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1.
Physiol Plant ; 176(2): e14260, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-38511471

RESUMEN

Bacosides are dammarane-type triterpenoidal saponins in Bacopa monnieri and have various pharmacological applications. All the bacosides are diversified from two isomers, i.e., jujubogenin and pseudojujubogenin. The biosynthetic pathway of bacoside is not well elucidated. In the present study, we characterized a UDP-glycosyltransferase, UGT79A18, involved in the glycosylation of pseudojujubogenin. UGT79A18 shows higher expression in response to 5 h of wounding, and 3 h of MeJA treatment. The recombinant UGT79A18 shows in vitro activity against a wide range of flavonoids and triterpenes and has a substrate preference for protopanaxadiol, a dammarane-type triterpene. Secondary metabolite analysis of overexpression and knockdown lines of UGT79A18 in B. monnieri identify bacopasaponin D, bacopaside II, bacopaside N2 and pseudojujubogenin glucosyl rhamnoside as the major bacosides that were differentially accumulated. In the overexpression lines of UGT79A18, we found 1.7-fold enhanced bacopaside II, 8-fold enhanced bacopasaponin D, 3-fold enhanced pseudojujubogenin glucosyl rhamnoside, and 1.6-fold enhanced bacopaside N2 content in comparison with vector control plant, whereas in the knockdown lines of UGT79A18, we found 1.4-fold reduction in bacopaside II content, 3-fold reduction in the bacopasaponin D content, 2-fold reduction in the pseudojujubogenin glucosyl rhamnoside content, and 1.5-fold reduction in bacopaside N2 content in comparison with vector control. These results suggest that UGT79A18 is a significant UDP glycosyltransferase involved in glycosylating pseudojujubogenin and enhancing the pseudojujubogenin-derived bacosides.


Asunto(s)
Acetatos , Bacopa , Ciclopentanos , Oxilipinas , Saponinas , Triterpenos , Bacopa/genética , Bacopa/química , Glicosiltransferasas/genética , Vías Biosintéticas , Triterpenos/química , Triterpenos/farmacología , Triterpenos/uso terapéutico , Damaranos , Uridina Difosfato , Extractos Vegetales/química
2.
Phys Chem Chem Phys ; 26(6): 5311-5322, 2024 Feb 07.
Artículo en Inglés | MEDLINE | ID: mdl-38268444

RESUMEN

To understand the physical phenomena responsible for radiation damage of the materials used in nuclear reactors, and thus study their operation life and/or efficiency, it is required to simulate the conditions by exposing the materials to energetic ions. Ceria (CeO2) has been proposed as one of the inert matrices for the transmutation of minor actinides in the futuristic inert matrix fuel (IMF) concept. The inert matrix should also contain burnable poison to compensate for the initial reactivity of fuel. In this context, gadolinium (Gd) is an excellent burnable poison with a high neutron absorption cross-section. In view of this, Gd2O3-CeO2 nano-powders were synthesized and sintered at 800 °C and 1300 °C to obtain different grain sizes and morphologies. FESEM and TEM were carried out to study the grain size of pristine pellets. The sintered pellets were irradiated with 80-MeV Ag ions (electronic energy loss (Se) regime) at room temperature to emulate the effect of fission fragments. For analysis of the effect of grain size on the irradiation-induced structural degradation at different fluences, GIXRD and Raman spectroscopy were performed. Significantly large damage has been observed for the smaller grain-sized samples (sintered at 800 °C) as compared to the large grain-sized sample (sintered at 1300 °C). Neither of the samples amorphized under the present experimental conditions as indicated by the presence of the Raman-active T2g mode (centred at 462 cm-1) and all the XRD peaks of fluorite cubic structure up to the highest fluence employed (1 × 1014 ions cm-2). X-ray photoelectron spectroscopy results demonstrate that Ce4+ to Ce3+ and vacancy-related isolated clusters are the main defects produced in the systems. The radiation tolerance behaviour of the samples is understood with the help of thermal spike simulation, which indicates higher transient lattice temperatures with longer duration in the smaller grain-sized sample upon irradiation. Gd-doped ceria thus possesses good radiation stability in the Se regime, indicating its potential for application in IMFs.

3.
Plant Cell Rep ; 43(4): 104, 2024 Mar 20.
Artículo en Inglés | MEDLINE | ID: mdl-38507094

RESUMEN

KEY MESSAGE: The present study reports differentially expressed transcripts in the waterlogging-induced adventitious root (AR) of Mentha arvensis; the identified transcripts will help to understand AR development and improve waterlogging stress response. Waterlogging notably hampers plant growth in areas facing waterlogged soil conditions. In our previous findings, Mentha arvensis was shown to adapt better in waterlogging conditions by initiating the early onset of adventitious root development. In the present study, we compared the transcriptome analysis of adventitious root induced after the waterlogging treatment with the control taproot. The biochemical parameters of total carbohydrate, total protein content, nitric oxide (NO) scavenging activity and antioxidant enzymes, such as catalase activity (CAT) and superoxide dismutase (SOD) activity, were enhanced in the adventitious root compared with control taproot. Analysis of differentially expressed genes (DEGs) in adventitious root compared with the control taproot were grouped into four functional categories, i.e., carbohydrate metabolism, antioxidant activity, hormonal regulation, and transcription factors that could be majorly involved in the development of adventitious roots. Differential expression of the upregulated and uniquely expressing thirty-five transcripts in adventitious roots was validated using qRT-PCR. This study has generated the resource of differentially and uniquely expressing transcripts in the waterlogging-induced adventitious roots. Further functional characterization of these transcripts will be helpful to understand the development of adventitious roots, leading to the resistance towards waterlogging stress in Mentha arvensis.


