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1.
Mol Cell ; 58(3): 453-67, 2015 May 07.
Artículo en Inglés | MEDLINE | ID: mdl-25866244

RESUMEN

Phosphoinositides (PI) are important signaling molecules in the nucleus that influence gene expression. However, if and how nuclear PI directly affects the transcriptional machinery is not known. We report that the lipid kinase PIP4K2B regulates nuclear PI5P and the expression of myogenic genes during myoblast differentiation. A targeted screen for PI interactors identified the PHD finger of TAF3, a TATA box binding protein-associated factor with important roles in transcription regulation, pluripotency, and differentiation. We show that the PI interaction site is distinct from the known H3K4me3 binding region of TAF3 and that PI binding modulates association of TAF3 with H3K4me3 in vitro and with chromatin in vivo. Analysis of TAF3 mutants indicates that TAF3 transduces PIP4K2B-mediated alterations in PI into changes in specific gene transcription. Our study reveals TAF3 as a direct target of nuclear PI and further illustrates the importance of basal transcription components as signal transducers.


Asunto(s)
Núcleo Celular/metabolismo , Proteínas de Homeodominio/metabolismo , Fosfatidilinositoles/metabolismo , Transcripción Genética , Secuencia de Aminoácidos , Animales , Diferenciación Celular/genética , Línea Celular , Núcleo Celular/genética , Electroforesis en Gel de Poliacrilamida , Perfilación de la Expresión Génica , Histonas/metabolismo , Proteínas de Homeodominio/genética , Lisina/metabolismo , Metilación , Ratones , Antígenos de Histocompatibilidad Menor , Datos de Secuencia Molecular , Mutación , Mioblastos/citología , Mioblastos/metabolismo , Análisis de Secuencia por Matrices de Oligonucleótidos , Fosfotransferasas (Aceptor de Grupo Alcohol)/genética , Fosfotransferasas (Aceptor de Grupo Alcohol)/metabolismo , Unión Proteica , Interferencia de ARN , Homología de Secuencia de Aminoácido , Factores Asociados con la Proteína de Unión a TATA , Factor de Transcripción TFIID/genética , Factor de Transcripción TFIID/metabolismo , Pez Cebra/embriología , Pez Cebra/genética , Pez Cebra/metabolismo
2.
Molecules ; 26(19)2021 Sep 25.
Artículo en Inglés | MEDLINE | ID: mdl-34641363

RESUMEN

Gui Zhen Cao is an herbal formulation that has been documented in Chinese traditional medicine as a remedy for diarrhea, dysentery, inflammation, and toxicity. The sources of this formulation (Bidens pilosa L., Bidens biternata (Lour.) Merr. & Sherff, Bidens bipinnata L.) are also listed in ethnomedicinal reports all over the world. In this study, all these plants are tested for in vitro anticandida activity. A quantitative evaluation of the phytochemicals in all these plants indicated that their vegetative parts are rich in tannins, saponins, oxalates, cyanogenic glycoside and lipids; moreover, the roots have high percentages of alkaloids, flavonoids, and phenols. The results indicated significant anticandida activity, especially for the hexane extract of B. bipinnata leaves which inhibited C. albicans (42.54%), C. glabrata (46.98%), C. tropicalis (50.89%), C. krusei (40.56%), and C. orthopsilosis (50.24%). The extract was subjected to silica gel chromatography and 220 fractions were obtained. Purification by High Performance Liquid Chromatography with Diode-Array Detection (HPLC-DAD) and Gas Chromatography tandem Mass Spectrometry (GC-MS/MS) analysis led to the identification of two anticandida compounds: dehydroabietic and linoleic acid having an inhibition of 85 and 92%, respectively.


