RESUMEN
Cadmium (Cd2+ ) is highly harmful to plant growth. Although Cd2+ induces programmed cell death (PCD) in plant cells, Cd2+ stress in whole plants during later developmental stages and the mechanism underlying Cd2+ -mediated toxicity are poorly understood. Here, we showed that Cd2+ limits plant growth, causes intense redness in leaf vein, leaf yellowing, and chlorosis during the R1 reproductive stage of soybean (Glycine max). These symptoms were associated with Cd2+ -induced PCD, as Cd2+ -stressed soybean leaves displayed decreased number of nuclei, enhanced cell death, DNA damage, and caspase 1 activity compared to unstressed leaves. Accordingly, Cd2+ -induced NRPs, GmNAC81, GmNAC30 and VPE, the DCD/NRP-mediated cell death signalling components, which execute PCD via caspase 1-like VPE activity. Furthermore, overexpression of the positive regulator of this cell death signalling GmNAC81 enhanced sensitivity to Cd2+ stress and intensified the hallmarks of Cd2+ -mediated PCD. GmNAC81 overexpression enhanced Cd2+ -induced H2 O2 production, cell death, DNA damage, and caspase-1-like VPE expression. Conversely, BiP overexpression negatively regulated the NRPs/GmNACs/VPE signalling module, conferred tolerance to Cd2+ stress and reduced Cd2+ -mediated cell death. Collectively, our data indicate that Cd2+ induces PCD in plants via activation of the NRP/GmNAC/VPE regulatory circuit that links developmentally and stress-induced cell death.
Asunto(s)
Apoptosis , Cadmio/efectos adversos , Glycine max/efectos de los fármacos , Células Vegetales/efectos de los fármacos , Hojas de la Planta/fisiología , Células Vegetales/fisiología , Glycine max/fisiologíaRESUMEN
The primordial TOR pathway, known to control growth and cell proliferation, has still not been fully described for plants. Nevertheless, in maize, an insulin-like growth factor (ZmIGF) peptide has been reported to stimulate this pathway. This research provides further insight into the TOR pathway in maize, using a biochemical approach in cultures of fast-growing (FG) and slow-growing (SG) calli, as a model system. Our results revealed that addition of either ZmIGF or insulin to SG calli stimulated DNA synthesis and increased the growth rate through cell proliferation and increased the rate of ribosomal protein (RP) synthesis by the selective mobilization of RP mRNAs into polysomes. Furthermore, analysis of the phosphorylation status of the main TOR and S6K kinases from the TOR pathway revealed stimulation by ZmIGF or insulin, whereas rapamycin inhibited its activation. Remarkably, a putative maize insulin-like receptor was recognized by a human insulin receptor antibody, as demonstrated by immunoprecipitation from membrane protein extracts of maize callus. Furthermore, competition experiments between ZmIGF and insulin for the receptor site on maize protoplasts suggested structural recognition of the putative receptor by either effector. These data were confirmed by confocal immunolocalization within the cell membrane of callus cells. Taken together, these data indicate that cell growth and cell proliferation in maize depend on the activation of the TOR-S6K pathway through the interaction of an insulin-like growth factor and its receptor. This evidence suggests that higher plants as well as metazoans have conserved this biochemical pathway to regulate their growth, supporting the conclusion that it is a highly evolved conserved pathway.
Asunto(s)
Receptor de Insulina/metabolismo , Proteínas Quinasas S6 Ribosómicas/metabolismo , Transducción de Señal , Somatomedinas/metabolismo , Serina-Treonina Quinasas TOR/metabolismo , Regulación hacia Arriba , Zea mays/metabolismo , Unión Competitiva , Membrana Celular/metabolismo , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Activación Enzimática/efectos de los fármacos , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Insulina/metabolismo , Insulina/farmacología , Fosforilación/efectos de los fármacos , Células Vegetales/efectos de los fármacos , Células Vegetales/enzimología , Células Vegetales/metabolismo , Proteínas de Plantas/agonistas , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Polirribosomas/efectos de los fármacos , Polirribosomas/metabolismo , Procesamiento Proteico-Postraduccional/efectos de los fármacos , Transporte de Proteínas/efectos de los fármacos , Receptor de Insulina/agonistas , Transducción de Señal/efectos de los fármacos , Regulación hacia Arriba/efectos de los fármacos , Zea mays/citología , Zea mays/crecimiento & desarrolloRESUMEN
Species of the genus Psychotria are used for multiple purposes in Brazilian folk medicine, either as water infusions, baths or poultices. This study was aimed to evaluate the genotoxic and antiproliferative effects of infusions of Psychotria brachypoda and P. birotula on the Allium cepa test. Exposure to distilled water was used as a negative control, while exposure to glyphosate was used as a positive control. The interaction of extracts (as a post-treatment) with the effects of glyphosate was also studied. Results showed that glyphosate and the extracts of both P. brachypoda and P. birotula reduced the mitotic index as compared with the negative control (distilled water). Surprisingly, however, both extracts from P. brachypoda and P. birotula caused a partial reversal of the antiproliferative effect of glyphosate when used as a post-treatment. Glyphosate also induced the highest number of cells with chromosomal alterations, which was followed by that of P. birotula extracts. However, the extracts from P. brachypoda did not show any significant genotoxic effect. Post-treatment of glyphosate-treated samples with distilled water allowed a partial recovery of the genotoxic effect of glyphosate, and some of the Psychotria extracts also did so. Notably, post-treatment of glyphosate-treated samples with P. brachypoda extracts induced a statistically significant apoptotic effect. It is concluded that P. brachypoda extracts show antiproliferative effects and are not genotoxic, while extracts of P. birotula show a less potent antiproliferative effect and may induce chromosomal abnormalities. The finding of a partial reversion of the effects of glyphosate by a post-treatment with extracts from both plants should be followed up.