Asunto(s)
Mentha , Mentha/genética , Mentha/metabolismo , Perfilación de la Expresión Génica , Raíces de Plantas/metabolismo
4.
Plant Cell Rep ; 43(6): 147, 2024 May 21.
Artículo en Inglés | MEDLINE | ID: mdl-38771491

RESUMEN

KEY MESSAGE: Thchit42 constitutive expression for fungal resistance showed synchronisation with leaf augmentation and transcriptome analysis revealed the Longifolia and Zinc finger RICESLEEPER gene is responsible for plant growth and development. Pelargonium graveolens essential oil possesses significant attributes, known for perfumery and aromatherapy. However, optimal yield and propagation are predominantly hindered by biotic stress. All biotechnological approaches have yet to prove effective in addressing fungal resistance. The current study developed transgenic geranium bridging molecular mechanism of fungal resistance and plant growth by introducing cassette 35S::Thchit42. Furthermore, 120 independently putative transformed explants were regenerated on kanamycin fortified medium. Primarily transgenic lines were demonstrated peak pathogenicity and antifungal activity against formidable Colletotrichum gloeosporioides and Fusarium oxysporum. Additionally, phenotypic analysis revealed ~ 2fold increase in leaf size and ~ 2.1fold enhanced oil content. To elucidate the molecular mechanisms for genotypic cause, de novo transcriptional profiles were analyzed to indicate that the auxin-regulated longifolia gene is accountable for augmentation in leaf size, and zinc finger (ZF) RICESLEEPER attributes growth upregulation. Collectively, data provides valuable insights into unravelling the mechanism of Thchit42-mediated crosstalk between morphological and chemical alteration in transgenic plants. This knowledge might create novel opportunities to cultivate fungal-resistant geranium throughout all seasons to fulfil demand.


Asunto(s)
Resistencia a la Enfermedad , Fusarium , Regulación de la Expresión Génica de las Plantas , Pelargonium , Hojas de la Planta , Plantas Modificadas Genéticamente , Pelargonium/genética , Fusarium/patogenicidad , Fusarium/fisiología , Resistencia a la Enfermedad/genética , Hojas de la Planta/genética , Hojas de la Planta/microbiología , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Enfermedades de las Plantas/microbiología , Enfermedades de las Plantas/genética , Colletotrichum/patogenicidad , Colletotrichum/fisiología , Aceites Volátiles/metabolismo , Aceites Volátiles/farmacología , Geranium/genética
5.
Plant Mol Biol ; 110(3): 235-251, 2022 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-35780285

RESUMEN

KEY MESSAGE: OscWRKY1 from Ocimum sanctum positively regulates phenylpropanoid pathway genes and rosmarinic acid content. OscWRKY1 overexpression promotes resistance against bacterial pathogen in Arabidopsis. WRKY transcription factor (TF) family regulates various developmental and physiological functions in plants. PAL genes encode enzymes which are involved in plant defense responses, but the direct regulation of PAL genes and phenylpropanoid pathway through WRKY TF's is not well characterized. In the present study, we have characterized an OscWRKY1 gene from Ocimum sanctum which shows induced expression by methyl jasmonate (MeJA), salicylic acid (SA), and wounding. The recombinant OscWRKY1 protein binds to the DIG-labeled (Digoxigenin) W-box cis-element TTGAC[C/T] and activates the LacZ reporter gene in yeast. Overexpression of OscWRKY1 enhances Arabidopsis resistance towards Pseudomonas syringae pv. tomato Pst DC3000. Upstream activator sequences of PAL and C4H have been identified to contain the conserved W-box cis-element (TTGACC) in both O. sanctum and Arabidopsis. OscWRKY1 was found to interact with W-box cis-element present in the PAL and C4H promoters. Silencing of OscWRKY1 using VIGS resulted in reduced expression of PAL, C4H, COMT, F5H and 4CL transcripts. OscWRKY1 silenced plants exhibit reduced PAL activity, whereas, the overexpression lines of OscWRKY1 in Arabidopsis exhibit increased PAL activity. Furthermore, the metabolite analysis of OscWRKY1 silenced plants showed reduced rosmarinic acid content. These results revealed that OscWRKY1 positively regulates the phenylpropanoid pathway genes leading to the alteration of rosmarinic acid content and enhances the resistance against bacterial pathogen in Arabidopsis.


