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Among the three active aldehyde oxidases in Arabidopsis thaliana leaves (AAO1-3), AAO3, which catalyzes the oxidation of abscisic-aldehyde to abscisic-acid, was shown recently to function as a reactive aldehyde detoxifier. Notably, aao2KO mutants exhibited less senescence symptoms and lower aldehyde accumulation, such as acrolein, benzaldehyde, and 4-hydroxyl-2-nonenal (HNE) than in wild-type leaves exposed to UV-C or Rose-Bengal. The effect of AAO2 expression absence on aldehyde detoxification by AAO3 and/or AAO1 was studied by comparing the response of wild-type plants to the response of single-functioning aao1 mutant (aao1S), aao2KO mutants, and single-functioning aao3 mutants (aao3Ss). Notably, aao3Ss exhibited similar aldehyde accumulation and chlorophyll content to aao2KO treated with UV-C or Rose-Bengal. In contrast, wild-type and aao1S exhibited higher aldehyde accumulation that resulted in lower remaining chlorophyll than in aao2KO leaves, indicating that the absence of active AAO2 enhanced AAO3 detoxification activity in aao2KO mutants. In support of this notion, employing abscisic-aldehyde as a specific substrate marker for AAO3 activity revealed enhanced AAO3 activity in aao2KO and aao3Ss leaves compared to wild-type treated with UV-C or Rose-Bengal. The similar abscisic-acid level accumulated in leaves of unstressed or stressed genotypes indicates that aldehyde detoxification by AAO3 is the cause for better stress resistance in aao2KO mutants. Employing the sulfuration process (known to activate aldehyde oxidases) in wild-type, aao2KO, and molybdenum-cofactor sulfurase (aba3-1) mutant plants revealed that the active AAO2 in WT employs sulfuration processes essential for AAO3 activity level, resulting in the lower AAO3 activity in WT than AAO3 activity in aao2KO.
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Nanotechnology is a rapidly developing discipline that has the potential to transform the way we approach problems in a variety of fields, including agriculture. The use of nanotechnology in sustainable agriculture has gained popularity in recent years. It has various applications in agriculture, such as the development of nanoscale materials and devices to boost agricultural productivity, enhance food quality and safety, improve the efficiency of water and nutrient usage, and reduce environmental pollution. Nanotechnology has proven to be very beneficial in this field, particularly in the development of nanoscale delivery systems for agrochemicals such as pesticides, fertilizers, and growth regulators. These nanoscale delivery technologies offer various benefits over conventional delivery systems, including better penetration and distribution, enhanced efficacy, and lower environmental impact. Encapsulating agrochemicals in nanoscale particles enables direct delivery to the targeted site in the plant, thereby reducing waste and minimizing off-target effects. Plants are fundamental building blocks of all ecosystems and evaluating the interaction between nanoparticles (NPs) and plants is a crucial aspect of risk assessment. This critical review therefore aims to provide an overview of the latest advances regarding the positive and negative effects of nanotechnology in agriculture. It also explores potential future research directions focused on ensuring the safe utilization of NPs in this field, which could lead to sustainable development. © 2024 Society of Chemical Industry.
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Agricultura , Nanotecnología , Agroquímicos/química , Agroquímicos/efectos adversos , Productos Agrícolas/crecimiento & desarrollo , Fertilizantes/análisis , Nanopartículas/química , Nanotecnología/métodos , Plaguicidas/química , Desarrollo SostenibleRESUMEN
Arsenic (As) is a major threat to the environment and human health due to its toxicity and carcinogenicity. Occurrence of alarming concentrations of As in water and soil leads to its bioaccumulation in crops which is a major health concern globally. Rice (Oryza sativa) is a staple food for a large population staying in As contaminated areas so, it is of utmost importance to reduce As levels in rice, especially grains. Amongst several strategies in practice, biotechnology may provide an effective option to reduce As accumulation in rice grains. Genetic engineering can be a viable approach to exploit potential genes playing roles in As metabolism pathway in plants. Besides, developing low As accumulating rice varieties through breeding is also an important area. Identifying genotypic variation in rice is a crucial step toward the development of a safe rice cultivar for growing in As-affected areas. Significant genotypic variation has been found in rice varieties for As accumulation in grains and that is attributable to differential expression of transporters, radial oxygen loss, and other regulators of As stress. This review provides recent updates on the research advances leading to transgenic and breeding approaches adopted to reduce As levels in rice, especially grains.
Arsenic (As) contamination in water, soil, and crops is creating a difficult situation for the large population across the globe. Various efforts are being made to reduce As levels in rice as it is a staple crop. This review presents recent biotechnological advances toward the development of low As accumulating rice. The review shall be an important information resource for the readers on the topic.
