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The present study examined the biosynthesis and characterization of selenium nanoparticles (SeNPs) using two contrasting endophytic selenobacteria, one Gram-positive (Bacillus sp. E5 identified as Bacillus paranthracis) and one Gram-negative (Enterobacter sp. EC5.2 identified as Enterobacter ludwigi), for further use as biofortifying agents and/or for other biotechnological purposes. We demonstrated that, upon regulating culture conditions and selenite exposure time, both strains were suitable “cell factories” for producing SeNPs (B-SeNPs from B. paranthracis and E-SeNPs from E. ludwigii) with different properties. Briefly, dynamic light scattering (DLS), transmission electron microscopy (TEM), and atomic force microscopy (AFM) studies revealed that intracellular E-SeNPs (56.23 ± 4.85 nm) were smaller in diameter than B-SeNPs (83.44 ± 2.90 nm) and that both formulations were located in the surrounding medium or bound to the cell wall. AFM images indicated the absence of relevant variations in bacterial volume and shape and revealed the existence of layers of peptidoglycan surrounding the bacterial cell wall under the conditions of biosynthesis, particularly in the case of B. paranthracis. Raman spectroscopy, Fourier transform infrared spectroscopy (FTIR), energy-dispersive X-ray (EDS), X-ray diffraction (XRD), and X-ray photoelectron spectroscopy (XPS) showed that SeNPs were surrounded by the proteins, lipids, and polysaccharides of bacterial cells and that the numbers of the functional groups present in B-SeNPs were higher than in E-SeNPs. Thus, considering that these findings support the suitability of these two endophytic stains as potential biocatalysts to produce high-quality Se-based nanoparticles, our future efforts must be focused on the evaluation of their bioactivity, as well as on the determination of how the different features of each SeNP modulate their biological action and their stability.

Background:Climate change challenges modern agriculture to develop alternative and eco-friendly solutions to alleviate (a)biotic stresses. The use of soil microbiomes from extreme environments opens new avenues to discover novel microorganisms and microbial functions to protect plants. In this study we confirm the ability of a bioinoculant, generated by natural engineering, to promote host development under water stress. Microbiome engineering was mediated through three factors i) Antarctic soil donation, ii) water deficit and iii) multigenerational tomato host selection. Results: We revealed that tomato plants growing in soils supplemented with Antarctic microbiota were tolerant to water deficit stress after 10 generations. Microbial community analysis using 16s rRNA amplicon sequencing data suggested that Candidatus Nitrocosmicus and Bacillusspp. were key taxa associated with the observed enhanced water deficit tolerance. Conclusions: We proposed that in situ microbiota engineering through the evolution of tridimensional factors (long-standing extreme climate adaption and host and stress selection) could represent a promising strategy for novel generation of microbial inoculants.

B-group vitamins are essential nutrients for living organisms. Despite that B-group vitamins are present in a multiplicity of foods, vitamin deficiency in human still occur in many countries, mainly by inadequate food intake and unbalanced diets. To overcome this global health problem, chemistry food supplementation is a widely used alternative. Moreover, in the specific case of folic acid, the biofortification with the natural folate form 5-methyltetrahydrofolate (5-MTHF), found in vegetables and produced by certain bacterial taxa such is the case of Lactic Acid Bacteria (LAB), would be a more efficient and secure alternative than the food supplementation of foods with folic acid. This review recommends the possibility to use LAB as an economically sustainable and environmentally friendly plant probiotic in order to produce vitamins biofortificated crops. This new approach could lead something interesting in two aspects: on one hand, the promotion of plant growth, and on the other hand, a biofortificated food crop for human nutrition and health safety to food security.

Simple Summary Scientific knowledge should transcend the barriers between the laboratory and the field to act in the service of humanity. Considering the enormous potential that soil offers for organic carbon (SOC) sequestration for the mitigation of greenhouse gas (GHG) emissions, and considering the recognized ecological importance of biological soil crusts (biocrusts) to be applied in the soil–plant continuum, we propose three perspectives to apply biocrusts to sustainable agriculture. Abstract The major priority of research in the present day is to conserve the environment by reducing GHG emissions. A proposed solution by an expert panel from 195 countries meeting at COP 21 was to increase global SOC stocks by 0.4% year−1 to compensate for GHG emissions, the ‘4 per 1000′ agreement. In this context, the application of biocrusts is a promising framework with which to increase SOC and other soil functions in the soil–plant continuum. Despite the importance of biocrusts, their application to agriculture is limited due to: (1) competition with native microbiota, (2) difficulties in applying them on a large scale, (3) a lack of studies based on carbon (C) balance and suitable for model parameterization, and (4) a lack of studies evaluating the contribution of biocrust weathering to increase C sequestration. Considering these four challenges, we propose three perspectives for biocrust application: (1) natural microbiome engineering by a host plant, using biocrusts; (2) quantifying the contribution of biocrusts to C sequestration in soils; and (3) enhanced biocrust weathering to improve C sequestration. Thus, we focus this opinion article on new challenges by using the specialized microbiome of biocrusts to be applied in a new environment to counteract the negative effects of climate change.

