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1.
J Exp Bot ; 2024 Apr 23.
Artículo en Inglés | MEDLINE | ID: mdl-38650362

RESUMEN

Seasonal bud dormancy in perennial woody plants is a crucial and intricate process that is vital for the survival and development of plants. Over the past few decades, significant advancements have been made in understanding many features of bud dormancy, particularly in model species, where certain molecular mechanisms underlying this process have been elucidated. In this review, we provide an overview of recent molecular progress in understanding bud dormancy in trees, with a specific emphasis on the integration of common signaling and molecular mechanisms identified across different tree species. Additionally, we address some challenges that have emerged in the in-depth understanding of bud dormancy and offer insights for future studies.

2.
Appl Opt ; 63(1): 104-111, 2024 Jan 01.
Artículo en Inglés | MEDLINE | ID: mdl-38175006

RESUMEN

In recent years, there has been a growing interest in the wideband propagation and control of terahertz (THz) radiation due to its potential for a variety of applications, such as 6G communication, sensing, and imaging. One promising approach in this area is the use of valley photonic crystals (VPCs), which exhibit properties like wider band gaps and robust propagation. In this paper, a two-dimensional dielectric silicon-air VPC is studied, which is constructed from a method of inversion symmetry breaking providing a band gap of 109.4 GHz at a mid-gap frequency of 0.376 THz. We employ an optimized bearded-stack interface to construct the VPC waveguide for wideband THz propagation along straight and Z-shaped paths. We demonstrate that a band-stop response can be achieved in a VPC by introducing periodic defects along the domain wall. Furthermore, the stop range can be tuned by varying the refractive index of the defects through incorporating liquid crystal along the domain wall of VPC. Our proposed structure and the techniques employed could be promising for the development of a band-stop filter (BSF) and other photonic components having potential applications in 6G communication and beyond.

3.
Planta ; 258(2): 26, 2023 Jun 24.
Artículo en Inglés | MEDLINE | ID: mdl-37354348

RESUMEN

MAIN CONCLUSION: LBD18 and IAA14 antagonistically interact with ARF7 through the electrostatic faces in the ARF7PB1 domain, modulating ARF7 transcriptional activity. Auxin Response Factor 7 (ARF7)/ARF19 control lateral root development by directly activating Lateral Organ Boundaries Domain 16 (LBD16)/LBD18 genes in Arabidopsis. LBD18 upregulates ARF19 expression by binding to the ARF19 promoter. It also interacts with ARF7 through the Phox and Bem1 (PB1) domain to enhance the ARF7 transcriptional activity, forming a dual mode of positive feedback loop. LBD18 competes with the repressor indole-3-acetic acid 14 (IAA14) for ARF7 binding through the PB1 domain. In this study, we examined the molecular determinant of the ARF7 PB1 domain for interacting with LBD18 and showed that the electronic faces in the ARF7 PB1 domain are critical for interacting with LBD18 and IAA14/17. We used a luminescence complementation imaging assay to determine protein-protein interactions. The results showed that mutation of the invariant lysine residue and the OPCA motif in the PB1 domain in ARF7 significantly reduces the protein interaction between ARF7 and LBD18. Transient gene expression assays with Arabidopsis protoplasts showed that IAA14 suppressed transcription-enhancing activity of LBD18 on the LUC reporter gene fused to the ARF19 promoter harboring an auxin response element, but mutation of the invariant lysine residue and OPCA motif in the PB1 domain of IAA14 reduced the repression capability of IAA14 for transcription-enhancing activity of LBD18. We further showed that the same mutation in the PB1 domain of IAA14 reduces its repression capability, thereby increasing the LUC activity induced by both ARF7 and LBD18 compared with IAA14. These results suggest that LBD18 competes with IAA14 for ARF7 binding via the electrostatic faces of the ARF7 PB1 domain to modulate ARF7 transcriptional activity.