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Arsénico , Oryza , Contaminantes del Suelo , Humanos , Arsénico/metabolismo , Oryza/genética , Contaminantes del Suelo/metabolismo , Biodegradación Ambiental , Biotecnología , SueloRESUMEN
Helianthus annuus is a potential metal accumulator plant, which can find application in cadmium (Cd) phytoremediation and provide economic gains in terms of oil yield. This study is focused on Cd accumulation analysis, physiological and biochemical responses of five varieties of H. annuus (DRSF-108, DRSF-113, LSFH-171, Phule Bhaskar and KBSH-44). Plantlets of all varieties were treated with various Cd concentrations (10, 50, 100, 300 and 500 mg kg-1) for 20, 40 and 60 days. DRSF-108 showed the maximum total Cd accumulation (430.52 mg kg-1) in whole plant while minimum accumulation was observed in KBSH-44 (150.66 mg kg-1) at 500 mg kg-1 Cd after 60 days. The highest level of proline and polyphenol in DRSF-108 were 27.206 µmol g-1 fw and 6.86 mg g-1 fw, respectively. Antioxidant enzymes (catalase, ascorbate peroxidase and glutathione reductase) also showed increased activity in response to Cd treatment. SEM-EDX analysis of potential accumulator genotype, DRSF-108, showed the distribution of intracellular Cd into plant tissues. Therefore, it is concluded that among five varieties, DRSF-108 was the most potential Cd accumulator and had a higher capacity for Cd tolerance compared to other varieties. Our findings may allow us to extend variety DRSF-108 for sustainable farming and Cd remediation.
Phytoremediation of environmental contaminants is a promising strategy, however, the success depends on the use of suitable plants. Helianthus annuus is a good phytoremediator plant and the use of most promising genotype is desirable. This work evaluated five genotypes of H. annuus for cadmium accumulation and identified the best variety, DRSF-108 that can be used for cadmium remediation. The work shall pave way for field application of sunflower plants in cadmium remediation.
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Helianthus , Contaminantes del Suelo , Cadmio , Helianthus/genética , Biodegradación Ambiental , Plantas , Agricultura , Raíces de Plantas/química , Contaminantes del Suelo/análisisRESUMEN
Unexpected bioaccumulation and biomagnification of heavy metal(loid)s (HMs) in the environment have become a predicament for all living organisms, including plants. The presence of these HMs in the plant system raised the level of reactive oxygen species (ROS) and remodeled several vital cellular biomolecules. These lead to several morphological, physiological, metabolic, and molecular aberrations in plants ranging from chlorosis of leaves to the lipid peroxidation of membranes, and degradation of proteins and nucleic acid including the modulation of the enzymatic system, which ultimately affects the plant growth and productivity. Plants are equipped with several mechanisms to counteract the HMs toxicity. Among them, seed priming (SP) technology has been widely tested with the use of several inorganic chemicals, plant growth regulators (PGRs), gasotransmitters, nanoparticles, living organisms, and plant leaf extracts. The use of these compounds has the potential to alleviate the HMs toxicity through the strengthening of the antioxidant defense system, generation of low molecular weight metallothionein's (MTs), and phytochelatins (PCs), and improving seedling vigor during early growth stages. This review presents an account of the sources, uptake and transport, and phytotoxic effects of HMs with special attention to different mechanism/s, occurring to mitigate the HMs toxicity in plants employing SP technology.Novelty statement: To the best of our knowledge, this review has delineated the consequences of HMs on the crucial plant processes, which ultimately affect plant growth and development. This review also compiled the up to dated information on phytotoxicity of HMs through the use of SP technology, this review discussed how different types of SP approaches help in diminishing the concentration HMs in plant systems. Also, we depicted mechanisms, represent how HMs transport and their actions on cellular levels, and emphasized, how diverse SP technology effectiveness in the mitigation of plants' phytotoxicity in unique ways.
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Metales Pesados , Biodegradación Ambiental , Metales Pesados/toxicidad , Metales Pesados/análisis , Semillas/química , Semillas/metabolismo , Productos Agrícolas/metabolismo , Especies Reactivas de Oxígeno/metabolismoRESUMEN
The Arabidopsis thaliana aldehyde oxidase 3 (AAO3) catalyzes the oxidation of abscisic aldehyde (ABal) to abscisic acid (ABA). Besides ABal, plants generate other aldehydes that can be toxic above a certain threshold. AAO3 knockout mutants (aao3) exhibited earlier senescence but equivalent relative water content compared with wild-type (WT) during normal growth or upon application of UV-C irradiation. Aldehyde profiling in leaves of 24-day-old plants revealed higher accumulation of acrolein, crotonaldehyde, 3Z-hexenal, hexanal and acetaldehyde in aao3 mutants compared with WT leaves. Similarly, higher levels of acrolein, benzaldehyde, crotonaldehyde, propionaldehyde, trans-2-hexenal and acetaldehyde were accumulated in aao3 mutants upon UV-C irradiation. Aldehydes application to plants hastened profuse senescence symptoms and higher accumulation of aldehydes, such as acrolein, benzaldehyde and 4-hydroxy-2-nonenal, in aao3 mutant leaves as compared with WT. The senescence symptoms included greater decrease in chlorophyll content and increase in transcript expression of the early senescence marker genes, Senescence-Related-Gene1, Stay-Green-Protein2 as well as NAC-LIKE, ACTIVATED-BY AP3/P1. Notably, although aao3 had lower ABA content than WT, members of the ABA-responding genes SnRKs were expressed at similar levels in aao3 and WT. Moreover, the other ABA-deficient mutants [aba2 and 9-cis-poxycarotenoid dioxygenase3-2 (nced3-2), that has functional AAO3] exhibited similar aldehydes accumulation and chlorophyll content like WT under normal growth conditions or UV-C irradiation. These results indicate that the absence of AAO3 oxidation activity and not the lower ABA and its associated function is responsible for the earlier senescence symptoms in aao3 mutant.
