RESUMO
The dose effect of radiation has long been a topic of concern, but the molecular mechanism behind it is still unclear. In this study, dried pea seeds were irradiated with 252Cf fission neutron source. Through analyzing the transcriptome and proteome of M1 generation pea (Pisum sativum L.) leaves, we studied the molecular rule and mechanism of neutron dose effect. Our results showed three important rules of global gene expression in the studied dose range. The rule closely related to the neutron absorbed dose at the transcription and translation levels is: the greater the difference in neutron absorbed dose between two radiation treatment groups, the greater the difference in differential expression between the two groups and the control group. We also obtained important sensitive metabolic pathways of neutron radiation, as well as related key genes. Furthermore, the overall molecular regulation mechanism of dose effect was revealed based on the main functional items obtained. Our research results can be applied to appropriate radiation dose estimation and agricultural production practice.
Assuntos
Nêutrons , Pisum sativum , Pisum sativum/efeitos da radiação , Pisum sativum/genética , Relação Dose-Resposta à Radiação , Transcriptoma/efeitos da radiação , Doses de Radiação , Folhas de Planta/efeitos da radiação , Folhas de Planta/metabolismo , Sementes/efeitos da radiação , Proteoma/efeitos da radiação , Proteoma/metabolismo , Regulação da Expressão Gênica de Plantas/efeitos da radiaçãoRESUMO
PURPOSE: Ionizing radiation is a harsh environmental factor that could induce plant senescence. We hypothesized that radiation-related senescence remodels proteome, particularly by triggering the accumulation of prion-like proteins in plant tissues. The object of this study, pea (Pisum sativum L.), is an agriculturally important legume. Research on the functional importance of amyloidogenic proteins was never performed on this species. MATERIALS AND METHODS: Pea seeds were irradiated in the dose range 5-50 Gy of X-rays. Afterward, Fourier-transform infrared spectroscopy (FTIR) was used to investigate changes in the secondary structure of proteins in germinated 3-day-old seedlings. Specifically, we evaluated the ratio between the amide I and II peaks. Next, we performed protein staining with Congo red to compare the presence of amyloids in the samples. In parallel, we profiled the detergent-resistant proteome fraction by ultrahigh-performance liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS). Differentially accumulated proteins were functionally analyzed in MapMan software, and the PLAAC tool was used to predict putative prion-like proteins. RESULTS: We showed a reduced germination rate but higher plant height and faster appearance of reproductive organs in the irradiated at dose of 50 Gy group compared with the control; furthermore, we demonstrated more ß-sheets and amyloid aggregates in the roots of stressed plants. We detected 531 proteins in detergent-resistant fraction extracted from roots, and 45 were annotated as putative prion-like proteins. Notably, 29 proteins were significantly differentially abundant between the irradiated and the control groups. These proteins belong to several functional categories: amino acid metabolism, carbohydrate metabolism, cytoskeleton organization, regulatory processes, protein biosynthesis, and RNA processing. Thus, the discovery proteomics provided deep data on novel aspects of plant stress biology. CONCLUSION: Our data hinted that protein accumulation stimulated seedlings' growth as well as accelerated ontogenesis and, eventually, senescence, primarily through translation and RNA processing. The increased abundance of primary metabolism-related proteins indicates more intensive metabolic processes triggered in germinating pea seeds upon X-ray exposure. The functional role of detected putative amyloidogenic proteins should be validated in overexpression or knockout follow-up studies.
Assuntos
Pisum sativum , Pisum sativum/efeitos da radiação , Pisum sativum/metabolismo , Pisum sativum/crescimento & desenvolvimento , Germinação/efeitos da radiação , Proteínas de Plantas/metabolismo , Radiação Ionizante , Amiloide/metabolismo , Amiloide/efeitos da radiação , Proteoma/efeitos da radiação , Proteoma/metabolismo , Sementes/efeitos da radiação , Sementes/metabolismo , Sementes/crescimento & desenvolvimentoRESUMO
Local damage induces a physiological response in higher plants by means of generation and propagation of variation potential (VP). The response includes changes in photosynthesis and respiration. The aim of the present study was to investigate the effect of these changes on adenosine triphosphate (ATP) content in pea leaves. VP was induced by local heating of the first mature leaf and registered using extracellular and intracellular electrodes. Photosynthesis and respiration were measured using Dual-PAM-100 and GFS-3000. ATP content was determined using a bioluminescence-based ATP determination kit. Two non-stimulated leaves (second and fourth) were investigated. We showed that heating induced VP that propagated into the second mature leaf, but only a slight electrical reaction was registered in the fourth mature leaf. VP-induced inactivation of photosynthesis developed in the second leaf and included two stages: short- and long-term inactivation. Local heating also caused a two-stage increase in ATP content in the second leaf, which was connected with the photosynthetic responses. Changes in photosynthesis and ATP content were not observed in the fourth leaf. The effect of VP on respiration was investigated under dark conditions. We found that variation potential induced short-term activation of respiration in the second leaf. Local heating induced ATP content increase which included only one stage under dark conditions. Changes in ATP and respiration were absent in the fourth leaf under dark conditions. Thus, VP-induced photosynthetic and respiratory changes are likely to increase ATP content in pea leaves.
