RESUMEN
PURPOSE: To provide an updated summary of recent advances in the application of gamma irradiation to elicit secondary metabolism and for induction of mutations in plant cell and organ cultures for the production of industrially important specialized metabolites (SMs). CONCLUSIONS: Research on the application of gamma radiation with plants has contributed a lot to microbial decontamination of seeds, and the promotion of physiological processes such as seed germination, seedling vigor, plant growth, and development. Various studies have demonstrated the influence of gamma rays on the morphology, physiology, and biochemistry of plants. Recent research efforts have also shown that low-dose gamma (5-100 Gy) irradiation can be utilized as an expedient solution to alleviate the deleterious effect of abiotic stresses and to obtain better yields of plants. Inducing mutagenesis using gamma irradiation has also evolved as a better option for inducing genetic variability in crops, vegetables, medicinal and ornamentals for their genetic improvement. Plant SMs are gaining increasing importance as pharmaceutical, therapeutic, cosmetic, and agricultural products. Plant cell, tissue, and organ cultures represent an attractive alternative to conventional methods of procuring useful SMs. Among the varied approaches the elicitor-induced in vitro culture techniques are considered an efficient tool for studying and improving the production of SMs. This review focuses on the utilization of low-dose gamma irradiation in the production of high-value SMs such as phenolics, terpenoids, and alkaloids. Furthermore, we present varied successful examples of gamma-ray-induced mutations in the production of SMs.
Asunto(s)
Rayos gamma , Células Vegetales , Metabolismo Secundario , Metabolismo Secundario/efectos de la radiación , Células Vegetales/metabolismo , Células Vegetales/efectos de la radiaciónRESUMEN
Fagonia indica (L.) is an important medicinal plant with multitude of therapeutic potentials. Such application has been attributed to the presence of various pharmacological important phytochemicals. However, the inadequate biosynthesis of such metabolites in intact plants has hampered scalable production. Thus, herein, we have established an in vitro based elicitation strategy to enhance such metabolites in callus culture of F. indica. Cultures were exposed to various doses of UV radiation (UV-C) and grown in different photoperiod regimes and their impact was evaluated on biomass accumulation, biosynthesis of phytochemicals along antioxidant expression. Cultures grown under photoperiod (16L/8D h) after exposure to UV-C (5.4 kJ/m2) accumulated optimal biomass (438.3 g/L FW; 16.4 g/L DW), phenolics contents (TPC: 11.8 µgGAE/mg) and flavonoids contents (TFC: 4.05 µgQE/mg). Similarly, HPLC quantification revealed that total production (6.967 µg/mg DW) of phytochemicals wherein kaempferol (1.377 µg/mg DW), apigenin (1.057 µg/mg DW), myricetin (1.022 µg/mg DW) and isorhamnetin (1.022 µg/mg DW) were recorded highly accumulated compounds in cultures at UV-C (5.4 kJ/m2) dose than other UV-C radiations and light regimes.. The antioxidants activities examined as DPPH (92.8%), FRAP (182.3 µM TEAC) and ABTS (489.1 µM TEAC) were also recorded highly expressed by cultures under photoperiod after treatment with UV-C dose 5.4 kJ/m2. Moreover, same cultures also expressed maximum % inhibition towards phospholipase A2 (sPLA2: 35.8%), lipoxygenase (15-LOX: 43.3%) and cyclooxygenases (COX-1: 55.3% and COX-2: 39.9%) with 1.0-, 1.3-, 1.3- and 2.8-fold increased levels as compared with control, respectively. Hence, findings suggest that light and UV can synergistically improve the metabolism of F. indica and could be used to produce such valuable metabolites on commercial scale.
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Antioxidantes/metabolismo , Luz , Fitoquímicos/metabolismo , Células Vegetales/metabolismo , Rayos Ultravioleta , Zygophyllaceae/metabolismo , Antioxidantes/efectos de la radiación , Técnicas In Vitro , Fitoquímicos/efectos de la radiación , Células Vegetales/efectos de la radiación , Zygophyllaceae/crecimiento & desarrollo , Zygophyllaceae/efectos de la radiaciónRESUMEN
Plants often encounter light intensities exceeding the capacity of photosynthesis (excessive light) mainly due to biotic and abiotic factors, which lower CO2 fixation and reduce light energy sinks. Under excessive light, the photosynthetic electron transport chain generates damaging molecules, hence leading to photooxidative stress and eventually to cell death. In this review, we summarize the mechanisms linking the excessive absorption of light energy in chloroplasts to programmed cell death in plant leaves. We highlight the importance of reactive carbonyl species generated by lipid photooxidation, their detoxification, and the integrating role of the endoplasmic reticulum in the adoption of phototolerance or cell-death pathways. Finally, we invite the scientific community to standardize the conditions of excessive light treatments.
