Isolation, Characterization and Targeted Metabolic Evaluation of Endophytic Fungi Harbored in 14 Seed-Derived Hypericum Species

Henzelyová, Jana; Antalová, Michaela; Nigutová, Katarína; Logoida, Mariia; Schreiberová, Andrea; Kusari, Souvik; Čellárová, Eva

doi:10.1055/a-1130-4703

Cited by 5 publications

(9 citation statements)

References 42 publications

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“…The studied fungal endophytes were isolated from hypericin-accumulating aerial tissues of several Hypericum representatives, except for the isolate 8RF1-3 from H. humifusum roots. Our previous results indicated that fungal endophytes are able to produce the same anthraquinones as the host plants [ 5 , 7 , 9 ]. The genome-mining of Hypericum- borne fungal endophytes led to the prediction of candidate biosynthetic gene clusters.…”

Section: Resultsmentioning

confidence: 99%

“…were selected based on their ability to produce anthraquinones and bisanthraquinones innate to the host plants. The experimental group comprised two Fusarium oxysporum isolates varying in metabolite production, Scedosporium apiospermum, Plectosphaerella cucumerina, and Diaporthe eres which were collected from plants acclimated to outdoor conditions in the Botanical Garden of Pavol Jozef Šafárik University in Košice, Slovakia [ 9 ] (Table 3 ). The cultures were identified by molecular markers ITS rDNA and protein-coding gene region of tef1α.…”

Section: Methodsmentioning

confidence: 99%

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In silico prediction of polyketide biosynthetic gene clusters in the genomes of Hypericum-borne endophytic fungi

Petijová,

Henzelyová,

Kuncová

et al. 2024

BMC Genomics

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Background The search for new bioactive natural compounds with anticancer activity is still of great importance. Even though their potential for diagnostics and treatment of cancer has already been proved, the availability is still limited. Hypericin, a naphthodianthrone isolated essentially from plant source Hypericum perforatum L. along with other related anthraquinones and bisanthraquinones belongs to this group of compounds. Although it has been proven that hypericin is synthesized by the polyketide pathway in plants, none of the candidate genes coding for key enzymes has been experimentally validated yet. Despite the rare occurrence of anthraquinones in plants, their presence in microorganisms, including endophytic fungi, is quite common. Unlike plants, several biosynthetic genes grouped into clusters (BGCs) in fungal endophytes have already been characterized. Results The aim of this work was to predict, identify and characterize the anthraquinone BGCs in de novo assembled and functionally annotated genomes of selected endophytic fungal isolates (Fusarium oxysporum, Plectosphaerella cucumerina, Scedosporium apiospermum, Diaporthe eres, Canariomyces subthermophilus) obtained from different tissues of Hypericum spp. The number of predicted type I polyketide synthase (PKS) BGCs in the studied genomes varied. The non-reducing type I PKS lacking thioesterase domain and adjacent discrete gene encoding protein with product release function were identified only in the genomes of C. subthermophilus and D. eres. A candidate bisanthraquinone BGC was predicted in C. subthermophilus genome and comprised genes coding the enzymes that catalyze formation of the basic anthraquinone skeleton (PKS, metallo-beta-lactamase, decarboxylase, anthrone oxygenase), putative dimerization enzyme (cytochrome P450 monooxygenase), other tailoring enzymes (oxidoreductase, dehydrogenase/reductase), and non-catalytic proteins (fungal transcription factor, transporter protein). Conclusions The results provide an insight into genetic background of anthraquinone biosynthesis in Hypericum-borne endophytes. The predicted bisanthraquinone gene cluster represents a basis for functional validation of the candidate biosynthetic genes in a simple eukaryotic system as a prospective biotechnological alternative for production of hypericin and related bioactive anthraquinones.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

In silico prediction of polyketide biosynthetic gene clusters in the genomes of Hypericum-borne endophytic fungi

Petijová,

Henzelyová,

Kuncová

et al. 2024

BMC Genomics

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“…Skyrin (7) and its precursors are not abundantly available in plants, but these compounds are produced by different classes of endophytic filamentous fungi [ 34 ]. It could be possible that Hypericum plant-associated endophytes produce these metabolites in planta and contribute to hypericin production, given that endophytes are known to produce secondary metabolites found in their host plants [ 23 , 35 ]. Besides, native endophytes might have acquired the skyrin (7) producing gene machinery through horizontal gene transfer in the course of co-evolution with Hypericum host plants.…”

Section: Discussionmentioning

confidence: 99%

“…Remarkably, skyrin (7) is typically biosynthesized by fungi belonging to different species and ecological niches and reported to have antimicrobial properties [ 19 , 20 ]. Thus far, the role of plant-associated microorganisms such as endophytes in the production of skyrin (7) is a plausible, open question, particularly since native endophytes harbored in H. perforatum can produce hypericin (6) [ 21 , 22 , 23 ]. Therefore, it is essential to identify the in planta site of localization and the role of skyrin (7) , as well as the related intermediates leading to the production of hypericin (6) .…”