Asunto(s)
Proteínas de Arabidopsis , Arabidopsis , Arabidopsis/metabolismo , Proteínas de Arabidopsis/metabolismo , Cinamatos , Depsidos , Digoxigenina/metabolismo , Resistencia a la Enfermedad/genética , Regulación de la Expresión Génica de las Plantas , Ocimum sanctum/metabolismo , Enfermedades de las Plantas/genética , Enfermedades de las Plantas/microbiología , Pseudomonas syringae/fisiología , Ácido Salicílico/metabolismo , Ácido Salicílico/farmacología , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Ácido Rosmarínico
6.
Plant Cell Rep ; 41(8): 1651-1671, 2022 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-35579713

RESUMEN

KEY MESSAGE: The present review highlights the regulatory roles of microRNAs in plant secondary metabolism and focuses on different bioengineering strategies to modulate secondary metabolite content in plants. MicroRNAs (miRNAs) are the class of small endogenous, essential, non-coding RNAs that riboregulate the gene expression involved in various biological processes in most eukaryotes. MiRNAs has emerged as important regulators in plants that function by silencing target genes through cleavage or translational inhibition. These miRNAs plays an important role in a wide range of plant biological and metabolic processes, including plant development and various environmental response controls. Several important plant secondary metabolites like alkaloids, terpenoids, and phenolics are well studied for their function in plant defense against different types of pests and herbivores. Due to the presence of a wide range of biological and pharmaceutical properties of plant secondary metabolites, it is important to study the regulation of their biosynthetic pathways. The contribution of miRNAs in regulating plant secondary metabolism is not well explored. Recent advancements in molecular techniques have improved our knowledge in understanding the molecular function of genes, proteins, enzymes, and small RNAs involved in different steps of secondary metabolic pathways. In the present review, we have discussed the recent progress made on miRNA biogenesis, its regulation, and highlighted the current research developed in the field of identification, analysis, and characterizations of various miRNAs that regulate plant secondary metabolism. We have also discussed how different bioengineering strategies such as artificial miRNA (amiRNA), endogenous target mimicry, and CRISPR/Cas9 could be utilized to enhance the secondary metabolite production in plants.


Asunto(s)
MicroARNs , ARN Pequeño no Traducido , Regulación de la Expresión Génica de las Plantas , MicroARNs/genética , MicroARNs/metabolismo , Desarrollo de la Planta , Plantas/genética , Plantas/metabolismo , ARN Pequeño no Traducido/metabolismo , Metabolismo Secundario/genética
7.
Plant Cell Physiol ; 62(5): 894-912, 2021 Oct 01.
Artículo en Inglés | MEDLINE | ID: mdl-34009389

RESUMEN

MicroRNAs (miRNAs) are small non-coding, endogenous RNAs containing 20-24 nucleotides that regulate the expression of target genes involved in various plant processes. A total of 1,429 conserved miRNAs belonging to 95 conserved miRNA families and 12 novel miRNAs were identified from Bacopa monnieri using small RNA sequencing. The Bm-miRNA target transcripts related to the secondary metabolism were further selected for validation. The Bm-miRNA expression in shoot and root tissues was negatively correlated with their target transcripts. The Bm-miRNA cleavage sites were mapped within the coding or untranslated region as depicted by the modified RLM-RACE. In the present study, we validate three miRNA targets, including asparagine synthetase, cycloartenol synthase and ferulate 5 hydroxylase (F5H) and elucidate the regulatory role of Bm-miR172c-5p, which cleaves the F5H gene involved in the lignin biosynthesis. Overexpression (OE) of Bm-miR172c-5p precursor in B. monnieri suppresses F5H gene, leading to reduced lignification and secondary xylem thickness under control and drought stress. By contrast, OE of endogenous target mimics (eTMs) showed enhanced lignification and secondary xylem thickness leading to better physiological response under drought stress. Taken together, we suggest that Bm-miRNA172c-5p might be a key player in maintaining the native phenotype of B. monnieri under control and different environmental conditions.


Asunto(s)
Bacopa/genética , Bacopa/metabolismo , Lignina/biosíntesis , MicroARNs/genética , Oxigenasas de Función Mixta/genética , Sequías , Regulación de la Expresión Génica de las Plantas , Lignina/genética , Oxigenasas de Función Mixta/metabolismo , Filogenia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Modificadas Genéticamente , Reproducibilidad de los Resultados , Análisis de Secuencia de ARN , Xilema/química , Xilema/metabolismo
8.
Planta ; 253(5): 89, 2021 Apr 05.
Artículo en Inglés | MEDLINE | ID: mdl-33818685