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Luz/efectos adversos , Células Vegetales/efectos de la radiación , Apoptosis/efectos de la radiación , Cloroplastos/efectos de la radiación , Retículo Endoplásmico/efectos de la radiación , Peroxidación de Lípido/efectos de la radiación , Reguladores del Crecimiento de las Plantas/fisiología , Hojas de la Planta/citología , Hojas de la Planta/efectos de la radiaciónRESUMEN
Solanum aculeatissimum Jacq. is a common plant in much of Brazil. Despite containing metabolites with a wide range of pharmacological applications, there are few tissue culture reports for this plant. The possibility of large-scale in vitro production of this material has significant biotechnological potential. Therefore, the objective of this study was to investigate the effect of light conditions on the growth of cells in suspension, observing the production and yield of biomass and bioactive compounds and the enzymatic behavior. Calli obtained from leaf segments were cultured in solid medium supplemented with 1 mg L-1 of 2,4-D, 2.5 mg L-1 kinetin, pH 5.7, in the dark. After 110 days of subculture, the calli were transferred to liquid medium. Cells were kept in the dark under agitation at 110 rpm and 25 °C and subcultured every 30 days. After 90 days of culture, 20 mL aliquots of cell suspension were added to flasks containing approximately 20 mL of medium (1:1) and cultured at different wavelengths (white, green, blue, red, and blue/red) under a photoperiod of 16 h with irradiance of 50 µmol m-2 s-1) and in the absence of light. The experiment was performed in a 6 × 6 factorial design (light condition × culture time). The cell cultures showed viability throughout the entire cycle, and chlorogenic and ferulic acids, orientin, quercitrin and, in higher amounts, quercetin, were detected in the first 7 days of culture. There was an increase in superoxide dismutase and catalase and a decrease in ascorbate peroxidase after exposure to different light conditions; for phenylalanine ammonia lyase, no differences were observed. The different light conditions were not sufficient to trigger responses in the concentrations of bioactive compounds, despite the detection of increased levels of the enzymes involved in cellular homeostasis.
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Luz , Solanum/metabolismo , Catalasa/metabolismo , Técnicas de Cultivo de Célula , Ácido Clorogénico/metabolismo , Conductividad Eléctrica , Flavonoides/metabolismo , Glucósidos/metabolismo , Concentración de Iones de Hidrógeno , Células Vegetales/metabolismo , Células Vegetales/efectos de la radiación , Hojas de la Planta/citología , Hojas de la Planta/metabolismo , Quercetina/análogos & derivados , Quercetina/metabolismo , Solanum/citología , Superóxido Dismutasa/metabolismoRESUMEN
Plasmodesmata (PD) are essential for plant development, but little is known about their regulation. Several studies have linked PD transport to chloroplast-centered signaling networks, but the physiological significance of this connection remains unclear. Here, we show that PD transport is strongly regulated by light and the circadian clock. Light promotes PD transport during the day, but light is not sufficient to increase rates of PD transport at night, suggesting a circadian gating mechanism. Silencing expression of the core circadian clock gene, LHY/CCA1, allows light to strongly promote PD transport during subjective night, confirming that the canonical plant circadian clock controls the PD transport light response. We conclude that PD transport is dynamically regulated during the day/night cycle. Due to the many roles of PD in plant biology, this discovery has strong implications for plant development, physiology, and pathogenesis.
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Arabidopsis/fisiología , Relojes Circadianos/efectos de la radiación , Luz , Nicotiana/fisiología , Células Vegetales/metabolismo , Células Vegetales/efectos de la radiación , Plasmodesmos/metabolismo , Plasmodesmos/efectos de la radiación , Arabidopsis/efectos de la radiación , Transporte Biológico/efectos de la radiación , Fotoperiodo , Hojas de la Planta/crecimiento & desarrollo , Hojas de la Planta/efectos de la radiación , Nicotiana/efectos de la radiaciónRESUMEN
Effect of metal oxide nanoparticles on calli of two wheat varieties: Parabola (stress tolerant) and Raweta (sensitive) was studied. ZnO induced 10% larger membrane damage in Raweta calli. TiO2, Al2O3, and ZrO2 caused nearly 30% greater lactate dehydrogenase leakage for Raweta compared to Parabola. UV-irradiation of samples containing ZnO particles intensified this effect. Membrane lipid peroxidation in ZnO treated Raweta calli was twice as high as in Parabola and further increased after UV-irradiation. TiO2, Al2O3, and ZrO2 nanoparticles caused a 4-fold increase in malondialdehyde concentration in Raweta calli in comparison to Parabola calli. The nanoparticles studied damaged the cellular defense system by inactivating the antioxidative enzymes.
