Diversity of endophytic bacteria with antimicrobial potential isolated from marine macroalgae from Yacila and Cangrejos beaches, Piura-Peru.

Endophytic bacteria found in marine macroalgae have been studied for their potential antimicrobial activity, consequently, they could serve as a valuable source of bioactive compounds to control pathogenic bacteria, yeasts, and fungi. Algae endophytic bacteria were isolated from Caulerpa sp., Ulva sp., Ahnfeltiopsis sp., and Chondracantus chamissoi from Yacila and Cangrejo Beaches (Piura, Peru). Antimicrobial assays against pathogenic bacteria were evaluated using cross-culture, over-plate, and volatile organic compound tests. Afterward, the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of selected crude extracts were determined, also ITS molecular analysis, antifungal activity, and PCR of iturin, fengycin, and surfactin genes were performed for bacteria strains exhibiting better activity. Forty-six algae endophytic bacteria were isolated from algae. Ten strains inhibited gram-positive pathogenic bacteria (Enterococcus faecalis, Staphylococcus epidermidis, S. aureus, and Listeria monocytogenes), and 12 inhibited gram-negative bacteria (Escherichia coli and Salmonella enteric sv typhimurium). Bacteria with better activity belong to Bacillus sp., Kluyvera ascorbata, Pantoea agglomerans, Leclercia adecarboxylata, and Enterobacter sp., which only four showed antifungal activities against Candida albicans, C. tropicalis, Colletotrichium sp., Fusarium sp., Fusarium oxysporum, and Alternaria sp. Furthermore, K. ascorbata YAFE21 and Bacillus sp. YCFE4 exhibited iturin and fengycin genes. The results indicate that the algae endophytic bacteria found in this study, particularly K. ascorbata YAFE21, Bacillus sp. YCFR6, L. adecarboxylata CUFE2, Bacillus sp. YUFE8, Enterobacter sp. YAFL1, and P. agglomerans YAFL6, could be investigated as potential producers of antimicrobial compounds due to their broad activity against various microorganisms.

Cordyceps cateniannulata: An endophyte of coffee, a parasite of coffee leaf rust and a pathogen of coffee pests.

Here, we report on a Cordyceps species entering into a multi-trophic, multi-kingdom association. Cordyceps cateniannulata, isolated from the stem of wild Coffea arabica in Ethiopia, is shown to function as an endophyte, a mycoparasite and an entomopathogen. A detailed polyphasic taxonomic study, including a multilocus phylogenetic analysis, confirmed its identity. An emended description of C. cateniannulata is provided herein. Previously, this species was known as a pathogen of various insect hosts in both the Old and New World. The endophytic status of C. cateniannulata was confirmed by re-isolating it from inoculated coffee plants. Inoculation studies have further shown that C. cateniannulata is a mycoparasite of Hemileia vastatrix, as well as an entomopathogen of major coffee pests; infecting and killing Hypothenemus hampei and Leucoptera coffeella. This is the first record of C. cateniannulata from Africa, as well as an endophyte and a mycoparasite. The implications for its use as a biocontrol agent are discussed.

Isolation and molecular identification of endophytic fungi associated with Campomanesia adamantium, a Brazilian Cerrado plant.

This work presents the isolation of endophytic fungi from the leaves of Campomanesia adamantium (Cambess.) O. Berg (Myrtaceae), a native species found in Brazil and popularly known as "guabiroba-do-campo", with abundant distribution in the Brazilian Cerrado. It has been popularly used for its anti-rheumatic, anti-inflammatory, antidiarrheal, blood cholesterol-reducing, urinary antiseptic, and depurative properties. Theese fungi are microorganisms that live inside higher plants, at least for a period of their life cycle, occupying the intercellular spaces of plant tissues such as leaves and stems. These fungi are harmless to the host plant, and their secondary metabolites promote protection, regulate growth, combat bacteria, viruses, and fungi, and promote resistance to abiotic stress, as well as insecticidal effects. Endophytic fungi associated with the leaves of C. adamantium were isolated using the culture medium isolation technique. After growth, the fungi were divided into groups based on morphotypes. Fungal genomic DNA was extracted, and a polymerase chain reaction (PCR) was conducted to amplify the ITS1-5.8S-ITS2 regions of rRNA, and the nucleotide sequences obtained were compared with those available in the GenBank database for molecular identification of the isolates. The phylogenetic tree was constructed using MEGA 11 software. The results showed representatives of the Ascomycota phylum, and it was possible to identify at the genus level 18 fungi of the genera Colletotrichum, Diaporthe, Epicoccum, and Neofusicoccum.

