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1.
Plant Mol Biol ; 106(1-2): 85-108, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-33629224

RESUMO

KEY MESSAGE: Overexpression of StCaM2 in tobacco promotes plant growth and confers increased salinity and drought tolerance by enhancing the photosynthetic efficiency, ROS scavenging, and recovery from membrane injury. Calmodulins (CaMs) are important Ca2+ sensors that interact with effector proteins and drive a network of signal transduction pathways involved in regulating the growth and developmental pattern of plants under stress. Herein, using in silico analysis, we identified 17 CaM isoforms (StCaM) in potato. Expression profiling revealed different temporal and spatial expression patterns of these genes, which were modulated under abiotic stress. Among the identified StCaM genes, StCaM2 was found to have the largest number of abiotic stress responsive promoter elements. In addition, StCaM2 was upregulated in response to some of the selected abiotic stress in potato tissues. Overexpression of StCaM2 in transgenic tobacco plants enhanced their tolerance to salinity and drought stress. Accumulation of reactive oxygen species was remarkably decreased in transgenic lines compared to that in wild type plants. Chlorophyll a fluorescence analysis suggested better performance of photosystem II in transgenic plants under stress compared to that in wild type plants. The increase in salinity stress tolerance in StCaM2-overexpressing plants was also associated with a favorable K+/Na+ ratio. The enhanced tolerance to abiotic stresses correlated with the increase in the activities of anti-oxidative enzymes in transgenic tobacco plants. Overall, our results suggest that StCaM2 can be a novel candidate for conferring salt and drought tolerance in plants.


Assuntos
Proteínas de Ligação ao Cálcio/metabolismo , Secas , Nicotiana/genética , Nicotiana/fisiologia , Proteínas de Plantas/metabolismo , Salinidade , Solanum tuberosum/metabolismo , Estresse Fisiológico , Antioxidantes/metabolismo , Proteínas de Ligação ao Cálcio/genética , Calmodulina/genética , Calmodulina/metabolismo , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Genoma de Planta , Germinação/efeitos dos fármacos , Germinação/genética , Íons , Membranas , Fotossíntese/efeitos dos fármacos , Reguladores de Crescimento de Plantas/farmacologia , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas , Prolina/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Sequências Reguladoras de Ácido Nucleico/genética , Plântula/efeitos dos fármacos , Plântula/genética , Plântula/crescimento & desenvolvimento , Estresse Fisiológico/efeitos dos fármacos , Estresse Fisiológico/genética , Nicotiana/enzimologia , Nicotiana/crescimento & desenvolvimento , Fatores de Transcrição/metabolismo , Água/metabolismo
2.
Physiol Mol Biol Plants ; 26(10): 2019-2033, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-33088046

RESUMO

Intracellular signaling is a critical determinant of the normal growth and development of plants. Signaling peptides, also known as peptide hormones, along with classical phytohormones, are the significant players of plant intracellular signaling. C-terminally encoded peptide (CEP), a 15-amino acid post-translationally peptide identified in Arabidopsis, plays a pivotal role in lateral root formation, nodulation, and act as long-distance root to shoot signaling molecule in N-starvation conditions. Expression of CEP gene members in Arabidopsis is perturbed by nitrogen starvation; however, not much is known regarding their role in other abiotic stress conditions. To gain a comprehensive insight into CEP biology, we identified CEP genes across diverse plant genera (Glycine max, Sorghum bicolor, Brassica rapa, Zea mays, and Oryza sativa) using bioinformatics tools. In silico promoter analysis revealed that CEP gene promoters show an abundance of abiotic stress-responsive elements suggesting a possible role of CEPs in abiotic stress signaling. Spatial and temporal expression patterns of CEP via RNA seq and microarray revealed that various CEP genes are transcriptionally regulated in response to abiotic stresses. Validation of rice CEP genes expression by qRT-PCR showed that OsCEP1, OsCEP8, OsCEP9, and OsCEP10 were highly upregulated in response to different abiotic stress conditions. Our findings suggest these CEP genes might be important mediators of the abiotic stress response and warrant further overexpression/knockout studies to delineate their precise role in abiotic stress response.