Assuntos
Trifosfato de Adenosina/metabolismo , Potenciais da Membrana , Fotossíntese , Pisum sativum/fisiologia , Folhas de Planta/fisiologia , Respiração Celular/efeitos da radiação , Temperatura Alta , Luz , Potenciais da Membrana/efeitos da radiação , Pisum sativum/efeitos da radiação , Fotossíntese/efeitos da radiação , Folhas de Planta/efeitos da radiação , Caules de Planta/fisiologia , Caules de Planta/efeitos da radiaçãoRESUMO
Comparative analysis of in vivo chlorophyll fluorescence imaging revealed that photosystem II (PSII) photochemical efficiency (Fv/Fm) of leaves of the Costata 2/133 pea mutant with altered pigment composition and decreased level of oligomerization of the light harvesting chlorophyll a/b-protein complexes (LHCII) of PSII (Dobrikova et al., 2000; Ivanov et al., 2005) did not differ from that of WT. In contrast, photosystem I (PSI) activity of the Costata 2/133 mutant measured by the far-red (FR) light inducible P700 (P700(+)) signal exhibited 39% lower steady state level of P700(+), a 2.2-fold higher intersystem electron pool size (e(-)/P700) and higher rate of P700(+) re-reduction, which indicate an increased capacity for PSI cyclic electron transfer (CET) in the Costata 2/133 mutant than WT. The mutant also exhibited a limited capacity for state transitions. The lower level of oxidizable P700 (P700(+)) is consistent with a lower amount of PSI related chlorophyll protein complexes and lower abundance of the PsaA/PsaB heterodimer, PsaD and Lhca1 polypeptides in Costata 2/133 mutant. Exposure of WT and the Costata 2/133 mutant to high light stress resulted in a comparable photoinhibition of PSII measured in vivo, although the decrease of Fv/Fm was modestly higher in the mutant plants. However, under the same photoinhibitory conditions PSI photochemistry (P700(+)) measured as ΔA820-860 was inhibited to a greater extent (50%) in the Costata 2/133 mutant than in the WT (22%). This was accompanied by a 50% faster re-reduction rate of P700(+) in the dark indicating a higher capacity for CET around PSI in high light treated mutant leaves. The role of chloroplast thylakoid organization on the stability of the PSI complex and its susceptibility to high light stress is discussed.
Assuntos
Complexos de Proteínas Captadores de Luz/química , Luz , Mutação , Complexo de Proteína do Fotossistema I/antagonistas & inibidores , Pisum sativum/genética , Pisum sativum/efeitos da radiação , Multimerização Proteica/genética , Clorofila/metabolismo , Clorofila A , Complexos de Proteínas Captadores de Luz/metabolismo , Pisum sativum/enzimologia , Pisum sativum/metabolismo , Complexo de Proteína do Fotossistema I/química , Complexo de Proteína do Fotossistema I/metabolismo , Estrutura Quaternária de Proteína , Espectrometria de FluorescênciaRESUMO
The effect of the exogenously added quercetin against the UV-B inhibition of the photosystem II (PSII) functions in isolated pea thylakoid membranes suspended at different pH of the medium (6.5, 7.6 and 8.4) was investigated. The data revealed that the interaction of this flavonoid with the membranes depends on the pH and influences the initial S0-S1 state distribution of PSII in the dark, the energy transfer between pigment-protein complexes of the photosynthetic apparatus and the membrane fluidity. Quercetin also displays a different UV-protective effect depending on its location in the membranes, as the effect is more pronounced at pH 8.4 when it is located at the membrane surface. The results suggest that quercetin induces structural changes in thylakoid membranes, one of the possible reasons for its protection of the photosynthetic apparatus.