Simple Summary In order to face the challenges posed by climate change, scientific research should be directed towards global needs while also keeping into account the need for increased plant productivity. In this sense, our scientific group from the Biocontrol Research Laboratory BIOREN (Temuco, Chile) and our collaborators, have been studying the enormous potential to enhance productivity by using suppressive soils. In this review, we highlight soil-suppressive microbiota as a natural source of biocontrol agents and we propose a strategy to create microbial assemblages, where the plant selects its own inoculants (when plants “cry for help”). This approach is based on the selection of specific taxa during the transition from a conducive to a suppressive soil. We hope that this strategy leads to generation of personalized bioinoculants to counteract the effects of climate change and increase agricultural sustainability. Abstract Crop migration caused by climatic events has favored the emergence of new soilborne diseases, resulting in the colonization of new niches (emerging infectious diseases, EIDs). Soilborne pathogens are extremely persistent in the environment. This is in large part due to their ability to reside in the soil for a long time, even without a host plant, using survival several strategies. In this regard, disease-suppressive soils, characterized by a low disease incidence due to the presence of antagonist microorganisms, can be an excellent opportunity for the study mechanisms of soil-induced immunity, which can be applied in the development of a new generation of bioinoculants. Therefore, here we review the main effects of climate change on crops and pathogens, as well as the potential use of soil-suppressive microbiota as a natural source of biocontrol agents. Based on results of previous studies, we also propose a strategy for the optimization of microbiota assemblages, selected using a host-mediated approach. This process involves an increase in and prevalence of specific taxa during the transition from a conducive to a suppressive soil. This strategy could be used as a model to engineer microbiota assemblages for pathogen suppression, as well as for the reduction of abiotic stresses created due to global climate change.

Considering that some members of the Proteaceae family are aluminum (Al) and manganese (Mn)–hyperaccumulating species, we aimed to evaluate the presence of the metal hyperaccumulation trait in the Proteaceae family inhabiting southern South America. We also questioned whether other metals (e.g. iron (Fe), copper (Cu) and zinc (Zn)) are hyperaccumulated in the leaves of these species. Mature and senesced leaves of the six Proteaceae species (Embothrium coccineum, Gevuina avellana, Orites myrtoidea, Lomatia hirsuta, L. ferruginea, and L. dentata) as well as soil samples were collected at nine sites along a very extensive latitudinal gradient in southern Chile. Leaves and soil samples were used for chemical analyses. The results showed that G. avellana, L. dentata and O. myrtoidea are Al hyperaccumulators, with G. avellana showing the highest leaf Al concentrations. These leaf Al concentrations were independent of percentage of Al saturation and soil pH. No other metal (Fe, Cu, Zn, Mn) was hyperaccumulated in Proteaceae leaves. However, high Mn concentrations in leaves were found, especially in the species with the lowest values of Al concentrations in their leaves. Three of six species evaluated showed Al hyperaccumulation in their leaves, suggesting that this trait is common in Proteaceae from southern South America.