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Ácido Abscísico/metabolismo , Aldehído Oxidasa/metabolismo , Aldehídos/toxicidad , Proteínas de Arabidopsis/metabolismo , Arabidopsis/genética , Reguladores del Crecimiento de las Plantas/metabolismo , Aldehído Oxidasa/genética , Aldehídos/metabolismo , Arabidopsis/fisiología , Proteínas de Arabidopsis/genética , Clorofila/metabolismo , Oxidación-Reducción , Hojas de la Planta/genética , Hojas de la Planta/fisiología , Senescencia de la PlantaRESUMEN
Drought, a major abiotic limiting factor, could be modulated with in-built reprogramming of plants at molecular level by regulating the activity of plant developmental processes, stress endurance and adaptation. The transgenic Arabidopsis thaliana over-expressing metallothionein 1 (MT1) gene of desi chickpea (Cicer arietinum L.) was subjected to transcriptome analysis. We evaluated drought tolerance of 7 days old plants of Arabidopsis thaliana in both wild-type (WT) as well as transgenic plants and performed transcriptome analysis. Our analysis revealed 24,737 transcripts representing 24,594 genes out of which 5,816 were differentially expressed genes (DEGs) under drought conditions and 841 genes were common in both genotypes. A total of 1251 DEGs in WT and 2099 in MT1 were identified in comparison with control. Out of the significant DEGs, 432 and 944 were upregulated, whereas 819 and 1155 were downregulated in WT and MT1 plants, respectively. The physiological and molecular parameters involving germination assay, root length measurements under different stress treatments and quantitative expression analysis of transgenic plants in comparison to wild-type were found to be enhanced. CarMT1 plants also demonstrated modulation of various other stress-responsive genes that reprogrammed themselves for stress adaptation. Amongst various drought-responsive genes, 24 DEGs showed similar quantitative expression as obtained through RNA sequencing data. Hence, these modulatory genes could be used as a genetic tool for understanding and delineating the mechanisms for fine-tuning of stress responses in crop plants.
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Arabidopsis , Cicer , Arabidopsis/genética , Arabidopsis/metabolismo , Cicer/genética , Cicer/metabolismo , Sequías , Perfilación de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Metalotioneína/genética , Metalotioneína/metabolismo , Plantas Modificadas Genéticamente/genética , Plantas Modificadas Genéticamente/metabolismo , Estrés Fisiológico/genética , Factores de Transcripción/genética , Factores de Transcripción/metabolismoRESUMEN
Purine degradation products have been shown to play roles in plant response to stresses such as drought, salinity, extended dark, nitrogen deficiency, and pathogen infection. In this study, we used Arabidopsis wild-type (WT) and an Atxdh1-knockout mutant defective in xanthine dehydrogenase1 (XDH1) to examine the role of degraded purine metabolites in the responses to wounding or UV-C stress applied to the middle leaves of the plant. Wounding or UV-C stress in the mutant resulted in lower fresh-weight, increased senescence symptoms, and increased cell death compared to WT plants. In addition, WT plants exhibited lower levels of oxidative stress indicators, reactive oxygen species, and malondialdehyde in their leaves than the mutant. Notably, transcripts and proteins functioning in the purine degradation pathway were regulated in such a way that it led to enhanced ureide levels in WT leaves 24h after applying the UV-C or wound stress. However, different remobilization of the accumulated ureides was observed after 72h of stress. In plants treated with UV-C, the concentration of allantoin was highest in young leaves, whereas in wounded plants it was lowest in these leaves and instead accumulated mainly in the middle leaves that had been wounded. These results indicated that in WT plants treated with UV-C, ureides were remobilized from the lower older and damaged leaves to support young leaf growth during the recovery period from stress. After wounding, however, whilst some ureides were remobilized to the young leaves, more remained in the wounded middle leaves to function as antioxidants and/or healing agents.
