Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 19 de 19
Filtrar
Mais filtros

Base de dados
Tipo de documento
Intervalo de ano de publicação
1.
Plant Cell ; 34(1): 209-227, 2022 01 20.
Artigo em Inglês | MEDLINE | ID: mdl-34623438

RESUMO

As the outermost layer of plants, the epidermis serves as a critical interface between plants and the environment. During leaf development, the differentiation of specialized epidermal cell types, including stomatal guard cells, pavement cells, and trichomes, occurs simultaneously, each providing unique and pivotal functions for plant growth and survival. Decades of molecular-genetic and physiological studies have unraveled key players and hormone signaling specifying epidermal differentiation. However, most studies focus on only one cell type at a time, and how these distinct cell types coordinate as a unit is far from well-comprehended. Here we provide a review on the current knowledge of regulatory mechanisms underpinning the fate specification, differentiation, morphogenesis, and positioning of these specialized cell types. Emphasis is given to their shared developmental origins, fate flexibility, as well as cell cycle and hormonal controls. Furthermore, we discuss computational modeling approaches to integrate how mechanical properties of individual epidermal cell types and entire tissue/organ properties mutually influence each other. We hope to illuminate the underlying mechanisms coordinating the cell differentiation that ultimately generate a functional leaf epidermis.


Assuntos
Diferenciação Celular , Desenvolvimento Vegetal , Epiderme Vegetal/fisiologia , Folhas de Planta/fisiologia
2.
Proc Natl Acad Sci U S A ; 117(27): 16027-16034, 2020 07 07.
Artigo em Inglês | MEDLINE | ID: mdl-32571946

RESUMO

Puzzle-shaped pavement cells provide a powerful model system to investigate the cellular and subcellular processes underlying complex cell-shape determination in plants. To better understand pavement cell-shape acquisition and the role of auxin in this process, we focused on the spirals of young stomatal lineage ground cells of Arabidopsis leaf epidermis. The predictability of lobe formation in these cells allowed us to demonstrate that the auxin response gradient forms within the cells of the spiral and fluctuates based on the particular stage of lobe development. We revealed that specific localization of auxin transporters at the different membranes of these young cells changes during the course of lobe formation, suggesting that these fluctuating auxin response gradients are orchestrated via auxin transport to control lobe formation and determine pavement cell shape.


Assuntos
Arabidopsis/metabolismo , Forma Celular/efeitos dos fármacos , Forma Celular/fisiologia , Ácidos Indolacéticos/metabolismo , Ácidos Indolacéticos/farmacologia , Proteínas de Arabidopsis , Transporte Biológico , Epiderme Vegetal/metabolismo , Folhas de Planta/metabolismo , Estômatos de Plantas/metabolismo
3.
Plant Physiol ; 187(1): 430-445, 2021 09 04.
Artigo em Inglês | MEDLINE | ID: mdl-34618142

