , Modelos Genéticos , Modelos Estatísticos , Filogenia , Especificidade da EspécieRESUMO
In vitro chloroplast import reactions and thylakoid association reactions have been performed with a series of C-terminal deletions and Cys-to-Ser substitution mutants of the pea NADPH:protochlorophyllide oxidoreductase (POR; EC 1.6.99). C-terminal deletions of the precursor POR (Delta362-400, Delta338-400, Delta315-400 and Delta300-400) were efficiently translocated across the chloroplast envelope. However, except the Delta396-400 mutant, no C-terminal deletion mutants or Cys-to-Ser substitution (Cys119, Cys281 and Cys309) mutants resisted post-treatment with thermolysin after the thylakoid association reactions. This suggests that these mutants were unable to properly associate to the thylakoids due to changes of the protein conformation of POR.
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Cisteína/metabolismo , Oxirredutases atuantes sobre Doadores de Grupo CH-CH , Oxirredutases/metabolismo , Sítios de Ligação , Membrana Celular/metabolismo , Cisteína/genética , Mutagênese , Oxirredutases/genética , Pisum sativum/enzimologia , Tilacoides/metabolismoRESUMO
Light-independent protochlorophyllide reduction leading to chlorophyll formation in the dark requires both chloroplast and nuclear gene expression in Chlamydomonas reinhardtii. Mutations in any one of the plastid (chlL, chlN, and chlB) or nuclear (y-1 to y-10) genes required for this process result in the phenotype of the yellow-in-the-dark or y mutants. Analysis of the chlL, chlN, and chlB transcript levels in both light- and dark-grown wild-type and y mutant cells showed that the y mutations have no effect on the transcription of these plastid genes. Protein gel blot analysis showed that the CHLN and CHLB proteins are present in similar amounts in light- and dark-grown wild-type cells, whereas CHLL is present only in wild-type cells grown in the dark or at light intensities < or =15 micromol m(-2) sec(-1). Analysis of chlL transcript distribution on polysome profiles and rates of protein turnover in chloramphenicol-treated cells suggested that CHLL formation is most probably blocked at translation initiation or elongation. Furthermore, treatment of cells with metabolic inhibitors and uncouplers of photosynthetic electron transport showed that regulation of CHLL formation is linked to the physiologic status of the chloroplast. Similar to wild-type cells, y mutants contain nearly identical amounts of CHLN and CHLB when grown in either light or darkness. However, no CHLL is present in any of the y mutants except y-7, which contains an immunoreactive CHLL smaller than the expected size. Our findings indicate that CHLL translation is negatively photoregulated by the energy state or redox potential within the chloroplast in wild-type cells and that nuclear y genes are required for synthesis or accumulation of the CHLL protein.