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1.
Physiol Plant ; 174(2): e13651, 2022 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-35174506

RESUMO

Root system architecture plays an important role in plant adaptation to drought stress. The root system architecture (RSA) consists of several structural features, which includes number and length of main and lateral roots along with the density and length of root hairs. These features exhibit plasticity under water-limited environments and could be critical to developing crops with efficient root systems for adaptation under drought. Recent advances in the omics approaches have significantly improved our understanding of the regulatory mechanisms of RSA remodeling under drought and the identification of genes and other regulatory elements. Plant response to drought stress at physiological, morphological, biochemical, and molecular levels in root cells is regulated by various phytohormones and their crosstalk. Stress-induced reactive oxygen species play a significant role in regulating root growth and development under drought stress. Several transcription factors responsible for the regulation of RSA under drought have proven to be beneficial for developing drought tolerant crops. Molecular breeding programs for developing drought-tolerant crops have been greatly benefitted by the availability of quantitative trait loci (QTLs) associated with the RSA regulation. In the present review, we have discussed the role of various QTLs, signaling components, transcription factors, microRNAs and crosstalk among various phytohormones in shaping RSA and present future research directions to better understand various factors involved in RSA remodeling for adaptation to drought stress. We believe that the information provided herein may be helpful in devising strategies to develop crops with better RSA for efficient uptake and utilization of water and nutrients under drought conditions.


Assuntos
Secas , Raízes de Plantas , Reguladores de Crescimento de Plantas , Raízes de Plantas/fisiologia , Fatores de Transcrição , Água
2.
Physiol Plant ; 173(4): 1765-1784, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-33665830

RESUMO

Aluminum (Al) toxicity is a major limiting factor for plant growth and productivity in acidic soil. At pH lower than 5.0 (pH < 5.0), the soluble and toxic form of Al (Al3+ ions) enters root cells and inhibits root growth and uptake of water and nutrients. The organic acids malate, citrate, and oxalate are secreted by the roots and chelate Al3+ to form a non-toxic Al-OA complex, which decreases the entry of Al3+ into the root cells. When Al3+ enters, it leads to the production of reactive oxygen species (ROS) in cells, which are toxic and cause damage to biomolecules like lipids, carbohydrates, proteins, and nucleic acids. When ROS levels rise beyond the threshold, plants activate an antioxidant defense system that comprises of superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), glutathione S-transferase (GST), ascorbic acid (ASA), phenolics and alkaloids etc., which protect plant cells from oxidative damage by scavenging and neutralizing ROS. Besides, ROS also play an important role in signal transduction and influence many molecular and cellular process like hormone signaling, gene expression, cell wall modification, cell cycle, programed cell death (PCD), and development. In the present review, the mechanisms of Al-induced ROS generation, ROS signaling, and crosstalk with other signaling pathways helping to combat Al toxicity have been summarized, which will help researchers to understand the intricacies of Al-induced plant response at cellular level and plan research for developing Al-toxicity tolerant crops for sustainable agriculture in acid soil-affected regions of the world.


Assuntos
Alumínio , Raízes de Plantas , Alumínio/toxicidade , Antioxidantes , Catalase/metabolismo , Estresse Oxidativo , Raízes de Plantas/metabolismo , Espécies Reativas de Oxigênio , Transdução de Sinais
3.
Plant Cell Rep ; 40(8): 1331-1343, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-34086069

RESUMO

Aluminium (Al) is one of the most abundant metals in earth crust, which becomes toxic to the plants growing in acidic soil. Phytohormones like ethylene, auxin, cytokinin, abscisic acid, jasmonic acid and gibberellic acid are known to play important role in regulating Al toxicity tolerance in plants. Exogenous applications of auxin, cytokinin and abscisic acid have shown significant effect on Al-induced root growth inhibition. Moreover, ethylene and cytokinin act synergistically with auxin in responding against Al toxicity. A number of studies showed that phytohormones play vital roles in controlling root responses to Al toxicity by modulating reactive oxygen species (ROS) signalling, cell wall modifications, organic acid exudation from roots and expression of Al responsive genes and transcription factors. This review provides a summary of recent studies related to involvement of phytohormone signalling and cross-talk with other pathways in regulating response against Al toxicity in plants.


