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1.
Biosci Biotechnol Biochem ; 88(10): 1164-1171, 2024 Sep 20.
Artigo em Inglês | MEDLINE | ID: mdl-39013611

RESUMO

SO2/H2SO3 can damage plants. However, its toxic mechanism has still been controversial. Two models have been proposed, cytosolic acidification model and cellular oxidation model. Here, we assessed the toxic mechanism of H2SO3 in three cell types of Arabidopsis thaliana, mesophyll cells, guard cells (GCs), and petal cells. The sensitivity of GCs of Chloride channel a (CLCa)-knockout mutants to H2SO3 was significantly lower than those of wildtype plants. Expression of other CLC genes in mesophyll cells and petal cells were different from GCs. Treatment with antioxidant, disodium 4,5-dihydroxy-1,3-benzenedisulfonate (tiron), increased the median lethal concentration (LC50) of H2SO3 in GCs indicating the involvement of cellular oxidation, while the effect was negligible in mesophyll cells and petal cells. These results indicate that there are two toxic mechanisms of SO2 to Arabidopsis cells: cytosolic acidification and cellular oxidation, and the toxic mechanism may vary among cell types.


Assuntos
Arabidopsis , Citosol , Oxirredução , Dióxido de Enxofre , Arabidopsis/metabolismo , Arabidopsis/genética , Arabidopsis/efeitos dos fármacos , Citosol/metabolismo , Dióxido de Enxofre/toxicidade , Dióxido de Enxofre/metabolismo , Concentração de Íons de Hidrogênio , Células do Mesofilo/metabolismo , Células do Mesofilo/efeitos dos fármacos , Proteínas de Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Canais de Cloreto/metabolismo , Canais de Cloreto/genética , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos
2.
Plant Physiol ; 180(2): 910-925, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-30910907

RESUMO

Abscisic acid (ABA) levels increase significantly in plants under stress conditions, and ABA is thought to serve as a key stress-response regulator. However, the direct effect of ABA on photosynthesis and the effect of mesophyll ABA on yield under both well-watered and drought conditions are still the subject of debate. Here, we examined this issue using transgenic Arabidopsis (Arabidopsis thaliana) plants carrying a dominant ABA-signaling inhibitor under the control of a mesophyll-specific promoter (FBPase::abi1-1, abbreviated to fa). Under normal conditions, fa plants displayed slightly higher stomatal conductance and carbon assimilation than wild-type plants; however, these parameters were comparable following ABA treatment. These observations suggest that ABA does not directly inhibit photosynthesis in the short term. The fa plants also exhibited a variety of altered phenotypes under optimal conditions, including more vigorous initial growth, earlier flowering, smaller flowers, and delayed chlorophyll degradation. Furthermore, under optimal conditions, fa plant seed production was less than a third of that observed for the wild type. However, under drought conditions, wild-type and fa seed yields were similar due to a significant reduction in wild-type seed and no reduction in fa seed. These findings suggest that endogenous basal ABA inhibits a stress-escape response under nonstressed conditions, allowing plants to accumulate biomass and maximize yield. The lack of a correlation between flowering time and plant biomass combined with delayed chlorophyll degradation suggests that this stress-escape behavior is regulated independently and upstream of other ABA-induced effects such as rapid growth and flowering.


Assuntos
Ácido Abscísico/farmacologia , Arabidopsis/fisiologia , Flores/fisiologia , Células do Mesofilo/fisiologia , Fotossíntese , Arabidopsis/efeitos dos fármacos , Arabidopsis/crescimento & desenvolvimento , Biomassa , Clorofila/metabolismo , Secas , Flores/anatomia & histologia , Gases/metabolismo , Células do Mesofilo/efeitos dos fármacos , Modelos Biológicos , Fotossíntese/efeitos dos fármacos , Estômatos de Plantas/efeitos dos fármacos , Estômatos de Plantas/fisiologia , Plantas Geneticamente Modificadas , Transgenes , Água
3.
Int J Mol Sci ; 21(16)2020 Aug 11.
Artigo em Inglês | MEDLINE | ID: mdl-32796535

RESUMO

The growing tips of plants grow sterile; therefore, disease-free plants can be generated from them. How plants safeguard growing apices from pathogen infection is still a mystery. The shoot apical meristem (SAM) is one of the three stem cells niches that give rise to the above ground plant organs. This is very well explored; however, how signaling networks orchestrate immune responses against pathogen infections in the SAM remains unclear. To reconstruct a transcriptional framework of the differentially expressed genes (DEGs) pertaining to various SAM cellular populations, we acquired large-scale transcriptome datasets from the public repository Gene Expression Omnibus (GEO). We identify here distinct sets of genes for various SAM cellular populations that are enriched in immune functions, such as immune defense, pathogen infection, biotic stress, and response to salicylic acid and jasmonic acid and their biosynthetic pathways in the SAM. We further linked those immune genes to their respective proteins and identify interactions among them by mapping a transcriptome-guided SAM-interactome. Furthermore, we compared stem-cells regulated transcriptome with innate immune responses in plants showing transcriptional separation among their DEGs in Arabidopsis. Besides unleashing a repertoire of immune-related genes in the SAM, our analysis provides a SAM-interactome that will help the community in designing functional experiments to study the specific defense dynamics of the SAM-cellular populations. Moreover, our study promotes the essence of large-scale omics data re-analysis, allowing a fresh look at the SAM-cellular transcriptome repurposing data-sets for new questions.


