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Engineering wheat (Triticum aestivum L.) for abiotic resilience by manipulating small ubiquitin-like modifiers

Thesis (MSc)--Stellenbosch University, 2016.

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Main Author: Le Roux, Marlon Luke
Other Authors: Botha-Oberholster, Anna-Maria
Format: Thesis
Language:en_ZA
Published: Stellenbosch : Stellenbosch University 2016
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access_status_str Open Access
author Le Roux, Marlon Luke
author2 Botha-Oberholster, Anna-Maria
author_browse Botha-Oberholster, Anna-Maria
Le Roux, Marlon Luke
author_facet Botha-Oberholster, Anna-Maria
Le Roux, Marlon Luke
author_sort Le Roux, Marlon Luke
collection Thesis
dc_rights_str_mv Stellenbosch University
description Thesis (MSc)--Stellenbosch University, 2016.
format Thesis
id oai:scholar.sun.ac.za:10019.1/98275
institution Stellenbosch University (South Africa)
language en_ZA
last_indexed 2026-06-10T12:42:38.497Z
license_str Other — see source repository
provenance_str_mv Harvested via OAI-PMH from SUNScholar — Stellenbosch University Repository
publishDate 2016
publishDateRange 2016
publishDateSort 2016
publisher Stellenbosch : Stellenbosch University
publisherStr Stellenbosch : Stellenbosch University
record_format dspace
source_str SUNScholar — Stellenbosch University Repository
spelling oai:scholar.sun.ac.za:10019.1/98275 Engineering wheat (Triticum aestivum L.) for abiotic resilience by manipulating small ubiquitin-like modifiers Le Roux, Marlon Luke Botha-Oberholster, Anna-Maria Van der Vyver, Christell Stellenbosch University. Faculty of AgriSciences. Dept. of Genetics. Wheat (Triticum aestivum L.) Wheat -- Breeding Wheat Genetic engineering Transgenic wheat Genetic modification of wheat UCTD Thesis (MSc)--Stellenbosch University, 2016. ENGLISH ABSTRACT: Research is becoming increasingly more focused on innovative strategies to improve wheat (Triticum aestivum L.) to meet current and future consumer demands. With the number of countries facing extreme climate variability increasing, drastic steps need to be taken to ensure local food security. Although genetic engineering of staple crops has been explored widely, they require challenging platforms with highly efficient explants with enhanced regenerative abilities for callus formation and somatic embryogenesis to consistently yield plantlets with altered attributes. Thus, this study describes the necessary steps to obtain transgenic wheat with relative ease, with the goal of improved abiotic stress tolerance. The developed in vitro regeneration and cryopreservation system was then used to introduce genes into bread wheat, namely Overly Tolerant to Salt 1 (OTS1) and OTS2 (both are Small Ubiquitin-like Modifier (SUMO) protease genes) and transcription factor known as Inducer of CBF expression 1 (ICE1). The class of molecules have emerged as an influential mechanism for targeted protein management, as SUMO proteases play a vital role in regulating pathway flux and are therefore ideal targets for manipulating stress-responsive SUMOylation. This study thus describes the isolation and cloning of three genes and its manipulation into constitutive and drought inducible vectors. The latter makes use of a unique promoter which was characterized in silico, after which it was then applied. Finally this study also demonstrated the compatibility of the vectors within wheat as it was confirmed that genetic modification of wheat was achieved by particle bombardment. AFRIKAANSE OPSOMMING: Navorsing lê toenemend meer klem op die ontwikkeling van innoverende strategieë met die oog op die verbetering van koring (Triticum aestivum L.) opbrengste om aan die huidige sowel as die toekomstige behoeftes te voldoen. Met die toenemende klimaat veranderinge wat talle lande in die gesig staar, moet drastiese stappe geneem word om plaaslike voedselsekuriteit te verseker. Alhoewel die genetiese modifisering van stapelvoedsel gewasse al wyd verken is, is daar ‘n behoeftes vir uitdagende platforms met die vermoë om begin material (bv. onvolwasse embrios) doeltreffend om te skakel in kallus en somatiese embrios om ten einde deurlopend plante met verbeterde eienskappe te lewer. Dus, die huidige studie beskryf die noodsaaklike stappe wat gevolg moet word om transgeniese koring met relatiewe gemak te verkry met die doel om strestolerasie te verbeter. Die ontwikkelde in vitro sisteem, was aangewend om bepaalde gene, naamlik “Overly Tolerant to Salt 1” (OTS1) and OTS2 (beide SUMO protease gene), in broodkoring uit te druk. Dieselfde stappe is gevolg met ‘n addisionele transkripsie factor genaamd “Inducer of CBF expression 1” (ICE1). Die SUMO (“Small Ubiquitin-like Modifier”) klas van molekule het na vore gekom as ‘n invloedryke meganisme vir geteikende proteinbestuur, aangesien SUMO protease ‘n noodsaaklike rol speel in die regulering van biochemiese pad verloop en is dus ideale teikens vir die manipulering van stress-verwante SUMO-lasie. Hierdie studie handel dus oor die isolasie en klonering van drie gene en hul manipulasie in konstitutiewe en droogte induserende vektore. Laasgenoemde maak gebruik van ‘n unieke promoter wat in silico gekarakteriseer is. Laastens word die bruikbaarheid van die vektore in koring gedemonstreer nadat suksesvolle genetiese modifisering bevestig is. Masters 2016-03-09T13:59:24Z 2018-05-31T03:00:05Z 2016-03 Thesis http://hdl.handle.net/10019.1/98275 en_ZA Stellenbosch University 161 pages : illustrations, maps application/pdf application/pdf Stellenbosch : Stellenbosch University
spellingShingle Wheat (Triticum aestivum L.)
Wheat -- Breeding
Wheat Genetic engineering
Transgenic wheat
Genetic modification of wheat
UCTD
Le Roux, Marlon Luke
Engineering wheat (Triticum aestivum L.) for abiotic resilience by manipulating small ubiquitin-like modifiers
title Engineering wheat (Triticum aestivum L.) for abiotic resilience by manipulating small ubiquitin-like modifiers
title_full Engineering wheat (Triticum aestivum L.) for abiotic resilience by manipulating small ubiquitin-like modifiers
title_fullStr Engineering wheat (Triticum aestivum L.) for abiotic resilience by manipulating small ubiquitin-like modifiers
title_full_unstemmed Engineering wheat (Triticum aestivum L.) for abiotic resilience by manipulating small ubiquitin-like modifiers
title_short Engineering wheat (Triticum aestivum L.) for abiotic resilience by manipulating small ubiquitin-like modifiers
title_sort engineering wheat triticum aestivum l for abiotic resilience by manipulating small ubiquitin like modifiers
topic Wheat (Triticum aestivum L.)
Wheat -- Breeding
Wheat Genetic engineering
Transgenic wheat
Genetic modification of wheat
UCTD
url http://hdl.handle.net/10019.1/98275
work_keys_str_mv AT lerouxmarlonluke engineeringwheattriticumaestivumlforabioticresiliencebymanipulatingsmallubiquitinlikemodifiers