Asunto(s)
Proteínas de Arabidopsis , Arabidopsis , Arabidopsis/metabolismo , Proteínas de Arabidopsis/metabolismo , Factor VII/genética , Factor VII/metabolismo , Regulación de la Expresión Génica de las Plantas , Ácidos Indolacéticos/metabolismo , Lisina/metabolismo , Raíces de Plantas/metabolismo , Factores de Transcripción/metabolismo
4.
Int J Mol Sci ; 23(23)2022 Dec 02.
Artículo en Inglés | MEDLINE | ID: mdl-36499546

RESUMEN

Neuropeptide B (NPB) and neuropeptide W (NPW) are neuropeptides, which constitute NPB/W signaling systems together with G-protein coupled receptors NPBWR1. The location and function of NPB/W signaling systems have been predominantly detected and mapped within the CNS, including their role in the modulation of inflammatory pain, neuroendocrine functions, and autonomic nervous systems. The aim of the study is to investigate the impact of diabetes on the neuropeptide B/W signaling system in different heart compartments and neurons which innervates it. In the RT-qPCR analysis, we observed the upregulation of mRNA for preproNPB in RV, for preproNPW in LA, and for NPBWR1 in DRG in diabetic rats. On the contrary, the expression of mRNA for NPBWR1 was downregulated in LV in diabetic rats. In the WB analysis, significant downregulation of NPBWR1 in LV (0.54-fold, p = 0.046) in diabetic rats was observed at the proteomic level. The presence of NPBWR1 was also confirmed in a dissected LCM section of cardiomyocytes and coronary arteries. The positive inotropic effect of NPW described on the diabetic cardiomyocytes in vitro could point to a possible therapeutic target for compensation of the contractile dysfunction in the diabetic heart. In conclusion, the NPB/W signaling system is involved in the regulation of heart functions and long-term diabetes leads to changes in the expression of individual members of this signaling system differently in each cardiac compartment, which is related to the different morphology and function of these cardiac chambers.


Asunto(s)
Diabetes Mellitus Experimental , Receptores de Neuropéptido , Ratas , Animales , Receptores de Neuropéptido/genética , Receptores de Neuropéptido/metabolismo , Proteómica , Diabetes Mellitus Experimental/genética , Receptores Acoplados a Proteínas G/genética , Receptores Acoplados a Proteínas G/metabolismo , ARN Mensajero/genética
5.
J Exp Bot ; 72(8): 2889-2902, 2021 04 02.
Artículo en Inglés | MEDLINE | ID: mdl-33595615

RESUMEN

Roots provide the plant with water and nutrients and anchor it in a substrate. Root development is controlled by plant hormones and various sets of transcription factors. Recently, various small peptides and their cognate receptors have been identified as controlling root development. Small peptides bind to membrane-localized receptor-like kinases, inducing their dimerization with co-receptor proteins for signaling activation and giving rise to cellular signaling outputs. Small peptides function as local and long-distance signaling molecules involved in cell-to-cell communication networks, coordinating root development. In this review, we survey recent advances in the peptide ligand-mediated signaling pathways involved in the control of root development in Arabidopsis. We describe the interconnection between peptide signaling and conventional phytohormone signaling. Additionally, we discuss the diversity of identified peptide-receptor interactions during plant root development.


Asunto(s)
Proteínas de Arabidopsis , Arabidopsis , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Regulación de la Expresión Génica de las Plantas , Meristema/metabolismo , Péptidos/metabolismo , Raíces de Plantas/metabolismo , Transducción de Señal
6.
Int J Mol Sci ; 22(16)2021 Aug 17.
Artículo en Inglés | MEDLINE | ID: mdl-34445532

RESUMEN

The specificity of a diagnostic assay depends upon the purity of the biomolecules used as a probe. To get specific and accurate information of a disease, the use of synthetic peptides in diagnostics have increased in the last few decades, because of their high purity profile and ability to get modified chemically. The discovered peptide probes are used either in imaging diagnostics or in non-imaging diagnostics. In non-imaging diagnostics, techniques such as Enzyme-Linked Immunosorbent Assay (ELISA), lateral flow devices (i.e., point-of-care testing), or microarray or LC-MS/MS are used for direct analysis of biofluids. Among all, peptide-based ELISA is considered to be the most preferred technology platform. Similarly, peptides can also be used as probes for imaging techniques, such as single-photon emission computed tomography (SPECT) and positron emission tomography (PET). The role of radiolabeled peptides, such as somatostatin receptors, interleukin 2 receptor, prostate specific membrane antigen, αß3 integrin receptor, gastrin-releasing peptide, chemokine receptor 4, and urokinase-type plasminogen receptor, are well established tools for targeted molecular imaging ortumor receptor imaging. Low molecular weight peptides allow a rapid clearance from the blood and result in favorable target-to-non-target ratios. It also displays a good tissue penetration and non-immunogenicity. The only drawback of using peptides is their potential low metabolic stability. In this review article, we have discussed and evaluated the role of peptides in imaging and non-imaging diagnostics. The most popular non-imaging and imaging diagnostic platforms are discussed, categorized, and ranked, as per their scientific contribution on PUBMED. Moreover, the applicability of peptide-based diagnostics in deadly diseases, mainly COVID-19 and cancer, is also discussed in detail.


