Metabolic engineering of essential oil composition in Lavendula latifolia by altering expression of borneol diphosphate synthase
department or school: Science, Irving K. Barber Faculty of (Okanagan)
department or school: Biology, Department of (Okanagan)
data provider: DSpace
degree campus: UBCO
creator: Najafianashrafi, Elaheh
issued: 2021-10-12T07:00:00Z
issued: 2021
related degree: Master of Science - MSc
degree grantor: University of British Columbia
description: The volatile monoterpene metabolites camphor and borneol are considered undesired constituents of high-end lavender essential oils (EO) produced for the cosmetic industry. In plants, these metabolites are derived from borneol diphosphate (BPP), which itself results from the rearrangement of geranyl diphosphate (GPP) via a reaction catalyzed by the enzyme borneol diphosphate synthase (BPPS). Mahmoud group has recently cloned a unique BPPS gene from Lavandula x intermedia (the LiBPPS). However, the in planta role of this gene has not been evaluated. In this study, we aimed to confirm the in planta function of LiBPPS through its constitutive expression in sense and antisense in spike lavender (L. latifolia). To achieve this goal, the coding sequence of LiBPPS was placed under the control of the Cauliflower Mosaic Virus 35S (CaMV35S) promoter in sense and antisense orientations, and stably expressed in transformed L. latifolia plants via Agrobacterium-mediated transformation using hygromycin as a selectable marker. We evaluated the effect of LiBPPS overexpression (in sense and antisense) on abundance of BPPS transcript in transformed plants. As anticipated, overexpression of LiBPPS in sense resulted in an increase in BPPS mRNA levels, while overexpression of the gene in antisense led to a decrease in BPPS mRNA levels in transgenic plants. To determine whether altering the expression of BPPS influenced EO yield, and abundances of camphor, borneol and other monoterpenes, we analyzed the EO of the transgenic plants by gas chromatography mass spectrometry (GC-MS). Manipulating BPPS expression did not have a significant effect on oil yield. However, we observed a significant reduction in borneol and camphor, and a significant increase in the production of other EO constituents such as 1,8 cineole and limonene in 90.9 % of the BPPS-antisense plants relative to wild-type plants. Furthermore, meeting our expectations, 60% of the BPPS-sense plants produced higher levels of camph…
aggregated CHO: https://circle.library.ubc.ca/rest/handle/2429/77846
note: METABOLIC ENGINEERING OF ESSENTIAL OIL COMPOSITION IN LAVENDULA LATIFOLIA BY ALTERING EXPRESSION OF BORNEOL DIPHOSPHATE SYNTHASE By Elaheh Najafianashrafi B.Sc., University of Tehran, 2008 A THESIS SUBMITTED IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF MASTER OF SCIENCE in THE COLLEGE OF GRADUATE STUDIES (Biology) THE UNIVERSITY OF BRITISH COLUMBIA (Okanagan) April 2021 © Elaheh Najafianashrafi, 2021 ii The following individuals certify that they have read, and recommend to the College of Graduate Studies for acceptance, a thesis/dissertation entitled: Metabolic engineering of essential oil composition in Lavendula latifolia by altering expression of borneol diphosphate synthase submitted by Elaheh Najafianashrafi in partial fulfillment of the requirements of the degree of Master of Science. Dr. Soheil Mahmoud, Biology, Irving K. Barber School of Science Supervisor Dr. Michel Deyholos, Biology, Irving K. Barber School of Science Supervisory Committee Member Dr. Thuy Dang, Chemistry, Irving K. Barber School of Science Supervisory Committee Member Dr. Hadi Mohammadi, Faculty of Applied Science, School of Engineering University Examiner iii Abstract The volatile monoterpene metabolites camphor and borneol are considered undesired constituents of high-end lavender essential oils (EO) produced for the cosmetic industry. In plants, these metabolites are derived from borneol diphosphate (BPP), which itself results from the rearrangement of geranyl diphosphate (GPP) via a reaction catalyzed by the enzyme borneol diphosphate synthase (BPPS). Mahmoud group has recently cloned a unique BPPS gene from Lavandula x intermedia (the LiBPPS). However, the in planta role of this gene has not been evaluated. In this study, we aimed to confirm the in planta function of LiBPPS through its constitutive expression in sense and antisense in spike lavender (L. latifolia). To achieve this goal, the coding sequence of LiBPPS was placed under the control of the Cauliflower Mosa…
has type: Thesis/Dissertation
date issued: 2021-05
is shown at: 10.14288/1.0396855
language: eng
degree discipline: Biology
provider: Vancouver : University of British Columbia Library
publisher: University of British Columbia
rights: Attribution-NonCommercial-NoDerivatives 4.0 International
rights URI: http://creativecommons.org/licenses/by-nc-nd/4.0/
scholar level: Graduate
type: Text
identifier URI: http://hdl.handle.net/2429/77846