Asunto(s)
Bidens/química , Candida/efectos de los fármacos , Cromatografía Líquida de Alta Presión/métodos , Medicamentos Herbarios Chinos/química , Flavonoides/farmacología , Cromatografía de Gases y Espectrometría de Masas/métodos , Extractos Vegetales/farmacología , Candida/crecimiento & desarrollo , Espectrometría de Masas en Tándem/métodos
3.
Curr Issues Mol Biol ; 21: 73-98, 2017.
Artículo en Inglés | MEDLINE | ID: mdl-27489344

RESUMEN

Various transcriptome studies have remained useful in unraveling the complexity of molecular pathways regulating the oil biochemical contents and fruit characteristics of agronomic value in olive. Genes networks associated with plant architect and abiotic stress tolerance have been constructed due to robust genomic data generated by the tools of genomics. This, familiarity will accelerate the breeding programmes in making the selection of high yielding olive genotypes promptly and efficiently. Moreover, comparative transcriptome studies for endogeneous enzymes at different expression sites explicate the contribution of various pathways in phenol and lipid oxidation in olive. Recently, non-targeted metabolomics and metabolic profiling techniques have not only made the understanding of metabolic changes easy but also elucidate biomarkers in fruits related to agronomic parameters and abiotic stresses. However, the alteration in the architectural build up of phenotypes auth-enticates the conservation of their potential genetic links that will invoke interest for future olive breeding.


Asunto(s)
Perfilación de la Expresión Génica , Olea/genética , Olea/metabolismo , Enzimas/genética , Enzimas/metabolismo , Regulación de la Expresión Génica de las Plantas , Aceite de Oliva/metabolismo , Fenoles/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Salinidad
4.
Curr Issues Mol Biol ; 22: 89-112, 2017.
Artículo en Inglés | MEDLINE | ID: mdl-27801354

RESUMEN

For human food security, the preservation of 7.4 million ex-situ germplasm is a global priority. However, ex-situ-conserved seeds are subject to aging, which reduces their viability and ultimately results in the loss of valuable genetic material over long periods. Recent progress in seed biology and genomics has revealed new opportunities to improve the long-term storage of ex-situ seed germplasm. This review summarizes the recent improvements in seed physiology and genomics, with the intention of developing genomic tools for evaluating seed aging. Several lines of seed biology research have shown promise in retrieving viability signal from various stages of seed germination. We conclude that seed aging is associated with mitochondrial alteration and programmed cell death, DNA and enzyme repair, anti-oxidative genes, telomere length, and epigenetic regulation. Clearly, opportunities exist for observing seed aging for developing genomic tools to increment the traditional germination test for effective conservation of ex-situ germplasm.


Asunto(s)
Conservación de los Recursos Naturales , Plantas Comestibles/fisiología , Semillas/fisiología , Envejecimiento/fisiología , Plantas Comestibles/genética
5.
Biochim Biophys Acta ; 1851(6): 898-910, 2015 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-25728392

RESUMEN

Phosphatidylinositol-5-phosphate (PtdIns5P)-4-kinases (PIP4Ks) are stress-regulated lipid kinases that phosphorylate PtdIns5P to generate PtdIns(4,5)P2. There are three isoforms of PIP4Ks: PIP4K2A, 2B and 2C, which localise to different subcellular compartments with the PIP4K2B isoform being localised predominantly in the nucleus. Suppression of PIP4K expression selectively prevents tumour cell growth in vitro and prevents tumour development in mice that have lost the tumour suppressor p53. p53 is lost or mutated in over 70% of all human tumours. These studies suggest that inhibition of PIP4K signalling constitutes a novel anti-cancer therapeutic target. In this review we will discuss the role of PIP4K in tumour suppression and speculate on how PIP4K modulates nuclear phosphoinositides (PPIns) and how this might impact on nuclear functions to regulate cell growth. This article is part of a Special Issue entitled Phosphoinositides.


Asunto(s)
1-Fosfatidilinositol 4-Quinasa/metabolismo , Núcleo Celular/enzimología , Regulación Leucémica de la Expresión Génica , Leucemia Mieloide Aguda/enzimología , Fosfatidilinositol 4,5-Difosfato/metabolismo , Fosfatos de Fosfatidilinositol/metabolismo , 1-Fosfatidilinositol 4-Quinasa/antagonistas & inhibidores , 1-Fosfatidilinositol 4-Quinasa/genética , Animales , Antineoplásicos/farmacología , Citoplasma/enzimología , Humanos , Isoenzimas/antagonistas & inhibidores , Isoenzimas/genética , Isoenzimas/metabolismo , Leucemia Mieloide Aguda/tratamiento farmacológico , Leucemia Mieloide Aguda/genética , Leucemia Mieloide Aguda/patología , Ratones , Inhibidores de Proteínas Quinasas/farmacología , Transducción de Señal , Proteína p53 Supresora de Tumor/deficiencia , Proteína p53 Supresora de Tumor/genética
6.
Biochem J ; 455(3): 347-58, 2013 Nov 01.
Artículo en Inglés | MEDLINE | ID: mdl-23909401

