The present study reports a screening for PGPR in a highly selective environment, the rhizosphere of rice plants, in southwestern of Spain. Among the 900 isolates, only 38% were positive for at least ...one of the biochemical activities to detect putative PGPR. The best 80 isolates were selected and identified by 16S rRNA partial sequencing. Among these, 13 strains were selected for growth promotion assays. Only one strain (BaC1-38) was able to significantly increase height, while nine strains significantly inhibited it. Five strains significantly increased dry weight, and only BaC1-21 significantly decreased it. Based on significant modifications in growth, three bacteria (BaC1-13, BaC1-21 and BaC1-38) were tested for systemic induction of resistance against stress challenge (salt and Xanthomonas campestris infection). Protection against salt stress and pathogen infection was similar; BaC1-38 protected by 80%, BaC1-13 by 50% and BaC1-21 only by 20%. Toxicity of salt stress to the plants was evaluated by photosynthetic efficiency of seedlings. Fv/Fm only decreased significantly in plants inoculated with BaC1-13. ΦPSII also decreased significantly in plants inoculated with BaC1-21, but increased significantly with BaC1-38. NPQ decreased significantly in plants inoculated with BaC1-21. The two strains able to induce systemic resistance against Xanthomonas campestris seem to work by different pathways. BaC1-13 primed enzymes related with the detoxification of reactive oxygen species (ROS). However, BaC1-38 primed pathogenesis-related proteins (PRs), and this pathway was more effective, both improved chlorophyll index confirming the priming state of the plant.
•Fluorescence emission of PSII is a very sensitive method to determine the ability of PGPR to induce resistance in the plants.•BaC1-13 induces biotic resistance primed enzymes related with the detoxification of ROS.•BaC1-38 induces biotic resistance primed PRs, and this pathway is more effective.•Chlorophyll index could be used as a rapid method for evaluating in primed plants pathogen attack resistance.•The rhizosphere is an ideal ecosystem to find highly specialized bacteria.
A screening for Plant Growth Promoting Rhizobacteria (PGPR) was carried out in the rhizosphere of wild populations of Nicotiana glauca Graham in south-eastern Spain. Nine hundred and sixty strains ...were isolated and grouped in four parataxonomic groups: Gram positive endospore forming bacilli, Gram positive non-endospore forming bacilli, Gram negative bacilli and others. Two groups were selected to continue the study: Gram negative bacilli since it was the most abundant, and Gram positive sporulated bacilli, seeking their sporulating capacity as an advantage for inoculants formulation. The ability of these to release siderophores and chitinases in vitro was evaluated. Ninety six isolates were siderophore producers, and 56 of them were also able to produce chitinases. Fifty percent of these were tested for growth promotion in tomato. The best results were obtained with 5 Gram negative bacilli and one Gram positive sporulated bacilli; 5 strains increased all growth parameters while one of them, N21.4, severely compromised plant growth. The ability of these 6 strains to induce systemic resistance against the leaf pathogen Xanthomonas campestris in tomato was evaluated. Five of them effectively reduced disease symptoms (up to 50%). The six strains were identified by 16s rDNA sequencing resulting in 3 Pseudomonas, 1 Bacillus and 2 Stenotrophomonas; it's striking that 2 Pseudomonas protected up to 50% while the other increased disease incidence. This indicates that systemic induction is strain specific and not necessarily related to production of siderophores and chitinases.
Nine plant growth-promoting rhizobacteria from different backgrounds were assayed on Glycine max var. Osumi to evaluate their potential as biotic elicitors to increase isoflavone (IF) levels. Strains ...were inoculated on 2 day old pregerminated seeds. Six days after inoculation, the seedlings were harvested. Biometric parameters were registered, and IFs were determined. Although only one strain (N21.4) increased total IF contents and only one (M84) caused significant decreases in total IF, five different behaviors were detected when the daidzein and genistein families were analyzed separately. All strains triggered IF metabolism so further studies have to be developed since the different beneficial effects of IF through the diet may be due to the different IF profiles. These are encouraging results from two points of view: (1) N21.4 increases IF in seedlings, and (2) all other beneficial strains trigger IF metabolism differentially; hence, both facts could be used to prepare food supplements or as enriched standardized foods after full development of the biotechnological procedure.