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Proteínas de Arabidopsis , Arabidopsis , Purinas/metabolismo , Rayos Ultravioleta/efectos adversos , Alantoína/metabolismo , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Sequías , Hojas de la Planta/metabolismo , Plantas Modificadas GenéticamenteRESUMEN
The roles of cytosolic O-acetylserine-(thiol)-lyase A (OASTLA), chloroplastic OASTLB, and mitochondrial OASTLC in plant selenate resistance were studied in Arabidopsis. Impairment in OASTLA and OASTLB resulted in reduced biomass, chlorophyll and soluble protein content compared with selenate-treated OASTLC-impaired and wild-type plants. The generally lower total selenium (Se), protein-Se, organic-sulfur and protein-sulfur (S) content in oastlA and oastlB compared with wild-type and oastlC leaves indicated that Se accumulation was not the main cause for the stress symptoms in these mutants. Notably, the application of selenate positively induced S-starvation markers and the OASTLs, followed by increased sulfite reductase, sulfite oxidase activities, and increased sulfite and sulfide concentrations. Taken together, our results indicate a futile anabolic S-starvation response that resulted in lower glutathione and increased oxidative stress symptoms in oastlA and oastlB mutants. In-gel assays of l-cysteine and l-seleno-cysteine, desulfhydrase activities revealed that two of the three OASTL activity bands in each of the oastl single mutants were enhanced in response to selenate, whereas the impaired proteins exhibited a missing activity band. The absence of differently migrated activity bands in each of the three oastl mutants indicates that these OASTLs are major components of desulfhydrase activity, degrading l-cysteine and l-seleno-cysteine in Arabidopsis.
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Arabidopsis , Liasas , Selenio , Arabidopsis/metabolismo , Liasas de Carbono-Oxígeno/metabolismo , Cisteína/metabolismo , Liasas/metabolismo , Ácido Selénico , Selenio/metabolismo , Serina/análogos & derivados , Compuestos de Sulfhidrilo/metabolismo , Sulfitos/metabolismo , Azufre/metabolismoRESUMEN
The present study assessed the utility of Allium cepa based cyto-genotoxicity bioassays in evaluating the arsenic toxicity and remediation potential of Pteris vittata on contaminated soil of Lakhimpur-Kheri district. Untreated and P. vittata treated soil extracts were used for cyto-genotoxicity tests in A. cepa. Results showed that P. vittata extracted high concentration of arsenic, which ranged from 220 to 1420 mgkg-1 in different soils. Cyto-genotoxic assessment of A. cepa showed that extract of P. vittata treated soil had lower cyto-genotoxic effects as compared to untreated soil. A higher mitotic index (10%) while lower mitotic depression (29%), relative abnormality rate (10%), chromosomal aberrations (1%) and micronuclei (2%) were detected in root meristematic cells of A. cepa exposed to remediated soil extract in comparison to untreated soil. The studies provide a simple, rapid and economic cyto-genotoxicity bioassay tool for evaluating toxicity of contaminated soils of contaminated soils as well as revealed the phytoremdiation property of P. vittata against arsenic toxicity.
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Arsénico , Pteris , Contaminantes del Suelo , Arsénico/análisis , Arsénico/toxicidad , Biodegradación Ambiental , Bioensayo , Cebollas , Suelo , Contaminantes del Suelo/análisis , Contaminantes del Suelo/toxicidadRESUMEN
The arsenic (As) contamination demands its remediation from the environment which is naturally possible by the application of Pteris vittata L. However, biomonitoring of phytoremediation potential of P. vittata at chromosomal and DNA level is still meager. The present study was designed to biomonitor the phytoremediation efficiency of P. vittata through phytotoxic and cyto-genotoxic biomarkers assessment using Trigonella foenum-graecum L. (Fenugreek; Methi) as test system. Study revealed hyperaccumulation potential of P. vittata which extracted arsenic in its tissues. Biomonitoring evaluation depicted that phytotoxic damage was reduced in Trigonella exposed to remediated soil, which was revealed through reduced electrolyte leakage, hydrogen peroxide and MDA content. Moreover, cyto-genetic endpoints like mitotic depression (44.03%), relative abnormality rate (16.6%) and chromosomal abnormality frequency (1.06%) were also lesser in test plants grown in remediated soil compared to those grown in non-remediated soil. Along with this various chromosomal aberrations like stickiness, breaks, laggards, bridges, fragmentations and micronuclei were also augmented in test plants exposed to non-remediated arsenic enriched soil. It was evident that arsenic enriched soil caused toxicity to plants in dose-dependent manner that was assessable through the analysis of biochemical parameters and cyto-genetic biomarkers. The cyto-genetic biomarkers are very efficient, simple and non-expensive tools to biomonitor arsenic toxicity at chromosomal as well as DNA level to assess the remediation potential of P. vittata in field conditions.