RESUMEN

The spatial and temporal regulation of the second messenger PtdIns(4,5)P2 has been shown to be crucial for regulating numerous processes in the cytoplasm and in the nucleus. Three isoforms of PIP5K1 (phosphatidylinositol 4-phosphate 5-kinase), A, B and C, are responsible for the regulation of the major pools of cellular PtdIns(4,5)P2. PIP5K1B is negatively regulated in response to oxidative stress although it remains unclear which pathways regulate its activity. In the present study, we have investigated the regulation of PIP5K1B by protein phosphorylation. Using MS analysis, we identified 12 phosphorylation sites on PIP5K1B. We developed a phospho-specific antibody against Ser413 and showed that its phosphorylation was increased in response to treatment of cells with phorbol ester, H2O2 or energy restriction. Using inhibitors, we define a stress-dependent pathway that requires the activity of the cellular energy sensor AMPK (AMP-activated protein kinase) and PKC (protein kinase C) to regulate Ser413 phosphorylation. Furthermore, we demonstrate that PKC can directly phosphorylate Ser413 in vitro. Mutation of Ser413 to aspartate to mimic serine phosphorylation decreased both PIP5K1B activity in vitro and PtdIns(4,5)P2 synthesis in vivo. Our studies show that collaboration between AMPK and PKC dictates the extent of Ser413 phosphorylation on PIP5K1B and regulates PtdIns(4,5)P2 synthesis.


Asunto(s)
Proteínas Quinasas Activadas por AMP/metabolismo , Metabolismo Energético , Estrés Oxidativo , Serina/genética , Células HEK293 , Células HeLa , Humanos , Peróxido de Hidrógeno/metabolismo , Mutación , Fosfatidilinositol 4,5-Difosfato/metabolismo , Fosforilación , Fosfotransferasas (Aceptor de Grupo Alcohol)/metabolismo , Serina/metabolismo
7.
Front Plant Sci ; 15: 1336571, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-38916034

RESUMEN

The tolerance to salinity stress is an intricate phenomenon at cellular and whole plant level that requires the knowledge of contributing physiological and biochemical processes and the genetic control of participating traits. In this context, present study was conducted with objective to evaluate the physiological, biochemical, and genetic responses of different wheat genotypes including bread wheat (BW) and synthetic hexaploids (SHs) under saline and control environment. The experiment was conducted in two factorial arrangement in randomized complete block design (RCBD), with genotypes as one factor and treatments as another factor. A significant decline in physiological traits (chlorophyll, photosynthesis, stomatal conductance, transpiration, and cell membrane stability) was observed in all genotypes due to salt stress; however, this decline was higher in BW genotypes as compared to four SH genotypes. In addition, the biochemical traits including enzymes [superoxide dismutase, catalase, and peroxidase (POD)] activity, proline, and glycine betaine (GB) illustrated significant increase along with increase in the expression of corresponding genes (TaCAT1, TaSOD, TaPRX2A, TaP5CS, and TaBADH-A1) due to salt stress in SHs as compared to BW. Correspondingly, highly overexpressed genes, TaHKT1;4, TaNHX1, and TaAKT1 caused a significant decline in Na+/K+ in SH as compared to BW genotypes under salt stress. Moreover, correlation analysis, principal component analysis (PCA), and heatmap analysis have further confirmed that the association and expression of physiological and biochemical traits varied significantly with salinity stress and type of genotype. Overall, the physiological, biochemical, and genetic evaluation proved SHs as the most useful stock for transferring salinity tolerance to other superior BW cultivars via the right breeding program.