Assuntos
Antioxidantes/farmacologia , Fotossíntese/efeitos da radiação , Pisum sativum/efeitos da radiação , Quercetina/farmacologia , Raios Ultravioleta , Concentração de Íons de Hidrogênio , Pisum sativum/efeitos dos fármacos , Pisum sativum/metabolismo , Fotossíntese/efeitos dos fármacos , Complexo de Proteína do Fotossistema II/metabolismo , Complexo de Proteína do Fotossistema II/efeitos da radiação , Tilacoides/metabolismoRESUMO
Plant responses to light involve a complex network of interactions among multiple plant hormones. In a screen for mutants showing altered photomorphogenesis under red light, we identified a mutant with dramatically enhanced leaf expansion and delayed petal senescence. We show that this mutant exhibits reduced sensitivity to ethylene and carries a nonsense mutation in the single pea (Pisum sativum) ortholog of the ethylene signaling gene ETHYLENE INSENSITIVE2 (EIN2). Consistent with this observation, the ein2 mutation rescues the previously described effects of ethylene overproduction in mature phytochrome-deficient plants. In seedlings, ein2 confers a marked increase in leaf expansion under monochromatic red, far-red, or blue light, and interaction with phytochromeA, phytochromeB, and long1 mutants confirms that ein2 enhances both phytochrome- and cryptochrome-dependent responses in a LONG1-dependent manner. In contrast, minimal effects of ein2 on seedling development in darkness or high-irradiance white light show that ethylene is not limiting for development under these conditions. These results indicate that ethylene signaling constrains leaf expansion during deetiolation in pea and provide further evidence that down-regulation of ethylene production may be an important component mechanism in the broader control of photomorphogenic development by phytochrome and cryptochrome.
Assuntos
Etilenos/metabolismo , Fitocromo/metabolismo , Pisum sativum/fisiologia , Reguladores de Crescimento de Plantas/metabolismo , Proteínas de Plantas/metabolismo , Criptocromos/metabolismo , Escuridão , Regulação para Baixo , Regulação da Expressão Gênica no Desenvolvimento , Regulação da Expressão Gênica de Plantas , Luz , Dados de Sequência Molecular , Mutação , Pisum sativum/genética , Pisum sativum/crescimento & desenvolvimento , Pisum sativum/efeitos da radiação , Folhas de Planta/genética , Folhas de Planta/crescimento & desenvolvimento , Folhas de Planta/fisiologia , Folhas de Planta/efeitos da radiação , Proteínas de Plantas/genética , Plântula/genética , Plântula/crescimento & desenvolvimento , Plântula/fisiologia , Plântula/efeitos da radiação , Transdução de SinaisRESUMO
Plants synthesize phenolic compounds in response to certain environmental signals or stresses. One large group of phenolics, flavonoids, is considered particularly responsive to ultraviolet (UV) radiation. However, here we demonstrate that solar blue light stimulates flavonoid biosynthesis in the absence of UV-A and UV-B radiation. We grew pea plants (Pisum sativum cv. Meteor) outdoors, in Finland during the summer, under five types of filters differing in their spectral transmittance. These filters were used to (1) attenuate UV-B; (2) attenuate UV-B and UV-A < 370 nm; (3) attenuate UV-B and UV-A; (4) attenuate UV-B, UV-A and blue light; and (5) as a control not attenuating these wavebands. Attenuation of blue light significantly reduced the flavonoid content in leaf adaxial epidermis and reduced the whole-leaf concentrations of quercetin derivatives relative to kaempferol derivatives. In contrast, UV-B responses were not significant. These results show that pea plants regulate epidermal UV-A absorbance and accumulation of individual flavonoids by perceiving complex radiation signals that extend into the visible region of the solar spectrum. Furthermore, solar blue light instead of solar UV-B radiation can be the main regulator of phenolic compound accumulation in plants that germinate and develop outdoors.
Assuntos
Flavonoides/metabolismo , Pisum sativum/efeitos da radiação , Folhas de Planta/efeitos da radiação , Cor , Pisum sativum/crescimento & desenvolvimento , Pisum sativum/metabolismo , Fenóis/metabolismo , Epiderme Vegetal/metabolismo , Epiderme Vegetal/efeitos da radiação , Folhas de Planta/metabolismo , Raios UltravioletaRESUMO
The acute effects of three typical polyaromatic hydrocarbons (PAHs): naphthalene (Naph), phenanthrene (Phen) and fluoranthene (Flu) on photochemical activity of photosystem II (PSII) in detached leaves of 3-week-old pea plants were studied. The leaves were exposed in water with PAHs under white light for 0.5-72 h. The activity of PSII was examined by prompt and delayed chlorophyll a (Chl a) fluorescence. The effects of PAHs depended on their concentration and exposure time. This dependency was more significant in the presence of chemical stressors (Triton X-100 or acetone) or under high intensity irradiance. Increased content of PAHs and long-term exposure (24-72 h) led to significant reduction of the maximum photochemical quantum efficiency (Fv/Fm) of PS II, changes in the polyphasic fluorescence induction (OJIP), and to decreasing amplitudes of fast and slow components of delayed Chl a fluorescence. The damage of PSII depended on water solubility of a given type of PAHs, their concentration and exposure time. During short-time exposure the compound with highest water-solubility - naphthalene - revealed the strongest effect. During long-time exposure the compounds with low water-solubility -Phen, Flu-revealed the strongest effect as the corresponding PAH accumulates in the thylakoids especially when the solution is oversaturated containing a solid phase. The reduction of PSII activity at the presence of naphthalene (30 mg L(-1)) was accompanied by transient generation of H2O2 as well as swelling of thylakoids and distortion of cell plasma membranes, which was indicated by electron microscopy images. Distortion of thylakoid membranes due to accumulation of PAHs as well as the development of oxidative stress seems to be the main pathways of PAHs influencing the photochemical activity of PS II.
