Development of molecular markers and cloning of genes involved in the biosynthesis of monoterpenes in Lavandula
department or school: Arts and Sciences, Irving K. Barber School of (Okanagan)
department or school: Biology, Department of (Okanagan)
data provider: DSpace
degree campus: UBCO
creator: Adal, Ayelign Mengesha
issued: 2019-08-16T16:03:16Z
issued: 2019
related degree: Doctor of Philosophy - PhD
degree grantor: University of British Columbia
description: Several species of the genus Lavandula (lavenders) are important plants grown for their essential oils (EO), which find extensive applications in cosmetics, personal care products and alternative medicines. The EO of these plants is mainly constituted of monoterpenes, the C₁₀ class of terpenoids. In this study, we investigated three aspects of lavenders using advanced genomic resources derived from EO-producing tissues of these plants. Firstly, we developed diagnostic simple sequence repeat (SSR) markers from expressed sequence tag (EST) databases to delineate closely related species. Over 250 SSRs (≥ 18 bp) were developed from L. angustifolia and L. x intermedia ESTs. Thirty-one EST-SSRs were validated, 24 of which had a strong discrimination power in donor plant species, and a considerable transferability into related species. Secondly, we identified and functionally characterized cDNAs encoding geranyl pyrophosphate synthase (GPPS), which produces GPP, the linear precursor to all regular monoterpenes. Lavender GPPS is a heterodimeric protein composed of a small subunit (LiGPPS.SSU1), and a large subunit (LiGPPS.LSU). Recombinant LiGPPS.SSU1 interacts with LiGPPS.LSU, catalyzing the condensation of isopentenyl pyrophosphate (IPP) and dimethylallyl pyrophosphate (DMAPP) to produce GPP. Recombinant LiGPPS.SSU1 also interacts with lavender and N. benthamiana geranylgeranyl pyrophosphate synthase (GGPPS; the enzyme responsible for synthesis of GGPP, the linear precursor to diterpenes), modifying its activity to form GPP. Transcript levels for LiGPPS.SSU1 and LiGPPS.LSU parallel total monoterpene concentration, with more transcripts present in flowers where most of lavender EO accumulates. Finally, we cloned and functionally characterized lavender cDNAs encoding two monoterpene synthase genes, 3-carene synthase (Li3CARS) and S-linalool synthase (LiS-LINS), in vitro. Recombinant Li3CARS and LiS-LINS proteins converted GPP into 3-carene and S-linalool, respectively. Li3C…
aggregated CHO: https://circle.library.ubc.ca/rest/handle/2429/71318
note: DEVELOPMENT OF MOLECULAR MARKERS AND CLONING OF GENES INVOLVED IN THE BIOSYNTHESIS OF MONOTERPENES IN LAVANDULA by Ayelign Mengesha Adal B.Sc., Jimma University, 2006 M.Sc., Leibniz University of Hannover, 2011 A THESIS SUBMITTED IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF DOCTOR OF PHILOSOPHY in THE COLLEGE OF GRADUATE STUDIES (Biology) THE UNIVERSITY OF BRITISH COLUMBIA (Okanagan) August 2019 © Ayelign Mengesha Adal, 2019 ii The following individuals certify that they have read, and recommend to the College of Graduate Studies for acceptance, a thesis/dissertation entitled: DEVELOPMENT OF MOLECULAR MARKERS AND CLONING OF GENES INVOLVED IN THE BIOSYNTHESIS OF MONOTERPENES IN LAVANDULA submitted by Ayelign Mengesha Adal in partial fulfillment of the requirements of the degree of Doctor of Philosophy Dr. Soheil S. Mahmoud, Biology, Irving K. Barber School of Arts and Sciences, UBC Supervisor Dr. Micheal Deyholos, Biology, Irving K. Barber School of Arts and Sciences, UBC Supervisory Committee Member Dr. Louise Nelson, Biology, Irving K. Barber School of Arts and Sciences, UBC Supervisory Committee Member Dr. Wesley Zandberg, Irving K. Barber School of Arts and Sciences, UBC University Examiner Dr. Jim Mattsson, Department of Biological Sciences, Simon Fraser University External Examiner Additional Committee Members include: Dr. Nasser Yalpani, Adjunct Professor, Biology, Irving K. Barber School of Arts and Sciences, UBC Supervisory Committee Member . iii Abstract Several species of the genus Lavandula (lavenders) are important plants grown for their essential oils (EO), which find extensive applications in cosmetics, personal care products and alternative medicines. The EO of these plants is mainly constituted of monoterpenes, the C10 class of terpenoids. In this study, we investigated three aspects of lavenders using advanced genomic resources derived from EO-producing tissues of these plants. Firstly, we developed diagnostic simple sequence repeat …
has type: Thesis/Dissertation
date issued: 2019-09
is shown at: 10.14288/1.0380463
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/2429/71318