Microbiome engineering optimized by Antarctic microbiota to support a plant host under water deficit.

Climate change challenges modern agriculture to develop alternative and eco-friendly solutions to alleviate abiotic and/or 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. We revealed that tomato plants growing in soils supplemented with Antarctic microbiota were tolerant to water deficit stress after 10 generations. A clear increase in tomato seedling tolerance against water deficit stress was observed in all soils over generations of Host Mediated Microbiome Engineering, being Fildes mixture the most representatives, which was evidenced by an increased survival time, plant stress index, biomass accumulation, and decreased leaf proline content. Microbial community analysis using 16s rRNA gene amplicon sequencing data suggested a microbiome restructuring that could be associated with increased tolerance of water deficit. Additionally, the results showed a significant increase in the relative abundance of Candidatus Nitrosocosmicus and Bacillus spp. which could be key taxa associated with the observed tolerance improvement. We proposed that in situ microbiota engineering through the evolution of three factors (long-standing extreme climate adaption and host and stress selection) could represent a promising strategy for novel generation of microbial inoculants.

Occurrence of Soil Fungi in Antarctic Pristine Environments.

The presence of fungi in pristine Antarctic soils is of particular interest because of the diversity of this microbial group. However, the extreme conditions that coexist in Antarctica produce a strong selective pressure that could lead to the evolution of novel mechanisms for stress tolerance by indigenous microorganisms. For this reason, in recent years, research on cold-adapted microorganisms has increased, driven by their potential value for applications in biotechnology. Cold-adapted fungi, in particular, have become important sources for the discovery of novel bioactive secondary metabolites and enzymes. In this study, we studied the fungal community structure of 12 soil samples from Antarctic sites, including King George Island (including Collins Glacier), Deception Island and Robert Island. Culturable fungi were isolated and described according to their morphological and phenotypical characteristics, and the richness index was compared with soil chemical properties to describe the fungal community and associated environmental parameters. We isolated 54 fungal strains belonging to the following 19 genera: Penicillium, Pseudogymnoascus, Lambertella, Cadophora, Candida, Mortierella, Oxygenales, Geomyces, Vishniacozyma, Talaromyces, Rhizopus, Antarctomyces, Cosmospora, Tetracladium, Leptosphaeria, Lecanicillium, Thelebolus, Bjerkandera and an uncultured Zygomycete. The isolated fungi were comprised of 70% Ascomycota, 10% Zygomycota, 10% Basidiomycota, 5% Deuteromycota and 5% Mucoromycota, highlighting that most strains were associated with similar genera grown in cold environments. Among the culturable strains, 55% were psychrotrophic and 45% were psychrophilic, and most were Ascomycetes occurring in their teleomorph forms. Soils from the Collins Glacier showed less species richness and greater species dominance compared with the rest of the sites, whereas samples 4, 7, and 10 (from Fildes Bay, Coppermine Peninsula and Arctowski Station, respectively) showed greater species richness and less species dominance. Species richness was related to the C/N ratio, whereas species dominance was inversely related to C and N content. Thus, the structure of the fungal community was mainly related to soil chemical parameters more than sample location and altitude.

Nutrient Use Efficiency of Southern South America Proteaceae Species. Are there General Patterns in the Proteaceae Family?

Plants from the Proteaceae family can thrive in old, impoverished soil with extremely low phosphorus (P) content, such as those typically found in South Western Australia (SWA) and South Africa. The South Western (SW) Australian Proteaceae species have developed strategies to deal with P scarcity, such as the high capacity to re-mobilize P from senescent to young leaves and the efficient use of P for carbon fixation. In Southern South America, six Proteaceae species grow in younger soils than those of SWA, with a wide variety of climatic and edaphic conditions. However, strategies in the nutrient use efficiency of Southern South (SS) American Proteaceae species growing in their natural ecosystems remain widely unknown. The aim of this study was to evaluate nutrient resorption efficiency and the photosynthetic nutrients use efficiency by SS American Proteaceae species, naturally growing in different sites along a very extensive latitudinal gradient. Mature and senescent leaves of the six SS American Proteaceae species (Embothrium coccineum, Gevuina avellana, Orites myrtoidea Lomatia hirsuta, L. ferruginea, and L. dentata), as well as, soil samples were collected in nine sites from southern Chile and were subjected to chemical analyses. Nutrient resorption (P and nitrogen) efficiency in leaves was estimated in all species inhabiting the nine sites evaluated, whereas, the photosynthetic P use efficiency (PPUE) and photosynthetic nitrogen (N) use efficiency (PNUE) per leaf unit were determined in two sites with contrasting nutrient availability. Our study exhibit for the first time a data set related to nutrient use efficiency in the leaves of the six SS American Proteaceae, revealing that for all species and sites, P and N resorption efficiencies were on average 47.7 and 50.6%, respectively. No correlation was found between leaf nutrient (P and N) resorption efficiency and soil attributes. Further, different responses in PPUE and PNUE were found among species and, contrary to our expectations, a higher nutrient use efficiency in the nutrient poorest soil was not found. We conclude that SS American Proteaceae species did not show a general pattern in the nutrient use efficiency among them neither with others Proteaceae species reported in the literature.

Understanding the Strategies to Overcome Phosphorus-Deficiency and Aluminum-Toxicity by Ryegrass Endophytic and Rhizosphere Phosphobacteria.

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.