Assuntos
Arabidopsis/genética , Arabidopsis/imunologia , Regulação da Expressão Gênica de Plantas , Meristema/genética , Meristema/imunologia , Imunidade Vegetal/genética , Transcrição Gênica , Arabidopsis/efeitos dos fármacos , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Flagelina/farmacologia , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Ontologia Genética , Meristema/efeitos dos fármacos , Células do Mesofilo/efeitos dos fármacos , Células do Mesofilo/metabolismo , Imunidade Vegetal/efeitos dos fármacos , Transcrição Gênica/efeitos dos fármacos , Transcriptoma/efeitos dos fármacos , Transcriptoma/genética
4.
Int J Mol Sci ; 21(5)2020 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-32121556

RESUMO

Nitric oxide (NO) plays an important role in stomata closure induced by environmental stimuli including pathogens. During pathogen challenge, nitric oxide (NO) acts as a second messenger in guard cell signaling networks to activate downstream responses leading to stomata closure. One means by which NO's action is achieved is through the posttranslational modification of cysteine residue(s) of target proteins. Although the roles of NO have been well studied in plant tissues and seedlings, far less is known about NO signaling and, more specifically, protein S-nitrosylation (SNO) in stomatal guard cells. In this study, using iodoTMTRAQ quantitative proteomics technology, we analyzed changes in protein SNO modification in guard cells of reference plant Arabidopsis thaliana in response to flg22, an elicitor-active peptide derived from bacterial flagellin. A total of 41 SNO-modified peptides corresponding to 35 proteins were identified. The proteins cover a wide range of functions, including energy metabolism, transport, stress response, photosynthesis, and cell-cell communication. This study creates the first inventory of previously unknown NO responsive proteins in guard cell immune responses and establishes a foundation for future research toward understanding the molecular mechanisms and regulatory roles of SNO in stomata immunity against bacterial pathogens.


Assuntos
Arabidopsis/citologia , Flagelina/farmacologia , Estômatos de Plantas/citologia , Estômatos de Plantas/metabolismo , Proteoma/metabolismo , Arabidopsis/efeitos dos fármacos , Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Sobrevivência Celular/efeitos dos fármacos , Análise por Conglomerados , Ontologia Genética , Células do Mesofilo/citologia , Células do Mesofilo/efeitos dos fármacos , Células do Mesofilo/metabolismo , Óxido Nítrico/metabolismo , Nitrosação , Estômatos de Plantas/efeitos dos fármacos , Estômatos de Plantas/fisiologia , Espécies Reativas de Oxigênio/metabolismo
5.
J Exp Bot ; 70(18): 4807-4818, 2019 09 24.
Artigo em Inglês | MEDLINE | ID: mdl-31056658

RESUMO

Decreases in photosynthetic rate, stomatal conductance (gs), and mesophyll conductance (gm) are often observed under elevated CO2 conditions. However, which anatomical and/or physiological factors contribute to the decrease in gm is not fully understood. Arabidopsis thaliana wild-type and carbon-metabolism mutants (gwd1, pgm1, and cfbp1) with different accumulation patterns of non-structural carbohydrates were grown at ambient (400 ppm) and elevated (800 ppm) CO2. Anatomical and physiological traits of leaves were measured to investigate factors causing the changes in gm and in the mesophyll resistance (expressed as the reciprocal of mesophyll conductance per unit chloroplast surface area facing to intercellular space, Sc/gm). When grown at elevated CO2, all the lines showed increases in cell wall mass, cell wall thickness, and starch content, but not in leaf thickness. gm measured at 800 ppm CO2 was significantly lower than at 400 ppm CO2 in all the lines. Changes in Sc/gm were associated with thicker cell walls rather than with excess starch content. The results indicate that the changes in gm and Sc/gm that occur in response to elevated CO2 are independent of non-structural carbohydrates, and the cell wall represents a greater limitation factor for gm than starch.


Assuntos
Arabidopsis/fisiologia , Dióxido de Carbono/metabolismo , Células do Mesofilo/efeitos dos fármacos , Cloroplastos/efeitos dos fármacos , Cloroplastos/metabolismo , Cloroplastos/ultraestrutura , Células do Mesofilo/metabolismo , Células do Mesofilo/ultraestrutura , Microscopia Eletrônica de Transmissão , Folhas de Planta/metabolismo
6.
Physiol Plant ; 165(3): 604-618, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-29744895

RESUMO

Mesophyll conductance to CO2 (gm ), a key photosynthetic trait, is strongly constrained by leaf anatomy. Leaf anatomical parameters such as cell wall thickness and chloroplast area exposed to the mesophyll intercellular airspace have been demonstrated to determine gm in species with diverging phylogeny, leaf structure and ontogeny. However, the potential implication of leaf anatomy, especially chloroplast movement, on the short-term response of gm to rapid changes (i.e. seconds to minutes) under different environmental conditions (CO2 , light or temperature) has not been examined. The aim of this study was to determine whether the observed rapid variations of gm in response to variations of light and CO2 could be explained by changes in any leaf anatomical arrangements. When compared to high light and ambient CO2 , the values of gm estimated by chlorophyll fluorescence decreased under high CO2 and increased at low CO2 , while it decreased with decreasing light. Nevertheless, no changes in anatomical parameters, including chloroplast distribution, were found. Hence, the gm estimated by analytical models based on anatomical parameters was constant under varying light and CO2 . Considering this discrepancy between anatomy and chlorophyll fluorescence estimates, it is concluded that apparent fast gm variations should be due to artefacts in its estimation and/or to changes in the biochemical components acting on diffusional properties of the leaf (e.g. aquaporins and carbonic anhydrase).