Gaeumannomyces graminis var. tritici is a soilborne pathogen that causes “take-all” disease, affecting cereal roots. In wheat, G. graminis var. tritici is the most important biotic factor, causing around 30 to 50% losses of yield. Chemical control of this fungal disease is difficult because G. graminis var. tritici is able to reside for a long time in soils. Therefore, the development of environmentally friendly biotechnological strategies to diminish the incidence of soilborne diseases is highly desirable. Natural products are a promising strategy for biocontrol of plant pathogens. A special emphasis is on medicinal plants due to their reported fungitoxic effects. Drimys winteri (canelo) is a medicinal plant that is widely used by the Mapuche ethnic group from Chile due to its anti-inflammatory activity. In addition, inhibitory effects of canelo against phytopathogenic fungi and pest insects have been reported. In this study, we isolated, purified, and identified six drimane sesquiterpenoid compounds from canelo (drimenin, drimenol, polygodial, isodrimeninol, valdiviolide, and drimendiol). Then, we evaluated their antimicrobial effects against G. graminis var. tritici. Compounds were identified by comparing Fourier-transform infrared spectroscopy (FTIR) data and the retention time in thin-layer chromatography (TLC) with those of pure standards. The putative antagonistic effects were confirmed by assessing hyphal cell wall damage using confocal microscopy and lipid peroxidation. Here, we reported the high potential of drimane sesquiterpenoids as natural antifungals against G. graminis var. tritici. Polygodial and isodrimeninol were the most effective, with 50% lethal concentrations (LC50s) between 7 and 10 μg ml−1 and higher levels of fungal lipid peroxidation seen. Accordingly, natural sesquiterpenoids purified from canelo are biologically active against G. graminis var. tritici and could be used as natural biofungicides for sustainable agriculture. IMPORTANCE More than two billion tons of pesticides are used every year worldwide. An interesting sustainable alternative to control plant pathogens is the use of natural products obtained from plants, mainly medicinal plants that offer secondary metabolites important to human/animal health. In this study, we isolated and identified six pure drimane sesquiterpenoids obtained from the bark of Drimys winteri. Additionally, we evaluated their antifungal activities against Gaeumannomyces graminis (the main biotic factor affecting cereal production, especially wheat) by assessing fungal cell wall damage and lipid peroxidation. The compounds obtained showed important antifungal properties against G. graminis var. tritici, mainly isodrimenol, which was the second-most-active compound after polygodial, with an LC50 against G. graminis var. tritici of around 9.5 μg ml−1. This information could be useful for the development of new natural or hemisynthetic antifungal agents against soilborne phytopathogens that could be used in green agriculture.

“Take-all” disease is the most important biotic factor affecting cereal productivity, causing 30–50% of crop losses. The causal agent is the ascomycete soil-borne pathogen Gaeumannomyces graminis var. tritici (Ggt). Current control measures are ineffective, because Ggt can remain saprophytic in soils for long periods. Therefore, the study of the microbiome residing in suppressive soils (SS) is a promising niche of Ggt biocontrol. Here, we evaluated the efficiency of Serratia sp., Bacillus sp., and Acinetobacter sp. isolated from SS against the incidence of Ggt on wheat. Our results demonstrated that plants inoculated with the bacterial consortium in both greenhouse and field conditions were highly efficient in Ggt biocontrol, more so than individual strains. The disease reduction was evidenced by higher biomass production, fewer copies of the Ggt genome with a concomitant curtailment of blackening of roots, a decrease of lipid peroxidation, and an increase of superoxide dismutase activity. The ability of the microbial consortium over that of single strains could be attributable to interspecies communication as a strategy to biocontrol, i.e., higher chitinase activity. In conclusion, bacterial consortia from SS are an important niche of Ggt biocontrol, serving as a model for other soil-borne pathogens.

Chile is among the largest avocado producers worldwide; however, currently, its production has diminished mainly attributed to abiotic stresses such as drought and soil salinity. Here, we evaluated the contribution of halotolerant bacterial consortia to water shortage and salt stress tolerance of avocado seedlings ( Persea americana Mill.) under field conditions. Inoculation trials were conducted in a commercial nursery to study the effects of two endophytic (C1 and C2) and two rhizosphere (C3 and C4) halotolerant bacterial consortia on growth, biomass, superoxide dismutase (SOD) and thiobarbituric acid reactive substances (TBARS) of avocado seedlings under salt (2% NaCl) stress and water shortage (50% less irrigation). Ours results revealed that avocado inoculation with C4 significantly ( P ≤ 0.05) increased aerial and root length; aerial and root fresh weight and chlorophyll content of salt-stressed seedlings; and the aerial length and root fresh weight of seedlings under water shortage. Similarly, the C4 significantly ( P ≤ 0.05) increased SOD activity in leaves of both the control and seedlings grown under salt stress and water shortage and also decreased TBARS content in leaves of control plants and of seedlings grown under salt stress. Whereas, C3 increased significantly ( P ≤ 0.05) aerial and root length and root fresh weight of salt-stressed seedlings; and also increased the trunk diameter and chlorophyll content of seedlings under water shortage. Similarly, C3 significantly ( P ≤ 0.05) stimulated SOD activity of leaves as compared to the control seedlings and also reduced the TBARS content of leaves and roots of avocado seedlings under salt stress. In contrast, the endophytic consortia were less efficient than rhizosphere consortia. Thus, C1 only increased the trunk diameter and chlorophyll content of salt–stressed seedlings and C2 increased the chlorophyll content of avocado seedlings under water shortage. Our study showed the favorable effect of bacterial inoculation on avocado nursey in nursery conditions under water shortage and salt stress, and identified consortia that potentially could be used as avocado biofertilizers.

