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Polyporoid fungi of Tanzania: Taxonomy, transcriptomics and biochemical analyses of Kusaghiporia usambarensis and Piptoporellus baudonii
Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Organismal Biology, Systematic Biology. Systematisk biologi, Systematic Biology. (Tibell)ORCID iD: 0000-0003-3442-9262
2020 (English)Doctoral thesis, comprehensive summary (Other academic)
Description
Abstract [en]

Polyporoid fungi refers to basidiomycetes with fruiting bodies with the hymenium located to the inner surfaces of pores or narrow tubes. The majority of polyporoids belongs to Polyporales. Most Polyporales are saprobes, but some are plant pathogens. The overall aim of this thesis was to study the taxonomy, systematics and chemistry of the two species Kusaghiporia usambarensis (saprobic) and Piptoporellus baudonii (a plant pathogen) collected from Tanzania, using morphological and molecular approaches, combined with transcriptomics and pharmacognostic investigations.

The main contribution of this thesis includes the description a new genus with the new species K. usambarensis from the Usambara Mountains, Tanzania; investigation of the chemical composition of volatile compounds from this medicinal mushroom; isolation and structure determination of a novel and most abundant peptide in K. usambarensis, and further to elucidate the phylogenetic position of Piptoporellus baudonii (formerly known as Laetiporus baudonii) by using a four molecular markers dataset.

Paper I was conducted applying a classical taxonomic approach, including both morphological and phylogenetic analyses, to describe a new genus and species K. usambarensis. Paper II, investigated volatiles and volatile derivatives in dichloromethane extracts of K. usambarensis analysed by GC-MS and NMR spectroscopy. The main elements were phenols, and esters, compounds that may explain the formerly reported antioxidant activity and traditional medicinal use of the mushroom. In paper III, screening of peptides in K. usambarensis revealed a novel cysteine-rich peptide, highly expressed at gene level and the most abundant compound in the fruiting body. Combined LC-MS and transcriptome analyses were used to determine the peptide sequence, and subsequently NMR spectroscopy to determine the 3D structure of the novel peptide, kusaghitide. In paper IV molecular techniques were used to elucidate the phylogenetic position of the parasitic Laetiporus baudonii. Phylogenetic analyses of combined 5.8S, nrLSU, nrSSU, and TEF1 gene sequences placed L. baudonii in the genus Piptoporellus, hence the new combination Piptoporellus baudonii was proposed. This thesis has contributed to build capacity in the fields of mycology, systematics and pharmacognosy in order to reinforce ecological knowledge and ethnopharmaceutical research for future drug discovery in Tanzania and Africa at large.

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis, 2020. , p. 56
Series
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology, ISSN 1651-6214 ; 1908
Keywords [en]
Systematics, Laetiporaceae, pharmacognosy, cysteine-rich peptide, baudonii
National Category
Biological Sciences
Research subject
Biology with specialization in Systematics
Identifiers
URN: urn:nbn:se:uu:diva-405548ISBN: 978-91-513-0881-4 (print)OAI: oai:DiVA.org:uu-405548DiVA, id: diva2:1404340
Public defence
2020-04-17, Ekmansalen, Evolutionsbiologiskt centrum (EBC), Norbyvägen 14-18, Uppsala, 10:00 (English)
Opponent
Supervisors
Available from: 2020-03-26 Created: 2020-02-28 Last updated: 2020-03-26
List of papers
1. Elucidating the phylogeny of the serious plant pathogen Piptoporellus baudonii using a multigene molecular dataset
Open this publication in new window or tab >>Elucidating the phylogeny of the serious plant pathogen Piptoporellus baudonii using a multigene molecular dataset
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(English)Manuscript (preprint) (Other academic)
Abstract [en]

Piptoporellus baudonii is proposed as a new combination for Laetiporus baudonii in Polyporales (Basidiomycota), based on morphological and molecular features. This parasitic macrofungus attacks cashew trees, Eucalyptus, cassava, Tectona and some indigenous trees in the Southern regions of Tanzania and poses a serious threat to agroforestry and livelihood conditions in the area. Maximum likelihood and Bayesian methods were applied to obtain phylogenetic trees for partial sequences of the 5.8S part of nuc rDNA internal transcribed spacer, parts of 28S nuc rDNA (28S) and 18S nuc rDNA (18S), in addition to parts of gene encoding the translation elongation factor 1-α (TEF1) of Laetiporus baudonii for comparisons with additional sequences from the antrodia clade. Our results reveal a strongly supported phylogenetic clade of L. baudonii with Piptoporellus in Fomitopsidaceae. Piptoporellus baudonii shares many morphological features with other members of Piptoporellus but differs in having broadly ellipsoid or rarely ovoid basidiospores, while in other Piptoporellus species the spores are cylindrical to oblong-ellipsoid. Both morphological and phylogenetic evidence, however, justify the incorporation of L. baudonii in Piptoporellus, to join the three other species in the genus.