Asunto(s)
Prueba de COVID-19/métodos , COVID-19/diagnóstico , Péptidos/análisis , COVID-19/virología , Bases de Datos Factuales , Ensayo de Inmunoadsorción Enzimática/métodos , Humanos , Tomografía de Emisión de Positrones/métodos , Receptores de Somatostatina , SARS-CoV-2/aislamiento & purificación , Espectrometría de Masas en Tándem/métodos , Tomografía Computarizada de Emisión de Fotón Único/métodos
7.
Int J Mol Sci ; 21(21)2020 Oct 22.
Artículo en Inglés | MEDLINE | ID: mdl-33105700

RESUMEN

Members of neuropeptide B/W signaling system have been predominantly detected and mapped within the CNS. In the rat, this system includes neuropeptide B (NPB), neuropeptide W (NPW) and their specific receptor NPBWR1. This signaling system has a wide spectrum of functions including a role in modulation of inflammatory pain and neuroendocrine functions. Expression of NPB, NPW and NPBWR1 in separate heart compartments, dorsal root ganglia (DRG) and stellate ganglia was proven by RT-qPCR, Western blot (WB) and immunofluorescence. Presence of mRNA for all tested genes was detected within all heart compartments and ganglia. The presence of proteins preproNPB, preproNPW and NPBWR1 was confirmed in all the chambers of heart by WB. Expression of preproNPW and preproNPB was proven in cardiac ganglionic cells obtained by laser capture microdissection. In immunofluorescence analysis, NPB immunoreactivity was detected in nerve fibers, some nerve cell bodies and smooth muscle within heart and both ganglia. NPW immunoreactivity was present in the nerve cell bodies and nerve fibers of heart ganglia. Weak nonhomogenous staining of cardiomyocytes was present within heart ventricles. NPBWR1 immunoreactivity was detected on cardiomyocytes and some nerve fibers. We confirmed the presence of NPB/W signaling system in heart, DRG and stellate ganglia by proteomic and genomic analyses.


Asunto(s)
Miocardio/metabolismo , Neuropéptidos/genética , Receptores Acoplados a Proteínas G/metabolismo , Receptores de Neuropéptido/metabolismo , Animales , Técnica del Anticuerpo Fluorescente , Ganglios Espinales/metabolismo , Expresión Génica , Masculino , Neuropéptidos/inmunología , Neuropéptidos/metabolismo , Ratas Zucker , Receptores Acoplados a Proteínas G/genética , Receptores Acoplados a Proteínas G/inmunología , Receptores de Neuropéptido/genética , Receptores de Neuropéptido/inmunología , Reproducibilidad de los Resultados , Transducción de Señal , Ganglio Estrellado/metabolismo
8.
Plant J ; 95(2): 233-251, 2018 07.
Artículo en Inglés | MEDLINE | ID: mdl-29681137

RESUMEN

A hierarchy of transcriptional regulators controlling lateral root formation in Arabidopsis thaliana has been identified, including the AUXIN RESPONSE FACTOR 7 (ARF7)/ARF19-LATERAL ORGAN BOUNDARIES DOMAIN 16 (LBD16)/LBD18 transcriptional network; however, their feedback regulation mechanisms are not known. Here we show that LBD18 controls ARF activity using the dual mode of a positive feedback loop. We showed that ARF7 and ARF19 directly bind AuxRE in the LBD18 promoter. A variety of molecular and biochemical experiments demonstrated that LBD18 binds a specific DNA motif in the ARF19 promoter to regulate its expression in vivo as well as in vitro. LBD18 interacts with ARFs including ARF7 and ARF19 via the Phox and Bem1 domain of ARF to enhance the transcriptional activity of ARF7 on AuxRE, and competes with auxin/indole-3-acetic acid (IAA) repressors for ARF binding, overriding the negative feedback loop exerted by Aux/IAA repressors. Taken together, these results show that LBD18 and ARFs form a double positive feedback loop, and that LBD18 uses the dual mode of a positive feedback loop by binding directly to the ARF19 promoter and through the protein-protein interactions with ARF7 and ARF19. This novel mechanism of feedback loops may constitute a robust feedback mechanism that ensures continued lateral root growth in response to auxin in Arabidopsis.


