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Halophytes-associated endophytic and rhizospheric bacteria: diversity,antagonism and metabolite production
Authors:Fehmida Bibi  Gary Allan Strobel  Muhammad Imran Naseer  Muhammad Yasir  Ahmed Abdullah Khalaf Al-Ghamdi  Esam Ibrahim Azhar
Affiliation:1. Special Infectious Agents Unit, King Fahd Medical Research Center, King Abdulaziz University, Jeddah, Saudi Arabiafehmeedaimran@yahoo.com"ORCIDhttp://orcid.org/0000-0002-5178-1471;2. Department of Plant Sciences, Montana State University, Bozeman, MT, USA;3. Center of Excellence in Genomic Medicine Research (CEGMR), King Abdulaziz University, Jeddah, Saudi Arabia;4. Special Infectious Agents Unit, King Fahd Medical Research Center, King Abdulaziz University, Jeddah, Saudi Arabia;5. Department of Medical Laboratory Technology, Faculty of Applied Medical Sciences, King Abdulaziz University, Jeddah, Saudi Arabia;6. Department of Medical Laboratory Technology, Faculty of Applied Medical Sciences, King Abdulaziz University, Jeddah, Saudi Arabia
Abstract:In Saudi Arabia, halophytes occupy tidal and intertidal forest ecosystems. They and their associated microflora have immense potential to yield novel and important useful natural products. Three halophytes (Avicennia marina, Halocnemum strobilaceum, Zygophyllum qatarense) were targeted for the isolation and identification of populations of endophytic and rhizospheric bacteria having antimicrobial potential. A total 554 bacterial isolates were initially screened against oomycetes fungal pathogens, Phytophthora capsici and Pythium ultimum. Of these, only 57 rhizospheric and endophytic bacteria exhibited inhibition against the targeted bioassay oomycetes. Tentative identification of the bacteria was on the basis of 16S rRNA gene sequences which revealed 92–100% sequence identity to type strains of related species and placed these organisms in six major classes: Actinobacteria, γ-Proteobacteria, Firmicutes, α-Proteobacteria, Flavobacteriia and β-Proteobacteria. When checked for lytic enzyme production, mostly the isolates of Actinobacteria and Firmicutes were potential enzyme producers. Detection of secondary metabolite biosynthetic genes – type I polyketide synthases, type II polyketide synthases and nonribosomal peptide synthetases – confirmed that 21 (35.5%) isolates were positive for at least one type of the biosynthetic gene. In order to identify metabolites, three isolates, Alteromonas australica (EA73), Aidingimonas halophila (EA105) and Halomonas zincidurans (EA127), were selected and subjected to chemical analyses using liquid chromatography–mass spectrometry and gas chromatography–mass spectrometry. Both analyses showed the presence of different bioactive compounds in the culture extracts of isolates some of which are already reported for their diverse biological activities such as 2, 4-Diacetylphloroglucinol. Our results demonstrated that halophytes represent an important source of potentially active bacteria producing antifungal metabolites of medical significance.
Keywords:Halophytes  antagonistic bacteria  enzyme production  16s rRNA gene sequence  metabolites identification
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