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This study aimed to evaluate the genomic profile of the Antarctic marine Curtobacterium sp. CBMAI 2942, as well as to optimize the conditions for chitinase production and antifungal potential for biological control. Assembly and annotation of the genome confirmed the genomic potential for chitinase synthesis, revealing two ChBDs of chitin binding (Chi C). The optimization enzyme production using an experimental design resulted in a 3.7-fold increase in chitinase production. The chitinase enzyme was identified by SDS-PAGE and confirmed through mass spectrometry analysis. The enzymatic extract obtained using acetone showed antifungal activity against the phytopathogenic fungus Aspergillus sp. series Nigri CBMAI 1846. The genetic capability of Curtobacterium sp. CBMAI 2942 for chitin degradation was confirmed through genomic analysis. The basal culture medium was adjusted, and the chitinase produced by this isolate from Antarctica showed significant inhibition against Aspergillus sp. Nigri series CBMAI 1846, which is a tomato phytopathogenic fungus. This suggests that this marine bacterium could potentially be used as a biological control of agricultural pests.

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A polyphasic approach was applied to characterize taxonomically a novel endophytic bacterial strain, designated as EP178T, which was previously isolated from Passiflora incarnata leaves and characterized as plant-growth promoter. The strain EP178T forms Gram stain-negative and rod-shaped cells, and circular and yellow-pigmented colonies. Its growth occurs at 10-37 °C, at pH 6.0-8.0, and tolerates up to 7% (w/v) NaCl. The major cellular fatty acids found were summed feature 8 (C18:1 ω7c), summed feature 3 (C16:1 ω6c /C16:1 ω7c), and C16:0, and the predominant ubiquinone was Q-9. The phylogenetic and nucleotide-similarity analysis with 16S rRNA gene sequences showed that strain EP178T belongs to Pseudomonas genus. The genomic-based G + C content was 65.5%. The average nucleotide identity and digital DNA-DNA hybridization values between strains EP178T and the closest type strain, P. oryzihabitans DSM 6835T, were 92.6% and 52.2%, respectively. Various genes associated with plant-growth promoting mechanisms were annotated from genome sequences. Based on the phenotypic, genomic, phylogeny and chemotaxonomic data, strain EP178T represents a new species of the genus Pseudomonas, for which the name Pseudomonas flavocrustae sp. nov. was proposed. The type strain is EP178T (= CBMAI 2609T = ICMP 24844T = MUM 23.01T).

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RESUMEN

This study aimed to evaluate the genomic profile of the Antarctic marine Curtobacterium sp. CBMAI 2942, as well as to optimize the conditions for chitinase production and antifungal potential for biological control. Assembly and annotation of the genome confirmed the genomic potential for chitinase synthesis, revealing two ChBDs of chitin binding (Chi C). The optimization enzyme production using an experimental design resulted in a 3.7-fold increase in chitinase production. The chitinase enzyme was identified by SDS-PAGE and confirmed through mass spectrometry analysis. The enzymatic extract obtained using acetone showed antifungal activity against the phytopathogenic fungus Aspergillus sp. series Nigri CBMAI 1846. The genetic capability of Curtobacterium sp. CBMAI 2942 for chitin degradation was confirmed through genomic analysis. The basal culture medium was adjusted, and the chitinase produced by this isolate from Antarctica showed significant inhibition against Aspergillus sp. Nigri series CBMAI 1846, which is a tomato phytopathogenic fungus. This suggests that this marine bacterium could potentially be used as a biological control of agricultural pests.

RESUMEN

Soybean crops are of great economic importance worldwide and in Brazil. This crop is a commodity that provides large amounts of financial resources to the country. Soybean productivity is influenced by several biotic and abiotic factors, and most of these factors cannot be controlled by agricultural practices. Due to the soybean cultivars used and their required yields, the soybean crop, similar to other agriculturally important crops, requires large amounts of mineral fertilizers. There are several microorganisms that colonize soybean plant roots without causing symptoms or damage. These microorganisms that colonize plant tissues are called endophytes and can often promote plant growth and development. Little is known about the factors that influence endophyticism. The aim of the present study was to evaluate whether Bacillus subtilis inoculant concentrations and levels of mineral fertilization recommended for the crop have any influence on the endophytic microbiome of soybean plant roots. The results show that B. subtilis inoculations did not affect the endophytic community of the roots; however, the evaluation of the microbial community structure according to the alpha diversity metrics observed richness, Chao1 index, Shannon index and Simpson index showed that microbial diversity of endophytes was higher at fertilization levels of 50 and 100%, with a significant difference (p < 0.05) between 0 and 50% and 0 and 100% fertilization.