Asunto(s)
Proteínas de Arabidopsis/fisiología , Arabidopsis/metabolismo , Regulación de la Expresión Génica de las Plantas , Factores de Transcripción/metabolismo , Factores de Transcripción/fisiología , Arabidopsis/genética , Arabidopsis/crecimiento & desarrollo , Proteínas de Arabidopsis/metabolismo , Retroalimentación Fisiológica , Raíces de Plantas/crecimiento & desarrollo , Raíces de Plantas/metabolismo , Regiones Promotoras Genéticas , Transcripción Genética
9.
BMC Plant Biol ; 19(1): 46, 2019 Jan 31.
Artículo en Inglés | MEDLINE | ID: mdl-30704405

RESUMEN

BACKGROUND: Adventitious root (AR) formation is a complex genetic trait, which is controlled by various endogenous and environmental cues. Auxin is known to play a central role in AR formation; however, the mechanisms underlying this role are not well understood. RESULTS: In this study, we showed that a previously identified auxin signaling module, AUXIN RESPONSE FACTOR(ARF)7/ARF19-LATERAL ORGAN BOUNDARIES DOMAIN(LBD)16/LBD18 via AUXIN1(AUX1)/LIKE-AUXIN3 (LAX3) auxin influx carriers, which plays important roles in lateral root formation, is involved in AR formation in Arabidopsis. In aux1, lax3, arf7, arf19, lbd16 and lbd18 single mutants, we observed reduced numbers of ARs than in the wild type. Double and triple mutants exhibited an additional decrease in AR numbers compared with the corresponding single or double mutants, respectively, and the aux1 lax3 lbd16 lbd18 quadruple mutant was devoid of ARs. Expression of LBD16 or LBD18 under their own promoters in lbd16 or lbd18 mutants rescued the reduced number of ARs to wild-type levels. LBD16 or LBD18 fused to a dominant SRDX repressor suppressed promoter activity of the cell cycle gene, Cyclin-Dependent Kinase(CDK)A1;1, to some extent. Expression of LBD16 or LBD18 was significantly reduced in arf7 and arf19 mutants during AR formation in a light-dependent manner, but not in arf6 and arf8. GUS expression analysis of promoter-GUS reporter transgenic lines revealed overlapping expression patterns for LBD16, LBD18, ARF7, ARF19 and LAX3 in AR primordia. CONCLUSION: These results suggest that the ARF7/ARF19-LBD16/LBD18 transcriptional module via the AUX1/LAX3 auxin influx carriers plays an important role in AR formation in Arabidopsis.


Asunto(s)
Proteínas de Arabidopsis/fisiología , Arabidopsis/crecimiento & desarrollo , Raíces de Plantas/crecimiento & desarrollo , Factores de Transcripción/fisiología , Arabidopsis/metabolismo , Proteínas de Arabidopsis/metabolismo , Regulación del Desarrollo de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Hipocótilo/crecimiento & desarrollo , Hipocótilo/metabolismo , Raíces de Plantas/metabolismo , Factores de Transcripción/metabolismo
10.
Planta ; 249(4): 1251-1258, 2019 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-30627888

RESUMEN

MAIN CONCLUSION: Lateral Organ Boundaries Domain 13 (LBD13), which is expressed in emerged lateral roots and encodes a transcriptional activator, plays an important role in lateral root formation in Arabidopsis. Lateral roots (LRs) are major determinants of root system architecture, contributing to the survival strategies of plants. Members of the LBD gene family encode plant-specific transcription factors that play key roles in plant organ development. Several LBD genes, such as LBD14, 16, 18, 29, and 33, have been shown to play important roles in regulating LR development in Arabidopsis. In the present study, we show that LBD13 is expressed in emerged LRs and LR meristems of elongated LRs and regulates LR formation in Arabidopsis. Transient gene expression assays with Arabidopsis protoplasts showed that LBD13 is localized to the nucleus and harbors transcription-activating potential. Knock-down of LBD13 expression by RNA interference resulted in reduced LR formation, whereas overexpression of LBD13 enhanced LR formation in transgenic Arabidopsis. Analysis of ß-glucuronidase (GUS) expression under the control of the LBD13 promoter showed that GUS staining was detected in LRs emerged from the primary root, but not in LR primordia. Moreover, both the distribution of LR primordium number and developmental kinetics of LR primordia were not affected either by knock-down or by overexpression of LBD13. Taken together, these results suggest that LBD13 is a nuclear-localized transcriptional activator and controls LR formation during or after LR emergence.