Assuntos
Dióxido de Carbono/farmacologia , Células do Mesofilo/metabolismo , Nicotiana/metabolismo , Folhas de Planta/metabolismo , Células do Mesofilo/efeitos dos fármacos , Fotossíntese/efeitos dos fármacos , Nicotiana/efeitos dos fármacos
7.
New Phytol ; 217(4): 1463-1474, 2018 03.
Artigo em Inglês | MEDLINE | ID: mdl-29220090

RESUMO

Mesophyll conductance (gm ) describes the movement of CO2 from the intercellular air spaces below the stomata to the site of initial carboxylation in the mesophyll. In contrast with C3 -gm , little is currently known about the intraspecific variation in C4 -gm or its responsiveness to environmental stimuli. To address these questions, gm was measured on five maize (Zea mays) lines in response to CO2 , employing three different estimates of gm . Each of the methods indicated a significant response of gm to CO2 . Estimates of gm were similar between methods at ambient and higher CO2 , but diverged significantly at low partial pressures of CO2 . These differences are probably driven by incomplete chemical and isotopic equilibrium between CO2 and bicarbonate under these conditions. Carbonic anhydrase and phosphoenolpyruvate carboxylase in vitro activity varied significantly despite similar values of gm and leaf anatomical traits. These results provide strong support for a CO2 response of gm in Z. mays, and indicate that gm in maize is probably driven by anatomical constraints rather than by biochemical limitations. The CO2 response of gm indicates a potential role for facilitated diffusion in C4 -gm . These results also suggest that water-use efficiency could be enhanced in C4 species by targeting gm .


Assuntos
Dióxido de Carbono/farmacologia , Produtos Agrícolas/fisiologia , Células do Mesofilo/fisiologia , Transpiração Vegetal/fisiologia , Zea mays/fisiologia , Anidrases Carbônicas/metabolismo , Produtos Agrícolas/efeitos dos fármacos , Células do Mesofilo/efeitos dos fármacos , Células do Mesofilo/enzimologia , Modelos Biológicos , Isótopos de Oxigênio , Fosfoenolpiruvato Carboxiquinase (ATP)/metabolismo , Fotossíntese/efeitos dos fármacos , Estômatos de Plantas/efeitos dos fármacos , Estômatos de Plantas/fisiologia , Transpiração Vegetal/efeitos dos fármacos , Ribulose-Bifosfato Carboxilase/metabolismo , Água , Zea mays/anatomia & histologia , Zea mays/efeitos dos fármacos , Zea mays/enzimologia
8.
New Phytol ; 219(4): 1421-1432, 2018 09.
Artigo em Inglês | MEDLINE | ID: mdl-29938800

RESUMO

The membrane-bound proton-pumping pyrophosphatase (V-PPase), together with the V-type H+ -ATPase, generates the proton motive force that drives vacuolar membrane solute transport. Transgenic plants constitutively overexpressing V-PPases were shown to have improved salinity tolerance, but the relative impact of increasing PPi hydrolysis and proton-pumping functions has yet to be dissected. For a better understanding of the molecular processes underlying V-PPase-dependent salt tolerance, we transiently overexpressed the pyrophosphate-driven proton pump (NbVHP) in Nicotiana benthamiana leaves and studied its functional properties in relation to salt treatment by primarily using patch-clamp, impalement electrodes and pH imaging. NbVHP overexpression led to higher vacuolar proton currents and vacuolar acidification. After 3 d in salt-untreated conditions, V-PPase-overexpressing leaves showed a drop in photosynthetic capacity, plasma membrane depolarization and eventual leaf necrosis. Salt, however, rescued NbVHP-hyperactive cells from cell death. Furthermore, a salt-induced rise in V-PPase but not of V-ATPase pump currents was detected in nontransformed plants. The results indicate that under normal growth conditions, plants need to regulate the V-PPase pump activity to avoid hyperactivity and its negative feedback on cell viability. Nonetheless, V-PPase proton pump function becomes increasingly important under salt stress for generating the pH gradient necessary for vacuolar proton-coupled Na+ sequestration.


Assuntos
Pirofosfatase Inorgânica/metabolismo , Nicotiana/enzimologia , Salinidade , Cloreto de Sódio/farmacologia , Vacúolos/enzimologia , Morte Celular/efeitos dos fármacos , Membrana Celular/efeitos dos fármacos , Membrana Celular/metabolismo , Difosfatos/metabolismo , Concentração de Íons de Hidrogênio , Isoenzimas/metabolismo , Potenciais da Membrana/efeitos dos fármacos , Células do Mesofilo/efeitos dos fármacos , Células do Mesofilo/enzimologia , Epiderme Vegetal/citologia , Epiderme Vegetal/efeitos dos fármacos , Bombas de Próton/metabolismo , Prótons , Estresse Fisiológico/efeitos dos fármacos , Nicotiana/efeitos dos fármacos , ATPases Vacuolares Próton-Translocadoras/metabolismo
9.
Plant Physiol ; 172(3): 1989-2001, 2016 11.
Artigo em Inglês | MEDLINE | ID: mdl-27663407