RESUMO

The plant hormone auxin, a master coordinator of development, regulates hypocotyl elongation during seedling growth. We previously identified the synthetic molecule RubNeddin 1 (RN1), which induces degradation of the AUXIN/INDOLE-3-ACETIC ACID (AUX/IAA) transcriptional repressors INDOLE-3-ACETIC ACID-INDUCIBLE3 (IAA3) and IAA7 in planta and strongly promotes hypocotyl elongation. In the present study, we show that despite the structural similarity of RN1 to the synthetic auxin 2,4-dichlorophenoxyacetic-acid (2,4-D), direct treatments with these compounds in Arabidopsis (Arabidopsis thaliana) result in distinct effects, possibly due to enhanced uptake of RN1 and low-level, chronic release of 2,4-D from RN1 in planta. We confirm RN1-induced hypocotyl elongation occurs via specific TRANSPORT INHIBITOR RESISTANT1 (TIR1)/AUXIN SIGNALING F-BOX (AFB) receptor-mediated auxin signaling involving TIR1, AFB2, and AFB5. Using a transcriptome profiling strategy and candidate gene approach, we identify the genes ZINC FINGER OF ARABIDOPSIS THALIANA10 (ZAT10), ARABIDOPSIS TOXICOS EN LEVADURA31 (ATL31), and WRKY DNA-BINDING PROTEIN33 (WRKY33) as being rapidly upregulated by RN1, despite being downregulated by 2,4-D treatment. RN1-induced expression of these genes also occurs via TIR1/AFB-mediated auxin signaling. Our results suggest both hypocotyl elongation and transcription of these genes are induced by RN1 via the promoted degradation of the AUX/IAA transcriptional repressor IAA7. Moreover, these three genes, which are known to be stress-related, act in an inter-dependent transcriptional regulatory network controlling hypocotyl elongation. Together, our results suggest ZAT10, ATL31, and WRKY33 take part in a common gene network regulating hypocotyl elongation in Arabidopsis downstream of a selective auxin perception module likely involving TIR1, AFB2, and AFB5 and inducing the degradation of IAA7.


Assuntos
Proteínas de Arabidopsis/genética , Arabidopsis/crescimento & desenvolvimento , Hipocótilo/crescimento & desenvolvimento , Ácidos Indolacéticos/metabolismo , Reguladores de Crescimento de Plantas/metabolismo , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/metabolismo , Hipocótilo/genética
4.
Proc Natl Acad Sci U S A ; 116(13): 6463-6472, 2019 03 26.
Artigo em Inglês | MEDLINE | ID: mdl-30850516

RESUMO

Auxin phytohormones control most aspects of plant development through a complex and interconnected signaling network. In the presence of auxin, AUXIN/INDOLE-3-ACETIC ACID (AUX/IAA) transcriptional repressors are targeted for degradation by the SKP1-CULLIN1-F-BOX (SCF) ubiquitin-protein ligases containing TRANSPORT INHIBITOR RESISTANT 1/AUXIN SIGNALING F-BOX (TIR1/AFB). CULLIN1-neddylation is required for SCFTIR1/AFB functionality, as exemplified by mutants deficient in the NEDD8-activating enzyme subunit AUXIN-RESISTANT 1 (AXR1). Here, we report a chemical biology screen that identifies small molecules requiring AXR1 to modulate plant development. We selected four molecules of interest, RubNeddin 1 to 4 (RN1 to -4), among which RN3 and RN4 trigger selective auxin responses at transcriptional, biochemical, and morphological levels. This selective activity is explained by their ability to consistently promote the interaction between TIR1 and a specific subset of AUX/IAA proteins, stimulating the degradation of particular AUX/IAA combinations. Finally, we performed a genetic screen using RN4, the RN with the greatest potential for dissecting auxin perception, which revealed that the chromatin remodeling ATPase BRAHMA is implicated in auxin-mediated apical hook development. These results demonstrate the power of selective auxin agonists to dissect auxin perception for plant developmental functions, as well as offering opportunities to discover new molecular players involved in auxin responses.


Assuntos
Ácidos Indolacéticos/metabolismo , Desenvolvimento Vegetal/fisiologia , Proteólise , Fatores de Transcrição/metabolismo , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Proteínas F-Box/metabolismo , Regulação da Expressão Gênica de Plantas , Proteína NEDD8/genética , Desenvolvimento Vegetal/genética , Reguladores de Crescimento de Plantas/genética , Reguladores de Crescimento de Plantas/metabolismo , Plantas Geneticamente Modificadas/genética , Receptores de Superfície Celular/metabolismo , Proteínas Ligases SKP Culina F-Box/metabolismo , Plântula/metabolismo , Transdução de Sinais , Transcrição Gênica/efeitos dos fármacos
5.
New Phytol ; 230(2): 535-549, 2021 04.
Artigo em Inglês | MEDLINE | ID: mdl-33438224

