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      Isolation, abundance and phylogenetic affiliation of endophytic actinomycetes associated with medicinal plants and screening for their in vitro antimicrobial biosynthetic potential

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          Abstract

          Microorganisms associated with medicinal plants are of interest as the producers of important bioactive compounds. To date, the diversity of culturable endophytic actinomycetes associated with medicinal plants is in its initial phase of exploration. In this study, 42 endophytic actinomycetes were isolated from different organs of seven selected medicinal plants. The highest number of isolates ( n = 22, 52.3%) of actinomycetes was isolated from roots, followed by stems ( n = 9, 21.4%), leaves ( n = 6, 14.2%), flowers ( n = 3, 7.1%), and petioles ( n = 2, 4.7%). The genus Streptomyces was the most dominant among the isolates (66.6%) in both the locations (Dampa TRF and Phawngpuii NP, Mizoram, India). From a total of 42 isolates, 22 isolates were selected for further studies based on their ability to inhibit one of the tested human bacterial or fungal pathogen. Selected isolates were identified based on 16S rRNA gene analysis and subsequently the isolates were grouped to four different genera; Streptomyces, Brevibacterium, Microbacterium, and Leifsonia. Antibiotic sensitivity assay was performed to understand the responsible antimicrobials present in the isolates showing the antimicrobial activities and revealed that the isolates were mostly resistant to penicillin G and ampicillin. Further, antimicrobial properties and antibiotic sensitivity assay in combination with the results of amplification of biosynthetic genes polyketide synthase (PKS-I) and non-ribosomal peptide synthetase (NRPS) showed that the endophytic actinomycetes associated with the selected medicinal plants have broad-spectrum antimicrobial activity. This is the first report of the isolation of Brevibacterium sp., Microbacterium sp., and Leifsonia xyli from endophytic environments of medicinal plants, Mirabilis jalapa and Clerodendrum colebrookianum. Our results emphasize that endophytic actinomycetes associated with medicinal plants are an unexplored resource for the discovery of biologically active compounds.

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          Most cited references33

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          Natural products from endophytic microorganisms.

          Endophytic microorganisms are to be found in virtually every plant on earth. These organisms reside in the living tissues of the host plant and do so in a variety of relationships ranging from symbiotic to pathogenic. Endophytes may contribute to their host plant by producing a plethora of substances that provide protection and ultimately survival value to the plant. Ultimately, these compounds, once isolated and characterized, may also have potential for use in modern medicine, agriculture, and industry. Novel antibiotics, antimycotics, immunosuppressants, and anticancer compounds are only a few examples of what has been found after the isolation and culturing of individual endophytes followed by purification and characterization of some of their natural products. The prospects of finding new drugs that may be effective candidates for treating newly developing diseases in humans, plants, and animals are great. Other applications in industry and agriculture may also be discovered among the novel products produced by endophytic microbes.
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            Biosynthesis of nonribosomal peptides1.

            Bacteria and fungi use large multifunctional enzymes, the so-called nonribosomal peptide synthetases (NRPSs), to produce peptides of broad structural and biological activity. Biochemical studies have contributed substantially to the understanding of the key principles of these modular enzymes that can draw on a much larger number of catalytic tools for the incorporation of unusual features compared with the ribosomal system. Several crystal structures of NRPS-domains have yielded deep insight into the catalytic mechanisms involved and have led to a better prediction of the products assembled and to the construction of hybrid enzymes. In addition to the structure-function relationship of the core- and tailoring-domains of NRPSs, which is the main focus of this review, different biosynthetic strategies and essential enzymes for posttranslational modification and editing are discussed.
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              Isolation and identification of actinobacteria from surface-sterilized wheat roots.

              This is the first report of filamentous actinobacteria isolated from surface-sterilized root tissues of healthy wheat plants (Triticum aestivum L.). Wheat roots from a range of sites across South Australia were used as the source material for the isolation of the endophytic actinobacteria. Roots were surface-sterilized by using ethanol and sodium hypochlorite prior to the isolation of the actinobacteria. Forty-nine of these isolates were identified by using 16S ribosomal DNA (rDNA) sequencing and found to belong to a small group of actinobacterial genera including Streptomyces, Microbispora, Micromonospora, and Nocardiodes spp. Many of the Streptomyces spp. were found to be similar, on the basis of their 16S rDNA gene sequence, to Streptomyces spp. that had been isolated from potato scabs. In particular, several isolates exhibited high 16S rDNA gene sequence homology to Streptomyces caviscabies and S. setonii. None of these isolates, nor the S. caviscabies and S. setonii type strains, were found to carry the nec1 pathogenicity-associated gene or to produce the toxin thaxtomin, indicating that they were nonpathogenic. These isolates were recovered from healthy plants over a range of geographically and temporally isolated sampling events and constitute an important plant-microbe interaction.
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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
                07 April 2015
                2015
                : 6
                : 273
                Affiliations
                [1] 1Molecular Microbiology and Systematics Laboratory, Department of Biotechnology, Mizoram University Aizawl, India
                [2] 2Biotechnology Division, CSIR-North East Institute of Science and Technology Jorhat, Assam, India
                [3] 3Molecular Glyco-biotechnology Group, Department of Biochemistry, National University of Ireland Galway Galway, Ireland
                Author notes

                Edited by: Andrea Genre, University of Turin, Italy

                Reviewed by: Marcela Claudia Pagano, Federal University of Minas Gerais, Brazil; Atul Pandey, Medical University of South Carolina, USA

                *Correspondence: Vijai K. Gupta, Molecular Glyco-biotechnology Group, Department of Biochemistry, National University of Ireland Galway, Galway, Ireland vijaifzd@ 123456gmail.com ;
                Bhim P. Singh, Molecular Microbiology and Systematics Laboratory, Department of Biotechnology, Mizoram University, Tanharil, Aizawl, Mizoram-796004, India bhimpratap@ 123456gmail.com

                This article was submitted to Plant-Microbe Interaction, a section of the journal Frontiers in Microbiology

                Article
                10.3389/fmicb.2015.00273
                4388002
                25904906
                f544c4c4-fcf4-4ea7-9b76-4642c82dc383
                Copyright © 2015 Passari, Mishra, Saikia, Gupta and Singh.

                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) or licensor 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
                : 03 December 2014
                : 19 March 2015
                Page count
                Figures: 6, Tables: 3, Equations: 0, References: 52, Pages: 13, Words: 8545
                Categories
                Plant Science
                Original Research

                Microbiology & Virology
                endophytic actinomycetes,16s rrna gene,antibiotic sensitivity,polyketide synthase (pks-i),non-ribosomal peptide synthetase (nrps)

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