Spatial and Temporal Shifts of Endophytic Bacteria in Conifer Seedlings of Abies religiosa (Kunth) Schltdl. & Cham.

Endophytes play an important role in plant development, survival, and establishment, but their temporal dynamics in young conifer plants are still largely unknown. In this study, the bacterial community was determined by metabarcoding of the 16S rRNA gene in the rhizoplane, roots, and aerial parts of 1- and 5-month-old seedlings of natural populations of Abies religiosa (Kunth) Schltdl. & Cham. In 1-month-old seedlings, Pseudomonas dominated aerial parts (relative abundance 71.6%) and roots (37.9%). However, the roots exhibited significantly higher bacterial species richness than the aerial parts, with the dissimilarity between these plant sections mostly explained by the loss of bacterial amplification sequence variants. After 5 months, Mucilaginibacter dominated in the rhizoplane (9.0%), Streptomyces in the roots (12.2%), and Pseudomonas in the aerial parts (18.1%). The bacterial richness and community structure differed significantly between the plant sections, and these variations were explained mostly by 1-for-1 substitution. The relative abundance of putative metabolic pathways significantly differed between the plant sections at both 1 and 5 months. All the dominant bacterial genera (e.g., Pseudomonas and Burkholderia-Caballeronia-Paraburkholderia) have been reported to have plant growth-promoting capacities and/or antagonism against pathogens, but what defines their role for plant development has still to be determined. This investigation improves our understanding of the early plant-bacteria interactions essential for natural regeneration of A. religiosa forest.

Endophytic diazotrophic communities from rice roots are diverse and weakly associated with soil diazotrophic community composition and soil properties.

AIM: Bacteria that promote plant growth, such as diazotrophs, are valuable tools for achieving a more sustainable production of important non-legume crops like rice. Different strategies have been used to discover new bacteria capable of promoting plant growth. This work evaluated the contribution of soil diazotrophs to the endophytic communities established in the roots of rice seedlings cultivated on seven representative soils from Uruguay. METHODS AND RESULTS: The soils were classified into two groups according to the C and clay content. qPCR, terminal restriction fragment length polymorphism (T-RFLP), and 454-pyrosequencing of the nifH gene were used for analyzing diazotrophs in soil and plantlets' roots grown from seeds of the same genotype for 25 days under controlled conditions. A similar nifH abundance was found among the seven soils, roots, or leaves. The distribution of diazotrophs was more uneven in roots than in soils, with dominance indices significantly higher than in soils (nifH T-RFLP). Dominant soils' diazotrophs were mainly affiliated to Alphaproteobacteria and Planctomycetota. Conversely, Alpha, Beta, Gammaproteobacteria, and Bacillota were predominant in different roots, though undetectable in soils. Almost no nifH sequences were shared between soils and roots. CONCLUSIONS: Root endophytic diazotrophs comprised a broader taxonomic range of microorganisms than diazotrophs found in soils from which the plantlets were grown and showed strong colonization patterns.

Endophytic bacterial diversity in the latex-bearing caulosphere of Hevea brasiliensis Müll. Arg.