RESUMO

In a world that will rely increasingly on efficient plant growth for sufficient food, it is important to learn about natural mechanisms of phytohormone action. In this work, the introduction of a fluorophore to an auxin molecule represents a sensitive and non-invasive method to directly visualise auxin localisation with high spatiotemporal resolution. The state-of-the-art multidisciplinary approaches of genetic and chemical biology analysis together with live cell imaging, liquid chromatography-mass spectrometry (LC-MS) and surface plasmon resonance (SPR) methods were employed for the characterisation of auxin-related biological activity, distribution and stability of the presented compounds in Arabidopsis thaliana. Despite partial metabolisation in vivo, these fluorescent auxins display an uneven and dynamic distribution leading to the formation of fluorescence maxima in tissues known to concentrate natural auxin, such as the concave side of the apical hook. Importantly, their distribution is altered in response to different exogenous stimuli in both roots and shoots. Moreover, we characterised the subcellular localisation of the fluorescent auxin analogues as being present in the endoplasmic reticulum and endosomes. Our work provides powerful tools to visualise auxin distribution within different plant tissues at cellular or subcellular levels and in response to internal and environmental stimuli during plant development.


Assuntos
Proteínas de Arabidopsis , Arabidopsis , Arabidopsis/metabolismo , Proteínas de Arabidopsis/metabolismo , Regulação da Expressão Gênica de Plantas , Ácidos Indolacéticos , Reguladores de Crescimento de Plantas , Raízes de Plantas/metabolismo
6.
Proc Natl Acad Sci U S A ; 114(18): 4597-4602, 2017 05 02.
Artigo em Inglês | MEDLINE | ID: mdl-28420793

RESUMO

The organic electronic ion pump (OEIP) provides flow-free and accurate delivery of small signaling compounds at high spatiotemporal resolution. To date, the application of OEIPs has been limited to delivery of nonaromatic molecules to mammalian systems, particularly for neuroscience applications. However, many long-standing questions in plant biology remain unanswered due to a lack of technology that precisely delivers plant hormones, based on cyclic alkanes or aromatic structures, to regulate plant physiology. Here, we report the employment of OEIPs for the delivery of the plant hormone auxin to induce differential concentration gradients and modulate plant physiology. We fabricated OEIP devices based on a synthesized dendritic polyelectrolyte that enables electrophoretic transport of aromatic substances. Delivery of auxin to transgenic Arabidopsis thaliana seedlings in vivo was monitored in real time via dynamic fluorescent auxin-response reporters and induced physiological responses in roots. Our results provide a starting point for technologies enabling direct, rapid, and dynamic electronic interaction with the biochemical regulation systems of plants.


Assuntos
Arabidopsis/crescimento & desenvolvimento , Eletrônica , Ácidos Indolacéticos/farmacologia , Fenômenos Fisiológicos Vegetais/efeitos dos fármacos , Plantas Geneticamente Modificadas/crescimento & desenvolvimento , Plântula/crescimento & desenvolvimento , Arabidopsis/genética , Ácidos Indolacéticos/metabolismo , Bombas de Íon , Fenômenos Fisiológicos Vegetais/genética , Plantas Geneticamente Modificadas/genética , Plântula/genética
7.
New Phytol ; 223(3): 1420-1432, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-31038751