8.
Front Plant Sci ; 14: 1107945, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-37123840

RESUMEN

Drought and heat stress are potential problems that can reduce wheat yield, particularly during the terminal growth stages in arid and semiarid regions of the world. The current study intended to examine the impact of individual and combined drought and heat stress on the biochemical contents (antioxidant enzymes, proline, soluble proteins, and soluble sugars), physiological parameters (chlorophyll content, cell membrane stability, photosynthesis, stomatal conductance, and transpiration), plant-water relations (relative water content, water potential, osmotic potential, and pressure potential), agronomic traits (flag leaf area, plant height, number of tillers per plant, spike length, grains per spike, and thousand-grain weight), and gene expression (TaHSF1a, TaWRKY-33, TaNAC2L, and TaGASR1) in four different thermostable and drought-tolerant wheat genotypes (i.e., Gold-16, HS-240, Suntop, and Hemai-13) collected from different countries. The tri-replicate experiment was conducted using two factorial arrangements in a randomized complete block design (RCBD). All measured traits, except total soluble sugars, proline, and cell membrane stability index, showed significant reduction under both combined and individual treatments. Furthermore, correlation analysis revealed a significant association between biochemical and physiological characteristics and crop agronomic productivity. Furthermore, principal component analysis (PCA) and heatmap analysis demonstrated significant levels of variation in traits according to the type of stress and nature of wheat genotype. The spectrographs and micrographs generated by scanning electron microscopy for the selected high- and low- tolerance samples revealed clear differences in mineral distribution and starch granulation. All studied genes showed comparatively high levels of relative expression under combined treatments of drought and heat stress in all wheat genotypes, but this expression was the highest in 'Gold-16' followed by 'HS-240', 'Suntop', and 'Hemai-13'. Overall, this study concluded that plants are proactive entities and they respond to stresses at all levels; however, the tolerant plants tend to retain the integrity of their biochemical, physiological, and molecular equilibrium.

9.
Front Plant Sci ; 14: 1332414, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-38379774

RESUMEN

Soybean is an important oilseed crop worldwide; however, it has a high sensitivity to temperature variation, particularly at the vegetative stage to the pod-filling stage. Temperature change affects physiochemical and genetic traits regulating the soybean agronomic yield. In this regard, the current study aimed to comparatively evaluate the effects of varying regimes of day and night temperatures (T1 = 20°C/12°C, T2 = 25°C/17°C, T3 = 30°C/22°C, T4 = 35°C/27°C, and T5 = 40°C/32°C) on physiological (chlorophyll, photosynthesis, stomatal conductance, transpiration, and membrane damage) biochemical (proline and antioxidant enzymes), genetic (GmDNJ1, GmDREB1G;1, GmHSF-34, GmPYL21, GmPIF4b, GmPIP1;6, GmGBP1, GmHsp90A2, GmTIP2;6, and GmEF8), and agronomic traits (pods per plant, seeds per plant, pod weight per plant, and seed yield per plant) of soybean cultivars (Swat-84 and NARC-1). The experiment was performed in soil plant atmosphere research (SPAR) units using two factorial arrangements with cultivars as one factor and temperature treatments as another factor. A significant increase in physiological, biochemical, and agronomic traits with increased gene expression was observed in both soybean cultivars at T4 (35°C/27°C) as compared to below and above regimes of temperatures. Additionally, it was established by correlation, principal component analysis (PCA), and heatmap analysis that the nature of soybean cultivars and the type of temperature treatments have a significant impact on the paired association of agronomic and biochemical traits, which in turn affects agronomic productivity. Furthermore, at corresponding temperature regimes, the expression of the genes matched the expression of physiochemical traits. The current study has demonstrated through extensive physiochemical, genetic, and biochemical analyses that the ideal day and night temperature for soybeans is T4 (35°C/27°C), with a small variation having a significant impact on productivity from the vegetative stage to the grain-filling stage.