RESUMO

Volatile compounds (VCs) emitted by phylogenetically diverse microorganisms (including plant pathogens and microbes that do not normally interact mutualistically with plants) promote photosynthesis, growth, and the accumulation of high levels of starch in leaves through cytokinin (CK)-regulated processes. In Arabidopsis (Arabidopsis thaliana) plants not exposed to VCs, plastidic phosphoglucose isomerase (pPGI) acts as an important determinant of photosynthesis and growth, likely as a consequence of its involvement in the synthesis of plastidic CKs in roots. Moreover, this enzyme plays an important role in connecting the Calvin-Benson cycle with the starch biosynthetic pathway in leaves. To elucidate the mechanisms involved in the responses of plants to microbial VCs and to investigate the extent of pPGI involvement, we characterized pPGI-null pgi1-2 Arabidopsis plants cultured in the presence or absence of VCs emitted by Alternaria alternata We found that volatile emissions from this fungal phytopathogen promote growth, photosynthesis, and the accumulation of plastidic CKs in pgi1-2 leaves. Notably, the mesophyll cells of pgi1-2 leaves accumulated exceptionally high levels of starch following VC exposure. Proteomic analyses revealed that VCs promote global changes in the expression of proteins involved in photosynthesis, starch metabolism, and growth that can account for the observed responses in pgi1-2 plants. The overall data show that Arabidopsis plants can respond to VCs emitted by phytopathogenic microorganisms by triggering pPGI-independent mechanisms.


Assuntos
Alternaria/química , Proteínas de Arabidopsis/metabolismo , Arabidopsis/enzimologia , Arabidopsis/microbiologia , Glucose-6-Fosfato Isomerase/metabolismo , Plastídeos/enzimologia , Compostos Orgânicos Voláteis/farmacologia , Alternaria/efeitos da radiação , Arabidopsis/crescimento & desenvolvimento , Arabidopsis/fisiologia , Parede Celular/metabolismo , Parede Celular/efeitos da radiação , Citocininas/metabolismo , Luz , Células do Mesofilo/efeitos dos fármacos , Células do Mesofilo/metabolismo , Células do Mesofilo/efeitos da radiação , Mutação/genética , Fotossíntese/efeitos da radiação , Plastídeos/efeitos dos fármacos , Proteoma/metabolismo , Amido/metabolismo
10.
Plant Physiol ; 171(1): 590-605, 2016 05.
Artigo em Inglês | MEDLINE | ID: mdl-26932234

RESUMO

Leaves are the plant's powerhouses, providing energy for all organs through sugar production during photosynthesis. However, sugars serve not only as a metabolic energy source for sink tissues but also as signaling molecules, affecting gene expression through conserved signaling pathways to regulate plant growth and development. Here, we describe an in vitro experimental assay, allowing one to alter the sucrose (Suc) availability during early Arabidopsis (Arabidopsis thaliana) leaf development, with the aim to identify the affected cellular and molecular processes. The transfer of seedlings to Suc-containing medium showed a profound effect on leaf growth by stimulating cell proliferation and postponing the transition to cell expansion. Furthermore, rapidly after transfer to Suc, mesophyll cells contained fewer and smaller plastids, which are irregular in shape and contain fewer starch granules compared with control mesophyll cells. Short-term transcriptional responses after transfer to Suc revealed the repression of well-known sugar-responsive genes and multiple genes encoded by the plastid, on the one hand, and up-regulation of a GLUCOSE-6-PHOSPHATE TRANSPORTER (GPT2), on the other hand. Mutant gpt2 seedlings showed no stimulation of cell proliferation and no repression of chloroplast-encoded transcripts when transferred to Suc, suggesting that GPT2 plays a critical role in the Suc-mediated effects on early leaf growth. Our findings, therefore, suggest that induction of GPT2 expression by Suc increases the import of glucose-6-phosphate into the plastids that would repress chloroplast-encoded transcripts, restricting chloroplast differentiation. Retrograde signaling from the plastids would then delay the transition to cell expansion and stimulate cell proliferation.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/crescimento & desenvolvimento , Cloroplastos/metabolismo , Proteínas de Transporte de Monossacarídeos/metabolismo , Folhas de Planta/crescimento & desenvolvimento , Sacarose/metabolismo , Arabidopsis/efeitos dos fármacos , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proliferação de Células/efeitos dos fármacos , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Células do Mesofilo/efeitos dos fármacos , Proteínas de Transporte de Monossacarídeos/genética , Mutação , Folhas de Planta/citologia , Folhas de Planta/metabolismo , Plântula/efeitos dos fármacos , Plântula/crescimento & desenvolvimento , Sacarose/farmacologia
11.
Plant Cell Environ ; 40(2): 203-215, 2017 02.
Artigo em Inglês | MEDLINE | ID: mdl-27861995

RESUMO

Stomata represent one resistor in a series of resistances for carbon and water exchange between the leaf and the atmosphere; the remaining resistors occurring within the leaf, commonly represented as mesophyll conductance to CO2 , gm , and leaf hydraulic conductance, kLeaf . Recent studies have proposed that gm and kLeaf may be coordinated across species because of shared pathways. We assessed the correlation between gm and kLeaf within cotton, under growth CO2 partial pressure and irradiance treatments and also with short-term variation in irradiance and humidity. gm was estimated using two isotopic techniques that allowed partitioning of total gm (Δ13 C-gm ) into cell wall plus plasma membrane conductance (Δ18 O-gm ) and chloroplast membrane conductance (gcm ). A weak correlation was found between Δ13 C-gm and kLeaf only when measured under growth conditions. However, Δ18 O-gm was related to kLeaf under both short-term environmental variation and growth conditions. Partitioning gm showed that gcm was not affected by short-term changes in irradiance or correlated with kLeaf , but was strongly reduced at high growth CO2 partial pressure. Thus, simultaneous measurements of gm , kLeaf and gcm suggest independent regulation of carbon and water transport across the chloroplast membrane with limited coordinated regulation across the cell wall and plasma membrane.


