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      Endophytes of Brazilian Medicinal Plants With Activity Against Phytopathogens

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          Abstract

          Plant diseases caused by phytopathogens are responsible for significant crop losses worldwide. Resistance induction and biological control have been exploited in agriculture due to their enormous potential. In this study, we investigated the antimicrobial potential of endophytic fungi of leaves and petioles of medicinal plants Vochysia divergens and Stryphnodendron adstringens located in two regions of high diversity in Brazil, Pantanal, and Cerrado, respectively. We recovered 1,304 fungal isolates and based on the characteristics of the culture, were assigned to 159 phenotypes. One isolate was selected as representative of each phenotype and studied for antimicrobial activity against phytopathogens. Isolates with better biological activities were identified based on DNA sequences and phylogenetic analyzes. Among the 159 representative isolates, extracts from 12 endophytes that inhibited the mycelial growth (IG) of Colletotrichum abscissum (≥40%) were selected to expand the antimicrobial analysis. The minimum inhibitory concentrations (MIC) of the extracts were determined against citrus pathogens, C. abscissum, Phyllosticta citricarpa and Xanthomonas citri subsp. citri and the maize pathogen Fusarium graminearum. The highest activity against C. abscissum were from extracts of Pseudofusicoccum stromaticum CMRP4328 (IG: 83% and MIC: 40 μg/mL) and Diaporthe vochysiae CMRP4322 (IG: 75% and MIC: 1 μg/mL), both extracts also inhibited the development of post-bloom fruit drop symptoms in citrus flowers. The extracts were promising in inhibiting the mycelial growth of P. citricarpa and reducing the production of pycnidia in citrus leaves. Among the isolates that showed activity, the genus Diaporthe was the most common, including the new species D. cerradensis described in this study. In addition, high performance liquid chromatography, UV detection, and mass spectrometry and thin layer chromatography analyzes of extracts produced by endophytes that showed high activity, indicated D. vochysiae CMRP4322 and P. stromaticum CMRP4328 as promising strains that produce new bioactive natural products. We report here the capacity of endophytic fungi of medicinal plants to produce secondary metabolites with biological activities against phytopathogenic fungi and bacteria. The description of the new species D. cerradensis, reinforces the ability of medicinal plants found in Brazil to host a diverse group of fungi with biotechnological potential.

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          MEGA7: Molecular Evolutionary Genetics Analysis Version 7.0 for Bigger Datasets.

          We present the latest version of the Molecular Evolutionary Genetics Analysis (Mega) software, which contains many sophisticated methods and tools for phylogenomics and phylomedicine. In this major upgrade, Mega has been optimized for use on 64-bit computing systems for analyzing larger datasets. Researchers can now explore and analyze tens of thousands of sequences in Mega The new version also provides an advanced wizard for building timetrees and includes a new functionality to automatically predict gene duplication events in gene family trees. The 64-bit Mega is made available in two interfaces: graphical and command line. The graphical user interface (GUI) is a native Microsoft Windows application that can also be used on Mac OS X. The command line Mega is available as native applications for Windows, Linux, and Mac OS X. They are intended for use in high-throughput and scripted analysis. Both versions are available from www.megasoftware.net free of charge.
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            MrBayes 3.2: Efficient Bayesian Phylogenetic Inference and Model Choice Across a Large Model Space

            Since its introduction in 2001, MrBayes has grown in popularity as a software package for Bayesian phylogenetic inference using Markov chain Monte Carlo (MCMC) methods. With this note, we announce the release of version 3.2, a major upgrade to the latest official release presented in 2003. The new version provides convergence diagnostics and allows multiple analyses to be run in parallel with convergence progress monitored on the fly. The introduction of new proposals and automatic optimization of tuning parameters has improved convergence for many problems. The new version also sports significantly faster likelihood calculations through streaming single-instruction-multiple-data extensions (SSE) and support of the BEAGLE library, allowing likelihood calculations to be delegated to graphics processing units (GPUs) on compatible hardware. Speedup factors range from around 2 with SSE code to more than 50 with BEAGLE for codon problems. Checkpointing across all models allows long runs to be completed even when an analysis is prematurely terminated. New models include relaxed clocks, dating, model averaging across time-reversible substitution models, and support for hard, negative, and partial (backbone) tree constraints. Inference of species trees from gene trees is supported by full incorporation of the Bayesian estimation of species trees (BEST) algorithms. Marginal model likelihoods for Bayes factor tests can be estimated accurately across the entire model space using the stepping stone method. The new version provides more output options than previously, including samples of ancestral states, site rates, site d N /d S rations, branch rates, and node dates. A wide range of statistics on tree parameters can also be output for visualization in FigTree and compatible software.
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              AMPLIFICATION AND DIRECT SEQUENCING OF FUNGAL RIBOSOMAL RNA GENES FOR PHYLOGENETICS

