RESUMO
Plant survival requires an ability to adapt to differing concentrations of nutrient and toxic soil ions, yet ion sensors and associated signaling pathways are mostly unknown. Aluminum (Al) ions are highly phytotoxic, and cause severe crop yield loss and forest decline on acidic soils which represent â¼30% of land areas worldwide. Here we found an Arabidopsis mutant hypersensitive to Al. The gene encoding a leucine-rich-repeat receptor-like kinase, was named Al Resistance1 (ALR1). Al ions binding to ALR1 cytoplasmic domain recruits BAK1 co-receptor kinase and promotes ALR1-dependent phosphorylation of the NADPH oxidase RbohD, thereby enhancing reactive oxygen species (ROS) generation. ROS in turn oxidatively modify the RAE1 F-box protein to inhibit RAE1-dependent proteolysis of the central regulator STOP1, thus activating organic acid anion secretion to detoxify Al. These findings establish ALR1 as an Al ion receptor that confers resistance through an integrated Al-triggered signaling pathway, providing novel insights into ion-sensing mechanisms in living organisms, and enabling future molecular breeding of acid-soil-tolerant crops and trees, with huge potential for enhancing both global food security and forest restoration.
Assuntos
Proteínas de Arabidopsis , Arabidopsis , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Alumínio/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Arabidopsis/genética , Arabidopsis/metabolismo , Íons , Solo , Regulação da Expressão Gênica de Plantas , Fatores de Transcrição/metabolismoRESUMO
Glucose (Glu) is involved in not only plant physiological and developmental events but also plant responses to abiotic stresses. Here, we found that the exogenous Glu improved root and shoot growth, reduced shoot cadmium (Cd) concentration, and rescued Cd-induced chlorosis in Arabidopsis thaliana (Columbia ecotype, Col-0) under Cd stressed conditions. Glucose increased Cd retained in the roots, thus reducing its translocation from root to shoot significantly. The most Cd retained in the roots was found in the hemicellulose 1. Glucose combined with Cd (Glu + Cd) treatment did not affect the content of pectin and its binding capacity of Cd while it increased the content of hemicelluloses 1 and the amount of Cd retained in it significantly. Furthermore, Leadmium Green staining indicated that more Cd was compartmented into vacuoles in Glu + Cd treatment compared with Cd treatment alone, which was in accordance with the significant upregulation of the expression of tonoplast-localized metal transporter genes, suggesting that compartmentation of Cd into vacuoles also contributes to the Glu-alleviated Cd toxicity. Taken together, we demonstrated that Glu-alleviated Cd toxicity is mediated through increasing Cd fixation in the root cell wall and sequestration into the vacuoles.
Assuntos
Arabidopsis/efeitos dos fármacos , Arabidopsis/metabolismo , Cádmio/metabolismo , Cádmio/toxicidade , Parede Celular/efeitos dos fármacos , Glucose/farmacologia , Vacúolos/metabolismo , Parede Celular/metabolismo , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/metabolismo , Vacúolos/efeitos dos fármacosRESUMO
Xyloglucan (XyG) has been reported to contribute to the aluminum (Al)-binding capacity of the cell wall in Arabidopsis (Arabidopsis thaliana). However, the influence of O-acetylation of XyG, accomplished by the putative O-acetyltransferase TRICHOME BIREFRINGENCE-LIKE27 (TBL27 [AXY4]), on its Al-binding capacity is not known. In this study, we found that the two corresponding TBL27 mutants, axy4-1 and axy4-3, were more Al sensitive than wild-type Columbia-0 plants. TBL27 was expressed in roots as well as in leaves, stems, flowers, and siliques. Upon Al treatment, even within 30 min, TBL27 transcript accumulation was strongly down-regulated. The mutants axy4-1 and axy4-3 accumulated significantly more Al in the root and wall, which could not be correlated with pectin content or pectin methylesterase activity, as no difference in the mutants was observed compared with the wild type when exposed to Al stress. The increased Al accumulation in the wall of the mutants was found to be in the hemicellulose fraction. While the total sugar content of the hemicellulose fraction did not change, the O-acetylation level of XyG was reduced by Al treatment. Taken together, we conclude that modulation of the O-acetylation level of XyG influences the Al sensitivity in Arabidopsis by affecting the Al-binding capacity in the hemicellulose.