Asunto(s)
Proteínas de Arabidopsis/fisiología , Arabidopsis/crecimiento & desarrollo , Genes de Plantas/genética , Proteínas Nucleares/fisiología , Raíces de Plantas/crecimiento & desarrollo , Factores de Transcripción/fisiología , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Regulación de la Expresión Génica de las Plantas , Genes de Plantas/fisiología , Proteínas Nucleares/genética , Plantas Modificadas Genéticamente , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Factores de Transcripción/genética
11.
Plant Physiol ; 174(1): 301-311, 2017 May.
Artículo en Inglés | MEDLINE | ID: mdl-28336771

RESUMEN

LATERAL ORGAN BOUNDARIES DOMAIN/ASYMMETRIC LEAVES2-LIKEs (hereafter referred to as LBD) are plant-specific transcription factors that play important roles in a plethora of plant growth and development. The leucine (Leu) zipper-like coiled-coil motif in the lateral organ boundaries domain of the class I LBD proteins has been proposed to mediate protein dimerization, but it has not been experimentally assessed yet. LBD16 and LBD18 have been well characterized to play important roles in lateral root development in Arabidopsis (Arabidopsis thaliana). Here, we investigated the role of the coiled-coil motif in the dimerization of LBD16 and LBD18 and in transcriptional regulation and biological function. We built the molecular models of the coiled coil of LBD16 and LBD18, providing the probable Leu zipper models of the helix dimer. Using a variety of molecular techniques, such as bimolecular fluorescence complementation, luciferase complementation imaging, GST pull down, and coimmunoprecipitation assays, we showed that the conserved Leu or valine residues in the coiled-coil motif are critical for the dimerization of LBD16 or LBD18. Using transgenic Arabidopsis plants that overexpress HA:LBD16 or HA:LBD16Q in lbd16 or HA:LBD18 or HA:LBD18Q in lbd18, we demonstrated that the homodimerization of LBD18 mediated by the coiled-coil motif is crucial for transcriptional regulation via promoter binding and for lateral root formation. In addition, we found that the carboxyl-terminal region beyond the coiled-coil motif in LBD18 acts as an additional dimerization domain. These results provide a molecular basis for homodimerization and heterodimerization among the 42 Arabidopsis LBD family members for displaying their biological functions.


Asunto(s)
Proteínas de Arabidopsis/genética , Arabidopsis/genética , Raíces de Plantas/genética , Factores de Transcripción/genética , Arabidopsis/citología , Arabidopsis/metabolismo , Proteínas de Arabidopsis/química , Proteínas de Arabidopsis/metabolismo , Sitios de Unión/genética , Regulación de la Expresión Génica de las Plantas , Modelos Moleculares , Mutación , Raíces de Plantas/metabolismo , Plantas Modificadas Genéticamente , Conformación Proteica , Multimerización de Proteína , Protoplastos/metabolismo , Factores de Transcripción/química , Factores de Transcripción/metabolismo
12.
Langmuir ; 34(22): 6315-6327, 2018 06 05.
Artículo en Inglés | MEDLINE | ID: mdl-29350535

RESUMEN

Contact charge electrophoresis (CCEP) uses steady electric fields to drive the oscillatory motion of conductive particles and droplets between two or more electrodes. In contrast to traditional forms of electrophoresis and dielectrophoresis, CCEP allows for rapid and sustained particle motions driven by low-power dc voltages. These attributes make CCEP a promising mechanism for powering active components for mobile microfluidic technologies. This Feature Article describes our current understanding of CCEP as well as recent strategies to harness it for applications in microfluidics and beyond.