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Óxido de Aluminio/toxicidad , Nanopartículas del Metal/química , Titanio/toxicidad , Triticum/efectos de los fármacos , Óxido de Zinc/toxicidad , Circonio/toxicidad , Óxido de Aluminio/química , Membrana Celular/metabolismo , Supervivencia Celular , Protección de Cultivos/métodos , L-Lactato Deshidrogenasa/antagonistas & inhibidores , Peroxidación de Lípido , Malondialdehído/metabolismo , Lípidos de la Membrana/química , Estrés Oxidativo , Tamaño de la Partícula , Peroxidasa/antagonistas & inhibidores , Células Vegetales/efectos de los fármacos , Células Vegetales/metabolismo , Células Vegetales/efectos de la radiación , Superóxido Dismutasa/antagonistas & inhibidores , Titanio/química , Triticum/citología , Triticum/enzimología , Rayos Ultravioleta/efectos adversos , Óxido de Zinc/química , Circonio/químicaRESUMEN
Cnidium officinale Makino is an important medicinal plant of oriental clinics and is considered as the main source of phthalides, polyphenols, and flavonoids. However, there is no available report regarding the effect of different light colors on the secondary metabolites composition of C. officinale. In this study different light (dark, white, blue, red and red: blue) conditions were arranged to raise callus on MS medium containing 0.5â¯mg·L-1 of each 2,4-D and BAP. Callus grown in dark condition showed maximum (2.0â¯g) fresh weight with lower total phenolic and flavonoids contents. Also, in dark condition callus faced higher catalase (CAT) and guaiacol peroxidase (GPX) activities to avoid free radicals. Mix (red: blue) light condition favored the synthesis of phenolics and flavonoids in callus at the cost of higher ascorbate peroxidase (APX) and superoxide dismutase (SOD) enzymes expression. However, DPPH free radical scavenging activity was less variable among the samples from the different light conditions. Interestingly, the HPLC profile showed higher (28.3⯵g·g-1 DW) phthalide accumulation in dark grown-cultures. Compared to other light conditions, 3-butyledinephthalide accumulation was higher (0.43⯵g·g-1 DW) in white light-grown callus. These findings suggest that light conditions play an important role in the regulation of in vitro callus growth and synthesis of important medicinal compounds of C. officinale.
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Antioxidantes/metabolismo , Cnidium/química , Flavonoides/metabolismo , Luz , Fenoles/metabolismo , Antioxidantes/química , Ascorbato Peroxidasas/metabolismo , Benzofuranos/análisis , Benzofuranos/metabolismo , Cromatografía Líquida de Alta Presión , Cnidium/crecimiento & desarrollo , Cnidium/metabolismo , Flavonoides/química , Fenoles/química , Células Vegetales/metabolismo , Células Vegetales/efectos de la radiación , Plantas Medicinales/química , Plantas Medicinales/metabolismo , Superóxido Dismutasa/metabolismoRESUMEN
Lepidium sativum L. is an important edible, herbaceous plant with huge medicinal value as cardio-protective, hepatoprotective and antitumor agent. This study was designed and performed to investigate biosynthesis of plant's active ingredients in callus cultures of L.â¯sativum in response to the exposure of multi spectral lights. Optimum biomass accumulation (15.36â¯g/L DW), total phenolic and flavonoid contents (TPC; 47.43â¯mg/g; TFC; 9.41â¯mg/g) were recorded in calli placed under white light (24â¯h) compared to rest of the treatments. Antioxidant enzymatic activities i.e. superoxide dismutase and peroxidase were found optimum in cultures exposed to green light (SOD; 0.054â¯nM/min/mg FW, POD; 0.501â¯nM/min/mg FW). Phytochemical analysis further confirmed the potential influence of white light exposure on enhanced production of plant's metabolites. Significant enhancement level of major metabolic compounds such as chlorogenic acid (7.20â¯mg/g DW), quercetin (22.08â¯mg/g DW), kaempferol (7.77â¯mg/g DW) and minor compounds including ferulic acid, sinapic acid, protocatechuic acid, vanillic acid and caffeic acid were recorded in white light compared to control (photoperiod), whereas blue light increased the p-coumaric acid accumulation. Moreover, callus cultures of this plant under white light (24â¯h) showed highest in vitro based anti-diabetic and antioxidant activities compared to other conditions. Finding of our current study revealed that multi spectral lights are proved to be an effective strategy for enhancing metabolic quantity of antioxidant and anti-diabetic bioactive compounds in callus cultures of L. sativum L.