Keywords
Africa, parasitic fungus, antrodia clade, Laeptiporus baudonii, macrofungus, Piptoporellus, 1 new taxon
National Category
Natural Sciences
Identifiers
urn:nbn:se:uu:diva-405385 (URN)
Available from: 2020-02-28 Created: 2020-02-28 Last updated: 2020-02-28
2. Phylogenetic position and taxonomy of Kusaghiporia usambarensis gen. et sp. nov. (Polyporales)
Open this publication in new window or tab >>Phylogenetic position and taxonomy of Kusaghiporia usambarensis gen. et sp. nov. (Polyporales)
2018 (English)In: Mycology, ISSN 2150-1203Article in journal (Refereed) Published
Abstract [en]

A large polyporoid mushroom from the West Usambara Mountains in North-eastern Tanzania produces dark brown, up to 60-cm large fruiting bodies that at maturity may weigh more than 10 kg. It has a high rate of mycelial growth and regeneration and was found growing on both dry and green leaves of shrubs; attached to the base of living trees, and it was also observed to degrade dead snakes and insects accidentally coming into contact with it. Phylogenetic analyses based on individual and concatenated data sets of nrLSU, nrSSU and the RPB2 and TEF1 genes showed it, together with Laetiporus, Phaeolus, Pycnoporellus and Wolfiporia, to form a monophyletic group in Polyporales. Based on morphological features and molecular data, it is described as Kusaghiporia usambarensis.

Place, publisher, year, edition, pages
Taylor & Francis, 2018
Keywords
Kusaghiporia; molecular phylogeny; polyporales; Tanzania; taxonomy; Usambara
National Category
Bioinformatics and Systems Biology
Research subject
Biology with specialization in Systematics
Identifiers
urn:nbn:se:uu:diva-348537 (URN)10.1080/21501203.2018.1461142 (DOI)
Funder
Sida - Swedish International Development Cooperation Agency, 2221
Available from: 2018-04-15 Created: 2018-04-15 Last updated: 2020-02-28Bibliographically approved
3. Chemical composition of the medicinal mushroom Kusaghiporia usambarensis
Open this publication in new window or tab >>Chemical composition of the medicinal mushroom Kusaghiporia usambarensis
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(English)Manuscript (preprint) (Other academic)
Abstract [en]

The mushroom Kusaghiporia usambarensis was recently described as a new species and the single member of a new genus. It appears endemic to Tanzania. The mushroom forms large, conspicuous fruiting bodies with an unusually high radial growth rate and an observed ability to attract insects. The mushroom is used by the local community both for food and in traditional medicine. In order to gain insight into both basic biology and the traditional use of this mushroom, we here report the first analyses of its chemical composition. A dichloromethane extract was prepared from the dried fruiting body and analysed by nuclear magnetic resonance and gas chromatography coupled to mass spectrometry. This extract of medium polar chemical constituents of Kusaghiporia usambarensis showed a high content of phenolic compounds and esters. The main elements were phenols, which account for ~22%, and esters ~20 %). These two groups of compounds may explain formerly reported antioxidant activity and traditional medicinal use of the mushroom. Revealing the chemistry also shed light on K. usambarensi as a sought-after delicacy: presence of 1-octen-3-one, octanal, 3-octen-2-one contribute to its fruity, cheesy and chicken flavour.

National Category
Natural Sciences
Identifiers
urn:nbn:se:uu:diva-405386 (URN)
Available from: 2020-02-28 Created: 2020-02-28 Last updated: 2020-02-28
4. Cysteine-rich peptide from the gigantic edible mushroom Kusaghiporia usambarensis (Laetiporaceae)
Open this publication in new window or tab >>Cysteine-rich peptide from the gigantic edible mushroom Kusaghiporia usambarensis (Laetiporaceae)
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(English)Manuscript (preprint) (Other academic)
Abstract [en]

Cysteine-rich peptides are produced by various organisms across all kingdoms and have triggered an interest in isolation of molecules for novel drug development. In this study, we report a novel cysteine-rich peptide, kusaghitide, isolated from the gigantic medicinal mushroom Kusaghiporia usambarensis. It is highly expressed in the K. usambarensis transcriptome and it is the most abundant compound in the methanol-water extract. The 54 amino acid residue long peptide was isolated through aqueous methanol 50% and a sample was reduced, alkylated and cleaved enzymatically. De novo sequencing was done by LC-MS/MS and obtained sequences were used for mining the transcriptome to search for the complete gene. The peptide was recombinantly expressed in One Shot BL21 Star Escherichia coli using lysogenic broth and minimal media. Its 3D NMR structure was determined using 2D and 3D NMR. Three hypothetical protein sequences similar to kusaghitide originate from Laetiporus sulphureusWolfiporia cocos and Sparassis crispa with per cent similarity of 76% and 58% and 53% respectively and were found by BLAST search in the NCBI database. Kusaghitide did not inhibit the growth of either Escherichia coli or Staphylococcus aureus. This is first report of a peptide from K. usambarensis in Laetiporaceae.

Keywords
Kusaghiporia, medicinal mushroom, recombinant protein production, cysteine-rich peptides, peptide structure.
National Category
Natural Sciences
Identifiers
urn:nbn:se:uu:diva-405387 (URN)
Available from: 2020-02-28 Created: 2020-02-28 Last updated: 2020-02-28

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