During the productive stage of Corylus avellana (hazelnut) in La Araucania, Chile (October-December 2018-19) we observed symptoms consisting of a brown-greyish spot occurring at the bottom of the nut and enlarging upwards to the…

Phosphobacteria, secreting organic acids and phosphatases, usually favor plant performance in acidic soils by increasing phosphorus (P) availability and aluminum (Al) complexing. However, it is not well-known how P-deficiency and Al-toxicity affect the phosphobacteria physiology. Since P and Al problems often co-occur in acidic soils, we have therefore proposed the evaluation of the single and combined effects of P-deficiency and Al-toxicity on growth, organic acids secretion, malate dehydrogenase (mdh) gene expression, and phosphatase activity of five Al-tolerant phosphobacteria previously isolated from ryegrass. These phosphobacteria were identified as Klebsiella sp. RC3, Stenotrophomona sp. RC5, Klebsiella sp. RCJ4, Serratia sp. RCJ6, and Enterobacter sp. RJAL6. The strains were cultivated in mineral media modified to obtain (i) high P in absence of Al–toxicity, (ii) high P in presence of Al–toxicity, (iii) low P in absence of Al–toxicity, and (iv) low P in presence of Al–toxicity. High and low P were obtained by adding KH2PO4 at final concentration of 1.4 and 0.05 mM, respectively. To avoid Al precipitation, AlCl3 × 6H2O was previously complexed to citric acid (sole carbon source) in concentrations of 10 mM. The secreted organic acids were identified and quantified by HPLC, relative mdh gene expression was determined by qRT-PCR and phosphatase activity was colorimetrically determined using p-nitrophenyl phosphate as substrate. Our results revealed that although a higher secretion of all organic acids was achieved under P–deficiency, the patterns of organic acids secretion were variable and dependent on treatment and strain. The organic acid secretion is exacerbated when Al was added into media, particularly in the form of malic and citric acid. The mdh gene expression was significantly up–regulated by the strains RC3, RC5, and RCJ6 under P–deficiency and Al–toxicity. In general, Al–tolerant phosphobacteria under P deficiency increased both acid and alkaline phosphatase activity with respect to the control, which was deepened when Al was present. The knowledge of this bacterial behavior in vitro is important to understand and predict the behavior of phosphobacteria in vivo. This knowledge is essential to generate smart and efficient biofertilizers, based in Al–tolerant phosphobacteria which could be expansively used in acidic soils.

Phosphorus (P) is an essential macronutrient for plant growth and is therefore an important component of agricultural fertilizers, especially for P–deficient soils. As a more environmentally friendly alternative to P–fertilization, phosphobacteria are regularly being proposed as bio–inoculants for plants that are cultivated in P–deficient soils. Although it is known that phosphobacteria inoculation can influence the growth and P nutrition of the host plant, the underlying responses at the molecular level have to date, not been fully elucidated. Here, we evaluated the single and combined effects of a phosphobacteria consortium inoculation and triple superphosphate fertilization on the biomass, P content, anti–oxidant capacity and phosphate–transporters genes of Lolium perenne. Plants were grown over 45 days in presence and absence of P–fertilization and phosphobacteria inoculation in two soils with different P bio–availabilities. Our results reveal that irrespective of P–availability in soils, the phosphobacteria inoculation provides an ‘enhancement effect’ of the P–fertilization on the development, nutrition and the alleviation of oxidative stress, in L. perenne. This was evidenced by higher biomass, increased P content, and lower cellular damage in tissues of the inoculated plants. The inoculation with phosphobacteria decreased the relative gene expression of superoxide dismutase isoforms in tissues of fertilized plants grown in the P–deficient soil. In addition, phosphobacteria inoculation was also associated with a lower relative expression of phosphate–transporter genes of P–fertilized plants. Therefore, instead of being an alternative to P–fertilization, phosphobacteria could rather be a promising complement to P–fertilization in P–deficient soils and potentially reduce fertilizer application rates. Based on our findings, an explicatory model of plant–soil–phosphobacteria interactions on the regulation of anti–oxidant and phosphate–transport capacities in P–deficient, is proposed.