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Antioxidantes/química , Hipoglucemiantes/metabolismo , Lepidium sativum/metabolismo , Luz , Polifenoles/metabolismo , Biomasa , Cromatografía Líquida de Alta Presión , Hipoglucemiantes/análisis , Hipoglucemiantes/química , Lepidium sativum/crecimiento & desarrollo , Peroxidasa/metabolismo , Fitoquímicos/análisis , Células Vegetales/metabolismo , Células Vegetales/efectos de la radiación , Proteínas de Plantas/metabolismo , Polifenoles/análisis , Polifenoles/química , Superóxido Dismutasa/metabolismoRESUMEN
Genome engineering in plants is highly dependent on the availability of effective molecular techniques. Despite vast quantities of research, genome engineering in plants is still limited in terms of gene delivery, which requires the use of infectious bacteria or harsh conditions owing to the difficulty delivering biomaterial into plant cells through the cell wall. Here, we describe a method that uses electroporation-mediated protein delivery into cultured Arabidopsis thaliana cells possessing an intact cell wall, and demonstrate Cre-mediated site-specific recombination. By optimizing conditions for the electric pulse, protein concentration, and electroporation buffer, we were able to achieve efficient and less-toxic protein delivery into Arabidopsis thaliana cells with 83% efficiency despite the cell wall. To the best of our knowledge, this is the first report demonstrating the electroporation-mediated protein delivery of Cre recombinase to achieve nucleic acid-free genome engineering in plant cells possessing an intact cell wall.
Asunto(s)
Arabidopsis/efectos de la radiación , Pared Celular/efectos de la radiación , Electroporación/métodos , Endocitosis , Integrasas/metabolismo , Células Vegetales/efectos de la radiación , Transporte de Proteínas , Arabidopsis/metabolismo , Pared Celular/metabolismo , Células Vegetales/metabolismoRESUMEN
Heat Shock Factor A2 (HsfA2) is part of the Heat Shock Factor (HSF) network, and plays an essential role beyond heat shock in environmental stress responses and cellular homeostatic control. Arabidopsis thaliana cell cultures derived from wild type (WT) ecotype Col-0 and a knockout line deficient in the gene encoding HSFA2 (HSFA2 KO) were grown aboard the International Space Station (ISS) to ascertain whether the HSF network functions in the adaptation to the novel environment of spaceflight. Microarray gene expression data were analyzed using a two-part comparative approach. First, genes differentially expressed between the two environments (spaceflight to ground) were identified within the same genotype, which represented physiological adaptation to spaceflight. Second, gene expression profiles were compared between the two genotypes (HSFA2 KO to WT) within the same environment, which defined genes uniquely required by each genotype on the ground and in spaceflight-adapted states. Results showed that the endoplasmic reticulum (ER) stress and unfolded protein response (UPR) define the HSFA2 KO cells' physiological state irrespective of the environment, and likely resulted from a deficiency in the chaperone-mediated protein folding machinery in the mutant. Results further suggested that additional to its universal stress response role, HsfA2 also has specific roles in the physiological adaptation to spaceflight through cell wall remodeling, signal perception and transduction, and starch biosynthesis. Disabling HsfA2 altered the physiological state of the cells, and impacted the mechanisms induced to adapt to spaceflight, and identified HsfA2-dependent genes that are important to the adaption of wild type cells to spaceflight. Collectively these data indicate a non-thermal role for the HSF network in spaceflight adaptation.
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Adaptación Biológica/genética , Diferenciación Celular , Factores de Transcripción del Choque Térmico/genética , Células Vegetales/metabolismo , Células Vegetales/efectos de la radiación , Vuelo Espacial , Diferenciación Celular/genética , Células Cultivadas , Perfilación de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Técnicas de Inactivación de Genes , Genotipo , Factores de Transcripción del Choque Térmico/metabolismo , Modelos Biológicos , IngravidezRESUMEN
Central to the building and reorganizing cytoskeletal arrays is creation of new polymers. Although nucleation has been the major focus of study for microtubule generation, severing has been proposed as an alternative mechanism to create new polymers, a mechanism recently shown to drive the reorientation of cortical arrays of higher plants in response to blue light perception. Severing produces new plus ends behind the stabilizing GTP-cap. An important and unanswered question is how these ends are stabilized in vivo to promote net microtubule generation. Here we identify the conserved protein CLASP as a potent stabilizer of new plus ends created by katanin severing in plant cells. Clasp mutants are defective in cortical array reorientation. In these mutants, both rescue of shrinking plus ends and the stabilization of plus ends immediately after severing are reduced. Computational modeling reveals that it is the specific stabilization of severed ends that best explains CLASP's function in promoting microtubule amplification by severing and array reorientation.