Rubber trees are a commercial cash crop, and the milky latex or polyisoprene they produce is the natural source of rubber. Little is known about the bacterial populations found in active zone of latex-bearing caulosphere. We employed a tailored cloud microbial bioinformatic approach for the identification and potential hypothetical ecological roles of an uncultured endophytic hidden bacterial community in the active zone of the latex-bearing caulosphere of Hevea brasiliensis. Small pieces of slivers were collected from healthy plant from the village: Belonia, South Tripura, rubber plantation in Northeastern India. These uncultured bacteria were identified using the V3-V4 hypervariable amplicon region of the 16 S rDNA gene. A total of 209,586 contigs have been generated. EasyMAP Version 1.0, a cloud-based microbial bioinformatics tool with an integrated QIIME2 pipeline, was used to analyze contigs. We detected 15 phyla and 91 OTUs (operational taxonomic units). Proteobacteria (73.5%) was the most enriched phylum, followed by Firmicutes (13.8%), Bacteroidetes (5.2%), and Actinobacteria (3.2%). Ammonia oxidizers, sulfate reducers, dehalogenation, chitin degradation, nitrite reducers, and aromatic hydrocarbon degraders were the most prevalent functional categories in the active zones of caulosphere. Furthermore, Gammaproteobacteria (49.2%) and Erwinia (29.19%) were the most abundant classes and genera of endophytic bacterial communities. Thus, the presence of a substantial amount of phosphate-solubilizing Gammaproteobacteria (PSB) may stimulate growth, increase plant resilience, suppress disease, and aid in the rubber and sugar breakdown. This is the first report of microbial endophytes associated with Hevea caulosphere.

Endosphere mycobiome in mature rice roots originate from both seedlings and soils.

Plant-fungus symbioses have functional relevance during plant growth and development. However, it is still unknown whether the endosphere fungi in mature plants originated from soils or seeds. To elucidate the origination of endosphere fungi in mature rice roots, the fungal communities in surface sterilized roots and shoots of mature rice plants germinated in soils, rhizosphere soils and seedlings germinated under sterile conditions were analyzed by Illumina-based sequencing and compared. Total 62 fungal OTUs shared in the seedlings, shoots and roots, 126 OTUs shared in the rhizosphere soils, shoots and roots. Fungal OTUs coexisted in the four types of samples belonged to genera of Rhizophagus, Trichoderma, Fusarium, Atractiella, Myrmecridium, Sporothrix, Microdochium, Massariosphaeria, and Phialemonium. The principle component analysis (PCA) and NMDS plot suggested that the fungal community structure in rhizosphere soils was different from that in seedlings significantly. Rhizosphere soil, shoot and root contained more similar fungal community. The fungal community in seedling was similar to that in shoot and root of mature plants. The results suggested that endophytic fungal communities in mature rice plants originated from both seedlings and rhizosphere soils, and more fungal taxa originated from rhizosphere soils. Mature rice plants contain mycobiome transmitted vertically from seeds, which suggests that inoculation of endophytic fungi isolated from seedlings might be an effective way to introduce beneficial fungal inoculants into rice plants successfully.

Antarctica's hidden mycoviral treasures in fungi isolated from mosses: A first genomic approach.

This study investigates the presence of mycoviruses in Antarctic fungi and elucidates their evolutionary relationships. To achieve this, we aligned mycoviral gene sequences with genomes of previously sequenced Antarctic endophytic fungi, made available by our research group and accessible via Joint Genome Institute. Our findings reveal that the most prevalent genetic regions in all endophytic fungi are homologous to Partitiviruses, Baculoviridae, and Phycodnaviridae. These regions display evidence of positive selection pressure, suggesting genetic diversity and the accumulation of nonsynonymous mutations. This phenomenon implies a crucial role for these regions in the adaptation and survival of these fungi in the challenging Antarctic ecosystems. The presence of mycoviruses in Antarctic endophytic fungi may indicate shared survival strategies between the virus and its host, shedding light on their evolutionary dynamics. This study underscores the significance of exploring mycoviruses within endophytic fungi and their contributions to genetic diversity. Future research avenues could delve into the functional implications of these conserved mycoviral genetic regions in Antarctic endophytic fungi, providing a comprehensive understanding of this intriguing association and genomic retention of viral region in fungi.

Endophytic species of Colletotrichum associated with cashew tree in northeastern Brazil.