RESUMEN

MAIN CONCLUSION: BmG10H-1 transcript from B. monnieri was functionally active. BmG10H-1 promoter drives GUS activity in response to MeJA and wounding. BmMYB35 regulates BmG10H-1 transcript by binding to its promoter. Geraniol 10-hydroxylase (G10H) is one of the important regulatory cytochrome P450 monooxygenase, which is involved in the biosynthesis of monoterpene alkaloids. However, G10H is not characterized at the enzymatic or at the regulatory aspect in B. monnieri. In the present study, we have identified two transcripts of BmG10H (BmG10H-1and BmG10H-2) and characterized the methyl jasmonate (MeJA) and wound responsive BmG10H-1 transcript from B. monnieri. BmG10H-1 showed induced expression after 3 h of MeJA and wounding treatment in the shoot. Yeast purified recombinant BmG10H-1 protein is enzymatically active, having Vmax of 0.16 µMsec-1 µg-1 protein and catalyzes the hydroxylation of geraniol to 10-hydroxy geraniol. The BmG10H-1 promoter was isolated by using the genome walking method. BmG10H-1 promoter can drive GUS expression in transgenic Arabidopsis thaliana. GUS activity of MeJA and wound-treated Arabidopsis seedlings were found to be increased as compared to the control untreated seedlings, whereas no GUS activity was found in deleted MeJA responsive and W-box cis-elements. This shows that the BmG10H-1 promoter contains functional MeJA (TGACG) and wound responsive (TGACCT) cis-elements. Further, shoot specific and MeJA responsive recombinant BmMYB35 protein was purified, which binds with the MYB recognition cis-element (TGGTTA) present in the BmG10H-1 promoter and transcriptionally activates the reporter gene in yeast. In conclusion, the characterization of MeJA and wound responsive BmG10H-1 provides novel information about its transcriptional regulation by binding with MYB transcription factor in B. monnieri.


Asunto(s)
Acetatos/metabolismo , Bacopa/genética , Bacopa/metabolismo , Ciclopentanos/metabolismo , Sistema Enzimático del Citocromo P-450/genética , Genes de Plantas/genética , Oxilipinas/metabolismo , Bacopa/enzimología , Secuencia de Bases , Regulación de la Expresión Génica de las Plantas , Plantas Modificadas Genéticamente/genética
9.
Plant Mol Biol ; 100(4-5): 351-365, 2019 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-31030374

RESUMEN

KEY MESSAGE: Present review describes the structure, evolution, transport mechanism and physiological functions of SWEETs. Their application using TALENs and CRISPR/CAS9 based genomic editing approach is discussed. Sugars Will Eventually be Exported Transporters (SWEET) proteins were first identified in plants as the novel family of sugar transporters which mediates the translocation of sugars across cell membranes. The SWEET family of sugar transporters is unique in terms of their structure which contains seven predicted transmembrane domains with two internal triple-helix bundles which possibly originate due to prokaryotic gene duplication. SWEETs perform diverse physiological functions such as pollen nutrition, nectar secretion, seed filling, phloem loading, and pathogen nutrition which we have discussed in the present review. We also discuss how transcriptional activator-like effector nucleases (TALENs) and CRISPR/CAS9 genome editing tools are used to engineer SWEET mutants which modulate pathogen resistance in plants and its applications in the field of agriculture. The expression of SWEETs promises to implement insights into many other cellular transport mechanisms. To conclude, the present review highlights the recent aspects which will further develop better understanding of molecular evolution, structure, and function of SWEET transporters in plants.


Asunto(s)
Proteínas de Transporte de Monosacáridos/fisiología , Proteínas de Plantas/fisiología , Membrana Celular/metabolismo , Resistencia a la Enfermedad , Evolución Molecular , Giberelinas/metabolismo , Modelos Moleculares , Proteínas de Transporte de Monosacáridos/química , Floema/metabolismo , Proteínas de Plantas/química , Plantas/metabolismo , Plantas/microbiología , Dominios Proteicos , Análisis de Secuencia de Proteína
10.
Planta ; 248(3): 519-544, 2018 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-29748819

RESUMEN

MAIN CONCLUSION: Steroidal saponins exhibited numerous pharmacological activities due to the modification of their backbone by different cytochrome P450s (P450) and UDP glycosyltransferases (UGTs). Plant-derived steroidal saponins are not sufficient for utilizing them for commercial purpose so in vitro production of saponin by tissue culture, root culture, embryo culture, etc, is necessary for its large-scale production. Saponin glycosides are the important class of plant secondary metabolites, which consists of either steroidal or terpenoidal backbone. Due to the existence of a wide range of medicinal properties, saponin glycosides are pharmacologically very important. This review is focused on important medicinal properties of steroidal saponin, its occurrence, and biosynthesis. In addition to this, some recently identified plants containing steroidal saponins in different parts were summarized. The high throughput transcriptome sequencing approach elaborates our understanding related to the secondary metabolic pathway and its regulation even in the absence of adequate genomic information of non-model plants. The aim of this review is to encapsulate the information related to applications of steroidal saponin and its biosynthetic enzymes specially P450s and UGTs that are involved at later stage modifications of saponin backbone. Lastly, we discussed the in vitro production of steroidal saponin as the plant-based production of saponin is time-consuming and yield a limited amount of saponins. A large amount of plant material has been used to increase the production of steroidal saponin by employing in vitro culture technique, which has received a lot of attention in past two decades and provides a way to conserve medicinal plants as well as to escape them for being endangered.