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Proteínas de Arabidopsis/genética , Arabidopsis/genética , Regulación de la Expresión Génica de las Plantas , Katanina/genética , Proteínas Asociadas a Microtúbulos/genética , Microtúbulos/metabolismo , Modelos Estadísticos , Arabidopsis/metabolismo , Arabidopsis/efectos de la radiación , Arabidopsis/ultraestructura , Proteínas de Arabidopsis/metabolismo , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Genes Reporteros , Katanina/metabolismo , Luz , Fototransducción , Proteínas Luminiscentes/genética , Proteínas Luminiscentes/metabolismo , Proteínas Asociadas a Microtúbulos/metabolismo , Microtúbulos/efectos de la radiación , Microtúbulos/ultraestructura , Mutación , Células Vegetales/metabolismo , Células Vegetales/efectos de la radiación , Células Vegetales/ultraestructura , Estabilidad Proteica , Procesos Estocásticos , Proteína Fluorescente RojaRESUMEN
Catechins, including catechin (C) and epicatechin (E), are the main type of flavonoids in cacao seeds. They play important roles in plant defense and have been associated with human health benefits. Although flavonoid biosynthesis has been extensively studied using in vitro and in vivo models, the regulatory mechanisms controlling their accumulation under light/dark conditions remain poorly understood. To identify differences in flavonoid biosynthesis (particularly catechins) under different light treatments, we used cacao cell suspensions exposed to white-blue light and darkness during 14 days. RNA-Seq was applied to evaluate differential gene expression. Our results indicate that light can effectively regulate flavonoid profiles, inducing a faster accumulation of phenolic compounds and shifting E/C ratios, in particular as a response to switching from white to blue light. The results demonstrated that HY5, MYB12, ANR and LAR were differentially regulated under light/dark conditions and could be targeted by overexpression aiming to improve catechin synthesis in cell cultures. In conclusion, our RNA-Seq analysis of cacao cells cultured under different light conditions provides a platform to dissect key aspects into the genetic regulatory network of flavonoids. These light-responsive candidate genes can be used further to modulate the flavonoid production in in vitro systems with value-added characteristics.
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Cacao/genética , Catequina/biosíntesis , Flavonoides/biosíntesis , Regulación de la Expresión Génica de las Plantas , Proteínas de Plantas/genética , Semillas/genética , Factores de Transcripción/genética , Cacao/citología , Cacao/metabolismo , Cacao/efectos de la radiación , Catequina/genética , Flavonoides/genética , Redes Reguladoras de Genes , Luz , Fotoperiodo , Células Vegetales/metabolismo , Células Vegetales/efectos de la radiación , Proteínas de Plantas/clasificación , Proteínas de Plantas/metabolismo , Semillas/citología , Semillas/metabolismo , Semillas/efectos de la radiación , Análisis de Secuencia de ARN , Factores de Transcripción/clasificación , Factores de Transcripción/metabolismo , TranscriptomaRESUMEN
UVB radiation (290-320 nm) causes diverse effects in plant cells that vary with the fluence rate of exposure. High fluence rates of UVB radiation cause damage to DNA and formation of reactive oxygen species in mitochondria and chloroplasts, which lead to oxidation of membrane proteins and lipids and inhibition of cellular functions. In response to oxidative stress, mitochondrial transmembrane potential dissipates, resulting in cytochrome c release and activation of metacaspases. This leads to the apoptosis-like cell death. The signaling mechanism based on UVB DNA damage includes checkpoint activation, cell-cycle arrest, and finally programmed cell death with characteristic DNA fragmentation and morphological hallmarks typical of apoptotic cells. Recently, it was shown that among the components of this signaling mechanism the transcriptional factor SOG1 (suppressor of gamma response 1) plays a key role in regulation of programmed cell death in plants. In contrast to its damaging effects, UVB radiation at low fluence rates can act as a regulatory signal that is specifically perceived by plants to promote acclimation and survival in sunlight. The protective action of UVB is based on expression of various genes, including those encoding flavonoid synthesis enzymes that provide a UVB-absorbing sunscreen in epidermal tissues and DNA photorepair enzymes. These processes are mediated by the UVB photoreceptor UVR8, which has been recently characterized at the molecular level. Now progress is made in uncovering the UVR8-mediated signaling pathway mechanism in the context of UVB photon perception and revealing the biochemical components of the early stages of light signal transduction. In this review, attention is focused on the achievements in studying these UVB-induced signaling processes.