Assuntos
Fluorenos/farmacologia , Naftalenos/farmacologia , Fenantrenos/farmacologia , Complexo de Proteína do Fotossistema II/efeitos dos fármacos , Pisum sativum/efeitos dos fármacos , Hidrocarbonetos Policíclicos Aromáticos/farmacologia , Clorofila/metabolismo , Clorofila A , Fluorenos/química , Fluorescência , Peróxido de Hidrogênio/metabolismo , Luz , Microscopia Eletrônica , Naftalenos/química , Estresse Oxidativo , Pisum sativum/metabolismo , Pisum sativum/efeitos da radiação , Pisum sativum/ultraestrutura , Fenantrenos/química , Fotossíntese/efeitos da radiação , Complexo de Proteína do Fotossistema II/metabolismo , Folhas de Planta/efeitos dos fármacos , Folhas de Planta/metabolismo , Folhas de Planta/efeitos da radiação , Folhas de Planta/ultraestrutura , Hidrocarbonetos Policíclicos Aromáticos/química , Tilacoides/efeitos dos fármacos , Tilacoides/metabolismoRESUMO
In this work, the effect of the exogenously added ascorbate (Asc) against the UV-B inhibition of the photosystem II (PSII) functions in isolated pea thylakoid membranes was studied. The results reveal that Asc decreases the UV-B induced damage of the donor and the acceptor side of PSII during short treatment up to 60 min. The exogenous Asc exhibits a different UV-protective effect on PSII centers in grana and stroma lamellae, as the effect is more pronounced on the PSIIß centers in comparison to PSIIα centers. Data also suggest that one of the possible protective roles of the Asc in photosynthetic membranes is the modification of the oxygen-evolving complex by influence on the initial S(0)-S(1) state distribution in the dark.
Assuntos
Ácido Ascórbico/farmacologia , Fotossíntese/efeitos dos fármacos , Fotossíntese/efeitos da radiação , Complexo de Proteína do Fotossistema II/efeitos da radiação , Pisum sativum/efeitos da radiação , Tilacoides/efeitos da radiação , Raios Ultravioleta , Oxigênio/metabolismo , Pisum sativum/efeitos dos fármacosRESUMO
The seeds with low level of room temperature phosphorescence (RTP) were selected from a lot of air-dry peas (Pisum sativum) with 62% germination. These strong seeds (95-97% germination percentage) in air-dry, imbibed or emerged states were exposed to 905 MHz GSM-band electromagnetic radiation (EMR). The following effects of EMR were observed. Fraction II with higher RTP level appeared in the air-dry seeds. The germination rate decreased 2-3 fold in the air-dry, swollen and sprouting seeds due to an increase in the ratio of the seedlings with morphological defects (from 3 to 38%) and suffocated seeds (from 1 to 15%). We suggest tentative mechanisms to account for the decreased fitness of peas under GSM-band EMR (905 MHz); also discussed is the role of non-enzymatic hydrolysis of carbohydrates and amino-carbonyl reaction in this process.
Assuntos
Telefone Celular , Radiação Eletromagnética , Pisum sativum/efeitos da radiação , Sementes/efeitos da radiação , Germinação/efeitos da radiação , Pisum sativum/fisiologia , Plântula , Sementes/fisiologiaRESUMO
The aim of this work was to determine which processes in air-dry seeds result in bimodal changes of the pea seed quality under the influence of low doses of gamma-radiation. Pea seeds (cv. "Nemchinovsky-85", harvest 2006, 82% germination persentage) were exposed to gamma-radiation at doses of 3, 10 and 100 Gy The germination percentage decreased to 45% four days after irradiation at the dose of 3 Gy, rised up to 87% at doses of 10 Gy, while the dose of 100 Gy killed the most part of seeds. Seed fractions differing in quality were selected using the metod of Room temperature phosphorecsence (RTP): strong seed frasction I from non-irradiated seeds; weak seed fraction II from the seeds irradiated at a dose of 3 Gy; dead seeds from the seeds irradiated at a dose of 100 Gy. ThermoChemiLuminecnsece (TCL) of seed powders and cotyledons was used. It was shown that the increase of the TCL level in the temperature range from 50 to 110 degreesC was associated with the lipid peroxidation products. The TCL level of seeds subjected to gamma-irradiation at a dose of 3 Gy was similar to that of non-irradiated seeds in the temperature range 50 to 100 degreesC. Therefore, lipid peroxidation was not the cause of the abnormal seedling appearance. The TCL level within this temperature range was increased only in seeds subjected to y-irradiation at a dose of 100 Gy. The TCL level at 150 degreesC was in proportion with the exogenous glucose amount. The increased TCL level of seeds subjected to y-irradiation at a dose of 3 Gy at 150 degreesC resulted from the increase of the glucose content. This means that the transition from the fraction of strong seeds into the fraction of weak ones was the result of the activation of hydrolysis processes. Decrease in the water content of seeds testified to utilization of bound water in this process. The decrease of the glucose content in the "improved" seeds subjected to gamma-irradiation at a dose of 10 Gy most probably indicates the participation of glucose in the amino carbonyl reaction. The latter could be the reason for the increased water content in the "improved" seeds and a decreased water permeability of cell membranes.