3.
J Contemp Dent Pract ; 21(7): 787-791, 2020 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-33020364

RESUMO

AIM: To evaluate the impact of oral health literacy (OHL) on the periodontal health among low-income-group workers of dental institutes. MATERIALS AND METHODS: A cross-sectional study was conducted on 137 class III and IV workers of dental college. Data collection was done by using a customized pro forma including demographics, REALD-30 for calculating OHL, and items on oral hygiene habits like toothpaste use, brushing frequency, and any other oral hygiene aid. Following which periodontal health status was determined, which was categorized into severe, moderate, and mild periodontitis (health). The analysis was done using the SPSS 11.5. Periodontal health status was associated with OHL scores, oral hygiene habits, and demographics using the chi-square test. The statistical significance level was set at 5% level. RESULTS: Among the 137 subjects, 25 participants reported health/mild periodontitis, 53 had moderate periodontitis, and 59 had severe periodontitis. Low OHL was observed in 52.5% and only 13.8% had high OHL. The participants who had low OHL, 56.94% (n = 41), were having severe disease, while the subjects who had better OHL, only 21.05% (n = 4) were found to have severe disease. CONCLUSION: The people with low socioeconomic classes can be reached effectively if the community involvement concept is used through the workers of dental institutions. But first efforts should be made to improve the OHL of these workers. CLINICAL SIGNIFICANCE: Improving OHL can be of great help to the clinicians and the community health workers because it helps them to make patient adherent to the treatment and the medications prescribed to them. Periodontitis affects people with low socioeconomic status and in the present study it was class III and IV workers. Improving OHL of this population will decrease the oral disease burden of India.


Assuntos
Letramento em Saúde , Estudos Transversais , Humanos , Índia/epidemiologia , Saúde Bucal , Pobreza
4.
Curr Genomics ; 17(3): 155-76, 2016 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-27252584

RESUMO

Rice, one of the most important cereal crops for mankind, feeds more than half the world population. Rice has been heralded as a model cereal owing to its small genome size, amenability to easy transformation, high synteny to other cereal crops and availability of complete genome sequence. Moreover, sequence wealth in rice is getting more refined and precise due to resequencing efforts. This humungous resource of sequence data has confronted research fraternity with a herculean challenge as well as an excellent opportunity to functionally validate expressed as well as regulatory portions of the genome. This will not only help us in understanding the genetic basis of plant architecture and physiology but would also steer us towards developing improved cultivars. No single technique can achieve such a mammoth task. Functional genomics through its diverse tools viz. loss and gain of function mutants, multifarious omics strategies like transcriptomics, proteomics, metabolomics and phenomics provide us with the necessary handle. A paradigm shift in technological advances in functional genomics strategies has been instrumental in generating considerable amount of information w.r.t functionality of rice genome. We now have several databases and online resources for functionally validated genes but despite that we are far from reaching the desired milestone of functionally characterizing each and every rice gene. There is an urgent need for a common platform, for information already available in rice, and collaborative efforts between researchers in a concerted manner as well as healthy public-private partnership, for genetic improvement of rice crop better able to handle the pressures of climate change and exponentially increasing population.

5.
Funct Integr Genomics ; 15(4): 395-412, 2015 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-25523384

RESUMO

Cyclophilins are a set of ubiquitous proteins present in all subcellular compartments, involved in a wide variety of cellular processes. Comparative bioinformatics analysis of the rice and Arabidopsis genomes led us to identify novel putative cyclophilin gene family members in both the genomes not reported previously. We grouped cyclophilin members with similar molecular weight and subtypes together in the phylogenetic tree which indicated their co-evolution in rice and Arabidopsis. We also characterized a rice cyclophilin gene, OsCyp2-P (Os02g0121300), isolated from a salinity-tolerant landrace, Pokkali. Publicly available massively parallel signature sequencing (MPSS) and microarray data, besides our quantitative real time PCR (qRT-PCR) data suggest that transcript abundance of OsCyp2-P is regulated under different stress conditions in a developmental and organ specific manner. Ectopic expression of OsCyp2-P imparted multiple abiotic stress tolerance to transgenic tobacco plants as evidenced by higher root length, shoot length, chlorophyll content, and K(+)/Na(+) ratio under stress conditions. Transgenic plants also showed reduced lipid peroxidase content, electrolyte leakage, and superoxide content under stress conditions suggesting better ion homeostasis than WT plants. Localization studies confirmed that OsCyp2-P is localized in both cytosol and nucleus, indicating its possible interaction with several other proteins. The overall results suggest the explicit role of OsCyp2-P in bestowing multiple abiotic stress tolerance at the whole plant level. OsCyp2-P operates via reactive oxygen species (ROS) scavenging and ion homeostasis and thus is a promising candidate gene for enhancing multiple abiotic stress tolerance in crop plants.