Assuntos
Células do Mesofilo/fisiologia , Folhas de Planta/fisiologia , Água/fisiologia , Dióxido de Carbono/farmacologia , Isótopos de Carbono , Difusão , Gossypium/anatomia & histologia , Gossypium/efeitos dos fármacos , Gossypium/crescimento & desenvolvimento , Luz , Células do Mesofilo/efeitos dos fármacos , Método de Monte Carlo , Folhas de Planta/anatomia & histologia , Folhas de Planta/efeitos dos fármacos , Especificidade da Espécie
12.
Plant Cell Environ ; 40(1): 138-149, 2017 01.
Artigo em Inglês | MEDLINE | ID: mdl-27761902

RESUMO

In Populus trichocarpa (black cottonwood), net photosynthesis (An ) varies with latitude and, in northern genotypes, is supported by higher stomatal conductance (gs ). We report here a parallel cline in mesophyll conductance (gm ) and link this variation to carbonic anhydrase (CA) activity. Using concurrent carbon isotope discrimination and chlorophyll fluorescence methods, we examined the effects of acetazolamide, an inhibitor of CA, on gm in six representative genotypes (three from either end of the north-south cline). Acetazolamide reduced CA activity, gm , gs , chloroplast CO2 concentration (Cc ) and An at normal CO2 (400 µmol mol-1 ), the latter being reversible at saturating CO2 . Absolute reductions in An , gm and CA activity were greater in northern genotypes than in southern genotypes (P < 0.025) but percent reductions were similar. In contrast, northern genotypes showed lower percent reduction in Cc compared to southern genotypes (P < 0.025). The northern genotypes had greater CA activity relative to both leaf area (two-fold) and mass (1.8-fold) (P < 0.016). The relationship between CA activity and gm was similar whether the variation was inherent or inhibitor induced. We suggest that greater CA activity contributes to higher gm in northern P. trichocarpa genotypes, but other diffusion pathway components may also be involved.


Assuntos
Anidrases Carbônicas/metabolismo , Células do Mesofilo/metabolismo , Populus/enzimologia , Acetazolamida/farmacologia , Genótipo , Células do Mesofilo/efeitos dos fármacos , Fotossíntese/efeitos dos fármacos , Estômatos de Plantas/efeitos dos fármacos , Estômatos de Plantas/fisiologia , Populus/efeitos dos fármacos , Populus/genética , Característica Quantitativa Herdável
13.
Plant Cell Environ ; 40(9): 1900-1915, 2017 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-28558173

RESUMO

Epidermal bladder cells (EBCs) have been postulated to assist halophytes in coping with saline environments. However, little direct supporting evidence is available. Here, Chenopodium quinoa plants were grown under saline conditions for 5 weeks. One day prior to salinity treatment, EBCs from all leaves and petioles were gently removed by using a soft cosmetic brush and physiological, ionic and metabolic changes in brushed and non-brushed leaves were compared. Gentle removal of EBC neither initiated wound metabolism nor affected the physiology and biochemistry of control-grown plants but did have a pronounced effect on salt-grown plants, resulting in a salt-sensitive phenotype. Of 91 detected metabolites, more than half were significantly affected by salinity. Removal of EBC dramatically modified these metabolic changes, with the biggest differences reported for gamma-aminobutyric acid (GABA), proline, sucrose and inositol, affecting ion transport across cellular membranes (as shown in electrophysiological experiments). This work provides the first direct evidence for a role of EBC in salt tolerance in halophytes and attributes this to (1) a key role of EBC as a salt dump for external sequestration of sodium; (2) improved K+ retention in leaf mesophyll and (3) EBC as a storage space for several metabolites known to modulate plant ionic relations.


Assuntos
Atriplex/fisiologia , Chenopodium quinoa/fisiologia , Epiderme Vegetal/citologia , Tolerância ao Sal/fisiologia , Plantas Tolerantes a Sal/fisiologia , Estresse Fisiológico , Atriplex/efeitos dos fármacos , Membrana Celular/efeitos dos fármacos , Membrana Celular/metabolismo , Chenopodium quinoa/efeitos dos fármacos , Cromatografia Gasosa-Espectrometria de Massas , Transporte de Íons/efeitos dos fármacos , Células do Mesofilo/efeitos dos fármacos , Células do Mesofilo/metabolismo , Metaboloma , Fenótipo , Epiderme Vegetal/efeitos dos fármacos , Folhas de Planta/fisiologia , Tolerância ao Sal/efeitos dos fármacos , Plantas Tolerantes a Sal/efeitos dos fármacos , Estresse Fisiológico/efeitos dos fármacos , Sacarose/farmacologia , Ácido gama-Aminobutírico/farmacologia
14.
Proc Natl Acad Sci U S A ; 111(29): E3015-23, 2014 Jul 22.
Artigo em Inglês | MEDLINE | ID: mdl-25002510

RESUMO

Plants, as sessile organisms, must coordinate various physiological processes to adapt to ever-changing surrounding environments. Stomata, the epidermal pores facilitating gas and water exchange, play important roles in optimizing photosynthetic efficiency and adaptability. Stomatal development is under the control of an intrinsic program mediated by a secretory peptide gene family--namely, EPIDERMAL PATTERNING FACTOR, including positively acting STOMAGEN/EPFL9. The phytohormone brassinosteroids and environment factor light also control stomatal production. However, whether auxin regulates stomatal development and whether peptide signaling is coordinated with auxin signaling in the regulation of stomatal development remain largely unknown. Here we show that auxin negatively regulates stomatal development through MONOPTEROS (also known as ARF5) repression of the mobile peptide gene STOMAGEN in mesophyll. Through physiological, genetic, transgenic, biochemical, and molecular analyses, we demonstrate that auxin inhibits stomatal development through the nuclear receptor TIR1/AFB-mediated signaling, and that MONOPTEROS directly binds to the STOMAGEN promoter to suppress its expression in mesophyll and inhibit stomatal development. Our results provide a paradigm of cross-talk between phytohormone auxin and peptide signaling in the regulation of stomatal production.