Assuntos
Alumínio/toxicidade , Reguladores de Crescimento de Plantas/metabolismo , Plantas/efeitos dos fármacos , Plantas/metabolismo , Ácido Abscísico/metabolismo , Ciclopentanos/metabolismo , Citocininas/metabolismo , Etilenos/metabolismo , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Ácidos Indolacéticos/metabolismo , Oxilipinas/metabolismo , Raízes de Plantas/citologia , Raízes de Plantas/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos
4.
Physiol Mol Biol Plants ; 27(12): 2859-2873, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-35035141

RESUMO

The quantitative real-time polymerase chain reaction (qRT-PCR) is the most sensitive and commonly used technique for gene expression studies in biological systems. However, the reliability of qRT-PCR results depends on the selection of reference gene(s) for data normalization. Horse gram (Macrotyloma uniflorum) is an important legume crop on which several molecular studies have been reported. However, the stability of reference genes has not been evaluated. In the present study, nine candidate reference genes were identified from horse gram RNA-seq data and evaluated in two horse gram genotypes, HPK4 and HPKM317 under six abiotic stresses viz. cold, drought, salinity, heat, abscisic acid and methyl viologen-induced oxidative stress. The results were evaluated using geNorm, Bestkeeper, Normfinder and delta-delta Ct methods and comprehensive ranking was assigned using RefFinder and RankAggreg software. The overall result showed that TCTP was one of the most stable genes in all samples and in genotype HPK4, while in HPKM317 profilin was most stably expressed. However, PSMA5 was identified as least stable in all the experimental conditions. Expression of target genes dehydrin and early response to dehydration 6 under drought stress was also validated using TCTP and profilin for data normalization, either alone or in combination, which confirmed their suitability for qRT-PCR data normalization. Thus, TCTP and profilin genes may be used for qRT-PCR data normalization for molecular and genomic studies in horse gram. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s12298-021-01104-0.

5.
Physiol Mol Biol Plants ; 25(1): 123-134, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30804635

RESUMO

Lentil (Lens culinaris) is one of the most important staple food crops of developing countries. Transcriptome based global gene expression profiling followed by validation of expression of important genes through quantitative real time-PCR (qRT-PCR) has achieved significance in recent years. However, there is a severe scarcity of information regarding stable reference genes in lentil, which is mandatory for qRT-PCR data normalisation. Hence, the present study was under-taken to identify the most stable reference gene(s) in lentil. Expression stability of eight candidate genes viz. ribulose 1,5-bisphosphate carboxylase large subunit (Rbcl), ribosomal protein L2 (RPL2), 18S rRNA, tubulin (Tub), elongation factor 1α (EF1α), glyceraldehydes-3-phosphate dehydrogenase (GAPDH), heat shock protein (HSP70), and Maturase (mat K) was evaluated in five varieties of lentil at three different stages of leaf development and abiotic stress conditions using qRT-PCR. The results were analysed using four types of statistical software viz., geNorm, BestKeeper, NormFinder and RefFinder; all softwares identified RPL2 as most stable under abiotic stress conditions and developmental stages followed by Tub and Rbcl; while, HSP70 was identified as least stable. Relative expression of the target genes, defensin and PR4, was evaluated under abiotic stress conditions and data normalisation was done using two stable reference genes, RPL2 and Tub, either alone or in combination and with two least stable genes, HSP70 and 18S. The present work provides a list of potential reference genes in lentil, which will help in selection of appropriate reference gene for qRT-PCR data normalization depending upon the experiment.