Assuntos
Ciclofilinas/genética , Nicotiana/genética , Oryza/genética , Proteínas de Plantas/genética , Potássio/metabolismo , Tolerância ao Sal , Sódio/metabolismo , Ciclofilinas/metabolismo , Homeostase , Pressão Osmótica , Proteínas de Plantas/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Nicotiana/metabolismo
6.
Ecotoxicol Environ Saf ; 101: 146-56, 2014 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-24507140

RESUMO

The present study was designed to study the growth and yield responses of a tropical potato variety (Solanum tuberosum L. cv. Kufri chandramukhi) to different levels of carbon dioxide (382 and 570ppm) and ozone (50 and 70ppb) in combinations using open top chambers (OTCs). Plants were exposed to three ozone levels in combination with ambient CO2 and two ozone levels at elevated CO2. Significant increments in leaf area and total biomass were observed under elevated CO2 in combination with ambient O3 (ECO2+AO3) and elevated O3 (ECO2+EO3), compared to the plants grown under ambient concentrations (ACO2+AO3). Yield measured as fresh weight of potato also increased significantly under ECO2+AO3 and ECO2+EO3. Yield, however, reduced under ambient (ACO2+AO3) and elevated ozone (ACO2+EO3) compared to ACO2 (filtered chamber). Number, fresh and dry weights of tubers of size 35-50mm and>50mm used for direct consumption and industrial purposes, respectively increased maximally under ECO2+AO3. Ambient as well as elevated levels of O3 negatively affected the growth parameters and yield mainly due to reductions in number and weight of tubers of sizes >35mm. The quality of potato tubers was also modified under different treatments. Starch content increased and K, Zn and Fe concentrations decreased under ECO2+AO3 and ECO2+EO3 compared to ACO2+AO3. Starch content reduced under ACO2+AO3 and ACO2+EO3 treatments compared to ACO2. These results clearly suggest that elevated CO2 has provided complete protection to ambient O3 as the potato yield was higher under ECO2+AO3 compared to ACO2. However, ambient CO2 is not enough to protect the plants under ambient O3 levels. Elevated CO2 also provided protection against elevated O3 by improving the yield. Quality of tubers is modified by both CO2 and O3, which have serious implications on human health at present and in future.


Assuntos
Poluentes Atmosféricos/toxicidade , Dióxido de Carbono/toxicidade , Ozônio/toxicidade , Solanum tuberosum/efeitos dos fármacos , Interações Medicamentosas , Crescimento/efeitos dos fármacos , Clima Tropical
7.
Acta Crystallogr D Biol Crystallogr ; 69(Pt 4): 555-63, 2013 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-23519664

RESUMO

Cyclophilins belong to a family of proteins that bind to the immunosuppressive drug cyclosporin A (CsA). Several members of this protein family catalyze the cis-trans isomerization of peptide bonds preceding prolyl residues. The present study describes the biochemical and structural characteristics of a cytosolic cyclophilin (TaCypA-1) cloned from wheat (Triticum aestivum L.). Purified TaCypA-1 expressed in Escherichia coli showed peptidyl-prolyl cis-trans isomerase activity, which was inhibited by CsA with an inhibition constant of 78.3 nM. The specific activity and catalytic efficiency (kcat/Km) of the purified TaCypA-1 were 99.06 ± 0.13 nmol s(-1) mg(-1) and 2.32 × 10(5) M(-1) s(-1), respectively. The structures of apo TaCypA-1 and the TaCypA-1-CsA complex were determined at 1.25 and 1.20 Šresolution, respectively, using X-ray diffraction. Binding of CsA to the active site of TaCypA-1 did not result in any significant conformational change in the apo TaCypA-1 structure. This is consistent with the crystal structure of the human cyclophilin D-CsA complex reported at 0.96 Å resolution. The TaCypA-1 structure revealed the presence of a divergent loop of seven amino acids (48)KSGKPLH(54) which is a characteristic feature of plant cyclophilins. This study is the first to elucidate the structure of an enzymatically active plant cyclophilin which shows peptidyl-prolyl cis-trans isomerase activity and the presence of a divergent loop.