Assuntos
Alumínio/metabolismo , Proteínas de Arabidopsis/metabolismo , Arabidopsis/enzimologia , Glucanos/metabolismo , Resolvases de Junção Holliday/metabolismo , Polissacarídeos/metabolismo , Xilanos/metabolismo , Acetilação , Proteínas de Arabidopsis/genética , Parede Celular/metabolismo , Resolvases de Junção Holliday/genética , Raízes de Plantas/metabolismoRESUMO
Previously, we reported that although the Arabidopsis (Arabidopsis thaliana) Xyloglucan Endotransglucosylase-Hydrolase31 (XTH31) has predominately xyloglucan endohydrolase activity in vitro, loss of XTH31 results in remarkably reduced in vivo xyloglucan endotransglucosylase (XET) action and enhanced Al resistance. Here, we report that XTH17, predicted to have XET activity, binds XTH31 in yeast (Saccharomyces cerevisiae) two-hybrid and coimmunoprecipitations assays and that this interaction may be required for XTH17 XET activity in planta. XTH17 and XTH31 may be colocalized in plant cells because tagged XTH17 fusion proteins, like XTH31 fusion proteins, appear to target to the plasma membrane. XTH17 expression, like that of XTH31, was substantially reduced in the presence of aluminum (Al), even at concentrations as low as 10 µm for 24 h or 25 µm for just 30 min. Agrobacterium tumefaciens-mediated transfer DNA insertion mutant of XTH17, xth17, showed low XET action and had moderately shorter roots than the wild type but was more Al resistant than the wild type. Similar to xth31, xth17 had low hemicellulose content and retained less Al in the cell wall. These data suggest a model whereby XTH17 and XTH31 may exist as a dimer at the plasma membrane to confer in vivo XET action, which modulates cell wall Al-binding capacity and thereby affects Al sensitivity in Arabidopsis.
RESUMO
Auxin is involved in not only plant physiological and developmental processes but also plant responses to abiotic stresses. In this study, cadmium (Cd(2+)) stress decreased the endogenous auxin level, whereas exogenous auxin (α-naphthaleneacetic acid, NAA, a permeable auxin analog) reduced shoot Cd(2+) concentration and rescued Cd(2+)-induced chlorosis in Arabidopsis thaliana. Under Cd(2+) stress conditions, NAA increased Cd(2+) retention in the roots and most Cd(2+) in the roots was fixed in hemicellulose 1 of the cell wall. NAA treatment did not affect pectin content and its binding capacity for Cd(2+), whereas it significantly increased the content of hemicellulose 1 and the amount of Cd(2+) retained in it. There were highly significant correlations between Cd(2+) concentrations in the root, cell wall and hemicellulose 1 when the plants were subjected to Cd(2+) or NAA+Cd(2+) treatment for 1 to 7d, suggesting that the increase in hemicellulose 1 contributes greatly to the fixation of Cd(2+) in the cell wall. Taken together, these results demonstrate that auxin-induced alleviation of Cd(2+) toxicity in Arabidopsis is mediated through increasing hemicellulose 1 content and Cd(2+) fixation in the root, thus reducing the translocation of Cd(2+) from roots to shoots.