RESUMO

distribution of auxin within plant tissues is of great importance for developmental plasticity, including root gravitropic growth. Auxin flow is directed by the subcellular polar distribution and dynamic relocalisation of auxin transporters such as the PIN-FORMED (PIN) efflux carriers, which can be influenced by the main natural plant auxin indole-3-acetic acid (IAA). Anthranilic acid (AA) is an important early precursor of IAA and previously published studies with AA analogues have suggested that AA may also regulate PIN localisation. Using Arabidopsis thaliana as a model species, we studied an AA-deficient mutant displaying agravitropic root growth, treated seedlings with AA and AA analogues and transformed lines to over-produce AA while inhibiting its conversion to downstream IAA precursors. We showed that AA rescues root gravitropic growth in the AA-deficient mutant at concentrations that do not rescue IAA levels. Overproduction of AA affects root gravitropism without affecting IAA levels. Treatments with, or deficiency in, AA result in defects in PIN polarity and gravistimulus-induced PIN relocalisation in root cells. Our results revealed a previously unknown role for AA in the regulation of PIN subcellular localisation and dynamics involved in root gravitropism, which is independent of its better known role in IAA biosynthesis.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Polaridade Celular , Gravitropismo/fisiologia , Ácidos Indolacéticos/metabolismo , Raízes de Plantas/fisiologia , ortoaminobenzoatos/metabolismo , Arabidopsis/efeitos dos fármacos , Polaridade Celular/efeitos dos fármacos , Ácidos Indolacéticos/química , Mutação/genética , Raízes de Plantas/anatomia & histologia , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/crescimento & desenvolvimento , Quinolonas/farmacologia , ortoaminobenzoatos/química , ortoaminobenzoatos/farmacologia
8.
Proc Natl Acad Sci U S A ; 112(7): E806-15, 2015 Feb 17.
Artigo em Inglês | MEDLINE | ID: mdl-25646449

RESUMO

Spatial regulation of the plant hormone indole-3-acetic acid (IAA, or auxin) is essential for plant development. Auxin gradient establishment is mediated by polarly localized auxin transporters, including PIN-FORMED (PIN) proteins. Their localization and abundance at the plasma membrane are tightly regulated by endomembrane machinery, especially the endocytic and recycling pathways mediated by the ADP ribosylation factor guanine nucleotide exchange factor (ARF-GEF) GNOM. We assessed the role of the early secretory pathway in establishing PIN1 polarity in Arabidopsis thaliana by pharmacological and genetic approaches. We identified the compound endosidin 8 (ES8), which selectively interferes with PIN1 basal polarity without altering the polarity of apical proteins. ES8 alters the auxin distribution pattern in the root and induces a strong developmental phenotype, including reduced root length. The ARF-GEF-defective mutants gnom-like 1 (gnl1-1) and gnom (van7) are significantly resistant to ES8. The compound does not affect recycling or vacuolar trafficking of PIN1 but leads to its intracellular accumulation, resulting in loss of PIN1 basal polarity at the plasma membrane. Our data confirm a role for GNOM in endoplasmic reticulum (ER)-Golgi trafficking and reveal that a GNL1/GNOM-mediated early secretory pathway selectively regulates PIN1 basal polarity establishment in a manner essential for normal plant development.


Assuntos
Proteínas de Arabidopsis/fisiologia , Arabidopsis/fisiologia , Fatores de Troca do Nucleotídeo Guanina/fisiologia , Arabidopsis/crescimento & desenvolvimento , Proteínas de Arabidopsis/metabolismo , Endocitose , Proteínas de Membrana Transportadoras/metabolismo , Transporte Proteico
9.
J Exp Bot ; 66(16): 4971-82, 2015 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-25911743

RESUMO

The establishment and maintenance of controlled auxin gradients within plant tissues are essential for a multitude of developmental processes. Auxin gradient formation is co-ordinated via local biosynthesis and transport. Cell to cell auxin transport is facilitated and precisely regulated by complex endomembrane trafficking mechanisms that target auxin carrier proteins to their final destinations. In turn, auxin and cross-talk with other phytohormones regulate the endomembrane trafficking of auxin carriers. Dissecting such rapid and complicated processes is challenging for classical genetic experiments due to trafficking pathway diversity, gene functional redundancy, and lethality in loss-of-function mutants. Many of these difficulties can be bypassed via the use of small molecules to modify or disrupt the function or localization of proteins. Here, we will review examples of the knowledge acquired by the use of such chemical tools in this field, outlining the advantages afforded by chemical biology approaches.