Asunto(s)
Saponinas/biosíntesis , Esteroides/biosíntesis , Perfilación de la Expresión Génica , Secuenciación de Nucleótidos de Alto Rendimiento , Técnicas In Vitro , Redes y Vías Metabólicas , Plantas/enzimología , Plantas/metabolismo , Plantas Medicinales/metabolismo , Técnicas de Cultivo de Tejidos
11.
Plant Biotechnol J ; 16(1): 221-233, 2018 01.
Artículo en Inglés | MEDLINE | ID: mdl-28636266

RESUMEN

As waterlogging and successive events severely influence growth and development of economically important plants, we attempted to characterize the role of a waterlogging-responsive group I (A-6) ethylene response factor (MaRAP2-4) from Mentha arvensis. Waterlogging, ethylene and methyl jasmonate rapidly induced the expression of MaRAP2-4. MaRAP2-4 interacted with multiple cis-elements like dehydration response elements (DRE1/2), anoxia/jasmonic acid response element (JARE) and GCC box showing its involvement in multiple responses. MaRAP2-4 localizes in the nucleus and acts as a transcriptional activator. Truncation and internal deletion identified a 20 amino acids potential transactivation domain (PLPSSVDAKLEAICQSLAIN) in MaRAP2-4. MaRAP2-4 transgenic Arabidopsis showed enhanced waterlogging and subsequent oxidative stress tolerance. Microarray analysis revealed that within up-regulated genes 483, 212 and 132 promoters carry either single or multiple copies of DRE, JARE and GCC cis-element/s, respectively. Within these promoters, a large section belongs to carbohydrate metabolism/transport, including many SWEET transporters. Further analysis showed MaRAP2-4 specifically targets two positions in AtSWEEET10 promoter carrying DRE and/or GCC box that might regulate carbohydrate availability and waterlogging tolerance. These results demonstrate that MaRAP2-4 is a positive regulator of waterlogging tolerance, and as energy-consuming processes such as carbohydrate biosynthesis are reduced under waterlogging-induced hypoxia, sugar transport through SWEETs may be the primary option to make sugar available to the required tissue.


Asunto(s)
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Mentha/metabolismo , Plantas Modificadas Genéticamente/metabolismo , Acetatos/metabolismo , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Ciclopentanos/metabolismo , Regulación de la Expresión Génica de las Plantas/genética , Regulación de la Expresión Génica de las Plantas/fisiología , Oxilipinas/metabolismo , Plantas Modificadas Genéticamente/genética , Regiones Promotoras Genéticas/genética
12.
BMC Genomics ; 18(1): 490, 2017 06 28.
Artículo en Inglés | MEDLINE | ID: mdl-28659188

RESUMEN

BACKGROUND: Bacopa monnieri commonly known as Brahmi is utilized in Ayurveda to improve memory and many other human health benefits. Bacosides enriched standardized extract of Bacopa monnieri is being marketed as a memory enhancing agent. In spite of its well known pharmacological properties it is not much studied in terms of transcripts involved in biosynthetic pathway and its regulation that controls the secondary metabolic pathway in this plant. The aim of this study was to identify the potential transcripts and provide a framework of identified transcripts involved in bacosides production through transcriptome assembly. RESULTS: We performed comparative transcriptome analysis of shoot and root tissue of Bacopa monnieri in two independent biological replicate and obtained 22.48 million and 22.0 million high quality processed reads in shoot and root respectively. After de novo assembly and quantitative assessment total 26,412 genes got annotated in root and 18,500 genes annotated in shoot sample. Quality of raw reads was determined by using SeqQC-V2.2. Assembled sequences were annotated using BLASTX against public database such as NR or UniProt. Searching against the KEGG pathway database indicated that 37,918 unigenes from root and 35,130 unigenes from shoot were mapped to 133 KEGG pathways. Based on the DGE data we found that most of the transcript related to CYP450s and UDP-glucosyltransferases were specifically upregulated in shoot tissue as compared to root tissue. Finally, we have selected 43 transcripts related to secondary metabolism including transcription factor families which are differentially expressed in shoot and root tissues were validated by qRT-PCR and their expression level were monitored after MeJA treatment and wounding for 1, 3 and 5 h. CONCLUSIONS: This study not only represents the first de novo transcriptome analysis of Bacopa monnieri but also provides information about the identification, expression and differential tissues specific distribution of transcripts related to triterpenoid sapogenin which is one of the most important pharmacologically active secondary metabolite present in Bacopa monnieri. The identified transcripts in this study will establish a foundation for future studies related to carrying out the metabolic engineering for increasing the bacosides biosynthesis and its regulation for human health benefits.


Asunto(s)
Bacopa/genética , Bacopa/metabolismo , Perfilación de la Expresión Génica , Raíces de Plantas/genética , Brotes de la Planta/genética , Saponinas/biosíntesis , Triterpenos/química , Ontología de Genes , Propanoles/metabolismo , Saponinas/química , Factores de Transcripción/metabolismo
13.
BMC Genomics ; 18(1): 54, 2017 01 09.
Artículo en Inglés | MEDLINE | ID: mdl-28068903

