Effect of Plant Growth-Promoting Rhizobacteria on Salicornia ramosissima Seed Germination under Salinity, CO2 and Temperature Stress

In a scenario of climate change and growing population, halophyte root microbiota interactions may be a sustainable solution to improve alternative crop production while combating abiotic stress. In this work, seeds of the cash crop halophyte Salicornia ramosissima were inoculated with five different plant growth-promoting rhizobacteria consortia, isolated from the rhizosphere of five halophytes in southwestern Spain salt marshes. For the first time, we recorded seed germination response to three interactive abiotic stressors, CO2 (400 and 700 ppm), temperature (25 and 29 ℃) and salinity (171, 510 and 1030 mM NaCl), all of them related to climate change. Salinity played a decisive role, as no significant differences were registered between treatments at 171 mM NaCl and no germination took place at 1030 mM NaCl. At 510 mM NaCl, one rhizobacterial consortium improved seed parameters notably, increasing up to 114% germination percentage and 65% seedlings biomass. These first findings encourage us to think that cash crop halophytes like S. ramosissima and halophyte root microbiota may be valuable resources for human or animal feeding in a future climate reality.Ministerio de Economía y Competitividad (CGL2016-75550-R)Ministerio de Educación, Cultura y Deporte (FPU014/03987

Coastal Ecosystems as Sources of Biofertilizers in Agriculture: From Genomics to Application in an Urban Orchard

Pantoea agglomerans RSO7, a rhizobacterium previously isolated from Spartina maritima grown on metal polluted saltmarshes, had demonstrated good plant growth promoting activity for its host halophyte, but was never tested in crops. The aims of this study were: (1) testing PGP activity on a model plant (alfalfa) in vitro; (2) testing a bacterial consortium including RSO7 as biofertilizer in a pilot experiment in urban orchard; and (3) identifying the traits related to PGP activities. RSO7 was able to enhance alfalfa growth in vitro, particularly the root system, besides improving plant survival and protecting plants against fungal contamination. In addition, in a pilot experiment in urban orchard, a consortium of three bacteria including RSO7 was able to foster the growth and yield of several winter crops between 1.5 and 10 fold, depending on species. Moreover, the analysis of chlorophyll fluorescence revealed that photosynthesis was highly ameliorated. Genome analysis of RSO7 depicted the robustness of this bacterial strain which showed resilience to multiple stresses (heat, cold, UV radiation, several xenobiotics). Together with wide metabolic versatility, genes conferring resistance to oxidative stress were identified. Many genes involved in metal resistance (As, Cu, Ni, Co, Zn, Se, Te) and in tolerance toward high osmolality (production of a battery of osmoprotectans) were also found. Regarding plant growth promoting properties, traits for phosphate solubilization, synthesis of a battery of siderophores and production of IAA were detected. In addition, the bacterium has genes related to key processes in the rhizosphere including flagellar motility, chemotaxis, quorum sensing, biofilm formation, plant-bacteria dialog, and high competitiveness in the rhizosphere. Our results suggest the high potential of this bacterium as bioinoculant for an array of crops. However, the classification in biosecurity group 2 prevents its use according to current European regulation. Alternative formulations for the application of the bioinoculant are discussed.Junta de Andalucía P11-RNM-7274-MOFIUS, University of Seville FIUS19/0065FEDER, Junta de Andalucía US-1262036University of Seville AE2-1

Safe Cultivation of Medicago sativa in Metal-Polluted Soils from Semi-Arid Regions Assisted by Heatand Metallo-Resistant PGPR

Soil contamination with heavy metals is a constraint for plant establishment and development for which phytoremediation may be a solution, since rhizobacteria may alleviate plant stress under these conditions. A greenhouse experiment was conducted to elucidate the effect of toxic metals on growth, the activities of ROS (reactive oxygen species)-scavenging enzymes, and gene expression of Medicago sativa grown under different metal and/or inoculation treatments. The results showed that, besides reducing biomass, heavy metals negatively affected physiological parameters such as chlorophyll fluorescence and gas exchange, while increasing ROS-scavenging enzyme activities. Inoculation of M. sativa with a bacterial consortium of heat- and metallo-resistant bacteria alleviated metal stress, as deduced from the improvement of growth, lower levels of antioxidant enzymes, and increased physiological parameters. The bacteria were able to effectively colonize and form biofilms onto the roots of plants cultivated in the presence of metals, as observed by scanning electron microscopy. Results also evidenced the important role of glutathione reductase (GR), phytochelatin synthase (PCS), and metal transporter NRAMP1 genes as pathways for metal stress management, whereas the gene coding for cytochrome P450 (CP450) seemed to be regulated by the presence of the bacteria. These outcomes showed that the interaction of metal-resistant rhizobacteria/legumes can be used as an instrument to remediate metal-contaminated soils, while cultivation of inoculated legumes on these soils is still safe for animal grazing, since inoculation with bacteria diminished the concentrations of heavy metals accumulated in the aboveground parts of the plants to below toxic levelsMarruecos. Centre National pour la Recherche Scientifique et Technique (CNRST)-España, Ministerio de Economía y Competitividad (MINECO)-PPR2 /2016/42Unión Europea (FEDER)-CGL2016-75550-