Assuntos
Ciclofilina A/química , Triticum/química , Cristalografia por Raios X , Ciclofilina A/metabolismo , Ciclosporina/metabolismo , Citosol/química , Imunossupressores/metabolismo , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , Estrutura Secundária de Proteína
8.
BMC Plant Biol ; 12: 107, 2012 Jul 10.
Artigo em Inglês | MEDLINE | ID: mdl-22780875

RESUMO

BACKGROUND: Metallothioneins (MT) are low molecular weight, cysteine rich metal binding proteins, found across genera and species, but their function(s) in abiotic stress tolerance are not well documented. RESULTS: We have characterized a rice MT gene, OsMT1e-P, isolated from a subtractive library generated from a stressed salinity tolerant rice genotype, Pokkali. Bioinformatics analysis of the rice genome sequence revealed that this gene belongs to a multigenic family, which consists of 13 genes with 15 protein products. OsMT1e-P is located on chromosome XI, away from the majority of other type I genes that are clustered on chromosome XII. Various members of this MT gene cluster showed a tight co-regulation pattern under several abiotic stresses. Sequence analysis revealed the presence of conserved cysteine residues in OsMT1e-P protein. Salinity stress was found to regulate the transcript abundance of OsMT1e-P in a developmental and organ specific manner. Using transgenic approach, we found a positive correlation between ectopic expression of OsMT1e-P and stress tolerance. Our experiments further suggest ROS scavenging to be the possible mechanism for multiple stress tolerance conferred by OsMT1e-P. CONCLUSION: We present an overview of MTs, describing their gene structure, genome localization and expression patterns under salinity and development in rice. We have found that ectopic expression of OsMT1e-P enhances tolerance towards multiple abiotic stresses in transgenic tobacco and the resultant plants could survive and set viable seeds under saline conditions. Taken together, the experiments presented here have indicated that ectopic expression of OsMT1e-P protects against oxidative stress primarily through efficient scavenging of reactive oxygen species.


Assuntos
Regulação da Expressão Gênica de Plantas , Metalotioneína/metabolismo , Família Multigênica , Nicotiana/fisiologia , Oryza/genética , Espécies Reativas de Oxigênio/metabolismo , Adaptação Fisiológica , Cromossomos de Plantas/genética , Cromossomos de Plantas/metabolismo , Biologia Computacional/métodos , Sequência Conservada , Cisteína/metabolismo , Sequestradores de Radicais Livres/metabolismo , Genes de Plantas , Metalotioneína/genética , Oryza/metabolismo , Oryza/fisiologia , Estresse Oxidativo , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , Plantas Geneticamente Modificadas/fisiologia , Plantas Tolerantes a Sal/genética , Plantas Tolerantes a Sal/metabolismo , Plantas Tolerantes a Sal/fisiologia , Sementes/genética , Sementes/metabolismo , Sementes/fisiologia , Cloreto de Sódio/farmacologia , Nicotiana/metabolismo
9.
Anesth Essays Res ; 16(1): 42-48, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36249140

