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Endophytic fungal assemblage in Achyranthes aspera Linn. revealed by internal transcribed spacer region of nuclear ribosomal RNA genes.

ABSTRACT: Only scanty reports are available on endophytic fungal associations in Achyranthes aspera Linn. Hence in this study a total of 504 isolates belonging to ten different species of fungi were isolated from asymptomatic, surface sterilised segments of leaf, stem and root of A. aspera collected from different locations of Kerala, India. Among the isolates ascomycetes were most prevalent. Colonisation rate of fungal endophytes was high in leaf tissue (95%) followed by stem (77.75%) and root segments (33.33%). The most frequent and dominant coloniser of the host plant were Colletotrichum sp., which was isolated from all locations. Scanning electron microscopy demonstrated the presence of hyphae in the intra and intercellular spaces of the plant tissue. Morphological and phylogenetic analyses using internal transcribed spacer (ITS) sequences of nuclear rRNA genes showed that the fungi recovered belonged to the lineages of Sordariomycetes, Dothideomycetes, Eurotiomycetes and Tremellomycetes. A maximum likelihood tree revealed the relationship between the obtained sequence data and the closest sequences retrieved from the GenBank.

SUBMITTER: John R

PROVIDER: S-EPMC5451360 | biostudies-literature | 2017 Jun

REPOSITORIES: biostudies-literature

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Endophytic fungal assemblage in Achyranthes aspera Linn. revealed by internal transcribed spacer region of nuclear ribosomal RNA genes.

John Reshma R Mathew Linu L

3 Biotech 20170531 2

Only scanty reports are available on endophytic fungal associations in Achyranthes aspera Linn. Hence in this study a total of 504 isolates belonging to ten different species of fungi were isolated from asymptomatic, surface sterilised segments of leaf, stem and root of A. aspera collected from different locations of Kerala, India. Among the isolates ascomycetes were most prevalent. Colonisation rate of fungal endophytes was high in leaf tissue (95%) followed by stem (77.75%) and root segments ( ...[more]

PMID: 28567620

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Project description:Achyranthes aspera Linn. (Amaranthaceae), commonly known as the Prickly Chaff flower, is used as herbal medicine in the Ivorian's culture, Africa. Nonetheless, there is currently a paucity of scientific information on A. aspera from the Ivory Coast. Herein, the antioxidant activity of A. aspera extracts (methanol, dichloromethane, ethyl acetate and infusion) as well as the enzymatic inhibitory potentials towards key enzymes in human diseases, namely Alzheimer's disease, (cholinesterases: AchE and BChE), type 2 diabetes (?-glucosidase and ?-amylase) and hyperpigmentation (tyrosinase) were assessed. The total phenolic (TPC) and flavonoid (TFC) content was determined using colorimetric methods and the individual compounds were characterized using ultra-high performance liquid chromatography coupled with hybrid quadrupole-Orbitrap high resolution mass spectrometry (UHPLC-HRMS). Furthermore, a network pharmacology analysis was conducted to predict putative targets of identified phenolic compounds. The highest TPC was observed in the infused extract (28.86 ± 0.12 mg GAE/g), while the dichloromethane extract (38.48 ± 1.48 mg RE/g) showed the highest level of TFC. UHPLC-HRMS analysis has revealed an abundance of fatty acids, flavonoids, phenols and acylquinic acids. Among tested extracts, the infused extract displayed the highest free radical quenching, reducing and metal-chelating ability. The extracts (except infusion) were effective as enzyme inhibitors against AChE, while only methanolic and infused extracts showed noteworthy anti-BChE effects. The methanolic extract showed a remarkable antityrosinase effect (56.24 ± 5.05 mg KAE/g), as well. Modest to moderate inhibitory activity was observed against ?-amylase (all extracts) and ?-glucosidase (only dichloromethane extract). Finally, the network pharmacology analysis suggested the carbonic anhydrase II enzyme as a putative target for explaining, at least in part, the traditional use of A. aspera preparations as diuretic and blood clotting agent. Data amassed herein tend to validate the use of A. aspera in traditional medicine, as well as act as a stepping stone for further studies in the quest for novel phytopharmaceuticals. In this context, it is desirable that this study will contribute to the validation of the traditional uses of this plant in the African herbal medicine, and to the valorization of the whole chain production of A. aspera, as a local and sustainable botanical resource.

Project description:BackgroundDNA barcoding offers an efficient way to determine species identification and to measure biodiversity. For dinoflagellates, an ancient alveolate group of about 2000 described extant species, DNA barcoding studies have revealed large amounts of unrecognized species diversity, most of which is not represented in culture collections. To date, two mitochondrial gene markers, Cytochrome Oxidase I (COI) and Cytochrome b oxidase (COB), have been used to assess DNA barcoding in dinoflagellates, and both failed to amplify all taxa and suffered from low resolution. Nevertheless, both genes yielded many examples of morphospecies showing cryptic speciation and morphologically distinct named species being genetically similar, highlighting the need for a common marker. For example, a large number of cultured Symbiodinium strains have neither taxonomic identification, nor a common measure of diversity that can be used to compare this genus to other dinoflagellates.Methodology/principal findingsThe purpose of this study was to evaluate the Internal Transcribed Spacer units 1 and 2 (ITS) of the rDNA operon, as a high resolution marker for distinguishing species dinoflagellates in culture. In our study, from 78 different species, the ITS barcode clearly differentiated species from genera and could identify 96% of strains to a known species or sub-genus grouping. 8.3% showed evidence of being cryptic species. A quarter of strains identified had no previous species identification. The greatest levels of hidden biodiversity came from Scrippsiella and the Pfiesteriaceae family, whilst Heterocapsa strains showed a high level of mismatch to their given species name.Conclusions/significanceThe ITS marker was successful in confirming species, revealing hidden diversity in culture collections. This marker, however, may have limited use for environmental barcoding due to paralogues, the potential for unidentifiable chimaeras and priming across taxa. In these cases ITS would serve well in combination with other markers or for specific taxon studies.

Dataset Information

Endophytic fungal assemblage in Achyranthes aspera Linn. revealed by internal transcribed spacer region of nuclear ribosomal RNA genes.

Publications

Endophytic fungal assemblage in Achyranthes aspera Linn. revealed by internal transcribed spacer region of nuclear ribosomal RNA genes.

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