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Chloroplast-localized adenosine-5'-phosphosulphate reductase (APR) generates sulfite and plays a pivotal role in reduction of sulfate to cysteine. The peroxisome-localized sulfite oxidase (SO) oxidizes excess sulfite to sulfate. Arabidopsis wild type, SO RNA-interference (SO Ri) and SO overexpression (SO OE) transgenic lines infiltrated with sulfite showed increased water loss in SO Ri plants, and smaller stomatal apertures in SO OE plants compared with wild-type plants. Sulfite application also limited sulfate and abscisic acid-induced stomatal closure in wild type and SO Ri. The increases in APR activity in response to sulfite infiltration into wild type and SO Ri leaves resulted in an increase in endogenous sulfite, indicating that APR has an important role in sulfite-induced increases in stomatal aperture. Sulfite-induced H2O2 generation by NADPH oxidase led to enhanced APR expression and sulfite production. Suppression of APR by inhibiting NADPH oxidase and glutathione reductase2 (GR2), or mutation in APR2 or GR2, resulted in a decrease in sulfite production and stomatal apertures. The importance of APR and SO and the significance of sulfite concentrations in water loss were further demonstrated during rapid, harsh drought stress in root-detached wild-type, gr2 and SO transgenic plants. Our results demonstrate the role of SO in sulfite homeostasis in relation to water consumption in well-watered plants.
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Proteínas de Arabidopsis , Arabidopsis , Oxidorreductasas actuantes sobre Donantes de Grupos Sulfuro , Sulfito-Oxidasa , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Glutatión Reductasa , Peróxido de Hidrógeno , Oxidorreductasas actuantes sobre Donantes de Grupos Sulfuro/genética , Sulfito-Oxidasa/genética , Sulfitos , AguaRESUMEN
Ulmus pumila L. is an excellent afforestation and biofuel tree that produces high-quality wood, rich in starch. In addition, U. pumila is highly adaptable to adverse environmental conditions, which is conducive to its utilization for vegetating saline soils. However, little is known about the physiological responses and transcriptional regulatory network of U. pumila under salt stress. In this study, we exposed five main cultivars in saline-alkali land (Upu2, 5, 8, 11, and 12) to NaCl stress. Of the five cultivars assessed, Upu11 exhibited the highest salt resistance. Growth and biomass accumulation in Upu11 were promoted under low salt concentrations (<150 mM). However, after 3 months of continuous treatment with 150 mM NaCl, growth was inhibited, and photosynthesis declined. A transcriptome analysis conducted after 3 months of treatment detected 7009 differentially expressed unigenes (DEGs). The gene annotation indicated that these DEGs were mainly related to photosynthesis and carbon metabolism. Furthermore, PHOTOSYNTHETIC ELECTRON TRANSFERH (UpPETH), an important electron transporter in the photosynthetic electron transport chain, and UpWAXY, a key gene controlling amylose synthesis in the starch synthesis pathway, were identified as hub genes in the gene coexpression network. We identified 25 and 62 unigenes that may interact with PETH and WAXY, respectively. Overexpression of UpPETH and UpWAXY significantly increased the survival rates, net photosynthetic rates, biomass, and starch content of transgenic Arabidopsis plants under salt stress. Our findings clarify the physiological and transcriptional regulators that promote or inhibit growth under environmental stress. The identification of salt-responsive hub genes directly responsible for photosynthesis and starch synthesis or metabolism will provide targets for future genetic improvements.
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Redes Reguladoras de Genes , Fotosíntesis , Proteínas de Plantas/metabolismo , Estrés Salino , Tolerancia a la Sal , Almidón/biosíntesis , Ulmus/fisiología , Perfilación de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Proteínas de Plantas/genética , Almidón/genética , Ulmus/genéticaRESUMEN
In December 2019, the outbreak of viral disease labeled as Novel Coronavirus started in Wuhan, China, which later came to be known as Covid-19. The disease has spread in almost every part of the world and has been declared a global pandemic in March 2020 by World Health Organization (WHO). The corona virus outbreak has emerged as one of the deadliest pandemics of all time in human history. The ongoing pandemic of COVID-19 has forced several countries of the world to observe complete lockdown forcing people to live in their homes. India also faced the phase of total lockdown for 21 days (in first phase) to avoid the spread of coronavirus to the maximum possible extent. This lockdown impacted the pollution levels of environment and improved air and water quality in the short span owing to very less human activities. The present work scientifically analyzed the available data for primary air pollutants (PM2.5, NO2, SO2 and CO) from two major Indian cities, Lucknow and New Delhi. The analysis was based on air quality data for before lockdown and after lockdown (first phase of 21 days) periods of 21 days each. The results showed significant decline in the studied air pollution indices and demonstrated improvement of air quality in both the cities. The major impact was seen in the levels of PM2.5, NO2 and CO. The levels of SO2 showed less significant decline during the lockdown period. The results are presented with future perspectives to mitigate air pollution in near future by adopting the short and periodical lockdown as a tool.