Assuntos
Proteínas de Transporte/metabolismo , Ácidos Indolacéticos/metabolismo , Reguladores de Crescimento de Plantas/metabolismo , Proteínas de Plantas/metabolismo , Transdução de Sinais , Transporte Proteico
10.
Biochem J ; 460(2): 177-85, 2014 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-24654932

RESUMO

Auxin gradients are established and maintained by polarized distribution of auxin transporters that undergo constitutive endocytic recycling from the PM (plasma membrane) and are essential for the gravitropic response in plants. The present study characterizes an inhibitor of endomembrane protein trafficking, TE1 (trafficking and endocytosis inhibitor 1/TENin1) that reduces gravitropic root bending in Arabidopsis thaliana seedlings. Short-term TE1 treatment causes accumulation of PM proteins, including the BR (brassinosteroid) receptor BRI1 (BR insensitive 1), PIP2a (PM intrinsic protein 2a) and the auxin transporter PIN2 (PIN-FORMED 2) in a PVC (pre-vacuolar related compartment), which is sensitive to BFA (Brefeldin A). This compound inhibits endocytosis from the PM and promotes trafficking to the vacuole, consistent with inhibition of retrieval of proteins to the TGN (trans-Golgi network) from the PVC and the PM. However, trafficking of newly synthesized proteins to the PM is unaffected. The short-term protein trafficking inhibition and long-term effect on plant growth and survival caused by TE1 were fully reversible upon drug washout. Structure-activity relationship studies revealed that only minor modifications were possible without loss of biological activity. Diversity in Arabidopsis ecotypes was also exploited to identify two Arabidopsis accessions that display reduced sensitivity to TE1. This compound and the resistant Arabidopsis accessions may be used as a resource in future studies to better understand endomembrane trafficking in plants.


Assuntos
Proteínas de Arabidopsis/fisiologia , Endocitose/efeitos dos fármacos , Gravitropismo/efeitos dos fármacos , Transporte Proteico/efeitos dos fármacos , Arabidopsis/genética , Arabidopsis/crescimento & desenvolvimento , Proteínas de Arabidopsis/metabolismo , Membrana Celular/metabolismo , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/fisiologia , Compostos de Piridínio/metabolismo , Compostos de Amônio Quaternário/metabolismo , Plântula/efeitos dos fármacos , Vacúolos/metabolismo , Rede trans-Golgi/metabolismo
11.
Annu Rev Plant Biol ; 72: 525-550, 2021 06 17.
Artigo em Inglês | MEDLINE | ID: mdl-34143651

RESUMO

The plant epidermis serves many essential functions, including interactions with the environment, protection, mechanical strength, and regulation of tissue and organ growth. To achieve these functions, specialized epidermal cells develop into particular shapes. These include the intriguing interdigitated jigsaw puzzle shape of cotyledon and leaf pavement cells seen in many species, the precise functions of which remain rather obscure. Although pavement cell shape regulation is complex and still a long way from being fully understood, the roles of the cell wall, mechanical stresses, cytoskeleton, cytoskeletal regulatory proteins, and phytohormones are becoming clearer. Here, we provide a review of this current knowledge of pavement cell morphogenesis, generated from a wealth of experimental evidence and assisted by computational modeling approaches. We also discuss the evolution and potential functions of pavement cell interdigitation. Throughout the review, we highlight some of the thought-provoking controversies and creative theories surrounding the formation of the curious puzzle shape of these cells.