6.
Reprod Health ; 11(1): 74, 2014 Oct 21.
Artigo em Inglês | MEDLINE | ID: mdl-25330906

RESUMO

BACKGROUND: Virtually all the evidence on the relationship between women's empowerment and use of contraception comes from cross-sectional studies that have emphasized macrosocial factors.This analysis tested whether literate and illiterate women are empowered by an intervention designed to provide information addressing technical and gender concerns and expand contraceptive choice, and evaluated the effects of women's decision-making power on contraceptive behavior. METHODS: The data came from a three-year quasi-experiment conducted in two comparable, yet not equivalent, rural blocks in Jharkhand, India. At the intervention block, a new contraceptive method was introduced at Ministry of Health health centers, providers were trained to offer family planning information and services which took into consideration gender power dynamics, and promotional messages and information about contraception were disseminated community-wide. Married women ages 15-49 who lived in the intervention and control blocks were sampled and interviewed before and after the intervention by a professional research firm. Data analyses included generalized linear models with interactions and covariate control. RESULTS: Women's normative beliefs concerning wives' power in decisions regarding money earned and visits to relatives and friends vis-à-vis their husbands' power were increased by the intervention; similar was the case among illiterate, but not literate, women regarding decisions related to childbearing. Concerning met need for contraception, the change for women with relatively more power who were illiterate was greater in the intervention than in the control area. CONCLUSION: The findings suggest that women were empowered by outreach visits that addressed gender dynamics and that their empowerment contributed to their met need for contraception. Generalizations to other settings, however, may be limited by cultural differences.


Assuntos
Comportamento Contraceptivo , Anticoncepção/estatística & dados numéricos , Necessidades e Demandas de Serviços de Saúde , Poder Psicológico , Adolescente , Adulto , Anticoncepção/psicologia , Tomada de Decisões , Escolaridade , Serviços de Planejamento Familiar/organização & administração , Feminino , Humanos , Índia , Pessoa de Meia-Idade , Avaliação das Necessidades , Saúde da População Rural , Fatores Socioeconômicos , Adulto Jovem
7.
BMC Genomics ; 14: 748, 2013 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-24182234

RESUMO

BACKGROUND: The Himalayan or Indian Mayapple (Podophyllum hexandrum Royle) produces podophyllotoxin, which is used in the production of semisynthetic anticancer drugs. High throughput transcriptome sequences or genomic sequence data from the Indian Mayapple are essential for further understanding of the podophyllotoxin biosynthetic pathway. RESULTS: 454 pyrosequencing of a P. hexandrum cell culture normalized cDNA library generated 2,667,207 raw reads and 1,503,232 high quality reads, with an average read length of 138 bp. The denovo assembly was performed by Newbler using default and optimized parameters. The optimized parameter generated 40, 380 assembled sequences, comprising 12,940 contigs and 27,440 singlets which resulted in better assembly as compared to default parameters. BLASTX analysis resulted in the annotation of 40,380 contigs/singlet using a cut-off value of ≤ 1E-03. High similarity to Medicago truncatula using optimized parameters and to Populus trichocarpa using default parameters was noted. The Kyoto encyclopedia of genes and genomes (KEGG) analysis using KEGG Automatic Annotation Server (KAAS) combined with domain analysis of the assembled transcripts revealed putative members of secondary metabolism pathways that may be involved in podophyllotoxin biosynthesis. A proposed schematic pathway for phenylpropanoids and podophyllotoxin biosynthesis was generated. Expression profiling was carried out based on fragments per kilobase of exon per million fragments (FPKM). 1036 simple sequence repeats were predicted in the P. hexandrum sequences. Sixty-nine transcripts were mapped to 99 mature and precursor microRNAs from the plant microRNA database. Around 961 transcripts containing transcription factor domains were noted. High performance liquid chromatography analysis showed the peak accumulation of podophyllotoxin in 12-day cell suspension cultures. A comparative qRT-PCR analysis of phenylpropanoid pathway genes identified in the present data was performed to analyze their expression patterns in 12-day cell culture, callus and rhizome. CONCLUSIONS: The present data will help the identification of the potential genes and transcription factors involved in podophyllotoxin biosynthesis in P. hexandrum. The assembled transcripts could serve as potential candidates for marker discovery and conservation, which should form the foundations for future endeavors.


