Isolation of endophytic fungi from South African plants, and screening for their antimicrobial and extracellular enzymatic activities and presence of type I polyketide synthases

Endophytes are bacteria or fungi which live inside the host plant and participate in many biological processes without causing disease or other adverse effects. Endophytes are recognised as a rich source of secondary metabolites with potentially useful pharmacological properties. Many South African medicinal plants are highly under-investigated sources of potentially useful endophytic microbes. In this report six endophytic fungi were obtained from the leaves, stems and roots of South African medicinal plants which are known for their traditional uses and pharmacological properties. The endophytic fungi were isolated from Cotyledon orbiculata L., Psychotria zombamontana (Kuntze) Petit, Tecomaria capensis (Thunb.) Lindl., Catha edulis (Vahl) Endl. and Melianthus comosus Vahl. The crude extracts of the isolated endophytic fungi were investigated for their antimicrobial potential, extracellular enzymatic activity and phosphate solubilization. Additionally, the present study used genetic screening to assess the ability of the endophytic fungi to synthesize bioactive compounds, indicated by the presence of the polyketide synthase type 1 (PKS 1) gene. In preliminary microbial inhibition screening the fungal extracts had promising antifungal activity against Cryptococcus neoformans and Candida albicans. Furthermore, the endophytic fungus Talaromyces funiculosus displayed extracellular enzymatic activity, namely xylanase and cellulase. Five fungal strains demonstrated ability to solubilize phosphate and three strains demonstrated the presence of polyketide synthase type 1 (PKS 1) gene. It is worth considering further investigation of their bioactive secondary metabolites.The University of Pretoria, South Africahttp://www.elsevier.com/locate/sajb2022-04-16hj2021Paraclinical Science

<em>In vitro</em> evaluation of antimicrobial activities and antibiotic susceptibility profiling of culturable actinobacteria from fresh water streams

665-673Actinobacteria are major producers of antibiotics, industrially significant enzymes and many pharmaceutically important biologically active compounds. Twenty two actinobacterial strains were isolated from fresh water stream sediment samples of Murlen National Park, Mizoram, India. The actinobacterial strains were screened against antifungal pathogens (Fusarium oxysporum, Fusarium udum, Fusarium proliferatum, Fusarium oxysporum ciceri and Fusarium graminearum), and antibacterial activities against five bacterial pathogens (Staphylococcus aureus, Pseudomonas aeruginosa, Micrococcus luteus, Bacillus subtilis and Escherichia coli) and a yeast pathogen Candida albicans. All strains showed antibacterial activity against E. coli and F. proliferatum. Based on the results of antagonistic, antibacterial and anti-yeast, two most potent strains Kocuria sp. and Streptomyces intermidus were further evaluated for their antibiotics susceptibility activity against 21 different antibiotics. Kocuria sp. showed resistance to 10 antibiotics whereas Streptomyces intermidus was resistance to 15 antibiotics. Modular genes Polyketide Synthase (PKS II) and Nonribosomal Peptide Synthetase (NRPS) were also detected in these two strains, which might be responsible for the production of secondary metabolites. Two volatile compounds, Di-N-octyl phthalate and 1-Bromo-3, 7-Dimethyloctane were identified from the extract of Streptomyces intermidus BPSWAC29 strain using Gas chromatography Mass spectrometry (GC-MS). This study highlights the promise of discovering novel actinobacteria with antimicrobial activity from underexplored niche biotopes such as fresh water stream sediments

Rhizospheric bacterial community of endemic rhododendron arboreum sm. ssp. delavayi along eastern himalayan slope in tawang

Information on rhizosphere microbiome of endemic plants from high mountain ecosystems against those of cultivated plantations is inadequate. Comparative bacterial profiles of endemic medicinal plant Rhododendron arboreum Sm. subsp. delavayi rhizosphere pertaining to four altitudinal zonation Pankang Thang (PTSO), Nagula, Y-junction and Bum La (Indo-China border; in triplicates each) along cold adapted Eastern slope of Himalayan Tawang region, India is described here. Significant differences in DGGE profile between below ground bulk vs. rhizospheric community profile associated with the plant was identified. Tagged 16S amplicon sequencing from PTSO (3912 m) to Burn La (4509 m), revealed that soil pH, total nitrogen (TN), organic matter (OM) significantly influenced the underlying bacterial community structure at different altitudes. The relative abundance of Acidobacteria was inversely related to pH, as opposed to TN which was positively correlated to Acidobacteria and Proteobacteria abundance. TN was also the significant predictor for less abundant taxonomic groups Chioroflexi, Gemmatimonadetes. and Nitrospirae. Bum La soil harbored less bacterial diversity compared to other sites at lower altitudes. The most abundant phyla at 3% genetic difference were Acidobacteria, Actinobacteria, and Proteobacteria amongst others. Analysis of similarity indicated greater similarity within lower altitudinal than higher altitudinal group (ANOSIM, R = 0.287, p = 0.02). Constraining the ordination with the edaphic factor explained 83.13% of variation. Unique phylotypes of Bradyrhizobium and uncultured Rhizobiales were found in significant proportions at the four regions. With over 1% relative abundance Actinobacteria (42.6%), Acidobacteria (24.02%), Proteobacteria (16.00%), AD3 (9.23%), WPS-2 (5.1%), and Chloroflexi (1.48%) dominated the core microbiome