11.
Saudi J Biol Sci ; 29(2): 1185-1190, 2022 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-35197785

RESUMEN

Medicinal plants have significant contribution in pharmaceutical industries being producers of compounds utilized as precursors for drug development. A plant of Lamiaceae family; Pseudocaryopteris foetida had not been investigated for its biomedical potential. Current research was aimed to investigate phytochemical analysis, cytotoxic potential and antioxidant activity of crude methanolic extract and fractions of Pseudocaryopteris foetida (leaves). The preliminary phytochemical analysis of crude methanolic extracts and fractions of Pseudocaryopteris foetida revealed that plant is rich in phenolic and flavonoid classes of secondary metabolites while presence of tannin was observed only in crude methanolic extract. The cytotoxicity was determined using brine shrimp lethality test. Different concentrations (25, 50, 100, 150, 200 and 250 µg/mL) of crude methanolic extract and fractions exhibited dose dependent cytotoxicity. However, The LD50 for all the extracts was more than 200 µg/mL indicating weak cytotoxic potential of Pseudocaryopteris foetida. The antioxidant capabilities of crude methanolic extract and fraction of Pseudocaryopteris foetida were analyzed by in vitro bio assays including DPPH, ABTS, Reducing power and phosphomolybdate antioxidant assays using ascorbic acid as standard. The crude methanolic extract showed IC50 (256.38 ± 0.6 and 314.95 ± 1.1 µg/mL) for DPPH and ABTS respectively, while total antioxidant capacity was calculated as 55.79 ± 0.5 µg/mL for crude methanolic extract of Pseudocaryopteris foetida while ascorbic acid indicated total antioxidant capacity of 71.89 ± 2.3 µg/mL. Study concluded that leaves of Pseudocaryopteris foetida were the rich source of antioxidant phytochemicals. Based on preliminary investigations further research should be focused to isolate bioactive phytochemicals as leading source of clinical medicines in future.

12.
PeerJ Comput Sci ; 7: e507, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-34013033

RESUMEN

Unmanned Aerial Systems (UAVs, Drones), initially known only for their military applications, are getting increasingly popular in the civil sector as well. Over the military canvas, drones have already proven themselves as a potent force multiplier through unmanned, round-the-clock, long-range and high-endurance missions for surveillance, reconnaissance, search and rescue, and even armed combat applications. With the emergence of the Internet of Things (IoT), commercial deployments of drones are also growing exponentially, ranging from cargo and taxi services to agriculture, disaster relief, risk assessment and monitoring of critical infrastructures. Irrespective of the deployment sector, drones are often entrusted to conduct safety, time and liability critical tasks, thus requiring secure, robust and trustworthy operations. In contrast, the rise in UAVs' demand, coupled with market pressure to reduce size, weight, power and cost (SwaP-C) parameters, has caused vendors to often ignore security aspects, thus inducing serious safety and security threats. As UAVs rely on Global Positioning System (GPS) for positioning and navigation, they can fall prey to GPS jamming and spoofing attacks. The vulnerability of GPS to spoofing has serious implications for UAVs, as victim drones using civil GPS can be misdirected or even completely hijacked for malicious intents, as already demonstrated in several academic research efforts using commercially available GPS spoofing hardware. Beside UAVs, GPS spoofing attacks are equally applicable to other GPS-dependent platforms, including manned aircraft, ground vehicles, and cellular systems. This paper conducts a comprehensive review of GPS spoofing threats, with a special focus on their applicability over UAVs and other GPS-dependent mobile platforms. It presents a novel taxonomy of GPS spoofing attacks and critically analyzes different spoofing techniques based upon placement of spoofing device, attack stealthiness, attack methodologies, and objectives of the attacker. We also discuss some of the recent experiments from open literature which utilized commercially available hardware for successfully conducting spoofing attacks.

