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The Iron Quadrangle (IQ) is one of the main iron ore producing regions of the world. The exploitation of its reserves jeopardizes the high biological endemism associated with this region. This work aimed to understand the diversity and bacterial potential associated with IQ caves. Floor and ceiling samples of seven ferruginous caves and one quartzite cave were collected, and their microbial relative abundance and diversity were established by 16S rRNA gene amplicon sequencing data. The results showed that ferruginous caves present higher microbial abundance and greater microbial diversity compared to the quartzite cave. Many species belonging to genera found in these caves, such as Pseudonocardia and Streptacidiphilus, are known to produce biomolecules of biotechnological interest as macrolides and polyketides. Moreover, comparative analysis of microbial diversity and metabolic potential in a biofilm in pendant microfeature revealed that the microbiota associated with this structure is more similar to the floor rather than ceiling samples, with the presence of genera that may participate in the genesis of these cavities, for instance, Ferrovum, Geobacter, and Sideroxydans. These results provide the first glimpse of the microbial life in these environments and emphasize the need of conservation programs for these areas, which are under intense anthropogenic exploration.

Serratia liquefaciens strain FG3 (SlFG3), isolated from the flower of Stachytarpheta glabra in the Brazilian ferruginous fields, has distinctive genomic, adaptive, and biotechnological potential. Herein, using a combination of genomics and molecular approaches, we unlocked the evolution of the adaptive traits acquired by S1FG3, which exhibits the second largest chromosome containing the largest conjugative plasmids described for Serratia. Comparative analysis revealed the presence of 18 genomic islands and 311 unique protein families involved in distinct adaptive features. S1FG3 has a diversified repertoire of genes associated with Nonribosomal peptides (NRPs/PKS), a complete and functional cluster related to cellulose synthesis, and an extensive and functional repertoire of oxidative metabolism genes. In addition, S1FG3 possesses a complete pathway related to protocatecuate and chloroaromatic degradation, and a complete repertoire of genes related to DNA repair and protection that includes mechanisms related to UV light tolerance, redox process resistance, and a laterally acquired capacity to protect DNA using phosphorothioation. These findings summarize that SlFG3 is well-adapted to different biotic and abiotic stress situations imposed by extreme conditions associated with ferruginous fields, unlocking the impact of the lateral gene transfer to adjust the genome for extreme environments, and providing insight into the evolution of prokaryotes. Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior -Brazil (CAPES)CAPES [001, CFP 51/2013, 3385/2013]; National Council of Technological and Scientific Development (CNPq)National Council for Scientific and Technological Development (CNPq) [481226/2013-3]; Foundation of Protection to Research of the State of Minas Gerais - FAPEMIG [APQ-02387-14, APQ-02357-17]; CNPqNational Council for Scientific and Technological Development (CNPq); Fundect-MS [TO 141/2016, TO 007/2015, 007/2015 SIAFEM 025139]; UFOP grants; Laboratory of Biochemistry and Molecular Biology of the Faculty of Agrarian and Veterinary Sciences UNESP, Jaboticabal campus Thanks to all members of the Laboratory of Biochemistry and Molecular Biology (LBBM, Federal University of Ouro Preto, UFOP) and to the Laboratory of Biochemistry and Molecular Biology of the Faculty of Agrarian and Veterinary Sciences UNESP, Jaboticabal campus, for their support. This study was financed in part by the Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior -Brazil (CAPES) - Finance Code 001 - (the BIGA Project, CFP 51/2013, process 3385/2013), National Council of Technological and Scientific Development (CNPq Process 481226/2013-3), and Foundation of Protection to Research of the State of Minas Gerais - FAPEMIG (process APQ-02387-14 and process APQ-02357-17). LMM, JCS, AMV, NFA and JAF have a research fellowship from CNPq. NFA has grants from Fundect-MS (TO 141/2016 and TO 007/2015) and Fundect-MS (007/2015 SIAFEM 025139). LAGM and LMM have UFOP grants. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