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                Author and article information

                Contributors
                Journal
                Front Microbiol
                Front Microbiol
                Front. Microbiol.
                Frontiers in Microbiology
                Frontiers Media S.A.
                1664-302X
                01 September 2021
                2021
                : 12
                : 714750
                Affiliations
                [1] 1Postgraduate Program of Microbiology, Parasitology and Pathology, Department of Pathology, Federal University of Paraná , Curitiba, Brazil
                [2] 2Department of Biomedicine, Centro Universitário Católica de Santa Catarina , Joinville, Brazil
                [3] 3Postgraduate Program of Genetics, Federal University of Paraná , Curitiba, Brazil
                [4] 4Department of General and Applied Biology, Biosciences Institute, State University of São Paulo , Rio Claro, Brazil
                [5] 5Department of Pharmaceutical Sciences, College of Pharmacy, University of Kentucky , Lexington, KY, United States
                [6] 6Center for Pharmaceutical Research and Innovation, College of Pharmacy, University of Kentucky , Lexington, KY, United States
                Author notes

                Edited by: Jia Liu, Chongqing University of Arts and Sciences, China

                Reviewed by: Sunil Kumar Deshmukh, The Energy and Resources Institute (TERI), India; Madhuree Kumari, Indian Institute of Science (IISc), India

                *Correspondence: Khaled A. Shaaban, khaled_shaaban@ 123456uky.edu

                ORCID: Chirlei Glienke, orcid.org/0000-0003-3394-0659; Jucélia Iantas, orcid.org/0000-0002-1203-7543; Jürgen Rohr, orcid.org/0000-0001-6447-5951; Jon S. Thorson, orcid.org/0000-0002-7148-0721; Khaled A. Shaaban, orcid.org/0000-0001-7638-4942

                This article was submitted to Microbe and Virus Interactions with Plants, a section of the journal Frontiers in Microbiology

                Article
                10.3389/fmicb.2021.714750
                8442585
                1b7de003-be94-4a00-b397-cee3e5c6854e
                Copyright © 2021 Iantas, Savi, Schibelbein, Noriler, Assad, Dilarri, Ferreira, Rohr, Thorson, Shaaban and Glienke.

                This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

                History
                : 25 May 2021
                : 03 August 2021
                Page count
                Figures: 6, Tables: 1, Equations: 0, References: 125, Pages: 18, Words: 0
                Funding
                Funded by: Conselho Nacional de Desenvolvimento Científico e Tecnológico 10.13039/501100003593
                Award ID: INCT 465440/2014-2
                Award ID: 309971/2016-0
                Award ID: 424738/2016-3
                Funded by: Fundação de Amparo à Pesquisa do Estado de São Paulo 10.13039/501100001807
                Award ID: 2015/50162-2
                Funded by: Foundation for the National Institutes of Health 10.13039/100000009
                Award ID: CA243529
                Award ID: R01 GM115261
                Funded by: National Center for Advancing Translational Sciences 10.13039/100006108
                Award ID: UL1TR000117
                Award ID: UL1TR001998
                Funded by: University of Kentucky 10.13039/100007472
                Funded by: Center for Pharmaceutical Research and Innovation, University of Kentucky 10.13039/100017502
                Award ID: CPRI, NIH P20 GM130456
                Categories
                Microbiology
                Original Research

                Microbiology & Virology
                phytopathogens,endophytes,secondary metabolites,natural product,endophytic fungi

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