13.
Plants (Basel) ; 10(11)2021 Nov 02.
Artículo en Inglés | MEDLINE | ID: mdl-34834723

RESUMEN

Wide spectrum medicinal significance augments plant utilization as the primary source of significant pharmaceutical agents. In vitro investigation of antioxidant and antimicrobial activity highlights the therapeutic potential of Otostegia limbata. Methanol extract of the plant (MEP) shows considerable dose dependent antioxidant ability at six concentrations (7.81 µg/mL to 250 µg/mL) in 2.2-diphenyl-1-picrylhydrazyl (DPPH) assay, phosphomolybdate assay (PMA) and reducing power assay (RPA). The plant capability to scavenge free radicals in the mixture ranged from 37.89% to 63.50% in a concentration-dependent manner. MEP was active against five tested bacterial strains in the agar-well diffusion method. Staphylococcus aureus, gram-positive bacteria was found to be most susceptible followed by S. epidermidis with 18.80 mm and 17.47 mm mean zone of inhibition. The mean inhibition zone against gram-negative strains Klebsiella pneumonia, Pseudomonas spp. and Escherichia coli were 15.07 mm, 14.73 mm, and 12.17 mm. MEP revealed potential against Alternaria spp. and Aspergillus terreus fungal strains evaluated through agar-tube dilution assay. Aspergillus terreus was more sensitive than Alternaria spp. with an average 78.45% and 68.0% inhibition. These findings can serve as a benchmark for forthcoming scrutiny such as bioactive components discovery and drug development.

14.
Saudi J Biol Sci ; 28(4): 2316-2322, 2021 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-33911946

RESUMEN

The usage of novel Plant Growth-Promoting Rhizobacteria (PGPR) as bioinoculant is a good opportunity for ecological farming practices to improve soil condition, quality of grain, crops' yield and biodiversity conservation. The purpose of recent research was focused to examine, isolate and characterize PGP bacteria that colonize the rhizosphere for the duration of the maize plant's seedling. For this purpose, 14 samples of soils and roots in the maize rhizosphere were collected from rock phosphate area of Hazara, Pakistan. Forty morphologically natural bacterial colonies have been extracted and tested for their PGP innovations and biocontrol residences and further recognized as plant production advancing rhizobacteria. To find the effective PGPR strains with numerous activities, an aggregate of 150 bacterial colonies were sequestered from different rhizospheric soils of the Hazara Pakistan rock phosphate area. These tested bacterial strains were subjected to biochemical description and in vitro screening for their plant growth-promoting qualities like generation of indole acetic acid (IAA), alkali (NH3), hydrogen cyanide (HCN), siderophores, catalases, proteases and pectinases. All the isolates of rhizobacteria showed IAA producing capacity, as well as found positive for catalase and HCN. The above results suggested that, in addition to biocontrol marketers, PGPR could be used as biofertilizers to substitute agro-chemicals in order to increase crop production. These microorganisms can therefore be further developed and used for greenhouse and discipline packages.

15.
J Virol Methods ; 276: 113760, 2020 02.
Artículo en Inglés | MEDLINE | ID: mdl-31712092

RESUMEN

Cotton production is widely effected by Cotton Leaf Curl Virus (CLCuV) in world posing serious losses to cotton yield.The CRT genes from CLCuV resistant G. arboreum and CLCuV susceptible G. hirsutum were cloned and sequenced to know the differences of protein composition in both species. Molecular techniques were used to isolate full length putative biotic stress resistance genes from G. arboreum besides the analysis of identified novel genes in model plant tobacco (Nicotiana tabacum) for resistance to cotton leaf curl disease complex. It was found that transgenic plants over expressing Hydroperoxidelyase (HPL) genes exhibited higher enzyme activity than wild type. In addition the genome sequence information was used for the purpose of gene isolation. Even for the enhanced expression of Calreticulin (CRT), AOS and HPL in G. hirsutum, it still showed susceptibility against CLCuV suggesting alternative genes and pathways involved for the expression of resistance.


Asunto(s)
Resistencia a la Enfermedad/genética , Genes de Plantas , Gossypium/genética , Nicotiana/virología , Enfermedades de las Plantas/virología , Gossypium/enzimología , Lipooxigenasa/genética , Luteoviridae/patogenicidad , Enfermedades de las Plantas/genética , Plantas Modificadas Genéticamente/enzimología , Plantas Modificadas Genéticamente/virología , Estrés Fisiológico
16.
Nucleic Acids Res ; 35(2): 664-77, 2007.
Artículo en Inglés | MEDLINE | ID: mdl-17179180