The ferruginous rupestrian grasslands (FRG) in the Iron Quadrangle (IQ) are ecosystems characterized by rocky soils with reduced availability of water and nutrients, but high levels of metals. In order to comprehend the interference of microorganisms on the adaptive process of endemic plant Mimosa calodendrum (Fabaceae), bacteria associated with its roots and rhizosphere were isolated. Fourteen isolates were obtained and subsequently grown in the presence of different concentrations of arsenic (As) species. The isolate Mc250, an Alcaligenes faecalis strain, resisted to 10 mM of As (III) and 800 mM of As (V). In the presence of this strain, atomic spectrometer detected a reduction of 55% for As (III) and 72% for As (V) respectively in 10 mM and 500 mM solution. Scanning electron microscopy of this isolate demonstrated morphological modification and EDX spectroscopy revealed the presence of both As species adsorbed on the membrane, justifying the removal observed in the in vitro assays. To validate this potential removal of As in vivo, tomato plants were used as grown model in the presence and absence of A. faecalis in soil previously contaminated with 5 mM of As (III). After 14 days, plants from contaminated soil had their growth improved when compared to untreated control plants. All these results suggest for the first time that plant-associated bacteria from FRG-IQ present potential for soil rhizoremediation and may benefit the adaptive processes of plants in extreme environments including application in recovering degraded areas.

Made available in DSpace on 2021-06-25T10:44:05Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-11-01 Here we present and analyze the complete genome of Alcaligenes faecalis strain Mc250 (Mc250), a bacterium isolated from the roots of Mimosa calodendron, an endemic plant growing in ferruginous rupestrian grasslands in Minas Gerais State, Brazil. The genome has 4,159,911 bp and 3,719 predicted protein-coding genes, in a single chromosome. Comparison of the Mc250 genome with 36 other Alcaligenes faecalis genomes revealed that there is considerable gene content variation among these strains, with the core genome representing only 39% of the protein-coding gene repertoire of Mc250. Mc250 encodes a complete denitrification pathway, a network of pathways associated with phenolic compounds degradation, and genes associated with HCN and siderophores synthesis; we also found a repertoire of genes associated with metal internalization and metabolism, sulfate/sulfonate and cysteine metabolism, oxidative stress and DNA repair. These findings reveal the genomic basis for the adaptation of this bacterium to the harsh environmental conditions from where it was isolated. Gene clusters associated with ectoine, terpene, resorcinol, and emulsan biosynthesis that can confer some competitive advantage were also found. Experimental results showed that Mc250 was able to reduce (∼60%) the virulence phenotype of the plant pathogen Xanthomonas citri subsp. citri when co-inoculated in Citrus sinensis, and was able to eradicate 98% of juveniles and stabilize the hatching rate of eggs to 4% in two species of agricultural nematodes. These results reveal biotechnological potential for the Mc250 strain and warrant its further investigation as a biocontrol and plant growth-promoting bacterium. Núcleo de Pesquisas em Ciências Biológicas (NUPEB) Universidade Federal de Ouro Preto Departamento de Ciências Biológicas (DECBI) Instituto de Ciências Exatas e Biológicas (ICEB) Universidade Federal de Ouro Preto (UFOP) Instituto Pristino Departamento de Bioquimica (DBQ) Instituto de Quimica (IQ) Universidade de São Paulo (USP) Instituto de Biotecnologia Aplicada a Agropecuária (BIOAGRO) Universidade Federal de Vicosa (UFV) Departamento de Tecnologia Faculdade de Ciências Agrárias e Veterinárias de Jaboticabal (FCAV) Universidade Estadual Paulista (UNESP) Faculdade de Computação (FACOM) Universidade Federal de Mato Grosso do Sul Departamento de Tecnologia Faculdade de Ciências Agrárias e Veterinárias de Jaboticabal (FCAV) Universidade Estadual Paulista (UNESP)