RESUMO

Background: The supraglottic airway device (SAD) has proved to be an appropriate alternative to endotracheal intubation in laparoscopic surgeries owing to various advantages, namely, decreased airway manipulation and stable hemodynamics. Aims: We compared the efficacy in terms of oropharyngeal leak pressure (OLP) and safety of laryngeal mask airway (LMA)-Supreme (LMA-S), i-gel, and Baska mask in patients undergoing laparoscopic cholecystectomy. Settings and Design: Ninety patients posted for elective laparoscopic cholecystectomy were selected for a prospective randomized comparative study in a tertiary care center. Materials and Methods: The study comprised three groups of 30 each based on the different SADs used Group-LS with LMA-Supreme, Group-IG with i-gel, and Group-BM with Baska mask. The secondary objectives were device insertion time, ease of insertion, changes in the peak airway pressure (PAP), heart rate, mean arterial pressure, and airway complications (sore throat, dysphagia. dysphonia, lip/tongue or dental injury, etc.) between three groups. Statistical Analysis: The quantitative data were analyzed using the one-way analysis of variance test and Bonferroni post hoc multiple comparison test. Qualitative data were compared using Chi.squared test. Results: OLP was significantly higher (P = 0.005) in the Baska mask than i-gel and LMA-S groups just after insertion and during carboperitonium. There was no significant difference in time for device insertion, number of attempts, ease of insertion, and use of manipulation (P > 0.05). However, the gastric tube insertion time was significantly lower in Group BM (9.59 ± 2.78) than Group IG with 12.79 ± 3.47 and Group LS with 10.84 ± 3.68 (P < 0.05). There were no significant differences between the groups with regard to changes in the PAP, heart rate, mean arterial pressure at different time intervals, and complications. Conclusion: Baska mask provided a significantly higher OLP compared to i-gel and LMA-S without significant airway morbidity in laparoscopic surgeries.

10.
PLoS One ; 15(7): e0236349, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32701993

RESUMO

Peptide signalling is an integral part of cell-to-cell communication which helps to relay the information responsible for coordinating cell proliferation and differentiation. Phytosulfokine Receptor (PSKR) is a transmembrane LRR-RLK family protein with a binding site for small signalling peptide, phytosulfokine (PSK). PSK signalling through PSKR promotes normal growth and development and also plays a role in defense responses. Like other RLKs, these PSKRs might have a role in signal transduction pathways related to abiotic stress responses. Genome-wide analysis of phytosulfokine receptor gene family has led to the identification of fifteen putative members in the Oryza sativa genome. The expression analysis of OsPSKR genes done using RNA-seq data, showed that these genes were differentially expressed in different tissues and responded specifically to heat, salt, drought and cold stress. Furthermore, the real-time quantitative PCR for fifteen OsPSKR genes revealed temporally and spatially regulated gene expression corresponding to salinity and drought stress. Our results provide useful information for a better understanding of OsPSKR genes and provide the foundation for additional functional exploration of the rice PSKR gene family in development and stress response.


Assuntos
Genoma de Planta/genética , Oryza/genética , Hormônios Peptídicos/genética , Peptídeos/genética , Proteínas de Plantas/genética , Secas , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas/genética , Peptídeos/classificação , Filogenia , Salinidade , Transdução de Sinais/genética , Cloreto de Sódio/metabolismo , Estresse Fisiológico/genética
11.
PLoS One ; 15(5): e0233493, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32453778

RESUMO

Glyoxalase pathway is the major pathway of methylglyoxal detoxification and is ubiquitously present in all organisms ranging from prokaryotes to eukaryotes. Glyoxalase I (GLYI) and Glyoxalase II (GLYII), the two core enzymes of this pathway work together to neutralize methylglyoxal (MG), a dicarbonyl molecule with detrimental cytotoxicity at higher concentrations. The first step towards the detoxification of MG is catalyzed by GLYI, a metalloenzyme that requires divalent metal ions (either Zn2+ as seen in eukaryotes or Ni2+ as in prokaryotes). However, both Zn2+ and Ni2+ dependent GLYIs have been shown to co-exist in a higher eukaryote i.e. Arabidopsis thaliana. In the present study, we determine the role of both Zn2+ dependent (AtGLYI2) and Ni2+ dependent (AtGLYI3, AtGLYI6) GLYIs from Arabidopsis in salinity stress tolerance. AtGLYI2 overexpressing Arabidopsis plants showed better growth rate while maintaining lower levels of MG under high saline conditions. They were taller with more number of silique formation with respect to their Ni2+ dependent counterparts. Further, lack in germination of Arabidopsis AtGLYI2 mutants in presence of exogenous MG indicates the direct involvement of Zn2+ dependent GLYI in MG detoxification, suggesting Zn2+ dependent GLYI as the main enzyme responsible for MG detoxification and salinity stress tolerance.