Assuntos
Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Arabidopsis/crescimento & desenvolvimento , Arabidopsis/genética , Proteínas de Ligação a DNA/metabolismo , Ácidos Indolacéticos/farmacologia , Células do Mesofilo/metabolismo , Estômatos de Plantas/crescimento & desenvolvimento , Fatores de Transcrição/metabolismo , Arabidopsis/efeitos dos fármacos , Pareamento de Bases/genética , Padronização Corporal/efeitos dos fármacos , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Genes de Plantas , Células do Mesofilo/efeitos dos fármacos , Modelos Biológicos , Peptídeos/genética , Peptídeos/metabolismo , Estômatos de Plantas/efeitos dos fármacos , Ligação Proteica/efeitos dos fármacos , Ligação Proteica/genética , Receptores Citoplasmáticos e Nucleares/metabolismo , Elementos de Resposta/genética , Plântula/efeitos dos fármacos , Plântula/crescimento & desenvolvimento , Transdução de Sinais/efeitos dos fármacos
15.
Plant Cell Physiol ; 57(11): 2334-2341, 2016 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-27519315

RESUMO

CO2-responsive CCT protein (CRCT) is the suggested positive regulator of starch synthesis in vegetative organs, particularly the leaf sheath of rice. In this study, we analyzed the effects of the starch level in the leaf sheath on the photosynthetic rate in the leaf blade using CRCT overexpression and RNA interference (RNAi) knockdown transgenic rice grown under ambient (38 Pa) or elevated (100 Pa) CO2 conditions. In leaf sheath, the starch content was markedly changed in relation to CRCT expression levels under both CO2 conditions. In contrast, the soluble sugar and starch contents of the leaf blade were markedly increased in the knockdown line grown under elevated CO2 conditions. The overexpression or RNAi knockdown of CRCT did not cause large effects on the photosynthetic rate of the transgenic lines grown under ambient CO2 condition. However, the photosynthetic rate of the overexpression line was enhanced, while that of the knockdown line was substantially decreased under elevated CO2 conditions. These photosynthetic rates were weakly correlated with the nitrogen contents and negatively correlated with the total non-structural carbohydrate contents. Thus, the capacity for starch synthesis in leaf sheath, which is controlled by CRCT, can indirectly affect the carbohydrate content, and then the photosynthetic rate in the leaf blade of rice grown under elevated CO2 conditions.


Assuntos
Dióxido de Carbono/farmacologia , Oryza/fisiologia , Fotossíntese , Folhas de Planta/metabolismo , Proteínas de Plantas/metabolismo , Amido/metabolismo , Biomassa , Metabolismo dos Carboidratos/efeitos dos fármacos , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Células do Mesofilo/efeitos dos fármacos , Células do Mesofilo/metabolismo , Células do Mesofilo/ultraestrutura , Nitrogênio/metabolismo , Oryza/genética , Oryza/crescimento & desenvolvimento , Fotossíntese/efeitos dos fármacos , Folhas de Planta/efeitos dos fármacos , Folhas de Planta/genética , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas , Solubilidade
16.
Plant Physiol ; 168(3): 1106-21, 2015 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-25944825

RESUMO

The membrane-bound Brassinosteroid insensitive1-associated receptor kinase1 (BAK1) is a common coreceptor in plants and regulates distinct cellular programs ranging from growth and development to defense against pathogens. BAK1 functions through binding to ligand-stimulated transmembrane receptors and activating their kinase domains via transphosphorylation. In the absence of microbes, BAK1 activity may be suppressed by different mechanisms, like interaction with the regulatory BIR (for BAK1-interacting receptor-like kinase) proteins. Here, we demonstrated that BAK1 overexpression in Arabidopsis (Arabidopsis thaliana) could cause detrimental effects on plant development, including growth arrest, leaf necrosis, and reduced seed production. Further analysis using an inducible expression system showed that BAK1 accumulation quickly stimulated immune responses, even under axenic conditions, and led to increased resistance to pathogenic Pseudomonas syringae pv tomato DC3000. Intriguingly, our study also revealed that the plasma membrane-associated BAK1 ectodomain was sufficient to induce autoimmunity, indicating a novel mode of action for BAK1 in immunity control. We postulate that an excess of BAK1 or its ectodomain could trigger immune receptor activation in the absence of microbes through unbalancing regulatory interactions, including those with BIRs. Consistently, mutation of suppressor of BIR1-1, which encodes an emerging positive regulator of transmembrane receptors in plants, suppressed the effects of BAK1 overexpression. In conclusion, our findings unravel a new role for the BAK1 ectodomain in the tight regulation of Arabidopsis immune receptors necessary to avoid inappropriate activation of immunity.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/imunologia , Autoimunidade , Imunidade Vegetal , Proteínas Quinases/metabolismo , Proteínas Serina-Treonina Quinases/química , Proteínas Serina-Treonina Quinases/metabolismo , Arabidopsis/genética , Arabidopsis/crescimento & desenvolvimento , Arabidopsis/microbiologia , Proteínas de Arabidopsis/química , Proteínas de Arabidopsis/genética , Autoimunidade/efeitos dos fármacos , Morte Celular/efeitos dos fármacos , Resistência à Doença/efeitos dos fármacos , Flagelina/farmacologia , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Genes Dominantes , Genes de Plantas , Células do Mesofilo/citologia , Células do Mesofilo/efeitos dos fármacos , Células do Mesofilo/metabolismo , Mutação/genética , Moléculas com Motivos Associados a Patógenos/metabolismo , Fenótipo , Doenças das Plantas/imunologia , Doenças das Plantas/microbiologia , Imunidade Vegetal/efeitos dos fármacos , Plantas Geneticamente Modificadas , Proteínas Quinases/genética , Estrutura Terciária de Proteína , Pseudomonas syringae/crescimento & desenvolvimento , Pseudomonas syringae/fisiologia , Plântula/citologia , Plântula/efeitos dos fármacos , Plântula/metabolismo
17.
Ann Bot ; 117(3): 401-19, 2016 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-26802013