Made available in DSpace on 2020-12-12T01:43:17Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-01-01 Fundo de Defesa da Citricultura Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG) Background. In Citrus cultures, three species of Xanthomonas are known to cause distinct diseases. X. citri subsp. citri patothype A, X. fuscans subsp. aurantifolii pathotypes B and C, and X. alfalfae subsp. citrumelonis, are the causative agents of cancrosis A, B, C, and citrus bacterial spots, respectively. Although these species exhibit different levels of virulence and aggressiveness, only limited alternatives are currently available for proper and early detection of these diseases in the fields. The present study aimed to develop a new molecular diagnostic method based on genomic sequences derived from the four species of Xanthomonas. Results. Using comparative genomics approaches, primers were synthesized for the identification of the four causative agents of citrus diseases. These primers were validated for their specificity to their target DNA by both conventional and multiplex PCR. Upon evaluation, their sensitivity was found to be 0.02 ng/µl in vitro and 1.5 × 104 CFU ml−1 in infected leaves. Additionally, none of the primers were able to generate amplicons in 19 other genomes of Xanthomonas not associated with Citrus and one species of Xylella, the causal agent of citrus variegated chlorosis (CVC). This denotes strong specificity of the primers for the different species of Xanthomonas investigated in this study. Conclusions. We demonstrated that these markers can be used as potential candidates for performing in vivo molecular diagnosis exclusively for citrus-associated Xanthomonas. The bioinformatics pipeline developed in this study to design specific genomic regions is capable of generating specific primers. It is freely available and can be utilized for any other model organism. Núcleo de Pesquisas em Ciências Biológicas Universidade Federal de Ouro Preto Departamento de Tecnologia Faculdade de Ciências Agrárias e Veterinárias de Jaboticabal Universidade Estadual Paulista-Unesp Departamento de Fitopatologia e Nematologia Escola Superior de Agricultura ‘‘Luiz de Queiroz’’ Departamento de Bioquímica Instituto de Química Universidade de São Paulo Instituto de Computação Universidade Estadual de Campinas Faculdade de Computação Universidade Federal de Mato Grosso do Sul Departamento de Ciências Biológicas Universidade Federal de Ouro Preto Departamento de Tecnologia Faculdade de Ciências Agrárias e Veterinárias de Jaboticabal Universidade Estadual Paulista-Unesp Fundo de Defesa da Citricultura: 007/2015 SIAFEM 025139 CAPES: 3385/2013 CNPq: 481226/2013-3 FAPEMIG: APQ-02387-14 CAPES: CFP 51/2013

Highlights • Cladosporium halotolerans removes 96 % of Mn2+ in two weeks by chelating mechanisms. • Hypocrea jecorina removes Mn2+ by oxidizing mechanisms with 56 % balance. • Laccases could be involved in Mn removal mechanisms played by C. halotolerans., Manganese (Mn) is toxic at higher concentrations requiring its removal before returning the wastewater to the environment. This article reported the Mn removal of two fungi strains isolated from mine wastewater. ITS rRNA region sequencing identified the fungi strains as Cladosporium halotolerans and Hypocrea jecorina. Mn2+ removal assays were performed in Sabouraud broth with 50 mg L−1 Mn2+ supplemented and bioleaching assays using MnO2 instead of MnSO4 at the same conditions. C. halotolerans removed 96 % of 50 mg L−1 Mn2+ at two weeks without MnO2 bioleaching with 649.9 mg of biomass and H. jecorina removed about 50 % of Mn2+ in 21 days from initial 50 mg of Mn2+ L-1 with 316.8 mg of biomass. Extracellular laccases were present in C. halotolerans agar regardless of the Mn addition. Mn adsorbed was detected on C. halotolerans hyphae. Mn oxidation was positive to H. jecorina by reaction of its medium with Leucoberbelin blue.