RESUMEN

Endophytes are considered one of the most important microbial resources for obtaining biomolecules of therapeutic use. Passiflora incarnata, widely employed by the pharmaceutical industry, shows therapeutic effects on anxiety, nervousness, constipation, dyspepsia and insomnia based on their antioxidant compounds. In this study, from 315 endophytic fungi isolated from P. incarnata leaves, 60 were selected to determinate presence of chemical constituents related with antioxidant activity, based on their production of soluble pigments. The promising fungi were evaluated specifically on their potential to produce phenolic compounds, flavonoids and for antioxidant activity. Five isolates significantly produced flavonoids and phenolic compounds in the ethyl acetate and n-Butanol extracts, also saponins and high antioxidant activity against the DPPH (2.2-diphenyl-1-picrylhydrazyl) free radical. A strain of Aspergillus nidulans var. dentatus (former Emericella dentata) was able to produce tannins as well; its butanolic extract was very similar than the BHT (butylated hydroxytoluene) (94.3% × 94.32%) and Rutin (95.8%) reference substances in the DPPH radical scavenging. Similarly, a Chaetomium strain exhibited 93.6% and 94.7% of antioxidant activity in their ethyl acetate and butanolic fractions, respectively. The chromatographic analysis of the ethyl acetate fraction from the Aspergillus strain revealed the production of orcinol (3.19%). Four-methoxymethylphenol (4.79%), sorbicillin (33.59%) and ergosterol (23.08%) was produced by Trichoderma longibrachiatum and isopropenyl-1,4-dimethyl-1,2,3,3a,4,5,6,7-octahydroazulene were found in two Fusarium oxysporum strains. The phytochemical screening showed that all analyzed fungi were able to produce a kind of secondary metabolite (phenols, flavonoids, tannins and/or saponins). The study shows a great underexplored potential for industrial application of P. incarnata endophytes.

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Bacteria exhibiting beneficial traits like increasing the bioavailability of essential nutrients and modulating hormone levels in plants are known as plant growth promoting (PGP) bacteria. The occurrence of this specific group of bacteria in the endophytic environment may reflect the decisive role they play in a particular condition. This study aimed to determine the taxonomical diversity of the culturable bacterial endophytes, isolated in the vegetative stage of passionflower (Passiflora incarnata), and assess its potential to promote plant growth by phenotypic and genotypic approaches. The sequencing and phylogenetic analysis of the 16S rRNA gene allowed us to classify 58 bacterial endophytes into nine genera. Bacillus (70.7%) was the most dominant genus, followed by Pseudomonas (8.6%) and Pantoea (6.9%). A few isolates belonged to Rhodococcus and Paenibacillus, whereas the genera Lysinibacillus, Microvirga, Xanthomonas, and Leclercia were represented by only one isolate. The strains were tested for nitrogen fixation, phosphate solubilization, indole-acetic-acid synthesis, and siderophore production. Moreover, PGP related genes (nifH, ipdC, asb, and AcPho) were detected by PCR-based screening. Most of the isolates (94.8%) displayed a potential for at least one of the PGP traits tested by biochemical assays or PCR-based screening. Nine strains were selected based on results from both approaches and were evaluated for boosting the Cape gooseberry (Physalis peruviana) germination and growth. All tested isolates improved germination in vitro, and the majority (78%) increased growth parameters in vivo. The results suggested that most of culturable bacteria inhabiting P. incarnata in the vegetative stage could be used as probiotics for agricultural systems. Besides, their occurrence may be associated with specific physiological needs typical of this development stage.