Assuntos
Arabidopsis/efeitos dos fármacos , Cádmio/análise , Ácidos Indolacéticos/química , Raízes de Plantas/efeitos dos fármacos , Brotos de Planta/efeitos dos fármacos , Polissacarídeos/química , Arabidopsis/metabolismo , Cádmio/química , Cádmio/toxicidade , Parede Celular/efeitos dos fármacos , Regulação da Expressão Gênica de Plantas , Raízes de Plantas/metabolismo , Brotos de Planta/metabolismo , Estresse Fisiológico , Ácidos Urônicos/químicaRESUMO
Whether aluminum toxicity is an apoplastic or symplastic phenomenon is still a matter of debate. Here, we found that three auxin overproducing mutants, yucca, the recessive mutant superroot2, and superroot1 had increased aluminum sensitivity, while a transfer DNA insertion mutant, xyloglucan endotransglucosylase/hydrolases15 (xth15), showed enhanced aluminum resistance, accompanied by low endogenous indole-3-acetic acid levels, implying that auxin may be involved in plant responses to aluminum stress. We used yucca and xth15 mutants for further study. The two mutants accumulated similar total aluminum in roots and had significantly reduced cell wall aluminum and increased symplastic aluminum content relative to the wild-type ecotype Columbia, indicating that altered aluminum levels in the symplast or cell wall cannot fully explain the differential aluminum resistance of these two mutants. The expression of Al sensitive1 (ALS1), a gene that functions in aluminum redistribution between the cytoplasm and vacuole and contributes to symplastic aluminum detoxification, was less abundant in yucca and more abundant in xth15 than the wild type, consistent with possible ALS1 function conferring altered aluminum sensitivity in the two mutants. Consistent with the idea that xth15 can tolerate more symplastic aluminum because of possible ALS1 targeting to the vacuole, morin staining of yucca root tip sections showed more aluminum accumulation in the cytosol than in the wild type, and xth15 showed reduced morin staining of cytosolic aluminum, even though yucca and xth15 had similar overall symplastic aluminum content. Exogenous application of an active auxin analog, naphthylacetic acid, to the wild type mimicked the aluminum sensitivity and distribution phenotypes of yucca, verifying that auxin may regulate aluminum distribution in cells. Together, these data demonstrate that auxin negatively regulates aluminum tolerance through altering ALS1 expression and aluminum distribution within plant cells, and plants must coordinate exclusion and internal detoxification to reduce aluminum toxicity effectively.
Assuntos
Transportadores de Cassetes de Ligação de ATP/metabolismo , Alumínio/farmacocinética , Proteínas de Arabidopsis/metabolismo , Arabidopsis/efeitos dos fármacos , Arabidopsis/metabolismo , Inativação Metabólica , Ácidos Indolacéticos/metabolismo , Transportadores de Cassetes de Ligação de ATP/genética , Alumínio/toxicidade , Arabidopsis/fisiologia , Proteínas de Arabidopsis/genética , Citosol/efeitos dos fármacos , Citosol/metabolismo , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Inativação Metabólica/genética , Meristema/efeitos dos fármacos , Meristema/metabolismo , Mutação , Ácidos Naftalenoacéticos/farmacologia , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/genética , Raízes de Plantas/crescimento & desenvolvimentoRESUMO
Here we report the function of a general regulatory factor, GENERAL REGULATORY FACTOR11 (GRF11), in terms of the iron (Fe) deficiency response. Physiological and molecular responses of the loss-of-function Arabidopsis thaliana grf11 mutant to Fe supply were investigated. Genes involved in posttranscriptional regulation of FER-LIKE FE DEFICIENCY-INDUCED TRANSCRIPTION FACTOR (FIT) were also analyzed. In addition, the molecular link between the signaling molecule nitric oxide (NO) and Fe deficiency responses was further dissected. Our results suggest that GRF11 is necessary for induction of Fe-deficiency-tolerance mechanisms. The FIT protein can bind to the promoter of GRF11, which contains an E-box motif. GRF11 also positively affects FIT transcription but has no influence on the genes involved in posttranscriptional regulation of FIT. Furthermore, NO positively regulates GRF11 induction upon the onset of Fe deficiency. We propose that, upon the onset of Fe deficiency, induction of FIT expression is dependent on GRF11, which acts downstream of NO to mediate Fe deficiency responses.