Assuntos
Glucose/metabolismo , Peroxidação de Lipídeos/efeitos da radiação , Medições Luminescentes/métodos , Pisum sativum/efeitos da radiação , Sementes/efeitos da radiação , Raios gama , Glucose/efeitos da radiação , Metabolismo dos Lipídeos/efeitos da radiação , Pisum sativum/metabolismo , Doses de RadiaçãoRESUMO
Enhanced level of UV-B radiation and heavy metals in irrigated soils due to anthropogenic activities are deteriorating the environmental conditions necessary for growth and development of plants. The present study was undertaken to study the individual and interactive effects of heavy metal nickel (NiCl(2)·6H(2)O; 0.01, 0.1, 1.0 mM) and UV-B exposure (0.4 W m(-2); 45 min corresponds to 1.08 KJ m(-2)) on growth performance and photosynthetic activity of pea (Pisum sativum L.) seedlings. Ni treatment at high doses (0.1 and 1.0 mM Ni) and UV-B alone reduced chlorophyll content and photosynthetic activity (oxygen yield, carbon fixation, photorespiration, and PSI, PSII, and whole chain electron transport activities), and declining trends continued with combined doses. In contrast to this, Ni at 0.01 mM appeared to be stimulatory for photosynthetic pigments and photosynthetic activity, thereby enhanced biomass was observed at this concentration. However, combined dose (UV-B + 0.01 mM Ni) caused inhibitory effects. Carotenoids showed different responses to each stress. Nickel at high doses strongly inhibited PSII activity and the inhibition was further intensified when chloroplasts were simultaneously exposed to UV-B radiation. PSI activity appeared to be more resistant to each stress. High doses of Ni (0.1 and 1.0 mM) and UV-B alone interrupted electron flow at the oxygen evolving complex. Similar damaging effects were caused by 0.01 and 0.1 mM Ni together with UV-B, but the damage extended to PSII reaction center in case of 1.0 mM Ni in combination with UV-B. In conclusion, the results demonstrate that low dose of Ni stimulated the growth performance of pea seedlings in contrast to its inhibitory role at high doses. However, UV-B alone and together with low as well as high doses of Ni proved to be toxic for P. sativum L.
Assuntos
Níquel/farmacologia , Fotossíntese , Pisum sativum/efeitos dos fármacos , Pisum sativum/efeitos da radiação , Plântula/crescimento & desenvolvimento , Estresse Fisiológico , Raios Ultravioleta , Dióxido de Carbono/metabolismo , Clorofila/metabolismo , Cloroplastos/metabolismo , Transporte de Elétrons , Níquel/metabolismo , Oxigênio/metabolismo , Pisum sativum/crescimento & desenvolvimento , Pisum sativum/metabolismo , Complexo de Proteína do Fotossistema I/metabolismo , Complexo de Proteína do Fotossistema II/metabolismo , Folhas de Planta/efeitos dos fármacos , Folhas de Planta/crescimento & desenvolvimento , Folhas de Planta/efeitos da radiação , Plântula/efeitos dos fármacos , Plântula/efeitos da radiaçãoRESUMO
The widespread use of NO(3)(-) fertilization has had a major ecological impact. NH(4)(+) nutrition may help to reduce this impact, although high NH(4)(+) concentrations are toxic for most plants. The underlying tolerance mechanisms are not yet fully understood, although they are thought to include the limitation of C, the disruption of ion homeostasis, and a wasteful NH(4)(+) influx/efflux cycle that carries an extra energetic cost for root cells. In this study, high irradiance (HI) was found to induce a notable tolerance to NH(4)(+) in the range 2.5-10mM in pea plants by inducing higher C availability, as shown by carbohydrate content. This capacity was accompanied by a general lower relative N content, indicating that tolerance is not achieved through higher net N assimilation on C-skeletons, and it was also not attributable to increased GS content or activity in roots or leaves. Moreover, HI plants showed higher ATP content and respiration rates. This extra energy availability is related to the internal NH(4)(+) content regulation (probably NH(4)(+) influx/efflux) and to an improvement of the cell ionic balance. The limited C availability at lower irradiance (LI) and high NH(4)(+) resulted in a series of metabolic imbalances, as reflected in a much higher organic acid content, thereby suggesting that the origin of the toxicity in plants cultured at high NH(4)(+) and LI is related to their inability to avoid large-scale accumulation of the NH(4)(+) ion.