Epigenetic manipulation for secondary metabolite activation in endophytic fungi: current progress and future directions

ABSTRACTFungal endophytes have emerged as a promising source of secondary metabolites with significant potential for various applications in the field of biomedicine. The biosynthetic gene clusters of endophytic fungi are responsible for encoding several enzymes and transcriptional factors that are involved in the biosynthesis of secondary metabolites. The investigation of fungal metabolic potential at genetic level faces certain challenges, including the synthesis of appropriate amounts of chemicals, and loss of the ability of fungal endophytes to produce secondary metabolites in an artificial culture medium. Therefore, there is a need to delve deeper into the field of fungal genomics and transcriptomics to explore the potential of fungal endophytes in generating secondary metabolites governed by biosynthetic gene clusters. The silent biosynthetic gene clusters can be activated by modulating the chromatin structure using chemical compounds. Epigenetic modification plays a significant role by inducing cryptic gene responsible for the production of secondary metabolites using DNA methyl transferase and histone deacetylase. CRISPR-Cas9-based genome editing emerges an effective tool to enhance the production of desired metabolites by modulating gene expression. This review primarily focuses on the significance of epigenetic elicitors and their capacity to boost the production of secondary metabolites from endophytes. This article holds the potential to rejuvenate the drug discovery pipeline by introducing new chemical compounds

In Vitro and In Vivo Plant Growth Promoting Activities and DNA Fingerprinting of Antagonistic Endophytic Actinomycetes Associates with Medicinal Plants.

Endophytic actinomycetes have shown unique plant growth promoting as well as antagonistic activity against fungal phytopathogens. In the present study forty-two endophytic actinomycetes recovered from medicinal plants were evaluated for their antagonistic potential and plant growth-promoting abilities. Twenty-two isolates which showed the inhibitory activity against at least one pathogen were subsequently tested for their plant-growth promoting activities and were compared genotypically using DNA based fingerprinting, including enterobacterial repetitive intergenic consensus (ERIC) and BOX repetitive elements. Genetic relatedness based on both ERIC and BOX-PCR generates specific patterns corresponding to particular genotypes. Exponentially grown antagonistic isolates were used to evaluate phosphate solubilization, siderophores, HCN, ammonia, chitinase, indole-3-acetic acid production, as well as antifungal activities. Out of 22 isolates, the amount of indole-3-acetic acid (IAA) ranging between 10-32 μg/ml was produced by 20 isolates and all isolates were positive for ammonia production ranging between 5.2 to 54 mg/ml. Among 22 isolates tested, the amount of hydroxamate-type siderophores were produced by 16 isolates ranging between 5.2 to 36.4 μg/ml, while catechols-type siderophores produced by 5 isolates ranging from 3.2 to 5.4 μg/ml. Fourteen isolates showed the solubilisation of inorganic phosphorous ranging from 3.2 to 32.6 mg/100ml. Chitinase and HCN production was shown by 19 and 15 different isolates, respectively. In addition, genes of indole acetic acid (iaaM) and chitinase (chiC) were successively amplified from 20 and 19 isolates respectively. The two potential strains Streptomyces sp. (BPSAC34) and Leifsonia xyli (BPSAC24) were tested in vivo and improved a range of growth parameters in chilli (Capsicum annuum L.) under greenhouse conditions. This study is the first published report that actinomycetes can be isolated as endophytes from within these plants and were shown to have antagonistic and plant growth promoting abilities. These results clearly suggest the possibility of using endophytic actinomycetes as bioinoculant for plant growth promotion, nutrient mobilization or as biocontrol agent against fungal phytopathogens for sustainable agriculture

Author Correction: Insights into the functionality of endophytic actinobacteria with a focus on their biosynthetic potential and secondary metabolites production

A correction to this article has been published and is linked from the HTML and PDF versions of this paper. The error has been fixed in the paper

Insights into the functionality of endophytic actinobacteria with a focus on their biosynthetic potential and secondary metabolites production

Abstract Endophytic actinobacteria play an important role in growth promotion and development of host plant by producing enormous quantities of novel bioactive natural products. In the present investigation, 169 endophytic actinobacteria were isolated from endospheric tissues of Rhynchotoechum ellipticum. Based on their antimicrobial potential, 81 strains were identified by 16rRNA gene analysis, which were taxonomically grouped into 15 genera. All identified strains were screened for their plant growth promoting attributes and, for the presence of modular polyketide synthases (PKSI, PKSII and nonribosomal peptide synthetase (NRPS) gene clusters to correlate the biosynthetic genes with their functional properties. Expression studies and antioxidant potential for four representative strains were evaluated using qRT-PCR and DPPH assay respectively. Additionally, six antibiotics (erythromycin, ketoconazole, fluconazole, chloramphenicol, rifampicin and miconazole) and nine phenolic compounds (catechin, kaempferol, chebulagic acid, chlorogenic acid, Asiatic acid, ferulic acid, arjunic acid, gallic acid and boswellic acid) were detected and quantified using UHPLC-QqQLIT-MS/MS. Furthermore, three strains (BPSAC77, 121 and 101) showed the presence of the anticancerous compound paclitaxel which was reported for the first time from endophytic actinobacteria. This study provides a holistic picture, that endophytic actinobacteria are rich bacterial resource for bioactive natural products, which has a great prospective in agriculture and pharmaceutical industries