RESUMEN

BACKGROUND: Pyrethrins are monoterpenoids and consist of either a chrysanthemic acid or pyrethric acid with a rethrolone moiety. Natural pyrethrins are safe and eco-friendly while possessing strong insecticidal properties. Despite such advantages of commercial value coming with the eco-friendly tag, most enzymes/genes involved in the pyrethrin biosynthesis pathway remain unidentified and uncharacterized. Since the flowers of Tanacetum cinerariifolium are rich in major pyrethrins, next generation transcriptome sequencing was undertaken to compare the flowers and the leaves of the plant de novo to identify differentially expressed transcripts and ascertain which among them might be involved in and responsible for the differential accumulation of pyrethrins in T. cinerariifolium flowers. RESULTS: In this first tissue specific transcriptome analysis of the non-model plant T. cinerariifolium, a total of 23,200,000 and 28,500,110 high quality Illumina next generation sequence reads, with a length of 101 bp, were generated for the flower and leaf tissue respectively. After functional enrichment analysis and GO based annotation using public protein databases such as UniRef, PFAM, SMART, KEGG and NR, 4443 and 8901 unigenes were identified in the flower and leaf tissue respectively. These could be assigned to 13344 KEGG pathways and the pyrethrin biosynthesis contextualized. The 2-C-methyl-D-erythritol 4-phosphate (MEP) pathway was involved in the biosynthesis of acid moiety of pyrethrin and this pathway predominated in the flowers as compared to the leaves. However, enzymes related to oxylipin biosynthesis were found predominantly in the leaf tissue, which suggested that major steps of pyrethrin biosynthesis occurred in the flowers. CONCLUSIONS: Transcriptome comparison between the flower and leaf tissue of T. cinerariifolium provided an elaborate list of tissue specific transcripts that was useful in elucidating the differences in the expression of the biosynthetic pathways leading to differential presence of pyrethrin in the flowers. The information generated on genes, pathways and markers related to pyrethrin biosynthesis in this study will be helpful in enhancing the production of these useful compounds for value added breeding programs. Related proteome comparison to overlay our transcriptome comparison can generate more relevant information to better understand flower specific accumulation of secondary metabolites in general and pyrethrin accumulation in particular.


Asunto(s)
Productos Biológicos/metabolismo , Chrysanthemum cinerariifolium/genética , Chrysanthemum cinerariifolium/metabolismo , Perfilación de la Expresión Génica , Genes de Plantas/genética , Insecticidas/metabolismo , Piretrinas/metabolismo , Ontología de Genes , Anotación de Secuencia Molecular , Proteómica , Análisis de Secuencia
14.
Brain Behav Immun ; 59: 173-189, 2017 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-27658543

RESUMEN

Studies have demonstrated separately that hypertension is associated with platelet activation in the periphery (resulting in accumulation and localized inflammatory response) and glial activation in the brain. We investigated the contribution of platelets in brain inflammation, particularly glial activation in vitro and in a rat model of hypertension. We found that HTN increased the expression of adhesion molecules like JAM-1, ICAM-1, and VCAM-1 on brain endothelium and resulted in the deposition of platelets in the brain. Platelet deposition in hypertensive rats was associated with augmented CD40 and CD40L and activation of astrocytes (GFAP expression) and microglia (Iba-1 expression) in the brain. Platelets isolated from hypertensive rats had significantly higher sCD40L levels and induced more prominent glial activation than platelets from normotensive rats. Activation of platelets with ADP induced sCD40L release and activation of astrocytes and microglia. Moreover, CD40L induced glial (astrocytes and microglia) activation, NFкB and MAPK inflammatory signaling, culminating in neuroinflammation and neuronal injury (increased apoptotic cells). Importantly, injection of ADP-activated platelets into normotensive rats strongly induced activation of astrocytes and microglia and increased plasma sCD40L levels compared with control platelets. On the contrary, inhibition of platelet activation by Clopidogrel or disruption of CD40 signaling prevented astrocyte and microglial activation and provided neuroprotection in both in vivo and in vitro conditions. Thus, we have identified platelet CD40L as a key inflammatory molecule for the induction of astrocyte and microglia activation, the major contributors to inflammation-mediated injury in the brain.


Asunto(s)
Astrocitos/inmunología , Plaquetas/química , Ligando de CD40/farmacología , Hipertensión/inmunología , Microglía/inmunología , Animales , Ligando de CD40/sangre , Proteínas de Unión al Calcio/biosíntesis , Proteínas de Unión al Calcio/genética , Moléculas de Adhesión Celular/metabolismo , Células Cultivadas , Citocinas/metabolismo , Proteína Ácida Fibrilar de la Glía/biosíntesis , Hipertensión/sangre , Inflamación/patología , Activación de Macrófagos/efectos de los fármacos , Masculino , Microcirculación , Proteínas de Microfilamentos/biosíntesis , Proteínas de Microfilamentos/genética , Proteínas Quinasas Activadas por Mitógenos/metabolismo , FN-kappa B/inmunología , Ratas , Ratas Sprague-Dawley
15.
Inorg Chem ; 56(14): 8363-8371, 2017 Jul 17.
Artículo en Inglés | MEDLINE | ID: mdl-28648048

RESUMEN

The effects of high pressure on the crystal structure of orthorhombic (Pnma) perovskite-type cerium scandate were studied in situ under high pressure by means of synchrotron X-ray powder diffraction, using a diamond-anvil cell. We found that the perovskite-type crystal structure remains stable up to 40 GPa, the highest pressure reached in the experiments. The evolution of unit-cell parameters with pressure indicated an anisotropic compression. The room-temperature pressure-volume equation of state (EOS) obtained from the experiments indicated the EOS parameters V0 = 262.5(3) Å3, B0 = 165(7) GPa, and B0' = 6.3(5). From the evolution of microscopic structural parameters like bond distances and coordination polyhedra of cerium and scandium, the macroscopic behavior of CeScO3 under compression was explained and reasoned for its large pressure stability. The reported results are discussed in comparison with high-pressure results from other perovskites.