Assuntos
Adaptação Fisiológica/efeitos dos fármacos , Trifosfato de Adenosina/metabolismo , Sulfato de Amônio/farmacologia , Carbono/metabolismo , Pisum sativum/metabolismo , Estresse Fisiológico/efeitos dos fármacos , Adaptação Fisiológica/efeitos da radiação , Trifosfato de Adenosina/análise , Sulfato de Amônio/análise , Sulfato de Amônio/metabolismo , Carboidratos/análise , Carbono/análise , Glutamato-Amônia Ligase/metabolismo , Nitratos/análise , Nitratos/metabolismo , Nitrogênio/análise , Nitrogênio/metabolismo , Pisum sativum/efeitos dos fármacos , Pisum sativum/efeitos da radiação , Fótons , Fotossíntese/efeitos dos fármacos , Folhas de Planta/enzimologia , Folhas de Planta/crescimento & desenvolvimento , Folhas de Planta/metabolismo , Raízes de Plantas/enzimologia , Raízes de Plantas/crescimento & desenvolvimento , Raízes de Plantas/metabolismo , Estresse Fisiológico/efeitos da radiaçãoRESUMO
The dose dependencies of growth and cytogenetical values have been built to determine the critical level of root apical meristem damage induced by cute irradiation in the range from 2 to 20 Gr. We have analyzed the frequencies of aberrant anaphases and the aberration distribution per cell, on the one hand, and the growth of biomass, the survival and regeneration of the root meristem, on the other hand. The critical level of damage to the stem apical meristem and root of seedlings was defined as 44-48% of aberrant anaphase. Exceeding of this level leads to the launch of suicidal program through induction of multiaberrant damages and interphase cell death. It appears that competition of clones of non-aberrant cells, the cells bearing 1 and 2 damages and multiaberrant cells plays the primary role in the mechanisms of recovery. The regeneration provides full or partial restoration of the main root apical meristem. However these local processes are insufficient to restore morphogenesis and survival of seedlings in excess of the critical level damage.
Assuntos
Aberrações Cromossômicas/efeitos da radiação , Raios gama , Meristema/efeitos da radiação , Pisum sativum/efeitos da radiação , Regeneração/efeitos da radiação , Biomassa , Cromossomos de Plantas/genética , Cromossomos de Plantas/ultraestrutura , Análise Citogenética , Relação Dose-Resposta à Radiação , Meristema/genética , Meristema/crescimento & desenvolvimento , Meristema/fisiologia , Meristema/ultraestrutura , Pisum sativum/genética , Pisum sativum/crescimento & desenvolvimento , Pisum sativum/fisiologia , Pisum sativum/ultraestrutura , Doses de Radiação , Regeneração/genéticaRESUMO
Ultraviolet-B radiation regulates plant growth and morphology at low and ambient fluence rates but can severely impact on plants at higher doses. Some plant UV-B responses are related to the formation of reactive oxygen species (ROS) and pyridoxine (vitamin B(6)) has been reported to be a quencher of ROS. UV-B irradiation of Arabidopsis Col-0 plants resulted in increased levels of PDX1 protein, compared with UV-A-exposed plants. This was shown by immunoblot analysis using specific polyclonal antibodies raised against the recombinant PDX1.3 protein and confirmed by mass spectrometry analysis of immunoprecipitated PDX1. The protein was located mainly in the cytosol but also to a small extent in the membrane fraction of plant leaves. Immunohistochemical analysis performed in pea revealed that PDX1 is present in UV-B-exposed leaf mesophyll and palisade parenchyma but not in epidermal cells. Pyridoxine production increased in Col-0 plants exposed to 3 days of UV-B, whereas in an Arabidopsis pdx1.3 mutant UV-B did not induce pyridoxine biosynthesis. In gene expression studies performed after UV-B exposure, the pdx1.3 mutant showed elevated transcript levels for the LHCB1*3 gene (encoding a chlorophyll a/b-binding protein of the photosystem II light-harvesting antenna complex) and the pathogenesis-related protein 5 (PR-5) gene, compared with wild type.