Endophytic cultivable bacteria of the metal bioaccumulator Spartina maritima improve plant growth but not metal uptake in polluted marshes soils

Endophytic bacterial population was isolated from Spartina maritima tissues, a heavy metal bioaccumulator cordgrass growing in the estuaries of Tinto, Odiel, and Piedras River (south west Spain), one of the most polluted areas in the world. Strains were identified and ability to tolerate salt and heavy metals along with plant growth promoting and enzymatic properties were analyzed. A high proportion of these bacteria were resistant toward one or several heavy metals and metalloids including As, Cu, and Zn, the most abundant in plant tissues and soil. These strains also exhibited multiple enzymatic properties as amylase, cellulase, chitinase, protease and lipase, as well as plant growth promoting properties, including nitrogen fixation, phosphates solubilization, and production of indole-3-acetic acid (IAA), siderophores and 1-aminocyclopropane-1-carboxylate (ACC) deaminase. The best performing strains (Micrococcus yunnanensis SMJ12, Vibrio sagamiensis SMJ18, and Salinicola peritrichatus SMJ30) were selected and tested as a consortium by inoculating S. maritima wild plantlets in greenhouse conditions along with wild polluted soil. After 30 days, bacterial inoculation improved plant photosynthetic traits and favored intrinsic water use efficiency. However, far from stimulating plant metal uptake, endophytic inoculation lessened metal accumulation in above and belowground tissues. These results suggest that inoculation of S. maritima with indigenous metal-resistant endophytes could mean a useful approach in order to accelerate both adaption and growth of this indigenous cordgrass in polluted estuaries in restorative operations, but may not be suitable for rhizoaccumulation purposes

Impact of Plant Growth Promoting Bacteria on Salicornia ramosissima Ecophysiology and Heavy Metal Phytoremediation Capacity in Estuarine Soils

Salicornia ramosissima is a C3 halophyte that grows naturally in South Western Spain salt marshes, under soil salinity and heavy metal pollution (mostly Cu, Zn, As, and Pb) caused by both natural and anthropogenic pressure. However, very few works have reported the phytoremediation potential of S. ramosissima. In this work, we studied a microbe-assisted phytoremediation strategy under greenhouse conditions. We inoculated plant growth promoting (PGP) and heavy metal resistant bacteria in pots with S. ramosissima and natural non-polluted and polluted sediments collected from Spanish estuaries. Then, we analyzed plant ecophysiological and metal phytoaccumulation response. Our data suggested that inoculation in polluted sediments improved S. ramosissima plant growth in terms of relative growth rate (RGR) (32%) and number of new branches (61%). S. ramosissima photosynthetic fitness was affected by heavy metal presence in soil, but bacteria inoculation improved the photochemical apparatus integrity and functionality, as reflected by increments in net photosynthetic rate (21%), functionality of PSII (Fm and Fv/Fm) and electron transport rate, according to OJIP derived parameters. Beneficial effect of bacteria in polluted sediments was also observed by augmentation of intrinsic water use efficiency (28%) and slightly water content (2%) in inoculated S. ramosissima. Finally, our results demonstrated that S. ramosissima was able to accumulate great concentrations of heavy metals, mostly at root level, up to 200 mg Kg–1 arsenic, 0.50 mg Kg–1 cadmium, 400 mg Kg–1 copper, 25 mg Kg–1 nickel, 300 mg Kg–1 lead, and 300 mg Kg–1 zinc. Bioaugmentation incremented S. ramosissima heavy metal phytoremediation potential due to plant biomass increment, which enabled a greater accumulation capacity. Thus, our results suggest the potential use of heavy metal resistant PGPB to ameliorate the capacity of S. ramosissima as candidate for phytoremediation of salty polluted ecosystems.Ministerio de Economía y Competitividad (CGL2016-75550-R, AEI/FEDER, UE

Improved Medicago sativa Nodulation under Stress Assisted by Variovorax sp. Endophytes

Legumes are the recommended crops to fight against soil degradation and loss of fertility because of their known positive impacts on soils. Our interest is focused on the identification of plant-growth-promoting endophytes inhabiting nodules able to enhance legume growth in poor and/or degraded soils. The ability of Variovorax paradoxus S110T and Variovorax gossypii JM-310T to promote alfalfa growth in nutrient-poor and metal-contaminated estuarine soils was studied. Both strains behaved as nodule endophytes and improved in vitro seed germination and plant growth, as well as nodulation in co-inoculation with Ensifer medicae MA11. Variovorax ameliorated the physiological status of the plant, increased nodulation, chlorophyll and nitrogen content, and the response to stress and metal accumulation in the roots of alfalfa growing in degraded soils with moderate to high levels of contamination. The presence of plant-growth-promoting traits in Variovorax, particularly ACC deaminase activity, could be under the observed in planta effects. Although the couple V. gossypii-MA11 reported a great benefit to plant growth and nodulation, the best result was observed in plants inoculated with the combination of the three bacteria. These results suggest that Variovorax strains could be used as biofertilizers to improve the adaptation of legumes to degraded soils in soil-recovery programs.España MCIN/AEI/10.13039/501100011033España Ministry of Science and Innovation UE “NextGenerationEU/PRTR (PDC2021-120951-I00