Anthracnose caused by Colletotrichum is the most severe and widely occurring cashew disease in Brazil. Colletotrichum species are commonly found as pathogens, endophytes and occasionally as saprophytes in a wide range of hosts. The endophytic species associated with cashew trees are poorly studied. In this study, we report the Colletotrichum endophytic species associated with cashew trees in two locations in the state of Pernambuco, their prevalence in different plant organs (leaves, veins, branches and inflorescences), and compare the species in terms of pathogenicity and aggressiveness using different inoculation methods (wounded × unwounded). Six species of Colletotrichum were identified according to multilocus phylogenetic analyses, including Colletotrichum asianum, Colletotrichum chrysophilum, Colletotrichum karsti, Colletotrichum siamense, Colletotrichum theobromicola, and Colletotrichum tropicale. There were differences in the percentage of isolation in relation to the prevalence of colonized tissues and collection locations. C. tropicale was the prevalent species in both geographic areas and plant tissues collected, with no pattern of distribution of species between areas and plant tissues. All isolates were pathogenic in injured tissues of cashew plants. The best method to test the pathogenicity of Colletotrichum species was utilizing the combination of leaves + presence of wounds + conidial suspension, as it better represents the natural infection process. C. siamense was the most aggressive species.

Diversity of fungal endophytes from mangrove plants of Santa Catarina Island, Brazil.

The mangrove ecosystem plays a crucial role in preserving the biodiversity of plants, animals, and microorganisms that are essential for materials cycles. However, the exploration of endophytic fungi isolated from mangroves, particulary in Santa Catarina (SC, Brazil), remains limited. Therefore, the purpose of this study was to assess the biodiversity of endophytic fungi found in Avicennia schaueriana, Laguncularia racemosa, Rhizophora mangle, and Spartina alterniflora from two mangroves on the Island of Santa Catarina: one impacted by anthropic action (Itacorubi mangrove) and the other environmentally preserved (Ratones mangrove). Samplings were carried out between January 2020 and May 2021. Fungi were isolated from leaves, stems, and roots, identified, and clustered into groups through morphological characteristics. Further, a representative strain of each group was identified through ITS1 sequencing. A total of 373 isolates were obtained from plant tissues, of which 96 and 277 isolates were obtained from Itacorubi and Ratones mangroves, respectively. Molecular identification showed that the endophytic fungal community comprised at least 19 genera. The data on fungal community diversity revealed comparable diversity indices for genera in both mangroves. However, we observed differences in the total frequency of fungal genera between impacted (27.38%) and non-impacted (72.62%) mangroves. These findings suggest that anthropic activities in and around the Santa Catarina mangroves have had negative impact on the frequency of endophytic fungi. This emphasizes the reinforcing the significance of preserving these environments to ensure the maintenance of fungal community diversity.

Antagonistic activity of endophytic actinobacteria from native potatoes (Solanum tuberosum subsp. tuberosum L.) against Pectobacterium carotovorum subsp. carotovorum and Pectobacterium atrosepticum.

BACKGROUND: The native potatoes (Solanum tuberosum subsp. tuberosum L.) grown in Chile (Chiloé) represent a new, unexplored source of endophytes to find potential biological control agents for the prevention of bacterial diseases, like blackleg and soft rot, in potato crops. RESULT: The objective of this study was the selection of endophytic actinobacteria from native potatoes for antagonistic activity against Pectobacterium carotovorum subsp. carotovorum and Pectobacterium atrosepticum, and their potential to suppress tissue maceration symptoms in potato tubers. This potential was determined through the quorum quenching activity using a Chromobacterium violaceaum ATCC 12472 Wild type (WT) bioassay and its colonization behavior of the potato plant root system (S. tuberosum) by means of the Double labeling of oligonucleotide probes for fluorescence in situ hybridization (DOPE-FISH) targeting technique. The results showed that although Streptomyces sp. TP199 and Streptomyces sp. A2R31 were able to inhibit the growth of the pathogens, only the Streptomyces sp. TP199 isolate inhibited Pectobacterium sp. growth and diminished tissue maceration in tubers (p ≤ 0.05). Streptomyces sp. TP199 had metal-dependent acyl homoserine lactones (AHL) quorum quenching activity in vitro and was able to colonize the root endosphere 10 days after inoculation. CONCLUSIONS: We concluded that native potatoes from southern Chile possess endophyte actinobacteria that are potential agents for the disease management of soft rot and blackleg.