Asunto(s)
Proliferación Celular/efectos de los fármacos , Glicina/análogos & derivados , Medicina Tradicional/métodos , Cebollas/citología , Células Vegetales/efectos de los fármacos , Extractos Vegetales/farmacología , Brasil , Interacciones Farmacológicas , Inhibidores Enzimáticos/farmacología , Inhibidores Enzimáticos/toxicidad , Glicina/farmacología , Glicina/toxicidad , Extractos Vegetales/toxicidad , GlifosatoRESUMEN
Species of the genus Psychotria are used for multiple purposes in Brazilian folk medicine, either as water infusions, baths or poultices. This study was aimed to evaluate the genotoxic and antiproliferative effects of infusions of Psychotria brachypoda and P. birotula on the Allium cepa test. Exposure to distilled water was used as a negative control, while exposure to glyphosate was used as a positive control. The interaction of extracts (as a post-treatment) with the effects of glyphosate was also studied. Results showed that glyphosate and the extracts of both P. brachypoda and P. birotula reduced the mitotic index as compared with the negative control (distilled water). Surprisingly, however, both extracts from P. brachypoda and P. birotula caused a partial reversal of the antiproliferative effect of glyphosate when used as a post-treatment. Glyphosate also induced the highest number of cells with chromosomal alterations, which was followed by that of P. birotula extracts. However, the extracts from P. brachypoda did not show any significant genotoxic effect. Post-treatment of glyphosate-treated samples with distilled water allowed a partial recovery of the genotoxic effect of glyphosate, and some of the Psychotria extracts also did so. Notably, post-treatment of glyphosate-treated samples with P. brachypoda extracts induced a statistically significant apoptotic effect. It is concluded that P. brachypoda extracts show antiproliferative effects and are not genotoxic, while extracts of P. birotula show a less potent antiproliferative effect and may induce chromosomal abnormalities. The finding of a partial reversion of the effects of glyphosate by a post-treatment with extracts from both plants should be followed up.
Asunto(s)
Proliferación Celular/efectos de los fármacos , Glicina/análogos & derivados , Medicina Tradicional/métodos , Cebollas/citología , Células Vegetales/efectos de los fármacos , Extractos Vegetales/farmacología , Brasil , Interacciones Farmacológicas , Inhibidores Enzimáticos/farmacología , Inhibidores Enzimáticos/toxicidad , Glicina/farmacología , Glicina/toxicidad , Extractos Vegetales/toxicidadRESUMEN
The toxicity of metronidazole (MTZ) in meristematic and elongation zones of Allium cepa roots was analyzed for 30 h of exposition. Toxic effects were evaluated by lipid peroxidation (content of thiobarbituric-reactive substances [TBARS]), reduced glutathione (GSH) levels, ascorbate acid and dehydroascorbate acid content, and enzymatic activities of superoxide dismutase and catalase. The root zones showed differentiated susceptibility to MTZ. In the elongation zone, MTZ induced an increase of TBARS content and a significant rise in GSH levels, whereas in the meristematic zone, lipid peroxidation was not observed and all antioxidant defense parameters analyzed were significantly increased. These results indicate that MTZ exposure induced oxidative stress in A. cepa roots, and that the antioxidant defenses in the meristematic zone are more efficient compared with the elongation zone, which is probably related to higher oxidative metabolism of meristematic tissue.