13.
Plant Cell Physiol ; 57(11): 2291-2299, 2016 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-27519310

RESUMEN

LATERAL ORGAN BOUNDARIES DOMAIN (LBD) transcription factor gene family members play key roles in diverse aspects of plant development. LBD10 and LBD27 have been shown to be essential for pollen development in Arabidopsis thaliana. From the previous RNA sequencing (RNA-Seq) data set of Arabidopsis pollen, we identified the mRNAs of LBD22, LBD25 and LBD36 in addition to LBD10 and LBD27 in Arabidopsis pollen. Here we conducted expression and cellular analysis using GFP:GUS (green fluorescent protein:ß-glucuronidase) reporter gene and subcellular localization assays using LBD:GFP fusion proteins expressed under the control of their own promoters in Arabidopsis. We found that these LBD proteins display spatially and temporally distinct and overlapping expression patterns during pollen development. Bimolecular fluorescence complementation and GST (glutathione S-transferase) pull-down assays demonstrated that protein-protein interactions occur among the LBDs exhibiting overlapping expression during pollen development. We further showed that LBD10, LBD22, LBD25, LBD27 and LBD36 interact with each other to form heterodimers, which are localized to the nucleus in Arabidopsis protoplasts. Taken together, these results suggest that combinatorial interactions among LBD proteins may be important for their function in pollen development in Arabidopsis.


Asunto(s)
Proteínas de Arabidopsis/metabolismo , Arabidopsis/crecimiento & desarrollo , Arabidopsis/metabolismo , Polen/crecimiento & desarrollo , Polen/metabolismo , Factores de Transcripción/metabolismo , Fluorescencia , Glucuronidasa/metabolismo , Proteínas Fluorescentes Verdes/metabolismo , Modelos Biológicos , Plantas Modificadas Genéticamente , Unión Proteica , Mapeo de Interacción de Proteínas , Protoplastos/metabolismo , Proteínas Recombinantes de Fusión/metabolismo , Fracciones Subcelulares/metabolismo
14.
Langmuir ; 32(49): 13167-13173, 2016 12 13.
Artículo en Inglés | MEDLINE | ID: mdl-27951714

RESUMEN

We investigate the dynamics of metallodielectric Janus particles moving via contact charge electrophoresis (CCEP) between two parallel electrodes. CCEP uses a constant voltage to repeatedly charge and actuate conductive particles within a dielectric fluid, resulting in rapid oscillatory motion between the electrodes. In addition to particle oscillations, we find that micrometer-scale Janus particles move perpendicular to the field at high speeds (up to 600 µm/s) and over large distances. We characterize particle motions and propose a mechanism based on the rotation-induced translation of the particle following charge transfer at the electrode surface. The propulsion mechanism is supported both by experiments with fluorescent particles that reveal their rotational motions and by simulations of CCEP dynamics that capture the relevant electrostatics and hydrodynamics. We also show that interactions among multiple particles can lead to repulsion, attraction, and/or cooperative motions depending on the position and phase of the respective particle oscillators. Our results demonstrate how particle asymmetries can be used to direct the motions of active colloids powered by CCEP.

15.
Plant Cell Rep ; 35(11): 2353-2367, 2016 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-27522520

RESUMEN

KEY MESSAGE: Functional characterization of two tobacco genes, one involved in xylan synthesis and the other, a positive regulator of secondary cell wall formation, is reported. Lignocellulosic secondary cell walls (SCW) provide essential plant materials for the production of second-generation bioethanol. Therefore, thorough understanding of the process of SCW formation in plants is beneficial for efficient bioethanol production. Recently, we provided the first proof-of-concept for using virus-induced gene silencing (VIGS) approach for rapid functional characterization of nine genes involved in cellulose, hemicellulose and lignin synthesis during SCW formation. Here, we report VIGS-mediated functional characterization of two tobacco genes involved in SCW formation. Stems of VIGS plants silenced for both selected genes showed increased amount of xylem formation but thinner cell walls than controls. These results were further confirmed by production of stable transgenic tobacco plants manipulated in expression of these genes. Stems of stable transgenic tobacco plants silenced for these two genes showed increased xylem proliferation with thinner walls, whereas transgenic tobacco plants overexpressing these two genes showed increased fiber cell wall thickness but no change in xylem proliferation. These two selected genes were later identified as possible members of DUF579 family involved in xylan synthesis and KNAT7 transcription factor family involved in positive regulation of SCW formation, respectively. Glycome analyses of cell walls showed increased polysaccharide extractability in 1 M KOH extracts of both VIGS-NbDUF579 and VIGS-NbKNAT7 lines suggestive of cell wall loosening. Also, VIGS-NbDUF579 and VIGS-NbKNAT7 lines showed increased saccharification rates (74.5 and 40 % higher than controls, respectively). All these properties are highly desirable for producing higher quantities of bioethanol from lignocellulosic materials of bioenergy plants.