Assuntos
Adaptação Fisiológica , Proteínas de Arabidopsis/metabolismo , Arabidopsis/enzimologia , Transferases de Grupos Nitrogenados/metabolismo , Pisum sativum/enzimologia , Piridoxina/biossíntese , Estresse Fisiológico , Raios Ultravioleta , Adaptação Fisiológica/genética , Arabidopsis/genética , Arabidopsis/efeitos da radiação , Proteínas de Arabidopsis/genética , Carbono-Nitrogênio Liases , Membrana Celular/efeitos da radiação , Citosol/efeitos da radiação , Expressão Gênica , Complexos de Proteínas Captadores de Luz/genética , Complexos de Proteínas Captadores de Luz/metabolismo , Mutação , Transferases de Grupos Nitrogenados/genética , Pisum sativum/efeitos da radiação , Complexo de Proteína do Fotossistema II/fisiologia , Complexo de Proteína do Fotossistema II/efeitos da radiação , Folhas de Planta/metabolismo , Folhas de Planta/efeitos da radiação , Folhas de Planta/ultraestrutura , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Proteínas Recombinantes/metabolismo , Estresse Fisiológico/genéticaRESUMO
Photosystem I (PSI) is severely damaged by chilling at 4 degrees C in low light, especially in the chilling sensitive plant cucumber. To investigate the early events in PSI photoinhibition, we examined structural changes in the level of pigment-protein complexes in cucumber leaves in comparison with pea leaves. The complexes were separated on a native green gel and an increase in the intensity of a band was observed only in light-chilled cucumber leaves. The 77 K fluorescence emission spectrum of this green band indicated that the band was mainly composed of PSI with light-harvesting complex I. Each lane was cut from the green gel and separated on a fully denaturing SDS-PAGE in the second dimension. The new green gel band observed after light-chilling in cucumber leaves lacked 19, 18, and 16.5 kDa polypeptides. These results suggest that light-chilling facilitates the release of three peripheral polypeptides as an early event of chilling stress in vivo, which results in the inactivation of PSI in intact cucumber leaves.
Assuntos
Cucumis sativus/efeitos da radiação , Peptídeos/efeitos da radiação , Complexo de Proteína do Fotossistema I/efeitos da radiação , Folhas de Planta/efeitos da radiação , Proteínas de Plantas/efeitos da radiação , Temperatura Baixa , Escuridão , Eletroforese em Gel Bidimensional , Luz , Pisum sativum/efeitos da radiação , Fotossíntese , Complexo de Proteína do Fotossistema I/isolamento & purificação , Proteínas de Plantas/isolamento & purificação , Espectrometria de FluorescênciaRESUMO
* Tetrahydrofolate derivatives are central cofactors of C1 metabolism. Using methotrexate as a specific inhibitor of folate biosynthesis, we altered the folate status in 10-d-old etiolated pea (Pisum sativum) leaves and followed the rate of chlorophyll synthesis upon illumination. * In our conditions, the folate concentration decreased only from 5.7 to 4.2 nmol g(-1) FW, but the amount of chlorophyll after 24 h of illumination was reduced 2.5 times. Folate status and rate of chlorophyll synthesis were apparently correlated through the methyl cycle. * Indeed, we observed that methyl-tetrahydrofolate was the folate derivative most affected by the treatment; the decrease of methyl-tetrahydrofolate was associated with a sharp rise in homocysteine and S-adenosylhomocysteine concentrations, which are normally maintained at very low values, shifting the methylation index (S-adenosylmethionine/S-adenosylhomocysteine ratio) from 7 to 1; the decrease of the methylation index reduced by a factor of 3 the activity of the Mg-protoporphyrin IX methyltransferase (CHLM), an essential enzyme for chlorophyll synthesis. CHLM gene expression and protein concentration remained unchanged, suggesting that this inhibition relied essentially on metabolic regulation. * These results point out that an even moderate change in the folate status may affect plant development and adaptation.
Assuntos
Arabidopsis/enzimologia , Carbono/metabolismo , Clorofila/biossíntese , Ácido Fólico/metabolismo , Metiltransferases/metabolismo , Pisum sativum/enzimologia , Arabidopsis/efeitos dos fármacos , Arabidopsis/efeitos da radiação , Luz , Metotrexato/farmacologia , Metilação/efeitos dos fármacos , Metilação/efeitos da radiação , Pisum sativum/efeitos dos fármacos , Pisum sativum/efeitos da radiação , Folhas de Planta/efeitos dos fármacos , Folhas de Planta/enzimologia , Folhas de Planta/efeitos da radiação , Tetra-Hidrofolatos/química , Tetra-Hidrofolatos/metabolismoRESUMO
Ultraviolet radiation (UV) though harmful but is an important and unavoidable component of terrestrial ecosystem to which plants have been exposed since their migration from aquatic to land habitat. Incoming UV-B radiation and heavy metals abundance in contaminated soils are significant environmental threat affecting metabolic functions of plants through generation of reactive oxygen species. Plants have evolved mechanisms to counteract these reactive radicals and to repair the damage caused by UV-B and heavy metals. This study describes the impact of supplemental UV-B (sUV-B) and nickel (Ni) singly and in combination on photosynthetic pigments, flavonoids, enzymatic and non-enzymatic antioxidants, metabolites and lipid peroxidation of Pisum sativum L. (pea) plants. Compared to the controls, both the stresses individually and in combination led to reductions in photosynthetic pigments, ascorbic acid, protein and catalase (CAT) activity whereas a reverse trend was observed for flavonoids, phenol, proline and thiol contents, superoxide dismutase (SOD) and peroxidase (POX) activities and lipid peroxidation (LPO). However, flavonoids increased significantly under individual exposure of sUV-B as compared to other treatments. An increase of LPO by 81% indicated the generation of reactive oxygen species under both the stress conditions. sUV-B and Ni in combination acted synergistically with stimulation of CAT activity by 51.6%, additively on SOD activity with increase of 16.4%, while other parameters showed antagonistic action of both the stresses.