Assuntos
Perfilação da Expressão Gênica , Genes de Plantas , Podophyllum/química , Mapeamento de Sequências Contíguas , Bases de Dados Genéticas , Biblioteca Gênica , Sequenciamento de Nucleotídeos em Larga Escala , Repetições de Microssatélites , Proteínas de Plantas/metabolismo , Podofilotoxina/genética , Podofilotoxina/metabolismo , Análise de Sequência de DNA , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Transcriptoma
8.
Proteome Sci ; 10(1): 34, 2012 May 23.
Artigo em Inglês | MEDLINE | ID: mdl-22621772

RESUMO

BACKGROUND: Podophyllotoxin (PTOX), the precursor for semi-synthesis of cancer therapeutics like etoposide, teniposide and etophos, is primarily obtained from an endangered medicinal herb, Podophyllum hexandrum Royle. PTOX, a lignan is biosynthetically derived from the phenylpropanoid pathway. The aim of this study is to investigate changes in the P. hexandrum cell proteome potentially related to PTOX accumulation in response to methyl jasmonate (MeJA) elicitation. High-resolution two-dimensional gel electrophoresis (2-DE) followed by colloidal Coomassie staining and mass spectrometric analysis was used to detect statistically significant changes in cell's proteome. RESULT: The HPLC analysis showed approximately 7-8 fold change in accumulation of PTOX, in the 12day old cell suspension culture (i.e. after 9days of elicitation) elicited with 100 µM MeJA as compared to the control. Using 2-DE a total of 233 spots was detected, out of which 105 spots were identified by MALDI TOF-TOF MS/MS. Data were subjected to functional annotation from a biological point of view through KEGG. The phenylpropanoid and monolignol pathway enzymes were identified, amongst these, chalcone synthase, polyphenol oxidase, caffeoyl CoA 3-O-methyltransferase, S-adenosyl-L-methionine-dependent methyltransferases, caffeic acid-O-methyl transferase etc. are noted as important. The relation of other differentially accumulated proteins with varied effects caused by elicitors on P. hexandrum cells namely stress and defense related protein, transcription and DNA replication and signaling are also discussed. CONCLUSIONS: Elicitor-induced PTOX accumulation in P. hexandrum cell cultures provides a responsive model system to profile modulations in proteins related to phenylpropanoid/monolignol biosynthesis and other defense responses. Present findings form a baseline for future investigation on a non-sequenced medicinal herb P. hexandrum at molecular level.

9.
Planta ; 233(5): 895-910, 2011 May.
Artigo em Inglês | MEDLINE | ID: mdl-21234598

RESUMO

The elaborate networks and the crosstalk of established signaling molecules like salicylic acid (SA), jasmonic acid (JA), ethylene (ET), abscisic acid (ABA), reactive oxygen species (ROS) and glutathione (GSH) play key role in plant defense response. To obtain further insight into the mechanism through which GSH is involved in this crosstalk to mitigate biotic stress, transgenic Nicotiana tabacum overexpressing Lycopersicon esculentum gamma-glutamylcysteine synthetase (LeECS) gene (NtGB lines) were generated with enhanced level of GSH in comparison with wild-type plants exhibiting resistance to pathogenesis as well. The expression levels of non-expressor of pathogenesis-related genes 1 (NPR1)-dependent genes like pathogenesis-related gene 1 (NtPR1), mitogen-activated protein kinase kinase (NtMAPKK), glutamine synthetase (NtGLS) were significantly enhanced along with NtNPR1. However, the expression levels of NPR1-independent genes like NtPR2, NtPR5 and short-chain dehydrogenase/reductase family protein (NtSDRLP) were either insignificant or were downregulated. Additionally, increase in expression of thioredoxin (NtTRXh), S-nitrosoglutathione reductase 1 (NtGSNOR1) and suppression of isochorismate synthase 1 (NtICS1) was noted. Comprehensive analysis of GSH-fed tobacco BY2 cell line in a time-dependent manner reciprocated the in planta results. Better tolerance of NtGB lines against biotrophic Pseudomonas syringae pv. tabaci was noted as compared to necrotrophic Alternaria alternata. Through two-dimensional gel electrophoresis (2-DE) and image analysis, 48 differentially expressed spots were identified and through identification as well as functional categorization, ten proteins were found to be SA-related. Collectively, our results suggest GSH to be a member in cross-communication with other signaling molecules in mitigating biotic stress likely through NPR1-dependent SA-mediated pathway.