Assuntos
Arabidopsis/crescimento & desenvolvimento , Lactoilglutationa Liase/genética , Lactoilglutationa Liase/metabolismo , Arabidopsis/enzimologia , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Regulação da Expressão Gênica de Plantas , Germinação , Mutação , Aldeído Pirúvico/metabolismo , Estresse Salino , Zinco/metabolismo
12.
Funct Integr Genomics ; 9(1): 109-23, 2009 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-18594887

RESUMO

Oryza sativa L. cv IR64 is a widely cultivated, salt-sensitive indica rice, while Pokkali is a well-known, naturally salt-tolerant relative. To understand the molecular basis of differences in their salinity tolerance, three subtractive cDNA libraries were constructed. A total of 1,194 salinity-regulated cDNAs are reported here that may serve as repositories for future individual gene-based functional genomics studies. Gene expression data using macroarrays and Northern blots gives support to our hypothesis that salinity tolerance of Pokkali may be due to constitutive overexpression of many genes that function in salinity tolerance and are stress inducible in IR64. Analysis of genome architecture revealed the presence of these genes on all the chromosomes with several distinct clusters. Notably, a few mapped on one of the major quantitative trait loci - Saltol - on chromosome 1 and were found to be differentially regulated in the two contrasting genotypes. The present study also defines a set of known abiotic stress inducible genes, including CaMBP, GST, LEA, V-ATPase, OSAP1 zinc finger protein, and transcription factor HBP1B, that were expressed at high levels in Pokkali even in the absence of stress. These proposed genes may prove useful as "candidates" in improving salinity tolerance in crop plants using transgenic approach.


Assuntos
Perfilação da Expressão Gênica , Oryza/genética , Oryza/fisiologia , Tolerância ao Sal/genética , Plântula/crescimento & desenvolvimento , Plântula/genética , Estresse Fisiológico/genética , Cromossomos de Plantas/genética , Clonagem Molecular , Etiquetas de Sequências Expressas , Regulação da Expressão Gênica de Plantas , Biblioteca Gênica , Genes de Plantas , Genótipo , Locos de Características Quantitativas/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Análise de Sequência de DNA
13.
Mol Biotechnol ; 42(2): 195-204, 2009 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-19214808

RESUMO

Cyclophilin 2 (OsCyp2) is a cytosolic member of immunophilin family from rice. We have isolated its full length cDNA (1,056 bp) with an open reading frame of 519 bp encoding a polypeptide of 172 amino acids and an estimated pI of 8.61. Peptidyl prolyl cis-trans isomerase activity of the protein was determined using N-succinyl-ala-ala-pro-phe-p-nitroanilidine as peptide substrate. It has a catalytic efficiency (K (cat)/K (m)) of 4.5 x 10(6)/(mol/l)/s, which is comparable to known cyclophilins from plants. Its activity is specifically inhibited by cyclosporin A, a macrolide drug inhibitor of cyclophilins. Transcript analysis showed it to be a developmentally and differentially regulated gene; showing changes in abundance at seedling, tillering and heading stage under non-stress and salinity stress conditions. Expression of OsCyp2 enhances the ability of Escherichia coli to survive under diverse abiotic stresses viz. salinity, high temperature, osmotic stress (mannitol) and oxidative stress (H(2)O(2)). OsCyp2 was able to complement the yeast mutant lacking native Cyp2 and also improved the growth of wild type yeast under above-mentioned stress conditions. Based on these results, we propose that OsCyp2 may serve as a 'suitable candidate' for raising transgenic plants for enhanced multiple abiotic stress tolerance.