Assuntos
Arabidopsis , Parede Celular , Células Epidérmicas , Células Vegetais , Epiderme Vegetal , Folhas de Planta
12.
J Exp Bot ; 61(2): 473-82, 2010.
Artigo em Inglês | MEDLINE | ID: mdl-19933317

RESUMO

Chloroplasts produce reactive oxygen species (ROS) during cellular stress. ROS are known to act as regulators of programmed cell death (PCD) in plant and animal cells, so it is possible that chloroplasts have a role in regulating PCD in green tissue. Arabidopsis thaliana cell suspension cultures are model systems in which to test this, as here it is shown that their cells contain well-developed, functional chloroplasts when grown in the light, but not when grown in the dark. Heat treatment at 55 degrees C induced apoptotic-like (AL)-PCD in the cultures, but light-grown cultures responded with significantly less AL-PCD than dark-grown cultures. Chloroplast-free light-grown cultures were established using norflurazon, spectinomycin, and lincomycin and these cultures responded to heat treatment with increased AL-PCD, demonstrating that chloroplasts affect AL-PCD induction in light-grown cultures. Antioxidant treatment of light-grown cultures also resulted in increased AL-PCD induction, suggesting that chloroplast-produced ROS may be involved in AL-PCD regulation. Cycloheximide treatment of light-grown cultures prolonged cell viability and attenuated AL-PCD induction; however, this effect was less pronounced in dark-grown cultures, and did not occur in antioxidant-treated light-grown cultures. This suggests that a complex interplay between light, chloroplasts, ROS, and nuclear protein synthesis occurs during plant AL-PCD. The results of this study highlight the importance of taking into account the time-point at which cells are observed and whether the cells are light-grown and chloroplast-containing or not, for any study on plant AL-PCD, as it appears that chloroplasts can play a significant role in AL-PCD regulation.


Assuntos
Apoptose , Arabidopsis/citologia , Arabidopsis/metabolismo , Cloroplastos/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Células Cultivadas
13.
Mol Plant ; 11(4): 553-567, 2018 04 02.
Artigo em Inglês | MEDLINE | ID: mdl-29288738

RESUMO

Brassinosteroid (BR) hormone signaling controls multiple processes during plant growth and development and is initiated at the plasma membrane through the receptor kinase BRASSINOSTEROID INSENSITIVE1 (BRI1) together with co-receptors such as BRI1-ASSOCIATED RECEPTOR KINASE1 (BAK1). BRI1 abundance is regulated by endosomal recycling and vacuolar targeting, but the role of vacuole-related proteins in BR receptor dynamics and BR responses remains elusive. Here, we show that the absence of two DUF300 domain-containing tonoplast proteins, LAZARUS1 (LAZ1) and LAZ1 HOMOLOG1 (LAZ1H1), causes vacuole morphology defects, growth inhibition, and constitutive activation of BR signaling. Intriguingly, tonoplast accumulation of BAK1 was substantially increased and appeared causally linked to enhanced BRI1 trafficking and degradation in laz1 laz1h1 plants. Since unrelated vacuole mutants exhibited normal BR responses, our findings indicate that DUF300 proteins play distinct roles in the regulation of BR signaling by maintaining vacuole integrity required to balance subcellular BAK1 pools and BR receptor distribution.


Assuntos
Proteínas Reguladoras de Apoptose/metabolismo , Proteínas de Arabidopsis/metabolismo , Arabidopsis/citologia , Arabidopsis/metabolismo , Brassinosteroides/metabolismo , Transdução de Sinais , Vacúolos/metabolismo , Proteínas Reguladoras de Apoptose/genética , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Mutação , Transporte Proteico
14.
Plant Sci ; 260: 50-59, 2017 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-28554474

RESUMO

The protoplast retracts during apoptosis-like programmed cell death (AL-PCD) and, if this retraction is an active component of AL-PCD, it should be used as a defining feature for this type of programmed cell death. We used an array of pharmacological and genetic tools to test if the rates of protoplast retraction in cells undergoing AL-PCD can be modulated. Disturbing calcium flux signalling, ATP synthesis and mitochondrial permeability transition all inhibited protoplast retraction and often also the execution of the death programme. Protoplast retraction can precede loss of plasma membrane integrity and cell death can be interrupted after the protoplast retraction had already occurred. Blocking calcium influx inhibited the protoplast retraction, reduced DNA fragmentation and delayed death induced by AL-PCD associated stresses. At higher levels of stress, where cell death occurs without protoplast retraction, blocking calcium flux had no effect on the death process. The results therefore strongly suggest that retraction of the protoplast is an active biological process dependent on an early Ca2+-mediated trigger rather than cellular disintegration due to plasma membrane damage. Therefore this morphologically distinct cell type is a quantifiable feature, and consequently, reporter of AL-PCD.