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Contaminantes Atmosféricos/análisis , Control de Enfermedades Transmisibles/estadística & datos numéricos , Infecciones por Coronavirus/prevención & control , Monitoreo del Ambiente , Pandemias/prevención & control , Neumonía Viral/prevención & control , COVID-19 , Ciudades/estadística & datos numéricos , Actividades Humanas/estadística & datos numéricos , Humanos , India , Material Particulado/análisisRESUMEN
The prokaryotic ßγ-crystallins are a large group of uncharacterized domains with Ca2+-binding motifs. We have observed that a vast number of these domains are found appended to other domains, in particular, the carbohydrate-active enzyme (CAZy) domains. To elucidate the functional significance of these prospective Ca2+ sensors in bacteria and this widespread domain association, we have studied one typical example from Clostridium beijerinckii, a bacterium known for its ability to produce acetone, butanol, and ethanol through fermentation of several carbohydrates. This novel glycoside hydrolase of family 64 (GH64), which we named glucanallin, is composed of a ßγ-crystallin domain, a GH64 domain, and a carbohydrate-binding module 56 (CBM56). The substrates of GH64, ß-1,3-glucans, are the targets for industrial biofuel production due to their plenitude. We have examined the Ca2+-binding properties of this protein, assayed its enzymatic activity, and analyzed the structural features of the ß-1,3-glucanase domain through its high-resolution crystal structure. The reaction products resulting from the enzyme reaction of glucanallin reinforce the mixed nature of GH64 enzymes, in contrast to the prevailing notion of them being an exotype. Upon disabling Ca2+ binding and comparing different domain combinations, we demonstrate that the ßγ-crystallin domain in glucanallin acts as a Ca2+ sensor and enhances the glycolytic activity of glucanallin through Ca2+ binding. We also compare the structural peculiarities of this new member of the GH64 family to two previously studied members.IMPORTANCE We have biochemically and structurally characterized a novel glucanase from the less studied GH64 family in a bacterium significant for fermentation of carbohydrates into biofuels. This enzyme displays a peculiar property of being distally modulated by Ca2+ via assistance from a neighboring ßγ-crystallin domain, likely through changes in the domain interface. In addition, this enzyme is found to be optimized for functioning in an acidic environment, which is in line with the possibility of its involvement in biofuel production. Multiple occurrences of a similar domain architecture suggest that such a "ßγ-crystallination"-mediated Ca2+ sensitivity may be widespread among bacterial proteins.
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Proteínas Bacterianas/química , Proteínas de Unión al Calcio/química , Calcio/química , Clostridium beijerinckii/enzimología , Glicósido Hidrolasas/química , beta-Cristalinas/química , gamma-Cristalinas/química , Secuencia de Aminoácidos , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Sitios de Unión , Calcio/metabolismo , Proteínas de Unión al Calcio/genética , Proteínas de Unión al Calcio/metabolismo , Clonación Molecular , Clostridium beijerinckii/química , Clostridium beijerinckii/genética , Cristalografía por Rayos X , Escherichia coli/genética , Escherichia coli/metabolismo , Fermentación , Expresión Génica , Vectores Genéticos/química , Vectores Genéticos/metabolismo , Glicósido Hidrolasas/genética , Glicósido Hidrolasas/metabolismo , Cinética , Modelos Moleculares , Unión Proteica , Conformación Proteica en Hélice alfa , Conformación Proteica en Lámina beta , Dominios y Motivos de Interacción de Proteínas , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Alineación de Secuencia , Homología de Secuencia de Aminoácido , Especificidad por Sustrato , beta-Cristalinas/genética , beta-Cristalinas/metabolismo , beta-Glucanos/química , beta-Glucanos/metabolismo , gamma-Cristalinas/genética , gamma-Cristalinas/metabolismoRESUMEN
The nitrogen (N)-rich ureides allantoin and allantoate, which are products of purine catabolism, play a role in N delivery in Leguminosae. Here, we examined their role as an N source in nonlegume plants using Arabidopsis (Arabidopsis thaliana) plants mutated in XANTHINE DEHYDROGENASE1 (AtXDH1), a catalytic bottleneck in purine catabolism. Older leaves of the Atxdh1 mutant exhibited early senescence, lower soluble protein, and lower organic N levels as compared with wild-type older leaves when grown with 1 mm nitrate but were comparable to the wild type under 5 mm nitrate. Similar nitrate-dependent senescence phenotypes were evident in the older leaves of allantoinase (Ataln) and allantoate amidohydrolase (Ataah) mutants, which also are impaired in purine catabolism. Under low-nitrate conditions, xanthine accumulated in older leaves of Atxdh1, whereas allantoin accumulated in both older and younger leaves of Ataln but not in wild-type leaves, indicating the remobilization of xanthine-degraded products from older to younger leaves. Supporting this notion, ureide transporter expression was enhanced in older leaves of the wild type in low-nitrate as compared with high-nitrate conditions. Elevated transcripts and proteins of AtXDH and AtAAH were detected in low-nitrate-grown wild-type plants, indicating regulation at protein and transcript levels. The higher nitrate reductase activity in Atxdh1 leaves compared with wild-type leaves indicated a need for nitrate assimilation products. Together, these results indicate that the absence of remobilized purine-degraded N from older leaves of Atxdh1 caused senescence symptoms, a result of higher chloroplastic protein degradation in older leaves of low-nitrate-grown plants.