Assuntos
Ciclofilinas/metabolismo , Escherichia coli/fisiologia , Melhoramento Genético/métodos , Oryza/metabolismo , Saccharomyces cerevisiae/fisiologia , Estresse Fisiológico/fisiologia , Ciclofilinas/genética , Oryza/genética , Engenharia de Proteínas/métodos , Proteínas Recombinantes/metabolismo
14.
Sci Rep ; 8(1): 5451, 2018 04 03.
Artigo em Inglês | MEDLINE | ID: mdl-29615695

RESUMO

Methylglyoxal(MG) is a potent cytotoxin that is produced as a byproduct of various metabolic reactions in the cell. The major enzymes for MG detoxification are Glyoxalase I(GLYI), Glyoxalase II(GLYII) and D-lactate dehydrogenase(D-LDH). These three enzymes work together and convert MG into D-pyruvate, which directly goes to TCA cycle. Here, a comparative study of the ability of MG detoxification of these three enzymes has been done in both E. coli and yeast. Ectopic expression of these three genes from Arabidopsis in E. coli in presence of different abiotic stress revealed the contribution of each of these genes in detoxifying MG. Yeast mutants of MG detoxification enzymes were also grown in different stress conditions to record the effect of each gene. These mutants were also used for complementation assays using the respective MG detoxifying genes from Arabidopsis in presence of various stress conditions. The MG content and the corresponding growth of cells was measured in all the bacterial as well as yeast strains. This study reveals differential contribution of MG detoxification enzymes in mitigating MG levels and alleviating stress in both prokaryotes as well as eukaryotes. GLYI and D-LDH were found to be key enzymes in MG detoxification under various abiotic stresses.


Assuntos
Inativação Metabólica , L-Lactato Desidrogenase/metabolismo , Lactoilglutationa Liase/metabolismo , Aldeído Pirúvico/metabolismo , Estresse Fisiológico , Escherichia coli/genética , Escherichia coli/fisiologia , L-Lactato Desidrogenase/genética , Lactoilglutationa Liase/genética , Estresse Oxidativo , Estresse Salino
15.
Front Plant Sci ; 8: 198, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28261251

RESUMO

Abiotic stress and climate change is the major concern for plant growth and crop yield. Abiotic stresses lead to enhanced accumulation of reactive oxygen species (ROS) consequently resulting in cellular damage and major losses in crop yield. One of the major scavengers of ROS is ascorbate (AA) which acts as first line of defense against external oxidants. An enzyme named ascorbate oxidase (AAO) is known to oxidize AA and deleteriously affect the plant system in response to stress. Genome-wide analysis of AAO gene family has led to the identification of five, three, seven, four, and six AAO genes in Oryza sativa, Arabidopsis, Glycine max, Zea mays, and Sorghum bicolor genomes, respectively. Expression profiling of these genes was carried out in response to various abiotic stresses and during various stages of vegetative and reproductive development using publicly available microarray database. Expression analysis in Oryza sativa revealed tissue specific expression of AAO genes wherein few members were exclusively expressed in either root or shoot. These genes were found to be regulated by both developmental cues as well as diverse stress conditions. The qRT-PCR analysis in response to salinity and drought stress in rice shoots revealed OsAAO2 to be the most stress responsive gene. On the other hand, OsAAO3 and OsAAO4 genes showed enhanced expression in roots under salinity/drought stresses. This study provides lead about important stress responsive AAO genes in various crop plants, which could be used to engineer climate resilient crop plants.

16.
PLoS One ; 11(7): e0159348, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27415831

RESUMO

The glyoxalase pathway is ubiquitously found in all the organisms ranging from prokaryotes to eukaryotes. It acts as a major pathway for detoxification of methylglyoxal (MG), which deleteriously affects the biological system in stress conditions. The first important enzyme of this system is Glyoxalase I (GLYI). It is a metalloenzyme which requires divalent metal ions for its activity. This divalent metal ion can be either Zn2+ as found in most of eukaryotes or Ni2+ as seen in prokaryotes. In the present study, we have found three active GLYI enzymes (AtGLYI2, AtGLYI3 and AtGLYI6) belonging to different metal activation classes coexisting in Arabidopsis thaliana. These enzymes have been found to efficiently complement the GLYI yeast mutants. These three enzymes have been characterized in terms of their activity, metal dependency, kinetic parameters and their role in conferring tolerance to multiple abiotic stresses in E. coli and yeast. AtGLYI2 was found to be Zn2+ dependent whereas AtGLYI3 and AtGLYI6 were Ni2+ dependent. Enzyme activity of Zn2+ dependent enzyme, AtGLYI2, was observed to be exceptionally high (~250-670 fold) as compared to Ni2+ dependent enzymes, AtGLYI3 and AtGLYI6. The activity of these GLYI enzymes correlated well to their role in stress tolerance. Heterologous expression of these enzymes in E. coli led to better tolerance against various stress conditions. This is the first report of a higher eukaryotic species having multiple active GLYI enzymes belonging to different metal activation classes.