Assuntos
Cálcio/metabolismo , Protoplastos/metabolismo , Transdução de Sinais/fisiologia , Morte Celular/genética , Morte Celular/fisiologia , Fragmentação do DNA , Necrose/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Transdução de Sinais/genética
15.
Methods Mol Biol ; 1242: 93-103, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25408447

RESUMO

Confocal live imaging of the amphiphilic styryl dye FM4-64 is a valuable technique to monitor organelle dynamics and in particular endocytic pathways. After application in plants, FM4-64 immediately stains the plasma membrane and is then integrated on vesicles following endomembrane system-dependent internalization processes. Over time, FM4-64 becomes distributed throughout the full vesicular network from the plasma membrane to the vacuole, including the components of the secretory pathways. Here we provide succinct examples of the many important developmental processes in plants that rely on endocytosis and describe two suitable methods to trace the endocytic pathways in Arabidopsis thaliana root cells based on the uptake of FM4-64.


Assuntos
Arabidopsis/citologia , Rastreamento de Células/métodos , Endocitose , Corantes Fluorescentes/farmacocinética , Compostos de Piridínio/farmacocinética , Compostos de Amônio Quaternário/farmacocinética , Arabidopsis/anatomia & histologia , Arabidopsis/química , Células Epiteliais/química , Microscopia Confocal/métodos , Células Vegetais/química , Raízes de Plantas/química , Raízes de Plantas/citologia
16.
Methods Mol Biol ; 1056: 51-62, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24306862

RESUMO

Chemical genomics is a highly effective approach for understanding complex and dynamic biological processes in plants. A chemical activity can be investigated by a reverse genetics strategy, for which a huge abundance and diversity of Arabidopsis thaliana mutants are readily available for exploitation. Here we present an approach to characterize a chemical of interest, as well as examples of studies demonstrating an effective combination of chemical genomics with reverse genetics strategies, drawn from recent literature on phytohormone signalling and auxin transport.


Assuntos
Arabidopsis/genética , Reguladores de Crescimento de Plantas/farmacologia , Genética Reversa , Arabidopsis/efeitos dos fármacos , Transporte Biológico , Avaliação Pré-Clínica de Medicamentos , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Ácidos Indolacéticos/metabolismo , Mutação , Plântula/efeitos dos fármacos , Plântula/genética , Transdução de Sinais
17.
Toxins (Basel) ; 6(2): 679-92, 2014 Feb 20.
Artigo em Inglês | MEDLINE | ID: mdl-24561479

RESUMO

The Fusarium mycotoxin deoxynivalenol (DON) can cause cell death in wheat (Triticum aestivum), but can also reduce the level of cell death caused by heat shock in Arabidopsis (Arabidopsis thaliana) cell cultures. We show that 10 µg mL(-1) DON does not cause cell death in Arabidopsis cell cultures, and its ability to retard heat-induced cell death is light dependent. Under dark conditions, it actually promoted heat-induced cell death. Wheat cultivars differ in their ability to resist this toxin, and we investigated if the ability of wheat to mount defense responses was light dependent. We found no evidence that light affected the transcription of defense genes in DON-treated roots of seedlings of two wheat cultivars, namely cultivar CM82036 that is resistant to DON-induced bleaching of spikelet tissue and cultivar Remus that is not. However, DON treatment of roots led to genotype-dependent and light-enhanced defense transcript accumulation in coleoptiles. Wheat transcripts encoding a phenylalanine ammonia lyase (PAL) gene (previously associated with Fusarium resistance), non-expressor of pathogenesis-related genes-1 (NPR1) and a class III plant peroxidase (POX) were DON-upregulated in coleoptiles of wheat cultivar CM82036 but not of cultivar Remus, and DON-upregulation of these transcripts in cultivar CM82036 was light enhanced. Light and genotype-dependent differences in the DON/DON derivative content of coleoptiles were also observed. These results, coupled with previous findings regarding the effect of DON on plants, show that light either directly or indirectly influences the plant defense responses to DON.