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Células Vegetales/efectos de la radiación , Transducción de Señal/efectos de la radiación , Rayos Ultravioleta , Animales , Muerte Celular/efectos de la radiación , Relación Dosis-Respuesta en la Radiación , Humanos , Células Vegetales/metabolismo , Proteínas de Plantas/metabolismoRESUMEN
The chloroplast relies on proteins encoded in the nucleus, synthesized in the cytosol and subsequently transported into chloroplast through the protein complexes Toc and Tic (Translocon at the outer/inner membrane of chloroplasts). A Tic complex member, Tic55, contains a redox-related motif essential for protein import into chloroplasts in peas. However, Tic55 is not crucial for protein import in Arabidopsis. Here, a tic55-II-knockout mutant of Arabidopsis thaliana was characterized for Tic55 localization, its relationship with other translocon proteins, and its association with plant leaf senescence when compared to the wild type. Individually darkened leaves (IDLs) obtained through dark-induced leaf senescence were used to demonstrate chlorophyll breakdown and its relationship with plant senescence in the tic55-II-knockout mutant. The IDLs of the tic55-II-knockout mutant contained higher chlorophyll concentrations than those of the wild type. Our microarray analysis of IDLs during leaf senescence identified seven senescence-associated genes (SAGs) that were downregulated in the tic55-II-knockout mutant: ASP3, APG7, DIN2, DIN11, SAG12, SAG13, and YLS9. Real-time quantitative PCR confirmed the reliability of microarray analysis by showing the same expression patterns with those of the microarray data. Thus, Tic55 functions in dark-induced aging in A. thaliana by indirectly regulating downstream SAGs expression. In addition, the expression of four NAC genes, including ANAC003, ANAC010, ANAC042, and ANAC075 of IDL treated tic55-II-knockout mutant appeared to be downregulated. Yeast one hybrid assay revealed that only ANAC003 promoter region can be bound by MYB108, suggesting that a MYB-NAC regulatory network is involved in dark-stressed senescence.
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Proteínas de Arabidopsis/genética , Clorofila/metabolismo , Regulación de la Expresión Génica de las Plantas , Proteínas de Transporte de Membrana/genética , Factores de Transcripción/genética , Secuencia de Aminoácidos , Arabidopsis/clasificación , Arabidopsis/genética , Arabidopsis/metabolismo , Arabidopsis/efectos de la radiación , Proteínas de Arabidopsis/metabolismo , Senescencia Celular , Cloroplastos/genética , Cloroplastos/metabolismo , Cloroplastos/efectos de la radiación , Oscuridad , Técnicas de Inactivación de Genes , Proteínas de Transporte de Membrana/deficiencia , Filogenia , Células Vegetales/metabolismo , Células Vegetales/efectos de la radiación , Hojas de la Planta/genética , Hojas de la Planta/metabolismo , Hojas de la Planta/efectos de la radiación , Regiones Promotoras Genéticas , Unión Proteica , Alineación de Secuencia , Homología de Secuencia de Aminoácido , Transducción de Señal , Factores de Transcripción/metabolismo , Técnicas del Sistema de Dos HíbridosRESUMEN
Ultraviolet B (UV-B) radiation induces the activation of MITOGEN-ACTIVATED PROTEIN KINASE PHOSPHATASE1 (MKP1) and its targets MPK3 and MPK6, but whether they participate in UV-B guard cell signaling is not clear. Here, evidence shows that UV-B-induced stomatal closure in Arabidopsis (Arabidopsis thaliana) is antagonistically regulated by MKP1 and MPK6 via modulating hydrogen peroxide (H2O2)-induced nitric oxide (NO) production in guard cells. The mkp1 mutant was hypersensitive to UV-B-induced stomatal closure and NO production in guard cells but not to UV-B-induced H2O2 production, suggesting that MKP1 negatively regulates UV-B-induced stomatal closure via inhibiting NO generation in guard cells. Moreover, MPK3 and MPK6 were activated by UV-B in leaves of the wild type and hyperactivated in the mkp1 mutant, but the UV-B-induced activation of MPK3 and MPK6 was largely inhibited in mutants for ATRBOHD and ATRBOHF but not in mutants for NIA1 and NIA2 mpk6 mutants showed defects of UV-B-induced NO production and stomatal closure but were normal in UV-B-induced H2O2 production, while stomata of mpk3 mutants responded to UV-B just like those of the wild type. The defect of UV-B-induced stomatal closure in mpk6 mutants was rescued by exogenous NO but not by exogenous H2O2 Furthermore, double mutant mkp1/mpk6 and the single mutant mpk6 showed the same responses to UV-B in terms of either stomatal movement or H2O2 and NO production. These data indicate that MPK6, but not MPK3, positively regulates UV-B-induced stomatal closure via acting downstream of H2O2 and upstream of NO, while MKP1 functions negatively in UV-B guard cell signaling via down-regulation of MPK6.