Asunto(s)
Pared Celular/genética , Silenciador del Gen , Genes de Plantas , Lignina/metabolismo , Virus de Plantas/fisiología , Metabolismo de los Hidratos de Carbono/genética , Fluoresceína-5-Isotiocianato/metabolismo , Regulación de la Expresión Génica de las Plantas , Vectores Genéticos/metabolismo , Glucosa/metabolismo , Glicómica , Glicosilación , Plantas Modificadas Genéticamente , Polisacáridos/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismo , Nicotiana/anatomía & histología , Nicotiana/genética , Nicotiana/virología , Xilema/genética
16.
Mol Cell Proteomics ; 12(2): 277-82, 2013 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-23139387

RESUMEN

Sophisticated approaches have recently led to the identification of novel autoantigens associated with Multiple Sclerosis (MuS), e.g. neurofascin, contactin, CNPase, and other T-cell receptor membrane anchored proteins. These putative antigens, although differing from the conventional myelin derivatives, are conceptually based on an animal model of experimental autoimmune encephalomyelitis. In this report we describe the identification of putative antigens based on their recognition by autoantibodies isolated from MuS patient serum. In a previous work from this laboratory we have shown that a peptide probe, named CSF114(Glc), specifically identifies serum autoantibodies in a subset of MuS patients, representing ∼30% of the patient population. The autoantibodies, purified from MuS patients' sera (six), through CSF114(Glc) affinity chromatography, detected three immunoreactive protein bands present in the rat brain. Proteomic analysis of the immunoreactive bands, involving MALDI and MS/MS techniques, revealed the presence of four proteins distinguishable by their mass: alpha fodrin, alpha actinin 1, creatine kinase, and CNPase. The immunoreactive profile of these rat brain proteins was compared with that of commercially available standard proteins by challenging against either CSF114(Glc) purified MuS autoantibodies, or monoclonal antibodies. Further discrimination among the rat brain proteins was provided by the following procedure: whereas monoclonal antibodies recognized all rat brain proteins, isolated MuS specific antibodies recognize only alpha actinin 1 as a putative antigen. In fact, alpha actinin 1 displayed a robust immunoreactive response against all MuS patients' sera examined, whereas the other three bands were not consistently detectable. Thus, alpha actinin 1, a cytoskeleton protein implicated in inflammatory/degenerative autoimmune diseases (lupus nephritis and autoimmune hepatitis) might be regarded as a novel MuS autoantigen, perhaps a prototypic biomarker for the inflammatory/degenerative process typical of the disease.


Asunto(s)
Actinina/inmunología , Autoanticuerpos/inmunología , Autoantígenos/inmunología , Esclerosis Múltiple/inmunología , Proteínas del Tejido Nervioso/inmunología , Péptidos/inmunología , 2',3'-Nucleótido Cíclico Fosfodiesterasas/sangre , 2',3'-Nucleótido Cíclico Fosfodiesterasas/inmunología , Actinina/sangre , Secuencia de Aminoácidos , Animales , Anticuerpos Monoclonales/sangre , Anticuerpos Monoclonales/inmunología , Autoanticuerpos/sangre , Autoantígenos/sangre , Encéfalo/inmunología , Encéfalo/metabolismo , Proteínas Portadoras/sangre , Proteínas Portadoras/inmunología , Forma BB de la Creatina-Quinasa/sangre , Forma BB de la Creatina-Quinasa/inmunología , Epítopos/sangre , Epítopos/inmunología , Glicosilación , Humanos , Proteínas de Microfilamentos/sangre , Proteínas de Microfilamentos/inmunología , Datos de Secuencia Molecular , Esclerosis Múltiple/sangre , Esclerosis Múltiple/patología , Proteínas del Tejido Nervioso/sangre , Péptidos/sangre , Ratas , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción , Espectrometría de Masas en Tándem
18.
Opt Express ; 22(3): 2868-80, 2014 Feb 10.
Artículo en Inglés | MEDLINE | ID: mdl-24663579

