Application of Lichen-Forming Fungi for Industrial Use

A special issue of Journal of Fungi (ISSN 2309-608X). This special issue belongs to the section "Environmental and Ecological Interactions of Fungi".

Deadline for manuscript submissions: closed (31 October 2021) | Viewed by 11035

Special Issue Editor


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Guest Editor
Korea Lichen Research Institute, Sunchon National University, Suncheon, Korea
Interests: biobank of lichen bioresources; comparative genomics of lichen-forming fungi for secondary metabolites genes; heterologous expression of lichen PKS genes; transformation of lichen-forming fungi; biodiversity of endolichenic fungi

Special Issue Information

Dear Colleagues,

Under the COVID-19 pendemic crisis, the exploration of pharmacologically useful and novel natural compounds originating from lichen-forming fungi (LFF) can provide a treatment option for patients. LFF produce very unique secondary metabolites (SM) possessing various pharmacological activities such as anti-microbial, anti-inflamation, anti-oxidant, and anti-cancer. Becuase of the limitations of research tools handling lichen secondary metabolite production at a large scale, noticeable progress of lichen SM commercialization has not been achieved yet. Nevertheless, certain researchers have attempted to overcome the inherent limitation of lichen-forming fungus with the aid of genomics and molecular biological approaches. The Special Issue is open to any research papers dealing with the recent progress of studying LFF, from fundamental biology to advanced and applied biotechology, including, but not limited to, lichen SM production.

Prof. Dr. Jae-Seoun Hur
Guest Editor

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Keywords

  • lichen bioresource
  • secondary metabolites
  • polyketide synthase (PKS) genes
  • biological activity
  • heterologous expression
  • biotechnology

Published Papers (4 papers)