RESUMEN

The genes for mitoribosomal protein S12 (Mrps12) and mitochondrial seryl-tRNA ligase (Sarsm and Sars2) are oppositely transcribed from a conserved promoter region of <200 bp in both human and mouse. Using a dual reporter vector we identified an array of 4 CCAAT box elements required for efficient transcription of the two genes in cultured mouse 3T3 cells, and for enforcing directionality in favour of Mrps12. Electrophoretic mobility shift assay (EMSA) and in vivo footprinting confirmed the importance of these promoter elements as sites of protein-binding, and EMSA supershift and chromatin immunoprecipitation (ChIP) assays identified NF-Y as the key transcription factor involved, revealing a common pattern of protein-DNA interactions in all tissues tested (liver, brain, heart, kidney and 3T3 cells). The inherently bidirectional activity of NF-Y makes it an especially suitable factor to govern promoters of this class, whose expression is linked to cell proliferation.


Asunto(s)
Factor de Unión a CCAAT/metabolismo , Regulación de la Expresión Génica , Genes Mitocondriales , Proteínas Mitocondriales/genética , Regiones Promotoras Genéticas , Proteínas Ribosómicas/genética , Serina-ARNt Ligasa/genética , Animales , Sitios de Unión , ADN Intergénico/química , Ensayo de Cambio de Movilidad Electroforética , Genes Reporteros , Ratones , Mitocondrias/genética , Mitocondrias/metabolismo , Proteínas Mitocondriales/metabolismo , Células 3T3 NIH , Biosíntesis de Proteínas , Proteínas Ribosómicas/metabolismo , Serina-ARNt Ligasa/metabolismo
17.
Plant Sci ; 267: 148-156, 2018 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-29362093

RESUMEN

Triterpenoid saponins are one of the most highly accumulated groups of functional components in soybean (Glycine max) and the oxidative reactions during their biosynthesis are required for their aglycone diversity. Natural mutants of soyasaponins in wild soybean (Glycine soja) are valuable resources for establishing the soyasaponin biosynthesis pathway and breeding new soybean varieties. In this study, we investigated the genetic mechanism behind the absence of group A saponins in a Korean wild soybean mutant, CWS5095. Whole genome sequencing (WGS) of CWS5095 identified four point mutations [Val6 → Asp, Ile231 → Thr, His294 → Gln, and Arg376 → Lys] in CYP72A69 (Glyma15g39090), which oxygenate the C-21 position of soyasapogenol B or other intermediates to produce soyasapogenol A, leading to group A saponin production. An in vitro enzyme activity assay of single-sited mutated clones indicated that the Arg376 > Lys mutation (a highly conserved mutation based on a nucleotide change from G → A at the 1,127th position) may lead to loss of gene function in the sg-5 mutant. A very high normalized expression value of 377 reads per kilo base per million (RPKM) of Glyma15g39090 in the hypocotyl axis at the early maturation seed-development stage confirmed their abundant presence in seed hypocotyls. A molecular dynamics analysis of the Arg376 > Lys mutation based on the CYP3A4 (a human CYP450) protein structure found that it was responsible for the increase in axis length toward the heme (active site), which is critically important for biological activity and ligand binding. Our results provide important information on how to eradicate bitter and astringent saponins in soybean by utilizing the reported mutation in Glyma15g39090, and its importance for seed hypocotyl development based on transcript abundance.


Asunto(s)
Sistema Enzimático del Citocromo P-450/genética , Glycine max/genética , Proteínas de Plantas/genética , Secuencia de Aminoácidos , Sistema Enzimático del Citocromo P-450/química , Sistema Enzimático del Citocromo P-450/metabolismo , Mutación , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , Saponinas/metabolismo , Alineación de Secuencia , Glycine max/metabolismo , Gusto , Secuenciación Completa del Genoma
18.
AoB Plants ; 10(6): ply067, 2018 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-30487965