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Proteínas de Arabidopsis/metabolismo , Arabidopsis/enzimología , Nitratos/metabolismo , Nitrógeno/metabolismo , Purinas/metabolismo , Xantina Deshidrogenasa/metabolismo , Alantoína/metabolismo , Amidohidrolasas/genética , Amidohidrolasas/metabolismo , Arabidopsis/genética , Arabidopsis/fisiología , Proteínas de Arabidopsis/genética , Proteína 5 Relacionada con la Autofagia/genética , Proteína 5 Relacionada con la Autofagia/metabolismo , Familia de las Proteínas 8 Relacionadas con la Autofagia/genética , Familia de las Proteínas 8 Relacionadas con la Autofagia/metabolismo , Mutación , Hojas de la Planta/enzimología , Hojas de la Planta/genética , Hojas de la Planta/fisiología , Factores de Tiempo , Ureohidrolasas/genética , Ureohidrolasas/metabolismo , Xantina Deshidrogenasa/genéticaRESUMEN
The study implements a periodical intermittent water cycle during rice cultivation providing insight potential in minimizing soil bio-available arsenic. Soil As concentrations were 34⯱â¯0.49 and 72.03⯱â¯0.54â¯mg kg-1 As respectively in two selected fields with rice cultivars gosai and satabdi, in comparison to 42.26⯱â¯0.37 and 83.69⯱â¯0.48â¯mg kg-1 in continuously flooded field soil, determined through ICP-MS. The study found higher translocation of silicon from soil to rice plant parts under intermittent irrigation having pH range of 7.6-9.4 and greater availability of soil organic content that in turn release more labile silicon from soil to aqueous phase for plant accumulation. This increased uptake of silicon strengthens rice shoots, nodes and leaf xylem-phloem integrity compared to conventional continuously flooded rice cultivation approach, suppressing the arsenic translocation, as observed under FE-SEM real-time imaging. Fresh plants were analysed for bioaccumulation and translocation factors of arsenic and silicon to justify the enhanced silicon uptake under proposed practice. Plant stress regulator enzymes viz. malondialdehyde (MDA), total protein, superoxide dismutase (SOD), guaiacol peroxidase (GPX) and ascorbate peroxidase (APX) from both conditions and found to be better in intermittent method over conventional practice with higher productivity.
Asunto(s)
Riego Agrícola/métodos , Arsénico/farmacocinética , Oryza/fisiología , Silicio/metabolismo , Contaminantes del Suelo/farmacocinética , Antioxidantes/metabolismo , Arsénico/análisis , Arsénico/toxicidad , Disponibilidad Biológica , India , Oryza/crecimiento & desarrollo , Oryza/metabolismo , Suelo/química , Contaminantes del Suelo/análisis , Contaminantes del Suelo/toxicidad , Estrés Fisiológico/efectos de los fármacosRESUMEN
The present study measured arsenic (As) concentrations in soil, groundwater and rice grain samples in two villages, Sarapur and Chinili, under Chakdaha block, Nadia district, West Bengal, India. This study also included a survey of the two villages to understand the knowledge among villagers about the As problem. Soil and groundwater samples were collected from fields in two villages while rice grain samples were collected from villagers' houses. The results revealed the presence of As in higher concentrations than the maximum permissible limit of As in drinking water (10 µg L-1 and 50 µg L-1 by WHO and Indian standard, respectively) in groundwater [124.50 ± 1.11 µg L-1 (Sarapur) and 138.20 ± 1.34 µg L-1 (Chinili)]. The level of As in soil was found to range from 47.7 ± 0.14 to 49.3 ± 0.19 mg Kg-1 in Sarapur and from 57.5 ± 0.25 to 62.5 ± 0.44 mg Kg-1 in Chinili which are also higher than European Union maximum acceptable limit in agricultural soil (i.e. 20 mg Kg-1). The analysis of As in rice grains of five varieties, collected from residents of two villages, showed the presence of higher than recommended safe level of As in rice by FAO/WHO (0.2 mg Kg-1). The As concentration order was Gosai (0.95 ± 0.044 mg kg-1), Satabdi (0.79 ± 0.038 mg kg-1), Banskathi (0.60 ± 0.026 mg kg-1), Kunti (0.47 ± 0.018 mg kg-1) and Ranjit (0.29 ± 0.021 mg kg-1). Importantly, Gosai and Satabdi were the most popular varieties being consumed by local people. The data of consumption of rice per day in the survey was used for the measurement of average daily dose and hazard quotient. It was seen that the As hazard was negatively correlated to the age of residents. Therefore, children and toddlers were at higher risk of As exposure than elderly people. In addition, people with skin related As toxicity symptoms were also cited in the two villages. The study emphasized the severity of As problem in remote areas of West Bengal, India where people consume As tainted rice due to lack of awareness about the As problem and associated health issues.