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Research

14 pages, 1764 KiB  
Article
Production and Activity of Cristazarin in the Lichen-Forming Fungus Cladonia metacorallifera
by Min-Hye Jeong, Chan-Ho Park, Jung A Kim, Eu Ddeum Choi, Soonok Kim, Jae-Seoun Hur and Sook-Young Park
J. Fungi 2021, 7(8), 601; https://doi.org/10.3390/jof7080601 - 26 Jul 2021
Cited by 8 | Viewed by 2092
Abstract
Lichens are a natural source of bioactive compounds. Cladonia metacorallifera var. reagens KoLRI002260 is a rare lichen known to produce phenolic compounds, such as rhodocladonic, thamnolic, and didymic acids. However, these metabolites have not been detected in isolated mycobionts. We investigated the effects [...] Read more.
Lichens are a natural source of bioactive compounds. Cladonia metacorallifera var. reagens KoLRI002260 is a rare lichen known to produce phenolic compounds, such as rhodocladonic, thamnolic, and didymic acids. However, these metabolites have not been detected in isolated mycobionts. We investigated the effects of six carbon sources on metabolite biosynthesis in the C. metacorallifera mycobiont. Red pigments appeared only in Lilly and Barnett’s media with fructose at 15 °C after 3 weeks of culture and decreased after 6 weeks. We purified these red pigments using preparative-scale high performance liquid chromatography and analyzed them via nuclear magnetic resonance. Results indicated that 1% fructose-induced cristazarin and 6-methylcristazarin production under light conditions. In total, 27 out of 30 putative polyketide synthase genes were differentially expressed after 3 weeks of culture, implying that these genes may be required for cristazarin production in C. metacorallifera. Moreover, the white collar genes Cmwc-1 and Cmwc-2 were highly upregulated at all times under light conditions, indicating a possible correlation between cristazarin production and gene expression. The cancer cell lines AGS, CT26, and B16F1 were sensitive to cristazarin, with IC50 values of 18.2, 26.1, and 30.9 μg/mL, respectively, which highlights the value of cristazarin. Overall, our results suggest that 1% fructose under light conditions is required for cristazarin production by C. metacorallifera mycobionts, and cristazarin could be a good bioactive compound. Full article
(This article belongs to the Special Issue Application of Lichen-Forming Fungi for Industrial Use)
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11 pages, 16845 KiB  
Article
Transcriptome Analysis Identifies a Gene Cluster for the Biosynthesis of Biruloquinone, a Rare Phenanthraquinone, in a Lichen-Forming Fungus Cladonia macilenta
by Wonyong Kim, Min-Hye Jeong, Sung-Hwan Yun and Jae-Seoun Hur
J. Fungi 2021, 7(5), 398; https://doi.org/10.3390/jof7050398 - 20 May 2021
Cited by 3 | Viewed by 2642
Abstract
Lichens are prolific producers of natural products of polyketide origin. We previously described a culture of lichen-forming fungus (LFF) Cladonia macilenta that produces biruloquinone, a purple pigment that is a phenanthraquinone rarely found in nature. However, there was no genetic information on the [...] Read more.
Lichens are prolific producers of natural products of polyketide origin. We previously described a culture of lichen-forming fungus (LFF) Cladonia macilenta that produces biruloquinone, a purple pigment that is a phenanthraquinone rarely found in nature. However, there was no genetic information on the biosynthesis of biruloquinone. To identify a biosynthetic gene cluster for biruloquinone, we mined polyketide synthase (PKS) genes from the genome sequence of a LFF isolated from thalli of C. macilenta. The 38 PKS in C. macilenta are highly diverse, many of which form phylogenetic clades with PKS previously characterized in non-lichenized fungi. We compared transcriptional profiles of the 38 PKS genes in two chemotypic variants, one producing biruloquinone and the other producing no appreciable metabolite in vitro. We identified a PKS gene (hereafter PKS21) that was highly upregulated in the LFF that produces biruloquinone. The boundaries of a putative biruloquinone gene cluster were demarcated by co-expression patterns of six clustered genes, including the PKS21. Biruloquinone gene clusters exhibited a high degree of synteny between related species. In this study we identified a novel PKS family responsible for the biosynthesis of biruloquinone through whole-transcriptome analysis. Full article
(This article belongs to the Special Issue Application of Lichen-Forming Fungi for Industrial Use)
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14 pages, 4206 KiB  
Article
Effect of Isolation Conditions on Diversity of Endolichenic Fungal Communities from a Foliose Lichen, Parmotrema tinctorum
by Ji Ho Yang, Seung-Yoon Oh, Wonyong Kim, Jung-Jae Woo, Hyeonjae Kim and Jae-Seoun Hur
J. Fungi 2021, 7(5), 335; https://doi.org/10.3390/jof7050335 - 26 Apr 2021
Cited by 11 | Viewed by 2672
Abstract
Endolichenic fungi (ELF) are emerging novel bioresources because their diverse secondary metabolites have a wide range of biological activities. Metagenomic analysis of lichen thalli demonstrated that the conventional isolation method of ELF covers a very limited range of ELF, and the development of [...] Read more.
Endolichenic fungi (ELF) are emerging novel bioresources because their diverse secondary metabolites have a wide range of biological activities. Metagenomic analysis of lichen thalli demonstrated that the conventional isolation method of ELF covers a very limited range of ELF, and the development of an advanced isolation method is needed. The influence of four variables were investigated in this study to determine the suitable conditions for the isolation of more diverse ELF from a radially growing foliose lichen, Parmotrema tinctorum. Four variables were tested: age of the thallus, severity of surface-sterilization of the thallus, size of a thallus fragment for the inoculation, and nutrient requirement. In total, 104 species (1885 strains) of ELF were isolated from the five individual thalli of P. tinctorum collected at five different places. Most of the ELF isolates belong to Sordariomycetes. Because each part of lichen thallus (of different age) has unique ELF species, the whole thallus of the foliose lichen is needed to isolate diverse ELF. Moderate sterilization is appropriate for the isolation of diverse ELF. Inoculation of small fragment (1 mm2) of lichen thallus resulted in the isolation of highest diversity of ELF species compared to larger fragments (100 and 25 mm2). Moreover, ELF species isolated from the small thallus fragments covered all ELF taxa detected from the medium and the large fragments in this study. The use of two media—Bold’s basal medium (nutrient poor) and potato dextrose agar (nutrient rich)—supported the isolation of diverse ELF. Among the tested variables, size of thallus fragment more significantly influenced the isolation of diverse ELF than other three factors. Species composition and richness of ELF communities from different lichen thalli differed from each other in this study. Full article
(This article belongs to the Special Issue Application of Lichen-Forming Fungi for Industrial Use)
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12 pages, 2195 KiB  
Article
Establishment of Agrobacterium tumefaciens-Mediated Transformation of Cladonia macilenta, a Model Lichen-Forming Fungus
by Rundong Liu, Wonyong Kim, Jaycee Augusto Paguirigan, Min-Hye Jeong and Jae-Seoun Hur
J. Fungi 2021, 7(4), 252; https://doi.org/10.3390/jof7040252 - 26 Mar 2021
Cited by 6 | Viewed by 2574
Abstract
Despite the fascinating biology of lichens, such as the symbiotic association of lichen-forming fungi (mycobiont) with their photosynthetic partners and their ability to grow in harsh habitats, lack of genetic tools manipulating mycobiont has hindered studies on genetic mechanisms underpinning lichen biology. Thus, [...] Read more.
Despite the fascinating biology of lichens, such as the symbiotic association of lichen-forming fungi (mycobiont) with their photosynthetic partners and their ability to grow in harsh habitats, lack of genetic tools manipulating mycobiont has hindered studies on genetic mechanisms underpinning lichen biology. Thus, we established an Agrobacterium tumefaciens-mediated transformation (ATMT) system for genetic transformation of a mycobiont isolated from Cladonia macilenta. A set of combinations of ATMT conditions, such as input biomass of mycobiont, co-cultivation period with Agrobacterium cells, and incubation temperature, were tested to identify an optimized ATMT condition for the C. macilenta mycobiont. As a result, more than 10 days of co-cultivation period and at least 2 mg of input biomass of the mycobiont were recommended for an efficient ATMT, owing to extremely slow growth rate of mycobionts in general. Moreover, we examined T-DNA copy number variation in a total of 180 transformants and found that 88% of the transformants had a single copy T-DNA insertion. To identify precise T-DNA insertion sites that interrupt gene function in C. macilenta, we performed TAIL-PCR analyses for selected transformants. A hypothetical gene encoding ankyrin repeats at its C-terminus was interrupted by T-DNA insertion in a transformant producing dark-brown colored pigment. Although the identification of the pigment awaits further investigation, this proof-of-concept study demonstrated the feasibility of use of ATMT in construction of a random T-DNA insertion mutant library in mycobionts for studying genetic mechanisms behind the lichen symbiosis, stress tolerance, and secondary metabolite biosynthesis. Full article
(This article belongs to the Special Issue Application of Lichen-Forming Fungi for Industrial Use)
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