RESUMEN

We describe the fabrication and characterization of plasmonic waveguides based on a periodic one-dimensional array of symmetric and asymmetric T-shaped structures. The devices are fabricated in a polymer resin using conventional 3D printing and subsequently overcoated with ~500 nm of Au. Using terahertz (THz) time-domain spectroscopy, we systematically measure the guided-wave transmission properties of the devices as a function of the different geometrical parameters. Through these measurements, we find that the resonance frequency associated with the lowest order mode depends primarily on the structure height and the cap width and appears to be independent of its lateral width. We also perform numerical simulations using the same geometrical parameters and find excellent agreement between experiment and simulation. We fabricate a waveguide in which the lateral width of the T-shaped structures is tapered in a linear fashion. While the spectrum of this device is similar to one without tapering, we observe relatively little reduction in the mode size, even as the structure width is reduced by a factor of eight.

19.
Plant Cell Rep ; 33(7): 1041-52, 2014 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-24585187

RESUMEN

KEY MESSAGE: We report a novel approach for enhanced accumulation of fatty acids and triacylglycerols for utilization as biodiesel in transgenic tobacco stems through xylem-specific expression of Arabidopsis DGAT1 and LEC2 genes. The use of plant biomass for production of bioethanol and biodiesel has an enormous potential to revolutionize the global bioenergy outlook. Several studies have recently been initiated to genetically engineer oil production in seeds of crop plants to improve biodiesel production. However, the "food versus fuel" issues have also sparked some studies for enhanced accumulation of oils in vegetative tissues like leaves. But in the case of bioenergy crops, use of woody stems is more practical than leaves. Here, we report the enhanced accumulation of fatty acids (FAs) and triacylglycerols (TAGs) in stems of transgenic tobacco plants expressing Arabidopsis diacylglycerol acyltransferase 1 (DGAT1) and leafy cotyledon2 (LEC2) genes under a developing xylem-specific cellulose synthase promoter from aspen trees. The transgenic tobacco plants accumulated significantly higher amounts of FAs in their stems. On an average, DGAT1 and LEC2 overexpression showed a 63 and 80% increase in total FA production in mature stems of transgenic plants over that of controls, respectively. In addition, selected DGAT1 and LEC2 overexpression lines showed enhanced levels of TAGs in stems with higher accumulation of 16:0, 18:2 and 18:3 TAGs. In LEC2 lines, the relative mRNA levels of the downstream genes encoding plastidic proteins involved in FA synthesis and accumulation were also elevated. Thus, here, we provide a proof of concept for our approach of enhancing total energy yield per plant through accumulation of higher levels of FAs in transgenic stems for biodiesel production.


Asunto(s)
Proteínas de Arabidopsis/genética , Diacilglicerol O-Acetiltransferasa/genética , Ácidos Grasos/metabolismo , Nicotiana/metabolismo , Tallos de la Planta/metabolismo , Factores de Transcripción/genética , Triglicéridos/metabolismo , Proteínas de Arabidopsis/metabolismo , Biocombustibles , Celulosa/metabolismo , Diacilglicerol O-Acetiltransferasa/metabolismo , Regulación de la Expresión Génica de las Plantas , Tallos de la Planta/genética , Plantas Modificadas Genéticamente , Regiones Promotoras Genéticas , ARN Mensajero/metabolismo , Nicotiana/genética , Nicotiana/crecimiento & desarrollo , Factores de Transcripción/metabolismo , Xilema/genética , Xilema/crecimiento & desarrollo
20.
Opt Express ; 21(21): 24422-30, 2013 Oct 21.
Artículo en Inglés | MEDLINE | ID: mdl-24150287

RESUMEN

We demonstrate that 3D printing, commonly associated with the manufacture of large objects, allows for the fabrication of high quality terahertz (THz) plasmonic structures. Using a commercial 3D printer, we print a variety of structures that include abrupt out-of-plane bends and continuously varying bends. The waveguides are initially printed in a polymer resin and then sputter deposited with ~500 nm of Au. This thickness of Au is sufficient to support low loss propagation of surface plasmon-polaritons with minimal impact from the underlying layer, thereby demonstrating a useful approach for fabricating a broad range of plasmonic structures that incorporate complex geometries. Using THz time-domain spectroscopy, we measure the guided-wave properties of these devices. We find that the propagation properties of the guided-wave modes are similar to those obtained in similar conventional metal-based waveguides, albeit with slightly higher loss. This additional loss is attributed to roughness associated with limitations that currently exist in commercial 3D printers.

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