Plant growth-promoting rhizobacteria (PGPR) are studied as complements/alternatives to chemical fertilizers used in agriculture. However, poor information exists on the potential of PGPR from undisturbed ecosystems. Here, we have evaluated the plant growth-promoting (PGP) effect of rhizobacterial consortia from undisturbed Chilean arid ecosystems (Consortium C1) and agro-ecosystems (Consortium C2) on plant biomass production. The PGP effects of C1 and C2 were assayed in wheat seedlings (Triticum aestivum L.) grown in pots under growth chamber conditions and in pots placed in an open greenhouse under natural conditions, using two different Chilean Andisols (Piedras Negras and Freire series) kept either at 30 or 60% of their maximum water holding capacity (MWHC). PGP effects depended on the soil type, MWHC and the growth conditions tested. Although both consortia showed PGB effects in artificial soils relative to controls in growth chambers, only C1 provoked a PGP effect at 60% MWHC in phosphorus-poor soil of the 'Piedras Negras' series. At natural conditions, however, only C1 exhibited statistically significant PGP effects at 30% MWHC in 'Piedras Negras', yet and most importantly allowed to maintain similar plant biomass as at 60% MWHC. Our results support possible applications of rhizobacterial consortia from arid ecosystems to improve wheat growth in Chilean Andisols under water shortage conditions.Wheat seedling inoculated with rhizobacterial consortia obtained from an undisturbed Chilean arid ecosystem showed improved growth in phosphorus-poor and partly dry soil. Arid ecosystems should be considered in further studies as an alternative source of microbial inoculants for agro-ecosystems subjected to stressful conditions by low nutrients and/or adverse climate events.

Soil microorganisms with phytase- and 1-aminocyclopropane-1-carboxylate (ACC) deaminase activities are widely studied as plant growth-promoting rhizobacteria (PGPR). Here, we explored the bacterial community structure and occurrence of putative PGPR in plants grown in agro-ecosystems and undisturbed ecosystems from northern, central, and southern Chile. Total rhizobacterial community structure was evaluated by denaturing gradient gel electrophoresis, and dominant bands present in diverse ecosystems were sequenced. Significant differences in total bacterial communities were shown with some bacterial orders (Enterobacteriales, Actinomycetales, and Rhizobiales) being highly similar to both ecosystems. Twenty-nine putative PGPR, showing phytate- and ACC-degrading activities and production of auxin, were selected from across the sites. Based on 16S rRNA gene sequencing, the putative PGPR were characterized as Enterobacteriales (Enterobacter, Serratia, Pantoea, Rahnella, Leclercia), Pseudomonas, and Bacillus, consistent with previously reported PGPR and endophytic bacteria. Beta-propeller phytase genes with similarity to Bacillus were also identified. PGPR from agro-ecosystems appeared to show higher auxin production compared to those from undisturbed ecosystems. This study demonstrates that putative PGPR are widely distributed across Chilean soils. Further understanding of their contribution to the growth and adaptation of plant hosts to local soil conditions may provide opportunity for development of new PGPR in Chilean agriculture.

Cereal production in southern Chile is based on ash-derived volcanic Andisols, which present suboptimal levels of available selenium (Se). Strategies are needed to improve Se content in cereal crops and concomitantly improve the nutritional quality of grain. Here, we investigated the occurrence of Se-tolerant bacteria (STB) in Andisols and evaluated Se tolerance and accumulation in STB. The inoculation of wheat with STB and the contributions of these bacteria to Se content in plants were also evaluated under greenhouse conditions. The results showed that Se amendment of Andisols stimulated some bacterial groups (Paenibacillaceae and Brucellaceae) but inhibited others (Clostridia, Burkholderiales, Chitinophagaceae and Oxalobacteraceae), as revealed by denaturing gradient gel electrophoresis. Furthermore, we found four STB isolates that displayed 1-aminocyclopropane-1-carboxylate deaminase (ACC deaminase activity) and that carried the acdS gene as revealed by PCR. The selected STB were characterised as Stenotrophomonas, Bacillus, Enterobacter and Pseudomonas according to partial sequencing of the 16S rRNA gene. After 24 h of culture in nutrient broth, the selected STB showed the ability to grow in high Se concentrations (5 and 10 mM) and to accumulate elemental Se in micro- and nanospherical deposits, transforming 50–80 % of the Se initially added. Greenhouse experiments with wheat showed that Se associated with STB (micro- and nanospheres of elemental Se and other intracellular forms) can be translocated into leaves of wheat plantlets.