RESUMO

BACKGROUND AND AIMS: This work investigates the involvement of local differentiation of cell wall matrix polysaccharides and the role of microtubules in the morphogenesis of mesophyll cells (MCs) of three types (lobed, branched and palisade) in the dicotyledon Vigna sinensis and the fern Asplenium nidus. METHODS: Homogalacturonan (HGA) epitopes recognized by the 2F4, JIM5 and JIM7 antibodies and callose were immunolocalized in hand-made leaf sections. Callose was also stained with aniline blue. We studied microtubule organization by tubulin immunofluorescence and transmission electron microscopy. RESULTS: In both plants, the matrix cell wall polysaccharide distribution underwent definite changes during MC differentiation. Callose constantly defined the sites of MC contacts. The 2F4 HGA epitope in V. sinensis first appeared in MC contacts but gradually moved towards the cell wall regions facing the intercellular spaces, while in A. nidus it was initially localized at the cell walls delimiting the intercellular spaces, but finally shifted to MC contacts. In V. sinensis, the JIM5 and JIM7 HGA epitopes initially marked the cell walls delimiting the intercellular spaces and gradually shifted in MC contacts, while in A. nidus they constantly enriched MC contacts. In all MC types examined, the cortical microtubules played a crucial role in their morphogenesis. In particular, in palisade MCs, cortical microtubule helices, by controlling cellulose microfibril orientation, forced these MCs to acquire a truncated cone-like shape. Unexpectedly in V. sinensis, the differentiation of colchicine-affected MCs deviated completely, since they developed a cell wall ingrowth labyrinth, becoming transfer-like cells. CONCLUSIONS: The results of this work and previous studies on Zea mays (Giannoutsou et al., Annals of Botany 2013; 112: : 1067-1081) revealed highly controlled local cell wall matrix differentiation in MCs of species belonging to different plant groups. This, in coordination with microtubule-dependent cellulose microfibril alignment, spatially controlled cell wall expansion, allowing MCs to acquire their particular shape.


Assuntos
Parede Celular/metabolismo , Embriófitas/metabolismo , Células do Mesofilo/metabolismo , Microtúbulos/metabolismo , Morfogênese , Polissacarídeos/metabolismo , Parede Celular/efeitos dos fármacos , Parede Celular/ultraestrutura , Colchicina/farmacologia , Embriófitas/efeitos dos fármacos , Epitopos/química , Epitopos/metabolismo , Glucanos/metabolismo , Células do Mesofilo/citologia , Células do Mesofilo/efeitos dos fármacos , Células do Mesofilo/ultraestrutura , Microtúbulos/efeitos dos fármacos , Morfogênese/efeitos dos fármacos , Coloração e Rotulagem , Tubulina (Proteína)/metabolismo
18.
Physiol Plant ; 157(1): 13-23, 2016 May.
Artigo em Inglês | MEDLINE | ID: mdl-26555406

RESUMO

The membranes of Zea mays (maize) mesophyll cell (MC) chloroplasts are more vulnerable to salinity stress than are those of bundle sheath cell (BSC) chloroplasts. To clarify the mechanism underlying this difference in salt sensitivity, we monitored changes in the glycerolipid and fatty acid compositions of both types of chloroplast upon exposure to salinity stress. The monogalactosyldiacylglycerol (MGDG) and digalactosyldiacylglycerol (DGDG) contents were higher in MC chloroplasts than in BSC chloroplasts, in both the presence and absence of salt treatment. Under salt conditions, the MGDG level in MC chloroplasts was significantly lower than under normal conditions, while it was unchanged in BSC chloroplasts. In both types of chloroplast, the contents of DGDG, phosphatidylglycerol and phosphatidylinositol remained at the same levels in control and salt-treated plants, whereas sulfoquinovosyldiacylglycerol and phosphatidylcholine were significantly lower and higher, respectively, upon salt treatment. In addition, the fatty acid composition and double bond index of individual lipid classes were changed by salt treatment in both BSC and MC chloroplasts, although these factors had no effect on glycerolipid content. These findings suggest that the difference in salt sensitivity of MC and BSC chloroplast membranes is related to differences in MGDG responses to salinity. Thus, we propose that the low MGDG content and the low sensitivity of MGDG to salinity in BSC chloroplasts render them more tolerant than MC chloroplasts to salinity stress.


