RESUMEN
Arabidopsis thaliana glutamate receptor-like (GLR) channels are amino acid-gated ion channels involved in physiological processes including wound signaling, stomatal regulation, and pollen tube growth. Here, fluorescence microscopy and genetics were used to confirm the central role of GLR3.3 in the amino acid-elicited cytosolic Ca2+ increase in Arabidopsis seedling roots. To elucidate the binding properties of the receptor, we biochemically reconstituted the GLR3.3 ligand-binding domain (LBD) and analyzed its selectivity profile; our binding experiments revealed the LBD preference for l-Glu but also for sulfur-containing amino acids. Furthermore, we solved the crystal structures of the GLR3.3 LBD in complex with 4 different amino acid ligands, providing a rationale for how the LBD binding site evolved to accommodate diverse amino acids, thus laying the grounds for rational mutagenesis. Last, we inspected the structures of LBDs from nonplant species and generated homology models for other GLR isoforms. Our results establish that GLR3.3 is a receptor endowed with a unique amino acid ligand profile and provide a structural framework for engineering this and other GLR isoforms to investigate their physiology.
Asunto(s)
Aminoácidos/metabolismo , Proteínas de Arabidopsis/ultraestructura , Arabidopsis/metabolismo , Dominios Proteicos/genética , Receptores de Glutamato/ultraestructura , Arabidopsis/genética , Proteínas de Arabidopsis/agonistas , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Sitios de Unión/genética , Calcio/metabolismo , Cristalografía por Rayos X , Citosol/metabolismo , Ligandos , Mutación , Raíces de Plantas/metabolismo , Plantas Modificadas Genéticamente , Receptores de Glutamato/genética , Receptores de Glutamato/metabolismo , Plantones/metabolismo , Relación Estructura-ActividadRESUMEN
The aim of the study was to characterize halotolerant bacteria and to evaluate their plant growth promotion potential on chia and quinoa seedlings under saline stress. Isolated microorganisms were evaluated for nitrogen fixation, phosphate solubilization, and production of siderophores and indole acetic acid. Three strains and two consortia were selected: Halomonas sp. (SFS), Micrococcus luteus (SA211), Bacillus sp. (HX11), C1 (SA211 + SFS), and C2 (SA211 + HX11). In vitro assays using water agar and half-strength Murashige-Skoog plates showed that an increase in salinity led to an increased seedlings mortality and a decrease in germination (lower than 40%), in total length (varying between 16% and 87% decreases), root length (from 60% to 92% lesser length) and dry weight (from 7% to 86% lower weight). Also, the relative growth index (RGI) decreased for both crops in most treatments, except those with HX11 and C2. These treatments had the highest growth parameters and RGI values in presence of high salinity in chia (50 and 100 mmol/L NaCl) and quinoa (200 and 400 mmol/L NaCl). SA211, the highest producer of indole acetic acid, showed a detrimental effect and anomalous phenotype on plants. Our results suggest that Bacillus sp. HX11, with multiple plant growth promotion traits and tolerance to saline stress, has a great potential as a bioinoculant in saline conditions and could be used as a biofertilizer for crop production.
RESUMEN
Environmental limitations influence food production and distribution, adding up to global problems like world hunger. Conditions caused by climate change require global efforts to be improved, but others like soil degradation demand local management. For many years, saline soils were not a problem; indeed, natural salinity shaped different biomes around the world. However, overall saline soils present adverse conditions for plant growth, which then translate into limitations for agriculture. Shortage on the surface of productive land, either due to depletion of arable land or to soil degradation, represents a threat to the growing worldwide population. Hence, the need to use degraded land leads scientists to think of recovery alternatives. In the case of salt-affected soils (naturally occurring or human-made), which are traditionally washed or amended with calcium salts, bio-reclamation via microbiome presents itself as an innovative and environmentally friendly option. Due to their low pathogenicity, endurance to adverse environmental conditions, and production of a wide variety of secondary metabolic compounds, members of the genus Streptomyces are good candidates for bio-reclamation of salt-affected soils. Thus, plant growth promotion and soil bioremediation strategies combine to overcome biotic and abiotic stressors, providing green management options for agriculture in the near future.
RESUMEN
Although the use of agrochemicals allowed increasing the crops productivity, in many cases led to soil deterioration. In this study, eight composite samples from different soils of two locations (San Martín and Anta) in Salta, Argentina, were collected and analyzed. All the samples were from loamy Entisols (0-20â¯cm depth) under reduced tillage without and with direct spray application of glyphosate. Twenty six variables were determined (physical, chemical, and biological soil quality indicators). From them, those of higher specificity and sensitivity to changes following glyphosate application were identified by a stepwise reduction of variables aided by statistical analysis. Samples were grouped regarding location and application of glyphosate, to identify differential effects upon variables, and glyphosate sensitive variables were selected by discarding those influenced by other factors. Thence, they were used to compose a first approximation to a combined soil quality indicator (CSQI) to assess the effect of glyphosate use in agriculture upon the soil. Overall, the set of physical variables showed the same discriminating structure as the biological set. Finally, two biological, two chemical, and two physical indicators resulted as the most specific to quality variations by the application of the herbicide, being the most sensitive the microbial biomass carbon and the (Aminomethyl)phosphonic acid concentration in soil. When these two were considered into a CSQI, it was possible to discriminate samples with the application of glyphosate (lower quality) from those without application (higher quality). To the best of our knowledge, this is the first attempt to propose a CSQI that could play an important role to prevent degradation in soils subjected to glyphosate application, as it could aid in the early detection of soil quality loss. This would provide to land managers a decision tool to let the land rest from glyphosate application, to ensure sustainable practices in agriculture.
Asunto(s)
Monitoreo del Ambiente/métodos , Glicina/análogos & derivados , Herbicidas/análisis , Contaminantes del Suelo/análisis , Glicina/análisis , Suelo/química , GlifosatoRESUMEN
Calcium imaging in plants requires a high-resolution microscope, able to perform volumetric acquisition in a few seconds, inducing as low photobleaching and phototoxicity as possible to the sample. Light sheet fluorescence microscopy offers these capabilities, with the further chance to mount the sample in vertical position, mimicking the plant's growth and physiological conditions.A protocol for plant preparation and mounting in a light sheet microscope is presented. First, the growth of Arabidopsis thaliana in a sample holder compatible with light sheet microscopy is described. Then, the requirements for sample alignment and image acquisition are detailed. Finally, the image processing steps to analyze calcium oscillations are discussed, with particular emphasis on ratiometric calcium imaging in Arabidopsis root hairs.