RESUMEN

Cotton leaf curl virus (CLCuV) disease is one of the major limiting factors in cotton production, particularly in widely cultivated Gossypium hirsutum varieties that are susceptible to attack by this virus. Several approaches have been employed to explore putative resistance genes in another cotton species, G. arboreum. However, the exact mechanisms conferring disease resistance in cotton are still unknown. In the current study, we used various approaches to identify possible resistance genes against CLCuV infection. We report the identification and isolation of a set of genes involved in the resistance response to viral infestation. PCR products containing genomic DNA gave multiple amplifications with a single primer in most reactions, and 38 fragments were cloned from G. arboreum and G. hirsutum. The sequences of cloned fragments belonged to various pathway genes and uncharacterized proteins. However, five amplified fragments (RM1, RM6, RM8, RM12 and RM31) showed similarity with R genes. Maximum homology (94 %) was observed with G. raimondii toll/interleukin receptor-like protein. BLAST search showed the homology of all resistance gene analogues (RGAs) with more than one chromosome, and multiple hits were observed on each chromosome for each RGA. Expression analysis through RT-PCR identified variable expression levels of the different RGAs in all tested genotypes. The expression level of RGAs differed between symptomatic and asymptomatic plants, with the exception of RGA 395, whose expression level was the same in both diseased and healthy plants. Knowledge of the interaction of these genes with various cotton pathogens could be utilized to improve the resistance of susceptible G. hirsutum and other plant species.

19.
Sci Rep ; 8(1): 1875, 2018 01 30.
Artículo en Inglés | MEDLINE | ID: mdl-29382843

RESUMEN

In plants, UGTs (UDP-glycosyltransferases) glycosylate various phytohormones and metabolites in response to biotic and abiotic stresses. Little is known about stress-responsive glycosyltransferases in plants. Therefore, it is important to understand the genomic and transcriptomic portfolio of plants with regard to biotic and abiotic stresses. Here, we identified 140, 154, and 251 putative UGTs in Brassica rapa, Brassica oleracea, and Brassica napus, respectively, and clustered them into 14 major phylogenetic groups (A-N). Fourteen major KEGG pathways and 24 biological processes were associated with the UGTs, highlighting them as unique modulators against environmental stimuli. Putative UGTs from B. rapa and B. oleracea showed a negative selection pressure and biased gene fractionation pattern during their evolution. Polyploidization increased the intron proportion and number of UGT-containing introns among Brassica. The putative UGTs were preferentially expressed in developing tissues and at the senescence stage. Differential expression of up- and down-regulated UGTs in response to phytohormone treatments, pathogen responsiveness and abiotic stresses, inferred from microarray and RNA-Seq data in Arabidopsis and Brassica broaden the glycosylation impact at the molecular level. This study identifies unique candidate UGTs for the manipulation of biotic and abiotic stress pathways in Brassica and Arabidopsis.


Asunto(s)
Arabidopsis/genética , Brassica/genética , Perfilación de la Expresión Génica/métodos , Genómica/métodos , Glicosiltransferasas/genética , Reguladores del Crecimiento de las Plantas/metabolismo , Estrés Fisiológico , Arabidopsis/enzimología , Brassica/enzimología , Regulación Enzimológica de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Glicosilación , Glicosiltransferasas/metabolismo , Filogenia
20.
Front Microbiol ; 9: 477, 2018.
Artículo en Inglés | MEDLINE | ID: mdl-29670582

RESUMEN

This study focused on rhizobacteria to promote sustainable crop production in arid regions of Saudi Arabia. The study isolated 17 tightly root-adhering rhizobacteria from various plants at Hada Al Sham in Saudi Arabia. All 17 rhizobacterial isolates were confirmed as plant growth promoting rhizobacteria by classical biochemical tests. Using 16S rDNA gene sequence analyses, the strains were identified as Bacillus, Acinetobacter and Enterobacter. Subsequently, the strains were assessed for their ability to improve the physiology, nutrient uptake, growth, and yield of alfalfa plants grown under desert agriculture conditions. The field trials were conducted in a randomized complete block design. Inoculation of alfalfa with any of these 17 strains improved the relative water content; chlorophyll a; chlorophyll b; carotenoid contents; nitrogen (N), phosphorus, and potassium contents; plant height; leaf-to-stem ratio; and fresh and dry weight. Acinetobacter pittii JD-14 was most effective to increase fresh and dry weight of alfalfa by 41 and 34%, respectively, when compared to non-inoculated control plants. Nevertheless, all strains enhanced crop traits when compared to controls plants, indicating that these desert rhizobacterial strains could be used to develop an eco-friendly biofertilizer for alfalfa and possibly other crop plants to enhance sustainable production in arid regions.

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