Metagenomic analyses, isolation and characterization of endophytic bacteria associated with Eucalyptus urophylla BRS07-01 in vitro plants.

Eucalyptus is the main species for the forestry industry in Brazil. Biotechnology and, more recently, gene editing offer significant opportunities for rapid improvements in Eucalyptus breeding programs. However, the recalcitrance of Eucalyptus species to in vitro culture is also a major limitation for commercial deployment of biotechnology techniques in Eucalyptus improvement. We evaluated various clones of Eucalyptus urophylla for their in vitro regeneration potential identified a clone, BRS07-01, with considerably higher regeneration rate (85%) in organogenesis, and significantly higher than most works described in literature. Endophytic bacteria are widely reported to improve in vitro plant growth and development. Hence, we believe that inclusion of endophytic plant growth promoting bacteria enhanced was responsible for the improved plantlets growth and development of this clone under in vitro culture. Metagenomic analysis was performed to isolate and characterize the prominent endophytic bacteria on BRS07-01 leaf tissue in vitro micro-cultures, and evaluate their impact on plant growth promotion. The analysis revealed the presence of the phyla Firmicutes (35%), Proteobacteria (30%) and much smaller quantities of Actinobacteria, Bacteroidetes, Gemmatimonadetes, Crenarchaeota, Euryarchaeota and Acidobacteria. Of the thirty endophytic bacterial strains isolated, eleven produced indole-3-acetic acid. Two of the isolates were identified as Enterobacter sp. and Paenibacillus polymyxa, which are nitrogen-fixing and capable of phosphate and produce ammonium. These isolates also showed similar positive effects on the germination of common beans (Phaseolus spp.). The isolates will now be tested as a growth promoter in Eucalyptus in vitro cultures. Graphical abstract for the methodology using cultivation independent and dependent methodologies to investigate the endophytic bacteria community from in vitro Eucalyptus urophylla BRS07-01.

Rhizobia and endophytic bacteria isolated from rainforest fragments within an iron ore mining site of the Eastern Brazilian Amazon.

The aim of the present study was to isolate and evaluate the diversity of rhizobial and endophytic bacterial strains from undisturbed native rainforests within an iron ore mining site of the Serra Norte de Carajás in the Eastern Brazilian Amazon region to assess their biotechnological utility in reclamation of areas. Experiments were conducted to capture strains from samples of the soil of these forests at the sites Arenito II, Noroeste II, and Sul IV using Macroptilium atropurpureum and Mimosa acutistipula var. ferrea as trap host plants. Only M. atropurpureum nodulated, and the different bacterial strains were isolated from its nodules. There was no difference in the number of nodules among the areas, but the Arenito II bacterial community was the most efficient, indicated by the aboveground biomass production and suitable shoot mass/root mass ratio. Fifty-two (52) bacterial isolates were obtained, distributed in five groups, including nodulating and endophytic bacteria: 32 from Arenito II, 12 from Noroeste II, and 8 from Sul IV. The nodulating Bradyrhizobium genus was common to the three areas, whereas Paraburkholderia was found only in Arenito II. The nodD1 gene was amplified in all the strains of both nodulating genera. Strains of the nodulating genus Methylobacterium were also isolated from the three areas; however, they did not nodulate the host of origin, and their nodD1 gene was not amplified. Endophytic strains were also isolated from the genera Paenibacillus, Pantoea, and Leifsonia in Arenito II, Leifsonia in Noroeste I, and Paenibacillus in Sul IV. The greater nodulation and rhizobial and endophytic bacterial diversity observed in Arenito II were probably due to the more suitable edaphic properties of the area. The isolated strains were incorporated in the collection of the Department of Soil Science of UFLA and will be investigated in relation to their symbiotic characteristics with native host plants, as well as their ability to perform other biological processes.