Assuntos
Proteínas 14-3-3/fisiologia , Proteínas de Arabidopsis/fisiologia , Arabidopsis/metabolismo , Ferro/metabolismo , Óxido Nítrico/metabolismo , Proteínas 14-3-3/genética , Proteínas 14-3-3/metabolismo , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Fatores de Transcrição Hélice-Alça-Hélice Básicos/metabolismo , Proteínas de Transporte de Cátions/metabolismo , FMN Redutase/metabolismo , Regulação da Expressão Gênica de Plantas , Técnicas de Inativação de Genes , Modelos Biológicos , Mutagênese Insercional , Nitrato Redutase/genética , Óxido Nítrico Sintase/genética , Fenótipo , Plantas Geneticamente Modificadas/metabolismo , ATPases Translocadoras de Prótons/metabolismoRESUMO
Xyloglucan endohydrolase (XEH) and xyloglucan endotransglucosylase (XET) activities, encoded by xyloglucan endotransglucosylase-hydrolase (XTH) genes, are involved in cell wall extension by cutting or cutting and rejoining xyloglucan chains, respectively. However, the physiological significance of this biochemical activity remains incompletely understood. Here, we find that an XTH31 T-DNA insertion mutant, xth31, is more Al resistant than the wild type. XTH31 is bound to the plasma membrane and the encoding gene is expressed in the root elongation zone and in nascent leaves, suggesting a role in cell expansion. XTH31 transcript accumulation is strongly downregulated by Al treatment. XTH31 expression in yeast yields a protein with an in vitro XEH:XET activity ratio of >5000:1. xth31 accumulates significantly less Al in the root apex and cell wall, shows remarkably lower in vivo XET action and extractable XET activity, has a lower xyloglucan content, and exhibits slower elongation. An exogenous supply of xyloglucan significantly ameliorates Al toxicity by reducing Al accumulation in the roots, owing to the formation of an Al-xyloglucan complex in the medium, as verified by an obvious change in chemical shift of (27)Al-NMR. Taken together, the data indicate that XTH31 affects Al sensitivity by modulating cell wall xyloglucan content and Al binding capacity.
Assuntos
Alumínio/toxicidade , Proteínas de Arabidopsis/metabolismo , Arabidopsis/enzimologia , Regulação Enzimológica da Expressão Gênica , Glucanos/metabolismo , Xilanos/metabolismo , Sequência de Aminoácidos , Arabidopsis/química , Arabidopsis/efeitos dos fármacos , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Parede Celular/metabolismo , Quelantes/análise , Quelantes/metabolismo , Regulação para Baixo , Regulação da Expressão Gênica de Plantas , Glucanos/análise , Glicosiltransferases/genética , Glicosiltransferases/metabolismo , Mutagênese Insercional , Especificidade de Órgãos , Fenótipo , Filogenia , Folhas de Planta/química , Folhas de Planta/efeitos dos fármacos , Folhas de Planta/enzimologia , Folhas de Planta/genética , Raízes de Plantas/química , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/enzimologia , Raízes de Plantas/genética , Plantas Geneticamente Modificadas , Polissacarídeos/análise , Polissacarídeos/metabolismo , Proteínas Recombinantes de Fusão , Plântula/química , Plântula/efeitos dos fármacos , Plântula/enzimologia , Plântula/genética , Análise de Sequência de DNA , Xilanos/análiseRESUMO
Gibberellic acid (GA) is involved in not only plant growth and development but also plant responses to abiotic stresses. Here it was found that treating the plants with GA concentrations from 0.1 to 5 µM for 24 h had no obvious effect on root elongation in the absence of cadmium (Cd), whereas in the presence of Cd2+, GA at 5 µM improved root growth, reduced Cd content and lipid peroxidation in the roots, indicating that GA can partially alleviate Cd toxicity. Cd2+ increased nitric oxide (NO) accumulation in the roots, but GA remarkably reduced it, and suppressed the up-regulation of the expression of IRT1. In contrary, the beneficial effect of GA on alleviating Cd toxicity was not observed in an IRT1 knock-out mutant irt1, suggesting the involvement of IRT1 in Cd2+ absorption. Furthermore, the GA-induced reduction of NO and Cd content can also be partially reversed by the application of a NO donor (S-nitrosoglutathione [GSNO]). Taken all these together, the results showed that GA-alleviated Cd toxicity is mediated through the reduction of the Cd-dependent NO accumulation and expression of Cd2+ uptake related gene-IRT1 in Arabidopsis.