Assuntos
Dipeptídeos/biossíntese , Nicotiana/fisiologia , Proteínas de Plantas/metabolismo , Ciclopentanos/metabolismo , Dipeptídeos/genética , Dipeptídeos/metabolismo , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Variação Genética , Glutationa/biossíntese , Glutationa/metabolismo , Solanum lycopersicum/enzimologia , Solanum lycopersicum/genética , Oxilipinas/metabolismo , Proteínas de Plantas/biossíntese , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas , Proteoma , Ácido Salicílico/metabolismo , Transdução de Sinais , Estresse Fisiológico , Nicotiana/genética , Nicotiana/metabolismo
10.
Plant Physiol Biochem ; 163: 15-25, 2021 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-33799014

RESUMO

Silicon (Si) is a beneficial macronutrient for plants. The Si supplementation to growth media mitigates abiotic and biotic stresses by regulating several physiological, biochemical and molecular mechanisms. The uptake of Si from the soil by root cells and subsequent transport are facilitated by Lsi1 (Low silicon1) belonging to nodulin 26-like major intrinsic protein (NIP) subfamily of aquaporin protein family, and Lsi2 (Low silicon 2) belonging to putative anion transporters, respectively. The soluble Si in the cytosol enhances the production of jasmonic acid, enzymatic and non-enzymatic antioxidants, secondary metabolites and induces expression of genes in plants under stress conditions. Silicon has been found beneficial in conferring tolerance against biotic and abiotic stresses by scavenging the reactive oxygen species (ROS) and regulation of different metabolic pathways. In the present review, Si transporters identified in various plant species and mechanisms of Si-mediated abiotic and biotic stress tolerance have been presented. In addition, role of Si in regulating gene expression under various abiotic and biotic stresses as revealed by transcriptome level studies has been discussed. This provides a deeper understanding of various mechanisms of Si-mediated stress tolerance in plants and may help in devising strategies for stress resilient agriculture.


Assuntos
Plantas , Silício , Agricultura , Transporte Biológico , Silício/farmacologia , Estresse Fisiológico
11.
Methods Mol Biol ; 2107: 99-126, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31893444

RESUMO

Legume crops offer a wide genetic diversity that can be exploited to raise improved crop varieties with higher tolerance against adverse climatic conditions. In order to achieve food and nutritional security, legume breeding programs should also incorporate advanced genomics tools. Genomes of many model and nonmodel legume crops have been sequenced, which provide opportunities to identify and characterize candidate genes to develop abiotic stress tolerant crops. Gene expression profiling is a powerful tool to identify candidate genes and understand their function. The present chapter describes two such strategies, that is, candidate gene expression profiling approach and global transcriptome profiling approach. The methods like RT-PCR and qRT-PCR that are being traditionally used to study expression of target genes under defined experimental conditions are discussed. In addition, global transcriptome analysis approach and its advancements are discussed. Details of next-generation sequencing (NGS) based RNA-sequencing (RNA-seq) and associated advanced bioinformatics tools to identify differentially expressing genes at a global level are also described.


Assuntos
Fabaceae/crescimento & desenvolvimento , Perfilação da Expressão Gênica/métodos , Proteínas de Plantas/genética , Produtos Agrícolas/genética , Produtos Agrícolas/crescimento & desenvolvimento , Fabaceae/genética , Regulação da Expressão Gênica de Plantas , Sequenciamento de Nucleotídeos em Larga Escala , Melhoramento Vegetal , Reação em Cadeia da Polimerase em Tempo Real , Análise de Sequência de RNA , Estresse Fisiológico , Sequenciamento do Exoma
12.
Methods Mol Biol ; 2107: 277-303, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31893454

RESUMO

Legume crops are subjected to a wide range of abiotic stresses, which stimulate an array of physiological, biochemical, and molecular responses. However, different genotypes may exhibit significant variations between individual responses, which can determine their tolerance or susceptibility to these stresses. The present chapter suggests a broad range of assays that can help in understanding stress perception by plants at cellular and molecular levels. The genotypes may be sorted depending on their tolerance potential, by broadly analysing morphological, physiological, biochemical, and enzyme kinetics parameters. These assays are very beneficial in revealing the mechanism of stress perception and response in varied plant types, and have helped in discriminating contrasting genotypes. Here, we have described detailed protocols of assays which may be carried out to assess tolerance or susceptibility to abiotic stresses. The analysis, as a whole, can help researchers understand the effect of abiotic stresses on plant biochemical pathways, be it photosynthesis, redox homeostasis, metabolite perturbation, signaling, transcription, and translation. These protocols may be beneficial in identification of suitable donors for breeding programs, as well as for identifying promising candidate genes or pathways for developing stress tolerant legume crops through genetic engineering.