Assuntos
Galactolipídeos/metabolismo , Glicolipídeos/metabolismo , Metabolismo dos Lipídeos/efeitos dos fármacos , Cloreto de Sódio/farmacologia , Zea mays/efeitos dos fármacos , Cloroplastos/efeitos dos fármacos , Cloroplastos/ultraestrutura , Membranas/efeitos dos fármacos , Membranas/ultraestrutura , Células do Mesofilo/efeitos dos fármacos , Células do Mesofilo/ultraestrutura , Salinidade , Estresse Fisiológico , Zea mays/metabolismo , Zea mays/ultraestrutura
19.
Physiol Plant ; 156(3): 351-66, 2016 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-26467664

RESUMO

Auxin is involved in a wide spectrum of physiological processes in plants, including responses controlled by the blue light photoreceptors phototropins: phototropic bending and stomatal movement. However, the role of auxin in phototropin-mediated chloroplast movements has never been studied. To address this question we searched for potential interactions between auxin and the chloroplast movement signaling pathway using different experimental approaches and two model plants, Arabidopsis thaliana and Nicotiana tabacum. We observed that the disturbance of auxin homeostasis by shoot decapitation caused a decrease in chloroplast movement parameters, which could be rescued by exogenous auxin application. In several cases, the impairment of polar auxin transport, by chemical inhibitors or in auxin carrier mutants, had a similar negative effect on chloroplast movements. This inhibition was not correlated with changes in auxin levels. Chloroplast relocations were also affected by the antiauxin p-chlorophenoxyisobutyric acid and mutations in genes encoding some of the elements of the SCF(TIR1)-Aux/IAA auxin receptor complex. The observed changes in chloroplast movement parameters are not prominent, which points to a modulatory role of auxin in this process. Taken together, the obtained results suggest that auxin acts indirectly to regulate chloroplast movements, presumably by regulating gene expression via the SCF(TIR1)-Aux/IAA-ARF pathway. Auxin does not seem to be involved in controlling the expression of phototropins.


Assuntos
Cloroplastos/metabolismo , Ácidos Indolacéticos/metabolismo , Citoesqueleto de Actina/efeitos dos fármacos , Citoesqueleto de Actina/metabolismo , Citoesqueleto de Actina/efeitos da radiação , Arabidopsis/efeitos dos fármacos , Arabidopsis/crescimento & desenvolvimento , Arabidopsis/efeitos da radiação , Transporte Biológico/efeitos dos fármacos , Cloroplastos/efeitos dos fármacos , Cloroplastos/efeitos da radiação , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Regulação da Expressão Gênica de Plantas/efeitos da radiação , Ácidos Indolacéticos/farmacologia , Luz , Células do Mesofilo/efeitos dos fármacos , Células do Mesofilo/metabolismo , Movimento , Mutação/genética , Fototropinas/genética , Fototropinas/metabolismo , Reguladores de Crescimento de Plantas/metabolismo , Folhas de Planta/efeitos dos fármacos , Folhas de Planta/metabolismo , Folhas de Planta/efeitos da radiação , Brotos de Planta/efeitos dos fármacos , Brotos de Planta/metabolismo , Brotos de Planta/efeitos da radiação , RNA Mensageiro/genética , RNA Mensageiro/metabolismo
20.
Ecotoxicol Environ Saf ; 133: 146-56, 2016 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-27448955

RESUMO

The auxin herbicide quinclorac is widely used for controlling weeds in transplanted and direct-seeded rice fields. However, its phytotoxic responses on rice are still unknown. Therefore, in the present investigation we studied the effects of different concentrations (0, 0.1 and 0.5g/L) of quinclorac herbicide on the physiological and biochemical changes of two rice cultivars (XS 134 and ZJ 88) and further analyzed the ameliorating role of salicylic acid (SA) on quinclorac toxicity in rice plants. The results revealed that exogenous application of SA significantly increased plant biomass and total chlorophyll contents in herbicide stressed plants. The lipid peroxidation and ROS (H2O2, O2(-.), (-)OH) production were significantly increased in roots and leaves of both rice cultivars under quinclorac stress, demonstrating an oxidative burst in rice plants. Whereas, application of SA significantly lowered ROS contents under quinclorac stress. Further, exogenous SA treatment significantly modulated antioxidant enzymes and enhanced GSH concentration in stress plants. Anatomical observations of leaf and root revealed that herbicide affected internal structures, while SA played a vital role in protection from toxic effects. Expression analysis of stress hormone ABA genes (OsABA8oxs, OsNCEDs) revealed that quinclorac application enhanced stress condition in cultivar ZJ 88, while SA treatment downregulated ABA genes more in cultivar XS 134, which correlated with the enhanced tolerance to quinclorac induced oxidative stress in this cultivar. The present study delineated that SA played a critical role under quinclorac stress in both rice cultivars by regulating antioxidant defense system, reducing ROS formation and preventing the degradation of internal cell organelles.


Assuntos
Ácido Abscísico/metabolismo , Antioxidantes/metabolismo , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Oryza/metabolismo , Quinolinas/toxicidade , Ácido Salicílico/farmacologia , Ácido Abscísico/genética , Clorofila/metabolismo , Glutationa Transferase/metabolismo , Peróxido de Hidrogênio/metabolismo , Peroxidação de Lipídeos/efeitos dos fármacos , Malondialdeído/metabolismo , Células do Mesofilo/efeitos dos fármacos , Células do Mesofilo/ultraestrutura , Oryza/crescimento & desenvolvimento , Estresse Oxidativo/efeitos dos fármacos , Estresse Oxidativo/fisiologia , Folhas de Planta/efeitos dos fármacos , Raízes de Plantas/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo
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