Fungal endophytes from leaves of Mandevilla catimbauensis (Apocynaceae): diversity and potential for L-asparaginase production.

In this study, we examined endophytic fungi in leaves of Mandevilla catimbauensis, an endemic plant species found in the Brazilian dry forest (Caatinga), and endophytic fungi's potential to produce L-asparaginase (L-ASNase). In total, 66 endophytes were isolated, and the leaf-fragment colonisation rate was 11.78%. Based on morphology, internal transcribed spacer (ITS), and partial large subunit (LSU) of ribosomal DNA sequencing, the endophytic fungi isolated belonged to six Ascomycota orders (Botryosphaeriales, Capnodiales, Diaporthales, Eurotiales, Marthamycetales, and Pleosporales). Phyllosticta species were the most frequent endophytes isolated (23 isolates [45.1%] from two species). The Shannon-Wiener and Fisher alpha index average values were 0.56 and 3.26, respectively. Twenty endophytes were randomly selected for the L-ASNase production test, of which fourteen isolates showed potential to produce the enzyme (0.48-2.22 U g-1), especially Phyllosticta catimbauensis URM 7672 (2.22 U g-1) and Cladosporium sp. G45 (2.11 U g-1). Phyllosticta catimbauensis URM 7672 was selected for the partial optimisation of L-ASNase production because of its ability to generate considerable amounts of enzyme. We obtained the highest L-ASNase activity (3.47 U g-1), representing an increase of 36.02% in enzymatic production, under the following experimental conditions: a pH of 4.2, 1.0% inoculum concentration, and 2.5% L-asparagine concentration. Our study showed that M. catimbauensis harbours an important diversity of endophytic fungi with biotechnological potential for L-ASNase production.

Antifungal potential against Sclerotinia sclerotiorum (Lib.) de Bary and plant growth promoting abilities of Bacillus isolates from canola (Brassica napus L.) roots.

Endophytic bacteria show important abilities in promoting plant growth and suppressing phytopathogens, being largely explored in agriculture as biofertilizers or biocontrol agents. Bacteria from canola roots were isolated and screened for different plant growth promotion (PGP) traits and biocontrol of Sclerotinia sclerotiorum. Thirty isolates belonging to Bacillus, Paenibacillus, Lysinibacillus, and Microbacterium genera were obtained. Several isolates produced auxin, siderophores, hydrolytic enzymes, fixed nitrogen and solubilized phosphate. Five isolates presented antifungal activity against S. sclerotiorum by the dual culture assay and four of them also inhibited fungal growth by volatile organic compounds production. All antagonistic isolates belonged to the Bacillus genus, and had their genomes sequenced for the search of biosynthetic gene clusters (BGC) related to antimicrobial metabolites. These isolates were identified as Bacillus safensis (3), Bacillus pumilus (1), and Bacillus megaterium (1), using the genomic metrics ANI and dDDH. Most strains showed several common BGCs, including bacteriocin, polyketide synthase (PKS), and non-ribosomal peptide synthetase (NRPS), related to pumilacidin, bacillibactin, bacilysin, and other antimicrobial compounds. Pumilacidin-related mass peaks were detected in acid precipitation extracts through MALDI-TOF analysis. The genomic features demonstrated the potential of these isolates in the suppression of plant pathogens; however, some aspects of plant-bacterial interactions remain to be elucidated.

Endophytic actinobacteria of Hymenachne amplexicaulis from the Brazilian Pantanal wetland produce compounds with antibacterial and antitumor activities.