Assuntos
Fabaceae/crescimento & desenvolvimento , Proteínas de Plantas/metabolismo , Produtos Agrícolas/crescimento & desenvolvimento , Produtos Agrícolas/metabolismo , Fabaceae/metabolismo , Regulação da Expressão Gênica de Plantas , Técnicas de Genotipagem , Homeostase , Oxirredução , Fotossíntese , Melhoramento Vegetal , Estresse Fisiológico
13.
Plant Signal Behav ; 9(8): e29246, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25763614

RESUMO

The contribution of Glutathione (GSH) in drought stress tolerance is an established fact. However, the proteins which are directly or indirectly related to the increased level of GSH in response to drought stress are yet to be known. To explore this, here, transgenic tobacco plants (NtGp11) overexpressing gamma-glutamylcysteine synthetase (γ-ECS) was tested for tolerance against drought stress. NtGp11 conferred tolerance to drought stress by increased germination rate, water retention, water recovery, chlorophyll, and proline content compared with wild-type plants. Semi-quantitative RT-PCR analysis revealed that the transcript levels of stress-responsive genes were higher in NtGp11 compared with wild-type in response to drought stress. Two-dimensional gel electrophoresis (2-DE) coupled with MALDI TOF-TOF MS/MS analysis has been used to identify 43 differentially expressed proteins in response to drought in wild-type and NtGp11 plants. The results demonstrated the up-accumulation of 58.1% of proteins among which 36%, 24%, and 20% of them were related to stress and defense, carbon metabolism and energy metabolism categories, respectively. Taken together, our results demonstrated that GSH plays an important role in combating drought stress in plants by inducing stress related genes and proteins like HSP70, chalcone synthase, glutathione peroxidase, thioredoxin peroxidase, ACC oxidase, and heme oxygenase I.


Assuntos
Adaptação Fisiológica , Secas , Glutamato-Cisteína Ligase/metabolismo , Glutationa/metabolismo , Nicotiana/metabolismo , Proteínas de Plantas/metabolismo , Estresse Fisiológico , Dipeptídeos/metabolismo , Expressão Gênica , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Glutamato-Cisteína Ligase/genética , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas , Reação em Cadeia da Polimerase , Proteômica , Nicotiana/genética , Água
14.
Eur J Med Chem ; 79: 102-9, 2014 May 22.
Artigo em Inglês | MEDLINE | ID: mdl-24727463

RESUMO

Flavonols are an important class of naturally occurring molecules and are known for their pharmacological activity. The activity is associated with the ability of flavonols to influence membrane-dependent processes. We have investigated the in-vitro anti-proliferative and anti-oxidant activity of galangin (GLN), fisetin (FTN) and quercetin (QTN), which possess variable number of phenolic hydroxyl groups. An attempt has been made to correlate the biological activity of these molecules with their interaction and localization in dipalmitoyl phosphatidyl choline (DPPC) bilayers, using differential dcanning calorimetry (DSC) and nuclear magnetic resonance (NMR) methods. Results indicate that GLN interacts to the alkyl chains of the lipid bilayer involving hydrophobic interactions. FTN and QTN interact with head region and sn-1-glycero region involving hydrogen bonding. Ring current induced chemical shifts of lipid protons, due to intermolecular interaction indicate that GLN acquires a parallel orientation with respect to the bilayer normal whereas FTN and QTN resume a mixed orientation. The membrane binding constants of these molecules are in the order GLN > QTN > FTN. It has been shown that the number and position of hydroxyl groups in these molecules play an important role in membrane binding and thereby in biological activity.