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Proteínas de Arabidopsis/metabolismo , Óxido Nítrico/metabolismo , Estomas de Plantas/fisiología , Arabidopsis/citología , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Regulación de la Expresión Génica de las Plantas , Peróxido de Hidrógeno/metabolismo , Quinasas de Proteína Quinasa Activadas por Mitógenos/genética , Quinasas de Proteína Quinasa Activadas por Mitógenos/metabolismo , Fosfatasas de la Proteína Quinasa Activada por Mitógenos/genética , Fosfatasas de la Proteína Quinasa Activada por Mitógenos/metabolismo , Proteínas Quinasas Activadas por Mitógenos/genética , Proteínas Quinasas Activadas por Mitógenos/metabolismo , Mutación , NADPH Oxidasas/genética , NADPH Oxidasas/metabolismo , Células Vegetales/metabolismo , Células Vegetales/efectos de la radiación , Estomas de Plantas/efectos de la radiación , Proteínas Tirosina Fosfatasas/genética , Proteínas Tirosina Fosfatasas/metabolismo , Transducción de Señal , Rayos UltravioletaRESUMEN
The present study describes the effects of light conditions, different kinds and concentrations of auxins [Naphthylacetic acid (NAA) and dichlorophenoxyacetic acid (2,4-D)] with cytokinin (Kin) in MS medium on callus induction and embryogenesis in Crataegus pseudoheterophylla, C. aronia and C.meyeri. At first leave explants sections were cultured on different combinations of plant growth regulators in dark and light for callus initiation and light conditions to evaluation the percentage and duration of survival, callus diameter, callus fresh weight and dry. Results of effects of plant growth regulators and light conditions on callus initiation revealed that highest percentage of callus initiation leaves in treatment (0.5 mg/l 2.4-D+0.5 mg/l KIN) for species C.pseudoheterophylla in dark conditions (100%). Dark conditions (100%) were more effective on callogenesis than light conditions (Photoperiodicity of 16-h and at light intensity of 40 µmol m-2 s-1). The callus induction of in vitro (64-100%) leaves was better than the ex vitro ones (0-100%). The combination of 2,4-D and Kin of in vitro leaves callogenesis has been indicated faster (one weeks) than the other combinations. The results also showed that the highest percentage (100%) and survival duration (6 months) was found in species C. pseudoheterophylla and C. meyeri in 0.1 mg/l 2,4.D + 0.5 mg/l KIN and 0.5 mg/l 2,4.D + 0.5 mg/l Kin. The minimum survival (0%) was absorbed in species C. aronia in 1 mg/l NAA. Maximum callus (10.63 and 10.00 mm respectively) was shown in 0.1 mg/l 2,4.D + 0.5 mg/l Kin and 0.5 mg/l 2,4.D + 0.5 mg/l Kin and was not significant differences after five week among species. The results showed that the highest fresh (1081.49 mg) and dry weight (506.88 and 506.98 mg respectively) was absorbed in species C. pseudoheterophylla in 0.1 mg/l 2,4.D + 0.5 mg/l Kin and 0.5 mg/l 2,4.D + 0.5 mg/l Kin. The embryogenesis was not occurred in any plant growth regulator combinations and species. The results of this study suggested that using 2,4-D with cytokinin (Kin) would be more beneficial for callogenesis.
Asunto(s)
Desdiferenciación Celular/efectos de los fármacos , Crataegus/química , Extractos Vegetales/farmacología , Crataegus/metabolismo , Luz , Células Vegetales/efectos de los fármacos , Células Vegetales/fisiología , Células Vegetales/efectos de la radiación , Extractos Vegetales/química , Reguladores del Crecimiento de las Plantas/farmacología , Hojas de la Planta/química , Hojas de la Planta/metabolismoRESUMEN
Global CO2 level presently recorded at 400 ppm is expected to reach 550 ppm in 2050, an increment likely to impact plant growth and productivity. Using targeted LC-MS and GC-MS platforms we quantified 229 and 29 metabolites, respectively in a time-course study to reveal short-term responses to different concentrations (1, 3, and 10 mM) of bicarbonate (HCO3-) under light and dark conditions. Results indicate that HCO3- treatment responsive metabolomic changes depend on the HCO3- concentration, time of treatment, and light/dark. Interestingly, 3 mM HCO3- concentration treatment induced more significantly changed metabolites than either lower or higher concentrations used. Flavonoid biosynthesis and glutathione metabolism were common to both light and dark-mediated responses in addition to showing concentration-dependent changes. Our metabolomics results provide insights into short-term plant cellular responses to elevated HCO3- concentrations as a result of ambient increases in CO2 under light and dark.
Asunto(s)
Arabidopsis/efectos de los fármacos , Arabidopsis/efectos de la radiación , Bicarbonatos/metabolismo , Factores Biológicos/análisis , Oscuridad , Luz , Metaboloma , Arabidopsis/química , Cromatografía de Gases , Cromatografía Liquida , Espectrometría de Masas , Metabolómica , Células Vegetales/química , Células Vegetales/efectos de los fármacos , Células Vegetales/efectos de la radiación , SuspensionesRESUMEN
The effect of the statistical properties of light on the value of the photoinduced reaction of the biological objects, which differ in the morphological and physiological characteristics, the optical properties, and the size of cells, was studied. The fruit of apple trees, the pollen of cherries, the microcuttings of blackberries in vitro, and the spores and the mycelium of fungi were irradiated by quasimonochromatic light fluxes with identical energy parameters but different values of coherence length and radius of correlation. In all cases, the greatest stimulation effect occurred when the cells completely fit in the volume of the coherence of the field, while both temporal and spatial coherence have a significant and mathematically certain impact on the physiological activity of cells. It was concluded that not only the spectral, but also the statistical (coherent) properties of the acting light play an important role in the photoregulation process.