Innovative compounds to battle multirresistance to antibiotics: use of pva-tannic acid nano- particles to inhibit staphylococcus pseudintermedius growth

Antibiotic resistance is an increasing public health problem that affects to numerous pathogens, including Staphylococcus pseudintermedius, which has a high prevalence of methicillin resistance and can be transmitted to humans. The development of new compounds to prevent the appearance of antibiotic resistances and find alternatives to classic therapies is essential to health protection. The main objective of the present study is to determine whether the nanoparticles of polyvinyl alcohol and tannic acid “PVA-TA NPs” are effective to inhibit Staphylococcus pseudintermedius and can be considered as an alternative therapy. The study includes increasing concentration essays of the nanoparticles and establishes, using the determination of turbidity by refractometry that could be a useful tool to inhibit bacterial growth, resulting successful. Nonetheless, further studies to identify the underlying action mechanisms of these nano-particles are going on in our group

Looking for new compounds to battle antibiotic resistance: optimization of organic solvents

A high number of pathogens have shown the capacity of resist to the action of antibiotics. This fact presents a very important problem to public health and highlights the necessity of looking for new substances as alternative to antibiotics. Since some of these substances have a hydrophobic nature they need to be solved in organic solvents. Some of these solvents are also toxic to bacteria, mainly affecting membranes. For these reasons it is necessary to perform a study of the toxicity of organic solvents to bacteria, which has been the aim of this work. Five organic solvents, DMF (dimethylformamide), TBME (tert-butyl methyl ether), THF (tetrahydrofuran), DMSO (dimethyl sulfoxide) and CH2Cl2 (dichloromethane), have been tested, using two of the most prevalent pathogens (Staphylococcus pseudintermedius and Pseudomonas aeruginosa,) both in animals and humans. The results show that, when possible, DMSO followed by TBME are the best options for testing new compounds in Staphylococcus pseudintermedius, whether DMF followed by TBME are the preferred solvents for testing hydrophobic compounds in Pseudomonas aeruginosa

Assessing the Biofortification of Wheat Plants by Combining a Plant Growth-Promoting Rhizobacterium (PGPR) and Polymeric Fe-Nanoparticles: Allies or Enemies?

Biofortification has been widely used to increase mineral nutrients in staple foods, such as wheat (Triticum aestivum). In this study, a new approach has been used by analyzing the effect of inoculation with a plant growth-promoting rhizobacterium (PGPR), namely, Bacillus aryabhattai RSO25 and the addition of 1% (v/v) of organometallic Fe-containing polymeric nanoparticles (FeNPs) alone and in combination. Previously, the minimal inhibitory concentration of FeNPs for the bacterium was determined in order not to inhibit bacterial growth. All treatments had minor effects on seed germination and plant survival. Considering the physiology of plants, several photosynthetic parameters were significantly improved in individual treatments with FeNPs or the bacterium, particularly the efficiency of the photosystem II and the electron transport rate, which is indicative of a better photosynthetic performance. However, at the end of the experiment, a significant effect on final plant growth was not observed in shoots or in roots. When using FeNPs alone, earlier spike outgrow was observed and the final number of spikes increased by 20%. Concerning biofortification, FeNPs increased the concentration of Fe in spikes by 35%. In fact, the total amount of Fe per plant base rose to 215% with regard to the control. Besides, several side effects, such as increased Ca and decreased Na and Zn in spikes, were observed. Furthermore, the treatment with only bacteria decreased Na and Fe accumulation in grains, indicating its inconvenience. On its side, the combined treatment led to intermediate Fe accumulation in spikes, since an antagonist effect between RSO25 and FeNPs was observed. For this reason, the combined treatment was discouraged. In conclusion, of the three treatments tested, FeNPs alone is recommended for achieving efficient Fe biofortification in wheat.Ministry of Science and Innovation of Spain grant PID2019-109371GB-I00Junta de Andalucía (Spain) Project FQM-135University of Seville, Spain PPIT-2020/0000109

Nanopartículas poliméricas de zinc y su potencial uso como agentes de biofortificación

En la actualidad el hambre y los malos hábitos alimenticios son un problema reconocido por la organización mundial de la salud (OMS). A pesar de todos los esfuerzos propuestos para solventar este problema, la OMS ha admitido que no se alcanzará el objetivo de hambre cero para 2030. Por todo esto y para aportar soluciones imaginativas a este problema, nos propusimos emplear nanopartículas poliméricas cargadas con zinc, junto con la bacteria Pantoea agglomerans, promotora de crecimiento de plantas (PGPR), para mejorar el valor nutricional de plantas como el trigo (Triticum aestivum), mediante bioforitficación. Las plantas de trigo fueron tratadas, tras su previa germinación, y se recogieron medidas de concentración de metales usando técnicas analíticas. Los resultados mostraron que los parámetros físicos y fisiológicos no se vieron afectados y se produjo una acumulación notoria del Zn en la parte aérea