Assuntos
Antioxidantes/metabolismo , Níquel/farmacologia , Pigmentos Biológicos/metabolismo , Pisum sativum/metabolismo , Raios Ultravioleta , Ácido Ascórbico/metabolismo , Carotenoides/metabolismo , Clorofila/metabolismo , Flavonoides/metabolismo , Peroxidação de Lipídeos/efeitos dos fármacos , Peroxidação de Lipídeos/efeitos da radiação , Pisum sativum/efeitos dos fármacos , Pisum sativum/efeitos da radiação , Fenóis/metabolismo , Proteínas de Plantas/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Estresse FisiológicoRESUMO
The impact of supplemental UV-B irradiation (sUV-B) and cadmium (Cd, 68 micromol kg(-1) soil) singly and in combination was studied on growth, pigments, enzymatic and non-enzymatic antioxidants, lipid peroxidation (LPO), uptake and translocation of Cd and yield of pea plants under field condition. Compared to the control, both the stresses individually and in combination led to in reductions in growth, photosynthetic pigments, ascorbic acid, catalase (CAT) activity and yield, whereas a reverse trend was observed for flavonoids, thiols and proline contents, superoxide dismutase (SOD) and peroxidase (POD) activities, and LPO. The uptake and translocation of Cd increased in different plant parts with duration of its treatment and also when applied with sUV-B. Combined treatment of sUV-B and Cd inhibited the growth and various metabolic processes antagonistically except the CAT activity which showed additive response. Further, CAT activity may be used as a bioindicator parameter to evaluate the individual and interactive effects of both the stresses.
Assuntos
Antioxidantes/metabolismo , Cádmio/toxicidade , Peroxidação de Lipídeos , Fotossíntese , Pisum sativum , Poluentes do Solo/toxicidade , Raios Ultravioleta , Cádmio/metabolismo , Catalase/metabolismo , Peroxidação de Lipídeos/efeitos dos fármacos , Peroxidação de Lipídeos/fisiologia , Peroxidação de Lipídeos/efeitos da radiação , Pisum sativum/efeitos dos fármacos , Pisum sativum/fisiologia , Pisum sativum/efeitos da radiação , Peroxidases/metabolismo , Fotossíntese/efeitos dos fármacos , Fotossíntese/efeitos da radiação , Prolina/metabolismo , Poluentes do Solo/metabolismo , Compostos de Sulfidrila/metabolismo , Superóxido Dismutase/metabolismoRESUMO
Cleaned, whole smooth green peas (Pisum sativum L.) were reconstituted by soaking in tap water of 40 degrees C (15, 20 or 25 min) and subsequently either toasted (100 degrees C during 1.5 min) and infrared (IR) radiated or just IR radiated. For IR radiation, a small-scale, propane-fired IR radiation plant was used with average residence times of 58 and 92 s respectively. After exiting the conveyor belt, peas were held for a pre-determined period (holding: 0, 15 or 30 min respectively) in a well-insulated container. Finally, all radiated peas were flaked (roll distance 0.75 mm) in a flaking mill located posterior to the IR plant and analysed for chemical and physical properties. Initial pea starch gelatinization degree (SGD) was 10.1% at a total starch content of 410.1 g/kg. Infrared processing during 92 s significantly improved the SGD (from 10.1% to 32.8%) of pea flakes compared to treatment during 58 s (SGD of 18.6%). The SGD was further improved with steam treatment of peas, prior to IR. For all determined parameters, no effect of holding time could be observed. Starch gelatinization degree can be improved by soaking, toasting and IR processing. The substantial improvement of SGD, however can be only obtained by a longer IR residence time as well as through toasting, prior to the IR processing. The improvements in SGD, however are paralleled by a lower durability of flakes (range 34.9% to 87.4%).