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Hongos/efectos de la radiación , Luz , Procesos Fotoquímicos , Células Vegetales/efectos de la radiación , Frutas/efectos de la radiación , Malus/efectos de la radiación , Micelio/efectos de la radiación , Polen/efectos de la radiación , Prunus avium/efectos de la radiación , Rubus/efectos de la radiaciónRESUMEN
Light-induced chloroplast movement and attachment to the plasma membrane are dependent on actin filaments. In Arabidopsis thaliana, the short actin filaments on the chloroplast envelope, cp-actin filaments, are essential for chloroplast movement and positioning. Furthermore, cp-actin-filament-mediated chloroplast movement is necessary for the strong-light-induced nuclear avoidance response. The proteins CHLOROPLAST UNUSUAL POSITIONING 1 (CHUP1), KINESIN-LIKE PROTEIN FOR ACTIN-BASED CHLOROPLAST MOVEMENT 1 (KAC1) and KAC2 are required for the generation and/or maintenance of cp-actin filaments in Arabidopsis. In land plants, CHUP1 and KAC family proteins play pivotal roles in the proper movement of chloroplasts and their attachment to the plasma membrane. Here, we report similar but distinct phenotypes in chloroplast and nuclear photorelocation movements between chup1 and kac1kac2 mutants. Measurement of chloroplast photorelocation movement indicated that kac1kac2, but not chup1, exhibited a clear strong-light-induced increase in leaf transmittance changes. The chloroplast movement in kac1kac2 depended on phototropin 2, CHUP1 and two other regulators for cp-actin filaments, PLASTID MOVEMENT IMPAIRED 1 and THRUMIN 1. Furthermore, kac1kac2 retained a weak but significant nuclear avoidance response although chup1 displayed a severe defect in the nuclear avoidance response. The kac1kac2chup1 triple mutant was completely defective in both chloroplast and nuclear avoidance responses. These results indicate that CHUP1 and the KACs function somewhat independently, but interdependently mediate both chloroplast and nuclear photorelocation movements.
Asunto(s)
Proteínas de Arabidopsis/genética , Arabidopsis/efectos de la radiación , Núcleo Celular/efectos de la radiación , Proteínas de Cloroplastos/genética , Cloroplastos/efectos de la radiación , Regulación de la Expresión Génica de las Plantas , Cinesinas/genética , Proteínas de Microfilamentos/genética , Citoesqueleto de Actina/metabolismo , Citoesqueleto de Actina/efectos de la radiación , Citoesqueleto de Actina/ultraestructura , Arabidopsis/genética , Arabidopsis/metabolismo , Arabidopsis/ultraestructura , Proteínas de Arabidopsis/metabolismo , Membrana Celular/metabolismo , Membrana Celular/efectos de la radiación , Membrana Celular/ultraestructura , Núcleo Celular/metabolismo , Núcleo Celular/ultraestructura , Proteínas de Cloroplastos/metabolismo , Cloroplastos/metabolismo , Cloroplastos/ultraestructura , Péptidos y Proteínas de Señalización Intracelular/genética , Péptidos y Proteínas de Señalización Intracelular/metabolismo , Cinesinas/metabolismo , Luz , Proteínas de Microfilamentos/metabolismo , Movimiento , Fototropinas/genética , Fototropinas/metabolismo , Células Vegetales/metabolismo , Células Vegetales/efectos de la radiación , Células Vegetales/ultraestructura , Hojas de la Planta/genética , Hojas de la Planta/metabolismo , Hojas de la Planta/efectos de la radiación , Hojas de la Planta/ultraestructuraRESUMEN
There is an abundant literature about the biological and physiological effects of UV-B light and the signaling and metabolic pathways it triggers and influences. Much less is known about UV-C light even though it seems to have a lot of potential for being effective in less time than UV-B light. UV-C light is known since long to exert direct and indirect inhibitory and damaging effects on living cells and is therefore commonly used for disinfection purposes. More recent observations suggest that UV-C light can also be exploited to stimulate the production of health-promoting phytochemicals, to extent shelf life of fruits and vegetables and to stimulate mechanisms of adaptation to biotic and abiotic stresses. Clearly some of these effects may be related to the stimulating effect of UV-C light on the production of reactive oxygen species (ROS) and to the stimulation of antioxidant molecules and mechanisms, although UV-C light could also trigger and regulate signaling pathways independently from its effect on the production of ROS. Our review clearly underlines the high potential of UV-C light in agriculture and therefore advocates for more work to be done to improve its efficiency and also to increase our understanding of the way UV-C light is perceived and influences the physiology of plants.