The increase in the number of deaths from infections caused by multidrug-resistant bacteria and cancer diseases highlights the need for new molecules with biological activity. Actinobacteria represent a potential source of new compounds, as these microorganisms have already produced a great diversity of clinically employed antibiotics. Endophytes from unexplored biomes, such as the Pantanal (the largest wetland in the world), can be a source of new molecules. Hymenachne amplexicaulis is among the unexplored native plants of the Pantanal in terms of its endophytic community. This plant is considered a weed in other countries due to its ability to adapt and compete with native plants, and there is evidence to suggest that the endophytic community of H. amplexicaulis plays an important role in this competitiveness. To explore its therapeutic potential, the present study isolated, identified (using partial sequence of the 16S rDNA) and bioprospected H. amplexicaulis endophytic actinobacteria. Ten isolates belonging to the genera Streptomyces, Microbispora, Leifsonia, and Verrucosispora were obtained from root fragments. The susceptibility profile of the isolates to the different classes of antibiotics was evaluated, with 80 % of the isolates showing resistance to the antibiotics Nalidixic Acid, Ampicillin, Chloramphenicol, Oxacillin, and Rifampicin. To assess antibacterial and antitumor activities, methanolic extracts were obtained by fermentation in SG culture medium at 36 °C at 180 rpm for 10 days. The extract produced from the S. albidoflavus CMRP4854 isolate was the only one to show activity against the Gram-negative bacterium Acinetobacter baumanii. Due to the great clinical importance of this pathogen and the difficulty in obtaining active compounds against it, the CMRP4854 isolate should be further investigated for the identification of active compounds and mode of action. We also emphasize the results obtained by the extract of the isolates Streptomyces albidoflavus CMRP4852 and Verrucosispora sp. CMRP4860 that presented antibacterial effect against Methicilin-resistant Staphylococcus aureus (MRSA) (MIC: 1.5 µg/mL and 13 µg/mL, respectively) and Vancomycin-resistant Enterococcus (VRE) (MIC: 40 µg/mL for both extracts). Extracts (200 µg/mL) of these two endophytes also showed selective cytotoxicity action against murine B16-F10 melanoma cells. However, the CMRP4852 extract also affected the density of normal cells. Due to these results, the crude extract of isolate CMRP4860 Verrucosispora sp., which was the only one that presented cytotoxicity and reduced cell density only in tumor cells, was selected for subsequent analysis involving scale-up fermentation of the CMRP4860 resulting in 9 fractions that were tested against both bacteria and tumor cells, with particular fractions showing promise and meriting further investigation. Taken together, the results of this study not only show for the first time that the endophytic community of H. amplexicaulis actinobacteria can produce secondary metabolites that potentially possess important antibacterial and cytotoxic properties, but also reinforce the pressing need to conserve biomes such as the Brazilian Pantanal.

Fungal endophytes in Peperomia obtusifolia and their potential as inhibitors of chickpea fungal pathogens.

Chickpea (Cicer arietinum L., Fabaceae) is the second most important legume after common bean (Phaseolus vulgaris L., Fabaceae) and third in production among the legumes grains worldwide. Ascochyta blight and Fusarium wilt are among the main fungal infections which cause the major losses of chickpea crop. In this work we report the phyto-pathogen controlling properties of 24 endophyte Phomopsis/Diaporthe isolates on the chickpea fungal pathogens Ascochyta rabiei, Fusarium oxysporum and Fusarium solani. The Phomopsis/Diaporthe strains were isolated amongst a total of 62 endophytic fungi from the aerial parts of the herbaceous perennial American plant Peperomia obtusifolia (Piperaceae) along with Fusarium, Septoria, Colletotrichum, Alternaria and Roussoella genera among others. Phomopsis/Diaporthe isolates were identified as Diaporthe infecunda (12 isolates), Diaporthe sackstoni (1 isolate), Diaporthe cf. brasiliensis (4 isolates) and Phomopsis cf. tuberivora (7 isolates). All the Phomopsis/Diaporthe strains antagonized A. rabiei strain AR2 with a mean of inhibition (% I) of 86.59 ± 1.49% in dual cultures. The metabolic characterization of the Phomopsis/Diaporthe strains showed groups in three clusters which were in agreement with the taxonomic identification. Bioautographic evaluation of organic extracts showed that those of D. cf. brasiliensis and D. infecunda were better as inhibitors. Strain Po 45 was one of the most active (cluster 1, 96.87% I), and its ethyl acetate extract inhibited A. rabiei growth in a bioautographic assay until at least 10 µg/mm applied showing a specific chromatographic band as the responsible of the A. rabiei inhibition.