Assuntos
Antioxidantes/farmacologia , Flavonoides/farmacologia , Membranas Artificiais , Quercetina/farmacologia , Antineoplásicos/síntese química , Antineoplásicos/química , Antineoplásicos/farmacologia , Antioxidantes/síntese química , Antioxidantes/química , Ciclo Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Cristalografia por Raios X , Relação Dose-Resposta a Droga , Flavonoides/síntese química , Flavonoides/química , Flavonóis , Células Hep G2 , Humanos , Células K562 , Células MCF-7 , Modelos Moleculares , Estrutura Molecular , Quercetina/síntese química , Quercetina/química , Relação Estrutura-Atividade
15.
Eur J Med Chem ; 80: 285-94, 2014 Jun 10.
Artigo em Inglês | MEDLINE | ID: mdl-24793879

RESUMO

The localization and interaction of six naturally occurring flavones (FLV, 5HF, 6HF, 7HF, CHY and BLN) in DPPC bilayers were studied using DSC and multi-nuclear NMR. DSC results indicate that FLV and 6HF interact with alkyl chains. The (1)H NMR shows interaction of flavones with the sn-glycero region. Ring current induced chemical shifts indicate that 6HF and BLN acquire parallel orientation in bilayers. 2D NOESY spectra indicate partitioning of the B-ring into the alkyl chain region. The DSC, NMR and binding studies indicate that 5HF and 7HF are located near head group region, while 6HF, CHY and BLN are located in the vicinity of sn-glycero region, and FLV is inserted deepest in the membrane.


Assuntos
Membrana Celular/metabolismo , Flavonas/metabolismo , Flavonas/farmacologia , Sequestradores de Radicais Livres/metabolismo , Sequestradores de Radicais Livres/farmacologia , Bicamadas Lipídicas/metabolismo , Compostos de Bifenilo/química , Varredura Diferencial de Calorimetria , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Flavonas/química , Sequestradores de Radicais Livres/química , Humanos , Hidróxidos/química , Espectroscopia de Ressonância Magnética , Picratos/química , Relação Estrutura-Atividade
16.
J Plant Physiol ; 171(11): 940-50, 2014 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-24913051

RESUMO

The role of glutathione (GSH) in plant defense is an established fact. However, the association of GSH with other established signaling molecules within the defense signaling network remains to be evaluated. Previously we have shown that GSH is involved in defense signaling network likely through NPR1-dependent salicylic acid (SA)-mediated pathway. In this study, to gain further insight, we developed chloroplast-targeted gamma-glutamylcysteine synthetase (γ-ECS) overexpressed transgenic Nicotiana tabacum (NtGp line) and constructed a forward subtracted cDNA (suppression subtractive hybridization (SSH)) library using NtGp line as a tester. Interestingly, in addition to SA-related transcripts like pathogenesis-related protein 1a (PR1a) and SAR8.2 m/2l, 1-aminocyclopropane-1-carboxylate oxidase (ACC oxidase), a key enzyme of ethylene (ET) biosynthesis, was identified in the SSH library. Besides, transcription factors like WRKY transcription factor 3 (WRKY3), WRKY1 and ethylene responsive factor 4 (ERF4), associated with SA and ET respectively, were also identified thus suggesting an interplay of GSH with ET and SA. Furthermore, proteomic profiling of NtGp line, performed by employing two-dimensional gel electrophoresis (2-DE), corroborated with the transcriptomic profile and several defense-related proteins like serine/threonine protein kinase, and heat shock 70 protein (HSP70) were identified with increased accumulation. Fascinatingly, induction of 1-aminocyclopropane-1-carboxylate synthase (ACC synthase) was also noted thus demonstrating the active involvement of GSH with ET. Protein gel blot analysis confirmed the enhanced accumulation of ACC oxidase in NtGp line. Together, our data revealed that GSH is involved in the synergistic multiple steps crosstalk through ET as well as SA to combat environmental stress.


Assuntos
Etilenos/metabolismo , Glutationa/metabolismo , Ácido Salicílico/metabolismo , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo
17.
J Biosci ; 38(2): 317-28, 2013 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-23660666