Made available in DSpace on 2019-10-06T15:59:04Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-10-01 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG) Microorganisms associated with plants have a great biotechnological potential, but investigations of these microorganisms associated with native plants in peculiar environments has been incipient. The objective of this study was to analyze the plant growth-promoting bacteria potential of cultivable bacteria associated with rare plants from the ferruginous rocky fields of the Brazilian Iron Quadrangle. The roots and rhizospheres of nine endemic plants species and samples of a root found in a lateritiric duricrust (canga) cave were collected, the culturable bacteria isolated and prospected for distinct biotechnological and ecological potentials. Out of the 148 isolates obtained, 8 (5.4%) showed potential to promote plant growth, whereas 4 (2.7%) isolates acted as biocontrol agents against Xanthomonas citri pathotype A (Xac306), reducing the cancrotic lesions by more than 60% when co-inoculated with this phytopathogen in Citrus sinensis plants. Moreover, other 4 (2.7%) isolates were classified as potential bioremediation agents, being able to withstand high concentrations of arsenite (5 mM As3+) and arsenate (800 mM As5+), by removing up to 35% and 15% of this metalloid in solution, respectively. These same four isolates had a positive influence on the growth of both the roots and the aerial parts when inoculated with tomato seeds in the soil contaminated with arsenic. This is the first time that an investigation highlights the potentialities of bacteria associated with rare plants of ferruginous rocky fields as a reservoir of microbiota of biotechnological and ecological interest, highlighting the importance of conservation of this area that is undergoing intense anthropic activity. Graphical abstract: [Figure not available: see fulltext.]. Programa de Pós-graduação em Biotecnologia Núcleo de Pesquisas em Ciências Biológicas Universidade Federal de Ouro Preto Departamento de Ciências Biológicas (DECBI) Instituto de Ciências Exatas e Biológicas (ICEB) Universidade Federal de Ouro Preto (UFOP) Departamento de Tecnologia Universidade Estadual Paulista UNESP, Campus de Jaboticabal Instituto Prístino Departamento de Tecnologia Universidade Estadual Paulista UNESP, Campus de Jaboticabal CAPES: 3385/2013 CNPq: 481226/2013-3 FAPEMIG: APQ-02387-14 CAPES: CFP 51/2013

Soil is a habitat where plant roots and microorganisms interact. In the region of the Brazilian Iron Quadrangle (IQ), studies involving the interaction between microbiota and plants have been neglected. Even more neglected are the studies involving the holoparasite plant Langsdorffia hypogaea Mart. (Balanophoraceae). The geomorphological peculiarities of IQ soil, rich in iron ore, as well as the model of interaction between L. hypogaea, its hosts and the soil provide a unique niche that acts as selective pressure to the evolution of plant growth-promoting bacteria (PGPB). The aim of this study was to prospect the bacterial microbiota of holoparasitic plant L. hypogaea, its plant host and corresponding rhizosphere of IQ soil, and to analyze the potential of these isolates as PGPB. We obtained samples of 11 individuals of L. hypogaea containing fragments of host and rhizosphere remnants, resulting in 81 isolates associated with Firmicutes and Proteobacteria phyla. The ability to produce siderophores, hydrocyanic acid (HCN), indole-3-acetic acid (IAA), nitrogen (N2) fixation, hydrolytic enzymes secretion and inhibition of enteropathogens, and phytopathogens were evaluated. Of the total isolates, 62, 86, and 93% produced respectively siderophores, IAA, and were able to fix N2. In addition, 27 and 20% of isolates inhibited the growth of enteropathogens and phytopathogens respectively, and 58% were able to produce at least one hydrolytic activity investigated. The high number of isolates that produce siderophores and indole-3-acetic acid suggests that this microbiota may be important for adaptation of plants to IQ. The results demonstrate for the first time the biological importance of Brazilian IQ species as reservoirs of specific microbiotas that might be used as plant growth-promoting bacteria on agricultural land or antropized soils that needs to be reforested.

The floral biology and reproductive phenology of Langsdorffia hypogaea were investigated. Flowering is annual and high synchronicity was observed along two years, with significant correlation with rainfall. The post-reproductive death of rhizomes suggests monocarpy. Flowers have anthers with extrorse dehiscence and a stigma exposed above the perigonium, and are arranged in dense inflorescences which produce extrafloral nectar. A total of 259 floral visitors were observed, mostly Hymenoptera/Formicidae (149 individuals, 17 ant species), and eight species of Araneae. Although ants were frequent floral visitors, a Coleoptera (Nitidulidae, Stelidota; 28% of total visits) species was the most abundant pollinator. In consequence of herbivory only 12.5% of all inflorescence clusters fruited, and we could not evaluate the percentage of pollinated plants due to insect damage. By being a rich resource in a season of scarcity, activity on inflorescences of L. hypogaea of both pollinators and herbivores was high.

Motivation Information about metabolic pathways in a comparative context is one of the most powerful tool to help the understanding of genome-based differences in phenotypes among organisms. Although several platforms exist that provide a wealth of information on metabolic pathways of diverse organisms, the comparison among organisms using metabolic pathways is still a difficult task. Results We present TabPath (Tables for Metabolic Pathway), a web-based tool to facilitate comparison of metabolic pathways in genomes based on KEGG. From a selection of pathways and genomes of interest on the menu, TabPath generates user-friendly tables that facilitate analysis of variations in metabolism among the selected organisms. Availability and implementation TabPath is available at http://200.239.132.160:8686.

