Testing of candidate genes for compound biosynthesis in Arnica montana

Mrakovčić, Nikolina

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master's thesis

Testing of candidate genes for compound biosynthesis in Arnica montana

Zagreb: University of Zagreb, Faculty of Science, 2022. urn:nbn:hr:217:279243

Mrakovčić, Nikolina

University of Zagreb
Faculty of Science
Department of Biology

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APA 6th Edition

Mrakovčić, N. (2022). Testing of candidate genes for compound biosynthesis in Arnica montana (Master's thesis). Zagreb: University of Zagreb, Faculty of Science. Retrieved from https://urn.nsk.hr/urn:nbn:hr:217:279243

MLA 8th Edition

Mrakovčić, Nikolina. "Testing of candidate genes for compound biosynthesis in Arnica montana." Master's thesis, University of Zagreb, Faculty of Science, 2022. https://urn.nsk.hr/urn:nbn:hr:217:279243

Chicago 17th Edition

Harvard

Mrakovčić, N. (2022). 'Testing of candidate genes for compound biosynthesis in Arnica montana', Master's thesis, University of Zagreb, Faculty of Science, accessed 21 January 2025, https://urn.nsk.hr/urn:nbn:hr:217:279243

Vancouver

Mrakovčić N. Testing of candidate genes for compound biosynthesis in Arnica montana [Master's thesis]. Zagreb: University of Zagreb, Faculty of Science; 2022 [cited 2025 January 21] Available at: https://urn.nsk.hr/urn:nbn:hr:217:279243

IEEE

N. Mrakovčić, "Testing of candidate genes for compound biosynthesis in Arnica montana", Master's thesis, University of Zagreb, Faculty of Science, Zagreb, 2022. Available at: https://urn.nsk.hr/urn:nbn:hr:217:279243

Cite this item: https://urn.nsk.hr/urn:nbn:hr:217:279243

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Metadata

Title	Testing of candidate genes for compound biosynthesis in Arnica montana
Title (croatian)	Testiranje gena kandidata uključenih u biosintezu metabolita biljke Arnica montana
Author	Nikolina Mrakovčić
Mentor	Gundula Noll (mentor)
Mentor	Nataša Bauer (sumentor)
Committee member	Nataša Bauer (predsjednik povjerenstva)
Committee member	Mirta Tkalec (član povjerenstva)
Committee member	Biljana Balen (član povjerenstva)
Committee member	Marin Ježić (član povjerenstva)
Granter	University of Zagreb Faculty of Science (Department of Biology) Zagreb
Defense date and country	2022-02-28, Croatia
Scientific / art field, discipline and subdiscipline	NATURAL SCIENCES Biology
Abstract	Arnica montana L. is a traditional medicinal plant used for centuries. It owes its medicinal properties to sesquiterpene lactones, helenalin and dihydrohelenalin. Traditionally, the plants are harvested from its natural habitat, but there is need for alternative ways to sustainably obtain bioactive compounds. To achieve alternative production of A. montana bioactive sesquiterpene lactones, their biosynthesis pathway must be elucidated. Functional characterization of candidate genes, that might be involved in sesquiterpene lactones biosynthesis, has been conducted in the heterologous expression system yeast Saccharomyces cerevisiae and compounds analysis via gas chromatography-mass spectrometry. Additionally, analysis of potential candidate genes expression patterns and sesquiterpene lactones content distribution in plants was carried out. Candidate genes AmGAS and AmGAO were verified to encode respective germacrene A synthase and germacrene A acid oxidase, and their enzymatic activity was confirmed. Candidate genes AmCYPs were functionally analysed to a certain extent, with candidate AmCYP7 showing great potential for further investigation. Correlation of expression patterns and content of bioactive compounds in A. montana tissues revealed that among tested, AmCYP7 gene has potential to be part of sesquiterpene lactones biosynthesis pathway and provided preliminary evidence for their biosynthesis at different developmental stages during the plant´s life cycle.
Abstract (croatian)	Arnica montana L. je biljka koja se stoljećima koristi u tradicionalnoj medicini. Za njezina ljekovita svojstva odgovorni su seskviterpenski laktoni, helenalin i dihidrohelenalin. Arnika se tradicionalno ubire s prirodnog staništa i postoji potreba za alternativnim i održivijim načinima sinteze bioaktivnih sastojaka. Kako bi alternativna proizvodnja bioaktivnih seskviterpenskih laktona bila moguća, potrebno je razjasniti put biosinteze. Provedena je funkcionalna karakterizacija gena kandidata iz A. montana, koji su potencijalno uključeni u biosintetski put seskviterpenskih laktona, u heterolognom ekspresijskom sustavu, kvascu Saccharomyces cerevisiae. Metaboliti sintetizirani u kvascu analizirani su plinskom kromatografijom i masenom spektrometrijom. Također, analizirani su ekspresijski obrasci gena kandidata te distribucija i sadržaj seskviterpenskih laktona u različitim tkivima biljke A. montana. Potvrđeno je da geni kandidati AmGAS i AmGAO kodiraju sintazu germakrina A, odnosno oksidazu germakrina A te je potvrđena njihova enzimska aktivnost. Geni AmCYP su djelomično funkcionalno okarakterizirani, a gen AmCYP7 pokazao je najveći potencijal za daljnje istraživanje. Korelacija ekspresijskih obrazaca gena kandidata i sadržaja bioaktivnih sastojaka u tkivima biljke A. montana pokazala je da je gen AmCYP7 vjerojatno uključen u biosintezu seskviterpenskih laktona i pružila preliminarne dokaze za biosintezu seskviterpenskih laktona u različitim razvojnim stadijima biljke A. montana.
Keywords
Keywords (croatian)
Language	english
URN:NBN	urn:nbn:hr:217:279243
Study programme	Title: Molecular Biology Study programme type: university Study level: graduate Academic / professional title: magistar/magistra molekularne biologije (magistar/magistra molekularne biologije)
Type of resource	Text
File origin	Born digital
Access conditions	Open access Embargo expiration date: 2024-02-28
Terms of use
Repository	Repository of the Faculty of Science
Created on	2022-03-16 10:21:08