Objective To assess organ damage, with emphasis on the cardiovascular system, over the different stages of the disease in a large SLE cohort. Methods Multicentre, longitudinal study of a cohort of ...4219 patients with SLE enrolled in the Spanish Society of Rheumatology Lupus Registry. Organ damage was ascertained using the Systemic Lupus International Collaborating Clinics/American College of Rheumatology Damage Index (SDI). We longitudinally analysed SDI (globally and for each domain) over time only in the 1274 patients whose dates of damage events had been recorded. Results During the first year after diagnosis of SLE, 20% of the 1274 patients presented with new damage manifestations. At years 2 and 3, new damage was recorded in 11% and 9% of patients. The annual percentage of patients with new damage after year 5 decreased to 5%. In the first year with the disease, most damage was accumulated in the musculoskeletal, neuropsychiatric and renal systems; in later stages, most damage was in the musculoskeletal, ocular and cardiovascular systems. Considering ‘cerebrovascular accident’ and ‘claudication for 6 months’ as cardiovascular items, the cardiovascular system was the second most affected system during the early stages of SLE, with 19% of the patients who presented with damage affected at first year after diagnosis. During the late stages, 20–25% of the patients presenting with new damage did so in this modified cardiovascular domain of the SDI. Conclusions New damage occurs mainly during the first year following diagnosis of SLE. Cardiovascular damage is relevant in both the early and the late stages of the disease. Strategies to prevent cardiovascular damage should be implemented early after diagnosis of SLE.
The aim of this study is to determine the involvement of the flavonol-anthocyanin pathway on plant adaptation to biotic stress using the B.amyloliquefaciens QV15 to trigger blackberry metabolism and ...identify target genes to improve plant fitness and fruit quality. To achieve this goal, field-grown blackberries were root-inoculated with QV15 along its growth cycle. At fruiting, a transcriptomic analysis by RNA-Seq was performed on leaves and fruits of treated and non-treated field-grown blackberries after a sustained mildew outbreak; expression of the regulating and core genes of the Flavonol-Anthocyanin pathway were analysed by qPCR and metabolomic profiles by UHPLC/ESI-qTOF-MS; plant protection was found to be up to 88%. Overexpression of step-controlling genes in leaves and fruits, associated to lower concentration of flavonols and anthocyanins in QV15-treated plants, together with a higher protection suggest a phytoanticipin role for flavonols in blackberry; kempferol-3-O-rutinoside concentration was strikingly high. Overexpression of RuF3H (Flavonol-3-hidroxylase) suggests a pivotal role in the coordination of committing steps in this pathway, controlling carbon flux towards the different sinks. Furthermore, this C demand is supported by an activation of the photosynthetic machinery, and boosted by a coordinated control of ROS into a sub-lethal range, and associated to enhanced protection to biotic stress.
Physiological, metabolic, and genetic changes produced by two plant growth promoting rhizobacteria (PGPR)
sp. (internal code of the laboratory: N 5.12 and N 21.24) inoculated in tomato plants ...subjected to moderate water stress (10% polyethylene glycol-6000; PEG) were studied. Photosynthesis efficiency, photosynthetic pigments, compatible osmolytes, reactive oxygen species (ROS) scavenging enzymes activities, oxidative stress level and expression of genes related to abscisic acid synthesis (ABA; 9-cis-epoxycarotenoid dioxygenase
gene), proline synthesis (Pyrroline-5-carboxylate synthase
gene), and plasma membrane ATPase (
gene) were measured. Photosynthetic efficiency was compromised by PEG, but bacterial-inoculated plants reversed the effects: while N5.12 increased carbon fixation (37.5%) maintaining transpiration, N21.24 increased both (14.2% and 31%), negatively affecting stomatal closure, despite the enhanced expression of NCDE1 and plasma membrane ATPase genes, evidencing the activation of different adaptive mechanisms. Among all parameters evaluated, photosynthetic pigments and antioxidant enzymes guaiacol peroxidase (GPX) and ascorbate peroxidase (APX) responded differently to both strains. N 5.12 increased photosynthetic pigments (70% chlorophyll a, 69% chlorophyll b, and 65% carotenoids), proline (33%), glycine betaine (4.3%), and phenolic compounds (21.5%) to a greater extent, thereby decreasing oxidative stress (12.5% in Malondialdehyde, MDA). Both bacteria have highly beneficial effects on tomato plants subjected to moderate water stress, improving their physiological state. The use of these bacteria in agricultural production systems could reduce the amount of water for agricultural irrigation without having a negative impact on food production.