Section: Introductionmentioning

confidence: 99%

MALDI-HRMS Imaging Maps the Localization of Skyrin, the Precursor of Hypericin, and Pathway Intermediates in Leaves of Hypericum Species

et al. 2020

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Hypericum perforatum and related species (Hypericaceae) are a reservoir of pharmacologically important secondary metabolites, including the well-known naphthodianthrone hypericin. However, the exact biosynthetic steps in the hypericin biosynthetic pathway, vis-à-vis the essential precursors and their localization in plants, remain unestablished. Recently, we proposed a novel biosynthetic pathway of hypericin, not through emodin and emodin anthrone, but skyrin. However, the localization of skyrin and its precursors in Hypericum plants, as well as the correlation between their spatial distribution with the hypericin pathway intermediates and the produced naphthodianthrones, are not known. Herein, we report the spatial distribution of skyrin and its precursors in leaves of five in vitro cultivated Hypericum plant species concomitant to hypericin, its analogs, as well as its previously proposed precursors emodin and emodin anthrone, using MALDI-HRMS imaging. Firstly, we employed HPLC-HRMS to confirm the presence of skyrin in all analyzed species, namely H. humifusum, H. bupleuroides, H. annulatum, H. tetrapterum, and H. rumeliacum. Thereafter, MALDI-HRMS imaging of the skyrin-containing leaves revealed a species-specific distribution and localization pattern of skyrin. Skyrin is localized in the dark glands in H. humifusum and H. tetrapterum leaves together with hypericin but remains scattered throughout the leaves in H. annulatum, H. bupleuroides, and H. rumeliacum. The distribution and localization of related compounds were also mapped and are discussed concomitant to the incidence of skyrin. Taken together, our study establishes and correlates for the first time, the high spatial distribution of skyrin and its precursors, as well as of hypericin, its analogs, and previously proposed precursors emodin and emodin anthrone in the leaves of Hypericum plants.

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“…Both techniques have been used to separate, identify and quantify series of structurally different SMs in plants include spectrophotometric [57,[73][74][75], GC-MS [56,76], LC-MS [77], LC-MS/MS [56,76,78], Liquid Chromatography-High Resolution Electrospray Ionization Mass Spectrometry (LC-HRESIMS) [57], Nuclear Magnetic Resonance (NMR) [44,79], Ultra-Fast Liquid Chromatography (UFLC-Q-TOF-MS/MS) [80][81], UFLC-triple-Q-TOF-MS/MS [82], Ultra-high performance Liquid Chromatography quadrupole-time of flight mass spectrometry [83], UHPLC-LTQ-Orbitrap [84], UHPLC/Q-Orbitrap-MS [85], HPLC-DAD [86], UHPLC-Q-TOF-MS [87], UHPLC-Q-TOF-MS/MS [88], HPLC-DAD-ESI-MS [89], UPLC-QTOF/MSE [90], UHPLC-MS/MS [91], HPLC-ESI-QTOF-MS [92], UPLC-QTOF-MS/MS [93], UHPLC-Q-TOF-MS/MS [94][95], UHPLC-ESI-MS/MS [96], UHPLC-ESI-Orbitrap-MS/MS [97], UPLC-IT-MS [98], UPLC-Q-TOF-MS [98], UPLC-PDA [45], HPLC-ESI-MS and MS 2 [99], reverse phase high performance liquid chromatography-diode array detector (RP-HPLC-DAD) [74], HPLC-HRMS [100]. been used to produce a yellow color that turn colorless on addition of dilute hydrochloric acid confirm flavonoid [102,[104][105]…”

Section: Techniques For Identify and Quantifying Secondary Metabolitesmentioning

confidence: 99%

Identification and Pharmacological Activities of Secondary Metabolites of Phytochemicals

Ovonramwen¹

2022

Preprint

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Plant secondary metabolites are organic compounds (alkaloids, flavonoids, phenolics, tannins, anthraquinones, saponins, steroids, lignins, terpenes) produced by plant for self-defense during growth and development in order to protect them from harmful agents. As population is increasing and quality of life is reducing couple with outbreak of some diseases that are posing threat on the economy, a lot of medicinal plants have attracted interest of researchers because of the usefulness of these metabolites in pharmaceuticals, food and cosmetic industries. In this review, the pharmacological activities of some medicinal plants that are rich in secondary metabolites were studied, means of identifying and quantifying using spectrophotometry and chromatography techniques were also discussed. However, this will assist to reduce the uses and dependent on the synthetic drug and the onset of the age related diseases.

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Isolation, Characterization and Targeted Metabolic Evaluation of Endophytic Fungi Harbored in 14 Seed-Derived Hypericum Species

Cited by 5 publications

References 42 publications

In silico prediction of polyketide biosynthetic gene clusters in the genomes of Hypericum-borne endophytic fungi

In silico prediction of polyketide biosynthetic gene clusters in the genomes of Hypericum-borne endophytic fungi

MALDI-HRMS Imaging Maps the Localization of Skyrin, the Precursor of Hypericin, and Pathway Intermediates in Leaves of Hypericum Species

Identification and Pharmacological Activities of Secondary Metabolites of Phytochemicals

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