Fundacao de Amparo a Pesquisa do Estado de Sao Paulo Fundacao de Amparo a Pesquisa do Estado de Minas Gerais BIGA grant-CAPES Background: Xanthomonas citri subsp. citri (Xac) is the causal agent of citrus canker. A proteomic analysis under in planta infectious and non-infectious conditions was conducted in order to increase our knowledge about the adaptive process of Xac during infection. Results: For that, a 2D-based proteomic analysis of Xac at 1, 3 and 5 days after inoculation, in comparison to Xac growth in NB media was carried out and followed by MALDI-TOF-TOF identification of 124 unique differentially abundant proteins. Among them, 79 correspond to up-regulated proteins in at least one of the three stages of infection. Our results indicate an important role of proteins related to biofilm synthesis, lipopolysaccharides biosynthesis, and iron uptake and metabolism as possible modulators of plant innate immunity, and revealed an intricate network of proteins involved in reactive oxygen species adaptation during Plants'Oxidative Burst response. We also identified proteins previously unknown to be involved in Xac-Citrus interaction, including the hypothetical protein XAC3981. A mutant strain for this gene has proved to be non-pathogenic in respect to classical symptoms of citrus canker induced in compatible plants. Conclusions: This is the first time that a protein repertoire is shown to be active and working in an integrated manner during the infection process in a compatible host, pointing to an elaborate mechanism for adaptation of Xac once inside the plant. Univ Fed Ouro Preto, Inst Ciencias Exatas & Biol, Dept Ciencias Biol DECBI, Ouro Preto, MG, Brazil Univ Fed Ouro Preto, Nucleo Pesquisas Ciencias Biol NUPEB, Ouro Preto, MG, Brazil Univ Fed Rio de Janeiro, Inst Quim, Dept Bioquim DBq, Rio De Janeiro, RJ, Brazil Univ Estadual Paulista, UNESP, Dept Tecnol, Fac Ciencias Agr & Vet Jaboticabal, Jaboticabal, SP, Brazil Univ Estadual Campinas, UNICAMP, Inst Quim, Campinas, SP, Brazil Univ Sao Paulo, Inst Quim, Dept Bioquim, Sao Paulo, SP, Brazil Univ Fed Sao Paulo UNIFESP, Dept Ciencias Biol, Diadema, SP, Brazil Virginia Tech, Biocomplex Inst, Blacksburg, VA USA Univ Sao Paulo, Inst Quim, Dept Bioquim, Sao Paulo, SP, Brazil Univ Fed Sao Paulo UNIFESP, Dept Ciencias Biol, Diadema, SP, Brazil CAPES FAPESP: 04/02006-7 Fundacao de Amparo a Pesquisa do Estado de Minas Gerais: CBB-APQ-04425-10 BIGA grant-CAPES: 3385/2013 Web of Science

The Xanthomonadaceae family consists of species of non-pathogenic and pathogenic γ-proteobacteria that infect different hosts, including humans and plants. In this study, we performed a comparative analysis using 69 fully sequenced genomes belonging to this family, with a focus on identifying proteins enriched in phytopathogens that could explain the lifestyle and the ability to infect plants. Using a computational approach, we identified seven phytopathogen-enriched protein families putatively secreted by type II secretory system: PheA (CM-sec), LipA/LesA, VirK, and four families involved in N-glycan degradation, NixE, NixF, NixL, and FucA1. In silico and phylogenetic analyses of these protein families revealed they all have orthologs in other phytopathogenic or symbiotic bacteria, and are involved in the modulation and evasion of the immune system. As a proof of concept, we performed a biochemical characterization of LipA from Xac306 and verified that the mutant strain lost most of its lipase and esterase activities and displayed reduced virulence in citrus. Since this study includes closely related organisms with distinct lifestyles and highlights proteins directly related to adaptation inside plant tissues, novel approaches might use these proteins as biotechnological targets for disease control, and contribute to our understanding of the coevolution of plant-associated bacteria.