16.
Crit Rev Biotechnol ; 36(5): 956-66, 2016 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-26177332

RESUMEN

Submergence, whether partial or complete, imparts some serious consequences on plants grown in flood prone ecosystems. Some plants can endure these conditions by embracing various survival strategies, including morphological adaptations and physiological adjustments. This review summarizes recent progress made in understanding of the stress and the acclimation responses of plants under waterlogged or submerged conditions. Waterlogging and submergence are often associated with hypoxia development, which may trigger various morphological traits and cellular acclimation responses. Ethylene, abscisic acid, gibberellic acid and other hormones play a crucial role in the survival process which is controlled genetically. Effects at the cellular level, including ATP management, starch metabolism, elemental toxicity, role of transporters and redox status have been explained. Transcriptional and hormonal interplay during this stress may provide some key aspects in understanding waterlogging and submergence tolerance. The level and degree of tolerance may vary depending on species or climatic variations which need to be studied for a proper understanding of waterlogging stress at the global level. The exploration of regulatory pathways and interplay in model organisms such as Arabidopsis and rice would provide valuable resources for improvement of economically and agriculturally important plants in waterlogging affected areas.


Asunto(s)
Adaptación Fisiológica , Fenómenos Fisiológicos de las Plantas , Plantas/anatomía & histología , Agua/fisiología , Inundaciones , Desarrollo de la Planta , Reguladores del Crecimiento de las Plantas/fisiología , Estrés Fisiológico , Transcripción Genética
17.
J Clin Densitom ; 19(2): 154-64, 2016.
Artículo en Inglés | MEDLINE | ID: mdl-25936482

RESUMEN

We evaluated functional measures of neuromuscular integrity and bone's resistance to fracture as a combined tool in discriminating osteoporosis patients with and without fractures. Functional aspects of neuromuscular integrity were quantified with a noninvasive measure of static and dynamic functional postural stability (FPS), and fracture resistance was obtained with bone shock absorption in patients with osteoporosis aged 65-85 and compared our measures with dual-energy X-ray absorptiometry and Fracture Risk Assessment Tool (FRAX [World Health Organization Collaborating Center for Metabolic Bone Diseases, Sheffield, UK]) in women with osteoporosis, some with and some without vertebral fractures. Patients with vertebral fracture showed larger static FPS (postural sway excursion) in the mediolateral and anterior-posterior directions, suggesting poorer balance. Most of the variables of dynamic FPS showed significant differences between fracture and no-fracture groups (e.g., the fracture group took significantly longer during turning, implying poorer dynamic balance control). Also, compared with healthy control subjects, all 4 dynamic FPS responses for osteoporosis patients with and without fracture were significantly poorer, suggesting potential risk for falls. In summary, patients with osteoporosis who have vertebral fractures (compared with patients with similarly low bone mineral density and other nonfracture risk fractures) have not only lower bone shock absorption damping (ζ) but also increased postural imbalance.


Asunto(s)
Densidad Ósea , Unión Neuromuscular/fisiopatología , Osteoporosis , Fracturas Osteoporóticas , Equilibrio Postural , Fracturas de la Columna Vertebral , Absorciometría de Fotón/métodos , Accidentes por Caídas/prevención & control , Anciano , Anciano de 80 o más Años , Femenino , Humanos , Masculino , Osteoporosis/diagnóstico , Osteoporosis/fisiopatología , Fracturas Osteoporóticas/diagnóstico , Fracturas Osteoporóticas/epidemiología , Fracturas Osteoporóticas/fisiopatología , Medición de Riesgo/métodos , Factores de Riesgo , Fracturas de la Columna Vertebral/diagnóstico , Fracturas de la Columna Vertebral/epidemiología , Fracturas de la Columna Vertebral/fisiopatología , Estados Unidos
18.
J Asian Nat Prod Res ; 18(2): 172-83, 2016.
Artículo en Inglés | MEDLINE | ID: mdl-26527419

RESUMEN

Saraca asoca bark has been used in the Ayurvedic system of medicine for female urino-genital disorders. We have recently reported the isolation and characterization of several compounds as markers to develop HPLC profiling of its methanol and aqueous methanol extracts. Now, a HPLC-PDA inactive compound, (+)-pinitol has been isolated and characterized from the bark of this medicinally important tree. Pinitol is a well known bioactive compound for a variety of biological activities, including hypoglycemic and anti-inflammatory activities. A process for the isolation of relatively good concentration of (+)-pinitol from S. asoca bark has been developed and its in vitro anti TNF-α and anti-inflammatory activities against carragenan-induced edema confirmed. While conducting experiments on the possible agonistic activity, it was found that (+)-pinitol showed up to eight fold reduction in the doses of ß-lactam antibiotics. The mechanism of its agonistic activity was studied by docking experiments which showed that different conformations of (+)-pinitol and antibiotics were actually in the same binding site with no significant change in the binding energy. These docking simulations, thus predict the possible binding mode of studied compounds and probable reason behind the synergistic effect of (+)-pinitol along with ß-lactam antibiotics.