Volatile organic compounds profile synthesized and released by endophytes of tomato (Solanum lycopersici L.) and their antagonistic role.

The endophytic microbiome uses mechanisms such as the secretion of diffusible antibiotic molecules, synthesis and release of volatile organic compounds, and/or toxins to protect plants. The aim of this research was to study the volatile organic compounds (VOCs) profile as well as the diffusible secondary metabolites produced and released by endophytic bacteria isolated from tomato plants that in in-vitro assays prevented growth of pathogenic fungi. Bacteria belonging to seven genera (Acinetobacter, Arthrobacter, Bacillus, Microbacterium, Pantoea, Pseudomonas, and Stenotrophomonas) were isolated from different tissues of tomato plants with and without symptoms of Gray leaf spot, a disease provoked by Stemphylium lycopersici. In vitro, antagonistic assays were performed and the effect of volatile and soluble compounds released by endophytic bacteria on the growth of pathogenic fungi was determined. The VOCs synthesized by the endophytes were extracted, identified and quantified. These isolates representatives of seven bacterial genera inhibited the growth of three fungal pathogens of tomato S. lycopersici, Alternaria alternata and Corynespora cassiicola, which was related to the synthesis of soluble compounds as well as VOCs. Endophytes synthesize and release different VOCs, probably due to the different type of interaction that each bacterium establishes with the fungus, presenting a range of fungal growth inhibition.

Endophytic fungus diversity in soybean plants submitted to conditions of elevated atmospheric CO2 and temperature.

Global climatic changes can have drastic impacts on plant species, including severe consequences for the agricultural species productivity. Many of these species present important mutualisms with endophytic fungi that positively influence their performance. The present study evaluated whether the increases in CO2 and temperature predicted for the year 2100 may cause changes in foliar carbon (C) and nitrogen (N) concentrations in soybean (Glycine max) and, consequently, the interactions with its endophytic fungi. The effects of elevated CO2 and temperature were evaluated in four treatments in open-top chambers: (i) control, (ii) increased temperature, (iii) increased CO2, and (iv) increased CO2 and temperature. Increased atmospheric CO2 resulted in decreased foliar N concentration, while increased temperature increased it. A total of 16 taxa of endophytic fungi were identified based on sequencing internal transcribed spacer regions of rRNA subunits. Increased atmospheric CO2 and temperature were observed to potentially modify the endophytic mycobiota of soybean plants. The results suggest that the fungi species substitution is a consequence of changes in foliar N concentration and C/N ratio. Predicted climatic changes shall affect the relationships between plant and endophytes, which in turn, will affect the performance and resistance of soybean, one of the most important crops in the world.

Multifunctional potential of endophytic bacteria from Anacardium othonianum Rizzini in promoting in vitro and ex vitro plant growth.

Anacardium othonianum Rizzini, a cashew tree native to the Brazilian Cerrado, is economically important due to its applications in the food, chemical and pharmaceutical industries. However, A. othonianum yields a crop with low productivity due to a number of factors, such as nutritionally poor soils, drought and losses due to pests and diseases. Brazil is one of the nine largest cashew nut producers worldwide, and sustainable technologies are needed to increase the productivity of this crop. In this context, the use of endophytic microorganisms could promote plant growth and provide protection against phytopathogens. In this study, the isolation of the root endophytic community of A. othonianum led to the characterization of 22 distinct bacterial strains with multifunctional traits for plant growth promotion. The results of in vitro assays to assess auxin synthesis, phosphate solubilization, phosphatase and siderophore production and biocontrol against Fusarium oxysporum led to the selection of Acinetobacter lwoffii Bac109 and Pantoea agglomerans Bac131 as the most promising strains. The reinoculation of the Bac109 and Bac131 strains onto A. othonianum seeds showed that the treatment containing a mixture of these strains was the most effective in promoting increases in the biometric parameters of early plant growth. Thus, this study highlights the biotechnological potential of a consortium of A. lwoffii Bac109 and P. agglomerans Bac131 for future applications in sustainable cashew cultivation.