RESUMO

Salicylic acid (SA) has been implicated in determining the outcome of interactions between many plants and their pathogens. Global changes in response to this phytohormone have been observed at the transcript level, but little is known of how it induces changes in protein abundance. To this end we have investigated the effect of 1 mM SA on soluble proteins of Arabidopsis thaliana leaves by proteomic analysis. An initial study at transcript level has been performed on temporal landscape, which revealed that induction of most of the SA-responsive genes occurs within 3 to 6 h post treatment (HPT) and the expression peaked within 24 HPT. Two-dimensional gel electrophoresis (2-DE) coupled with MALDI-TOF MS/MS analysis has been used to identify differentially expressed proteins and 63 spots have been identified successfully. This comparative proteomic profiling of SA treated leaves versus control leaves demonstrated the changes of many defence related proteins like pathogenesis related protein 10a (PR10a), diseaseresistance- like protein, putative late blight-resistance protein, WRKY4, MYB4, etc. along with gross increase in the rate of energy production, while other general metabolism rate is slightly toned down, presumably signifying a transition from 'normal mode' to 'defence mode'.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Reguladores de Crescimento de Plantas/fisiologia , Folhas de Planta/metabolismo , Proteoma/metabolismo , Ácido Salicílico/metabolismo , Arabidopsis/imunologia , Metabolismo dos Carboidratos , Resistência à Doença , Metabolismo Energético , Regulação da Expressão Gênica de Plantas , Anotação de Sequência Molecular , Reguladores de Crescimento de Plantas/farmacologia , Biossíntese de Proteínas , Ácido Salicílico/farmacologia , Estresse Fisiológico
18.
J Proteomics ; 93: 117-32, 2013 Nov 20.
Artigo em Inglês | MEDLINE | ID: mdl-23369890

RESUMO

The genus Mentha has been widely used in food, flavor, culinary, cosmetic and pharmaceutical industries. Substantial damage to this crop happened regularly due to environmental stresses like metal toxicity and pathogen attack. Here, an approach has been taken to raise transgenic mint over-expressing γ-glutamyl-cysteine synthetase (γ-ECS), the rate-limiting enzyme of GSH biosynthesis, resulted enhanced GSH content and its in planta expression confers significant tolerance towards abiotic/biotic stresses viz. metal toxicity - Cd, Zn as well as against infection of Alternaria alternata and Rhizoctonia solani. A differential proteomic analysis through 2-DE and MALDI TOF-TOF MSMS was performed to focus on the altered abundance of functionally important protein species in control and infected transgenic mint. Results showed a significant variation in the protein profile of the infected transgenic plant as compared to the wild/control transgenic counterpart. In addition to protein species related to stress and defense, redox regulation, transcription factors and energy & metabolism, protein species related to signaling and gene regulation as well as cell division also showed differential accumulation in infected transgenic. Hence, proteomics can be used as a tool to decipher the mechanism of action of GSH in providing tolerance against a necrotrophic fungus, A. alternata in transgenic mint. BIOLOGICAL SIGNIFICANCE: The reported work describes a comparative proteomics of non-model unsequenced plants like Mentha. There is a comparative protein profile between transgenic and its wild counterparts under control and infected condition. The work has an impact in crop proteomics and also tries to explain the application of proteomic approach to decipher the mechanism by which a foreign metabolite mediates stress tolerance in plant under control and infected condition. This article is part of a Special Issue entitled: Translational Plant Proteomics.


Assuntos
Glutamato-Cisteína Ligase/genética , Glutationa/metabolismo , Mentha/metabolismo , Folhas de Planta/metabolismo , Plantas Geneticamente Modificadas/metabolismo , Alternaria/genética , Alternaria/patogenicidade , Produtos Agrícolas/genética , Mentha/genética , Doenças das Plantas/microbiologia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Proteoma/genética , Proteômica , Estresse Fisiológico , Regulação para Cima
19.
Indian J Pharm Sci ; 75(2): 233-7, 2013 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-24019576

RESUMO

Quercetin is one of the most abundant naturally occurring flavonoid and is associated with a wide range of biological activities, such as antioxidant, antiinflammatory and anticancer activities. However, there are multiple problems associated with the bioavailability of quercetin, thereby restricting its use. Taking this into consideration, the structure of quercetin was modified by removal of multiple hydroxyl groups and introduction of substituents such as Cl, OCH3 and N (CH3)2 on the p-position of the B-ring. The effect of structural modification on the anticancer activity was studied using four different cell lines, including MCF-7, HepG2, HCT-15 and PC-3. Compound 1a has shown an activity better than quercetin in HepG2 cell lines, whereas 1c and 1e showed significant growth inhibition of the HCT-15 cell lines.

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