Asunto(s)
Antibacterianos/aislamiento & purificación , Antibacterianos/farmacología , Antiinflamatorios/aislamiento & purificación , Antiinflamatorios/farmacología , Fabaceae/química , Inositol/análogos & derivados , beta-Lactamas/aislamiento & purificación , beta-Lactamas/farmacología , Animales , Antibacterianos/química , Antiinflamatorios/química , Carragenina/farmacología , Cromatografía Líquida de Alta Presión , Edema/inducido químicamente , Edema/tratamiento farmacológico , Femenino , Humanos , India , Inositol/farmacología , Lipopolisacáridos/farmacología , Medicina Ayurvédica , Staphylococcus aureus Resistente a Meticilina/efectos de los fármacos , Pruebas de Sensibilidad Microbiana , Estructura Molecular , Resonancia Magnética Nuclear Biomolecular , Corteza de la Planta/química , Ratas , Estereoisomerismo , Factor de Necrosis Tumoral alfa/antagonistas & inhibidores , beta-Lactamas/química
19.
Plant Mol Biol ; 89(1-2): 173-86, 2015 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-26319514

RESUMEN

The AP2/ERFs are one of the most important family of transcription factors which regulate multiple responses like stress, metabolism and development in plants. We isolated PsAP2 a novel AP2/ERF from Papaver somniferum which was highly upregulated in response to wounding followed by ethylene, methyl jasmonate and ABA treatment. PsAP2 showed specific binding with both DRE and GCC box elements and it was able to transactivate the reporter genes in yeast. PsAP2 overexpressing transgenic tobacco plants exhibited enhanced tolerance towards both abiotic and biotic stresses . Real time transcript expression analysis showed constitutive upregulation of tobacco Alternative oxidase1a and Myo-inositol-1-phosphate synthase in PsAP2 overexpressing tobacco plants. Further, PsAP2 showed interaction with NtAOX1a promoter in vitro, it also specifically activated the NtAOX1a promoter in yeast and tobacco BY2 cells. The silencing of PsAP2 using VIGS lead to significant reduction in the AOX1 level in P. somniferum. Taken together PsAP2 can directly bind and transcriptionally activate NtAOX1a and its overexpression in tobacco imparted increased tolerance towards both abiotic and biotic stress.


Asunto(s)
Papaver/fisiología , Proteínas de Plantas/fisiología , Estrés Fisiológico/fisiología , Factores de Transcripción/fisiología , Secuencia de Aminoácidos , Secuencia de Bases , Regulación de la Expresión Génica de las Plantas/genética , Regulación de la Expresión Génica de las Plantas/fisiología , Silenciador del Gen/fisiología , Datos de Secuencia Molecular , Papaver/genética , Filogenia , Proteínas de Plantas/genética , Plantas Modificadas Genéticamente/genética , Plantas Modificadas Genéticamente/fisiología , Regiones Promotoras Genéticas/genética , Regiones Promotoras Genéticas/fisiología , Estrés Fisiológico/genética , Nicotiana/genética , Nicotiana/fisiología , Factores de Transcripción/genética
20.
Homeopathy ; 104(1): 24-8, 2015 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-25576268

RESUMEN

BACKGROUND: Lycopodium clavatum (Lyc) is a widely used homeopathic medicine for the liver, urinary and digestive disorders. Recently, acetyl cholinesterase (AchE) inhibitory activity has been found in Lyc alkaloid extract, which could be beneficial in dementia disorder. However, the effect of Lyc has not yet been explored in animal model of memory impairment and on cerebral blood flow. AIM: The present study was planned to explore the effect of Lyc on learning and memory function and cerebral blood flow (CBF) in intracerebroventricularly (ICV) administered streptozotocin (STZ) induced memory impairment in rats. MATERIALS AND METHODS: Memory deficit was induced by ICV administration of STZ (3 mg/kg) in rats on 1st and 3rd day. Male SD rats were treated with Lyc Mother Tincture (MT) 30, 200 and 1000 for 17 days. Learning and memory was evaluated by Morris water maze test on 14th, 15th and 16th day. CBF was measured by Laser Doppler flow meter on 17th day. RESULTS: STZ (ICV) treated rats showed impairment in learning and memory along with reduced CBF. Lyc MT and 200 showed improvement in learning and memory. There was increased CBF in STZ (ICV) treated rats at all the potencies of Lyc studied. CONCLUSION: The above study suggests that Lyc may be used as a drug of choice in condition of memory impairment due to its beneficial effect on CBF.


Asunto(s)
Circulación Cerebrovascular/efectos de los fármacos , Homeopatía , Lycopodium , Trastornos de la Memoria/tratamiento farmacológico , Memoria/efectos de los fármacos , Animales , Masculino , Aprendizaje por Laberinto/efectos de los fármacos , Ratas , Ratas Sprague-Dawley , Estreptozocina/farmacología
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