Assuntos
Arabidopsis/fisiologia , Expressão Ectópica do Gene/fisiologia , Escherichia coli/enzimologia , Lactoilglutationa Liase/metabolismo , Estresse Fisiológico/fisiologia , Arabidopsis/enzimologia , Proteínas de Arabidopsis/isolamento & purificação , Proteínas de Arabidopsis/fisiologia , Clonagem Molecular , Escherichia coli/fisiologia , Cinética , Lactoilglutationa Liase/fisiologia , Níquel/metabolismo , Zinco/metabolismo
17.
Environ Pollut ; 174: 279-88, 2013 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-23291007

RESUMO

The present study was conducted to assess morphological, biochemical and yield responses of palak (Beta vulgaris L. cv Allgreen) to ambient and elevated levels of CO(2) and O(3), alone and in combination. As compared to the plants grown in charcoal filtered air (ACO(2)), growth and yield of the plants increased under elevated CO(2) (ECO(2)) and decreased under combination of ECO(2) with elevated O(3) (ECO(2) + EO(3)), ambient O(3) (ACO(2) + AO(3)) and elevated O(3) (EO(3)). Lipid peroxidation, ascorbic acid, catalase and glutathione reductase activities enhanced under all treatments and were highest in EO(3.) Foliar starch and organic carbon contents increased under ECO(2) and ECO(2) + EO(3) and reduced under EO(3) and ACO(2) + AO(3.) Foliar N content declined in all treatments compared to ACO(2) resulting in alteration of C/N ratio. This study concludes that ambient level of CO(2) is not enough to counteract O(3) impact, but elevated CO(2) has potential to counteract the negative effects of future O(3) level.


Assuntos
Poluentes Atmosféricos/farmacologia , Beta vulgaris/fisiologia , Dióxido de Carbono/farmacologia , Ozônio/toxicidade , Poluentes Atmosféricos/toxicidade , Antioxidantes/metabolismo , Ácido Ascórbico/metabolismo , Beta vulgaris/efeitos dos fármacos , Beta vulgaris/metabolismo , Peroxidação de Lipídeos , Fotossíntese/efeitos dos fármacos , Folhas de Planta/crescimento & desenvolvimento , Folhas de Planta/metabolismo
18.
Plant Signal Behav ; 8(1): e22734, 2013 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-23123451

RESUMO

Cyclophilins constitute a subgroup of large family of proteins called immunophilins, which also include FKBPs and Parvulins. They are remarkably conserved in all genera, highlighting their pivotal role in important cellular processes. Most cyclophilins display PPIase enzymatic activity, multiplicity, diverse cellular locations and active role in protein folding which render them to be included in the class of diverse set of proteins called molecular chaperones. Due to their distinct PPIase function, besides protein disulfide isomerases and protein foldases, cyclophilins have been deemed necessary for in vivo chaperoning activity. Unlike other cellular chaperones, these proteins are specific in their respective targets. Not all cyclophilin proteins possess PPIase activity, indicating a loss of their PPIase activity during the course of evolution and gain of function independent of their PPIase activity. The PPIase function of cyclophilins is also compensated by their functional homologs, like FKBPs. Multiple cyclophilin members in plants like Arabidopsis and rice have been reported to be associated with diverse functions and regulatory pathways through their foldase, scaffolding, chaperoning or other unknown activities. Although many functions of plant cyclophilins were reported or suggested, the physiological relevance and molecular basis of stress-responsive expression of plant cyclophilins is still largely unknown. However, their wide distribution and ubiquitous nature signifies their fundamental importance in plant survival. Several of these members have also been directly linked to multiple stresses. This review attempts to deal with plant cyclophilins with respect to their role in stress response.


Assuntos
Adaptação Fisiológica , Ciclofilinas/metabolismo , Peptidilprolil Isomerase/metabolismo , Proteínas de Plantas/metabolismo , Plantas/metabolismo , Estresse Fisiológico
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