Asunto(s)
Arsénico/análisis , Contaminación de Alimentos/análisis , Oryza/química , Contaminantes del Suelo/análisis , Contaminantes Químicos del Agua/análisis , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Arsénico/toxicidad , Niño , Preescolar , Exposición Dietética/análisis , Femenino , Agua Subterránea/análisis , Conocimientos, Actitudes y Práctica en Salud , Encuestas Epidemiológicas , Humanos , India , Lactante , Masculino , Persona de Mediana Edad , Medición de Riesgo , Salud Rural , Suelo/química , Contaminantes Químicos del Agua/toxicidad , Adulto JovenRESUMEN
The Arabidopsis (Arabidopsis thaliana) aldehyde oxidases are a multigene family of four oxidases (AAO1-AAO4) that oxidize a variety of aldehydes, among them abscisic aldehyde, which is oxidized to the phytohormone abscisic acid. Toxic aldehydes are generated in plants both under normal conditions and in response to stress. The detoxification of such aldehydes by oxidation is attributed to aldehyde dehydrogenases but never to aldehyde oxidases. The feasibility of the detoxification of aldehydes in siliques via oxidation by AAO4 was demonstrated, first, by its ability to efficiently oxidize an array of aromatic and aliphatic aldehydes, including the reactive carbonyl species (RCS) acrolein, hydroxyl-2-nonenal, and malondialdehyde. Next, exogenous application of several aldehydes to siliques in AAO4 knockout (KO) Arabidopsis plants induced severe tissue damage and enhanced malondialdehyde levels and senescence symptoms, but not in wild-type siliques. Furthermore, abiotic stresses such as dark and ultraviolet C irradiation caused an increase in endogenous RCS and higher expression levels of senescence marker genes, leading to premature senescence of KO siliques, whereas RCS and senescence marker levels in wild-type siliques were hardly affected. Finally, in naturally senesced KO siliques, higher endogenous RCS levels were associated with enhanced senescence molecular markers, chlorophyll degradation, and earlier seed shattering compared with the wild type. The aldehyde-dependent differential generation of superoxide and hydrogen peroxide by AAO4 and the induction of AAO4 expression by hydrogen peroxide shown here suggest a self-amplification mechanism for detoxifying additional reactive aldehydes produced during stress. Taken together, our results indicate that AAO4 plays a critical role in delaying senescence in siliques by catalyzing aldehyde detoxification.
Asunto(s)
Aldehído Oxidasa/metabolismo , Aldehídos/metabolismo , Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Semillas/metabolismo , Ácido Abscísico/metabolismo , Aldehído Oxidasa/genética , Secuencia de Aminoácidos , Arabidopsis/enzimología , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Secuencia de Bases , Benzaldehídos/metabolismo , Biocatálisis , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Regulación de la Expresión Génica de las Plantas/efectos de la radiación , Técnicas de Inactivación de Genes , Peróxido de Hidrógeno/metabolismo , Peróxido de Hidrógeno/farmacología , Concentración de Iones de Hidrógeno , Cinética , Malondialdehído/metabolismo , Oxidantes/metabolismo , Oxidantes/farmacología , Oxidación-Reducción , Plantas Modificadas Genéticamente , Especies Reactivas de Oxígeno/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Semillas/enzimología , Semillas/genética , Homología de Secuencia de Aminoácido , Especificidad por Sustrato , Factores de Tiempo , Rayos UltravioletaRESUMEN
Salicornia and Sarcocornia are almost identical halophytes whose edible succulent shoots hold promise for commercial production in saline water. Enhanced sulfur nutrition may be beneficial to crops naturally grown on high sulfate. However, little is known about sulfate nutrition in halophytes. Here we show that Salicornia europaea (ecotype RN) exhibits a significant increase in biomass and organic-S accumulation in response to supplemental sulfate, whereas Sarcocornia fruticosa (ecotype VM) does not, instead exhibiting increased sulfate accumulation. We investigated the role of two pathways on organic-S and biomass accumulation in Salicornia and Sarcoconia: the sulfate reductive pathway that generates Cys and l-Cys desulfhydrase that degrades Cys to H2S, NH3, and pyruvate. The major function of O-acetyl-Ser-(thiol) lyase (OAS-TL; EC 2.5.1.47) is the formation of l-Cys, but our study shows that the OAS-TL A and OAS-TL B of both halophytes are enzymes that also degrade l-Cys to H2S. This activity was significantly higher in Sarcocornia than in Salicornia, especially upon sulfate supplementation. The activity of the sulfate reductive pathway key enzyme, adenosine 5'-phosphosulfate reductase (APR, EC 1.8.99.2), was significantly higher in Salicornia than in Sarcocornia These results suggest that the low organic-S level in Sarcocornia is the result of high l-Cys degradation rate by OAS-TLs, whereas the greater organic-S and biomass accumulation in Salicornia is the result of higher APR activity and low l-Cys degradation rate, resulting in higher net Cys biosynthesis. These results present an initial road map for halophyte growers to attain better growth rates and nutritional value of Salicornia and Sarcocornia.