An extensive microbiological study has been carried out in a membrane bioreactor fed with activated sludge and metal-working fluids. Functional diversity and dynamics of bacterial communities were ...studied with different approaches. Functional diversity of culturable bacterial communities was studied with different Biolog™ plates. Structure and dynamics of bacterial communities were studied in culturable and in non-culturable fractions using a 16S rRNA analysis. Among the culturable bacteria, Alphaproteobacteria and Gammaproteobacteria were the predominant classes. However, changes in microbial community structure were detected over time. Culture-independent analysis showed that Betaproteobacteria was the most frequently detected class in the membrane bioreactor (MBR) community with Zoogloea and Acidovorax as dominant genera. Also, among non-culturable bacteria, a process of succession was observed. Longitudinal structural shifts observed were more marked for non-culturable than for culturable bacteria, pointing towards an important role in the MBR performance. Microbial community metabolic abilities assessed with Biolog™ Gram negative, Gram positive and anaerobic plates also showed differences over time for Shannon's diversity index, kinetics of average well colour development, and the intensely used substrates by bacterial community in each plate.
Berries are a rich source of antioxidants compounds, among which is the catechin group. Determination of the monomers (catechin and epicatechin) in fruits is a first step in the way to establish a ...relationship between polyphenols and their effects on human health. The purpose of this work is to develop a method to determine free catechins in blackberry by MEKC and to characterize levels of catechins in fresh fruits of Rubus fruticosus var. Lochness throughout the annual production period. A methanolic extract was prepared from fresh fruit. Then, it was evaporated and the residue was extracted with diethyl ether. MEKC conditions: phosphoric acid, 30 mmol/L; SDS, 40 mmol/L and triethylamine, 0.1% v/v at pH 2.3; −15 kV of voltage; 10‐s hydrodynamic injection; 25°C temperature; and detection at 200 nm. Instrumental and interday precision were lower than 4.7 and 10% RSD, respectively. Only (−)‐epicatechin was quantified in blackberries and ranged from 120 to 620 mg/kg fresh weight, which were the lowest values in December and the highest in June. A solid–liquid extraction and an MEKC method were successfully applied to determine (−)‐epicatechins in blackberry for the first time. A strong dependence of (−)‐epicatechin on the annual average temperature was observed.
The relationship between air pollution and the allergenic capacity of pollen is widely accepted, with allergenicity being directly related to air pollution. To our knowledge, this is the first study ...comparing the differential expression of Lolium perenne pollen genes by RNAseq, in two wild populations with different levels of air pollution. The objective is to search for proteins that are expressed differentially in both situations and to establish a relationship with increased allergenic capacity. Two populations of L. perenne (Madrid and Ciudad Real) have been studied in two consecutive years, under the rationale that overexpressed genes in Madrid, with higher levels of NO2 and SO2, could be a cause for their greater allergenic capacity. Heat shock proteins (HSP), glycoside hydrolases, proteins with leucin-rich repeat motifs, and proteins with EF-HAND motifs were consistently overexpressed in Madrid pollen in the two years studied. Interestingly, some genes were overexpressed only in one of the years studied, such as pectinesterases in the first year, and lipid transfer proteins (LTPs) and thaumatin in the second. Despite the fact that the potential of all these proteins in relation to possible allergies has been reported, this is the first time they are cited as possible allergens of L. perenne. The results found can contribute decisively to the knowledge of the allergens of L. perenne and their relationship with atmospheric pollution, and to the development of much more effective vaccines.
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