Assuntos
Arabidopsis/efeitos dos fármacos , Cádmio/toxicidade , Poluentes Ambientais/toxicidade , Giberelinas/farmacologia , Reguladores de Crescimento de Plantas/farmacologia , Substâncias Protetoras/farmacologia , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Transporte/genética , Peroxidação de Lipídeos/efeitos dos fármacos , Óxido Nítrico/metabolismo , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/metabolismoRESUMO
The cell wall (CW) has been recognized as the major target of aluminum (Al) toxicity. However, the components responsible for Al accumulation and the mechanisms of Al-induced CW function disruption are still elusive. The contribution of different CW components (pectin, hemicellulose 1 [HC1], and HC2) to adsorb Al and the effect of Al on xyloglucan endotransglucosylase/hydrolyase activity were investigated in Arabidopsis (Arabidopsis thaliana) in this study. A fractionation procedure was optimized to effectively extract different CW components, especially to prevent the HC fraction from pectin contamination. When CW materials extracted from Al-treated roots (50 µm Al for 24 h) were fractionated, about 75% of CW Al accumulated in the HC1 fraction. A time-dependent kinetic study showed that only when the HC1 fraction was removed was the amount of Al adsorbed decreased sharply. In vivo localization of xyloglucan endotransglucosylase (XET) activity showed that Al greatly inhibited this enzyme activity within 30 min of exposure, which was concomitant with Al-induced callose deposition in roots. Results from real-time reverse transcription-polymerase chain reaction indicated that three genes may constitute the major contributors to XET activity and that the inhibition of XET activity by Al is caused by transcriptional regulation. These results, to our knowledge for the first time, demonstrate that HC is the major pool for Al accumulation. Furthermore, Al-induced reduction in XET activity could play an important role in Al-induced root growth inhibition.
Assuntos
Alumínio/metabolismo , Arabidopsis/metabolismo , Parede Celular/química , Raízes de Plantas/crescimento & desenvolvimento , Polissacarídeos/metabolismo , Adsorção , Arabidopsis/enzimologia , Arabidopsis/genética , Regulação da Expressão Gênica de Plantas , Glicosiltransferases/metabolismo , Raízes de Plantas/metabolismo , RNA de Plantas/genéticaRESUMO
It is important to research the rules about accumulation and distribution of arsenic and cadmium by tea plants, which will give us some scientific ideas about how to control the contents of arsenic and cadmium in tea. In this study, by field investigation and pot trial, we found that mobility of arsenic and cadmium in tea plants was low. Most arsenic and cadmium absorbed were fixed in feeding roots and only small amount was transported to the above-ground parts. Distribution of arsenic and cadmium, based on their concentrations of unit dry matter, in tea plants grown on un-contaminated soil was in the order: feeding roots>stems approximately main roots>old leaves>young leaves. When tea plants were grown on polluted soils simulated by adding salts of these two metals, feeding roots possibly acted as a buffer and defense, and arsenic and cadmium were transported less to the above-ground parts. The concentration of cadmium in soil significantly and negatively correlated with chlorophyll content, photosynthetic rate, transpiration rate and biomass production of tea plants.