Assuntos
Arabidopsis/efeitos dos fármacos , Arabidopsis/efeitos da radiação , Luz , Tricotecenos/toxicidade , Triticum/efeitos dos fármacos , Triticum/efeitos da radiação , Arabidopsis/citologia , Sobrevivência Celular/efeitos dos fármacos , Fusarium , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Genótipo , Temperatura Alta , Doenças das Plantas , Plântula/efeitos dos fármacos , Plântula/genética , Plântula/metabolismo , Plântula/efeitos da radiação , Tricotecenos/farmacologia , Triticum/genética , Triticum/metabolismo
18.
PLoS One ; 8(7): e69542, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23922734

RESUMO

The Fusarium genus of fungi is responsible for commercially devastating crop diseases and the contamination of cereals with harmful mycotoxins. Fusarium mycotoxins aid infection, establishment, and spread of the fungus within the host plant. We investigated the effects of the Fusarium mycotoxin deoxynivalenol (DON) on the viability of Arabidopsis cells. Although it is known to trigger apoptosis in animal cells, DON treatment at low concentrations surprisingly did not kill these cells. On the contrary, we found that DON inhibited apoptosis-like programmed cell death (PCD) in Arabidopsis cells subjected to abiotic stress treatment in a manner independent of mitochondrial cytochrome c release. This suggested that Fusarium may utilise mycotoxins to suppress plant apoptosis-like PCD. To test this, we infected Arabidopsis cells with a wild type and a DON-minus mutant strain of F. graminearum and found that only the DON producing strain could inhibit death induced by heat treatment. These results indicate that mycotoxins may be capable of disarming plant apoptosis-like PCD and thereby suggest a novel way that some fungi can influence plant cell fate.


Assuntos
Apoptose/efeitos dos fármacos , Arabidopsis/citologia , Arabidopsis/efeitos dos fármacos , Fusarium/química , Tricotecenos/toxicidade , Arabidopsis/microbiologia , Sobrevivência Celular/efeitos dos fármacos , Cicloeximida/farmacologia , Citocromos c/metabolismo , Etanol/farmacologia , Resposta ao Choque Térmico/efeitos dos fármacos , Temperatura Alta , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Suspensões
19.
Plant Signal Behav ; 5(4): 467-8, 2010 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-20383058

RESUMO

Plant programmed cell death (PCD) is a cell-controlled process that plays an essential role in development and stress responses. Apoptotic-like PCD (AL-PCD) results in a characteristic cell corpse containing a condensed cytoplasm. We recently showed that chloroplast-produced reactive oxygen species (ROS) can play a role in regulating AL-PCD. Here we show that ROS may play a variety of roles in AL-PCD regulation, depending on type and localisation of the ROS activity. Treatment of Arabidopsis thaliana cells with the antioxidants ascorbate and glutathione, which are not specific in the forms of ROS that they scavenge, resulted in increased heat stress-induced AL-PCD. However, treatment with catalase, which specifically scavenges hydrogen peroxide (H(2)O(2)) only, temporally promoted cell survival and suppressed AL-PCD after the heat treatment. These results suggest that H(2)O(2) functions as an important mobile signal that positively regulates AL-PCD in plants and that other ROS forms may play different roles in AL-PCD regulation, perhaps by acting as positive or negative regulators of components of AL-PCD signaling pathways.

SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA