There are many technologies being developed for crop breeding. Two interesting technologies are genetic modification and genetic editing. Competitive pressures and changing consumer preferences are forcing organizations to invest heavily in these two technologies. Organizations must decide which traits they want to target and must commit significant time a money to the project. Traditionally, firms would decide which project to embark on if the project is net present value positive. Throughout the research and development process managers have flexibility to abandon the project once new information is received. That flexibility has value and real option analysis must be performed to value that flexibility. Once the value of a GM and GE project is determined, how might an organization decide which project to do? The concept of minimum required acreage (MRA) is developed in this study, allowing organizations to compare GM and GE technologies and decide which project to invest it.

Wong Wai Mei. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2001. / Includes bibliographical references (leaves 175-192). / Abstracts in English and Chinese. / Declaration --- p.ii / Acknowledgements --- p.iii / Abstract --- p.iv / Abbreviation --- p.vi / Table of Contents --- p.vii / Chapter Chapter 1 --- Introduction --- p.1 / Chapter Section I --- The Making of Genetically Modified Organisms --- p.2 / Chapter 1.1 --- Conventional breeding in agriculture --- p.2 / Chapter 1.2 --- What is genetic engineering? --- p.4 / Chapter 1.3 --- Plant transformation --- p.5 / Chapter 1.3.1 --- Agrobacterium-mediated --- p.6 / Chapter 1.3.2 --- Direct gene transfer --- p.8 / Chapter 1.3.2.1 --- Microparticle bombardment --- p.8 / Chapter 1.3.2.2 --- Protoplasts --- p.9 / Chapter 1.3.3 --- Gene silencing --- p.10 / Chapter 1.4 --- Examples of genetically modified crops --- p.13 / Chapter 1.5 --- Foreign genes commonly found in transgenic plants --- p.14 / Chapter Section II --- Benefits and Environmental Concern of GMOs --- p.17 / Chapter 2.1 --- Mechanism of GMO --- p.17 / Chapter 2.1.1 --- Herbicide tolerant crops --- p.18 / Chapter 2.1.2 --- Insect resistant crops --- p.19 / Chapter 2.1.3 --- Delayed ripening crops --- p.20 / Chapter 2.1.4 --- Virus resistant crops --- p.20 / Chapter 2.2 --- Benefits of GMOs --- p.21 / Chapter 2.3 --- Impact of GM foods to human health and the environment --- p.22 / Chapter 2.3.1 --- Human health --- p.22 / Chapter 2.3.1.1 --- GM potatoes --- p.23 / Chapter 2.3.1.2 --- CaMV risks? --- p.24 / Chapter 2.3.1.3 --- Food allergy --- p.25 / Chapter 2.3.2 --- Environmental concerns --- p.26 / Chapter 2.3.2.1 --- Horizontal gene transfer --- p.27 / Chapter 2.3.2.1.1 --- Selectable marker genes --- p.27 / Chapter 2.3.2.1.2 --- Herbicide resistant genes --- p.29 / Chapter 2.3.2.1.3 --- Insect resistant genes --- p.29 / Chapter 2.3.2.2 --- Ecology --- p.30 / Chapter 2.3.2.2.1 --- Monarch butterfly --- p.30 / Chapter Section III --- Future developments of GMO --- p.32 / Chapter 3.1 --- Designer Food and engineered plants --- p.32 / Chapter 3.1.1 --- Insect resistance --- p.33 / Chapter 3.1.2 --- Viral resistance --- p.33 / Chapter 3.1.3 --- Fungal resistance --- p.34 / Chapter 3.1.4 --- Nutritional quality --- p.34 / Chapter 3.1.5 --- Modifications of oil composition --- p.35 / Chapter 3.1.6 --- Medical applications --- p.37 / Chapter 3.1.7 --- Environmental applications --- p.40 / Chapter 3.1.7.1 --- Tolerance to high salinity and drought --- p.40 / Chapter 3.1.7.2 --- Tolerance to frost --- p.41 / Chapter 3.1.7.3 --- Bioremediation --- p.42 / Chapter 3.1.7.4 --- Biodegradable products --- p.43 / Chapter Section IV --- Regulation of GMO --- p.44 / Chapter 4.1 --- The question of labeling --- p.44 / Chapter 4.1.1 --- Moral and ethical issues --- p.44 / Chapter 4.1.2 --- Animal welfare --- p.45 / Chapter 4.2 --- International practice in GMO labeling --- p.46 / Chapter 4.2.1 --- United States of America --- p.46 / Chapter 4.2.2 --- Canada --- p.48 / Chapter 4.2.3 --- European Union --- p.49 / Chapter 4.2.4 --- Australia and New Zealand --- p.50 / Chapter 4.2.5 --- Japan --- p.51 / Chapter 4.2.6 --- Republic of Korea --- p.52 / Chapter 4.2.7 --- China --- p.53 / Chapter 4.2.8 --- Taiwan --- p.53 / Chapter 4.2.9 --- Hong Kong --- p.54 / Chapter Section V --- Uses of crops --- p.56 / Chapter 5.1 --- Uses of crops --- p.56 / Chapter 5.1.1 --- Soybean --- p.56 / Chapter 5.1.2 --- Corn --- p.57 / Chapter 5.1.3 --- Tomato --- p.58 / Chapter 5.1.4 --- Potato --- p.59 / Chapter 5.1.5 --- Rice --- p.60 / Chapter 5.1.6 --- Rapeseed --- p.61 / Chapter 5.1.7 --- Oil --- p.62 / Chapter 5.2 --- "Food additives, hormones and flavourings" --- p.63 / Chapter Chapter 2 --- Materials & Methods --- p.65 / Chapter 2.1 --- Materials --- p.66 / Chapter 2.1.1 --- Growth media & agar --- p.66 / Chapter 2.1.2 --- Reagents for agarose gel electrophoresis --- p.67 / Chapter 2.1.3 --- Reagents for preparation of competent cells --- p.67 / Chapter 2.1.4 --- Reagents for measurement of DNA concentration --- p.68 / Chapter 2.1.4.1 --- Measurement of DNA concentration by PicoGreen --- p.68 / Chapter 2.1.5 --- Reagents for Southern hybridization --- p.68 / Chapter 2.2 --- Methods --- p.70 / Chapter 2.2.1 --- Restriction endonuclease digestion --- p.70 / Chapter 2.2.2 --- Agarose gel electrophoresis of DNA --- p.70 / Chapter 2.2.3 --- DNA recovery from agarose gel --- p.71 / Chapter 2.2.3.1 --- QIAquick® gel extraction --- p.71 / Chapter 2.2.4 --- Ligation of purified DNA fragment into vector --- p.72 / Chapter 2.2.5 --- Transformation --- p.72 / Chapter 2.2.6 --- Rubidium chloride method for making competent cells --- p.12 / Chapter 2.2.7 --- Plasmid DNA preparation --- p.73 / Chapter 2.2.7.1 --- Concert Rapid Mini Prep --- p.73 / Chapter 2.2.7.2 --- QIAprep® Miniprep --- p.74 / Chapter 2.2.8 --- Extraction of plant genomic DNA --- p.75 / Chapter 2.2.8.1 --- Qiagen DNeasy´ёØ Plant Mini Kit --- p.75 / Chapter 2.2.9 --- Southern Hybridization --- p.75 / Chapter 2.2.9.1 --- Denaturation --- p.76 / Chapter 2.2.9.2 --- Blot transfer --- p.76 / Chapter 2.2.9.3 --- Pre-hybridization --- p.77 / Chapter 2.2.9.4 --- Synthesis of radiolabelled probe --- p.77 / Chapter 2.2.9.5 --- Hybridization of radiolabelled probe on filter --- p.77 / Chapter 2.2.9.6. --- Detection of hybridized probes --- p.78 / Chapter 2.2.10 --- Measurement of DNA concentration --- p.78 / Chapter 2.2.10.1 --- Determination of DNA on EtBr stained gel --- p.78 / Chapter 2.2.10.2 --- Determination of DNA by UV spectrophotometer --- p.78 / Chapter 2.2.10.3 --- Determination of DNA by PicoGreen --- p.79 / Chapter 2.2.11 --- DNA sequencing --- p.80 / Chapter 2.2.11.1 --- Automated sequencing by ABI Prism 377 --- p.80 / Chapter Chapter 3 --- PCR Diagnostics --- p.81 / Chapter 3.1 --- Applications of PCR to processed foods --- p.82 / Chapter 3.1.1 --- DNA quality --- p.82 / Chapter 3.1.2 --- PCR & Multiplex PCR --- p.83 / Chapter 3.1.3 --- Choice of primers --- p.84 / Chapter 3.1.4 --- Inhibitors --- p.84 / Chapter 3.2 --- Materials & Methods --- p.85 / Chapter 3.2.1 --- Selection of primers --- p.85 / Chapter 3.2.2 --- Amplification of target sequences --- p.86 / Chapter 3.2.3 --- Multiple amplification of target sequences --- p.87 / Chapter 3.3 --- Results --- p.88 / Chapter 3.4 --- Discussion --- p.93 / Chapter Chapter 4 --- Quality Control in GMO detection --- p.95 / Chapter 4.1 --- Standardization of pre- and post- PCR analysis --- p.96 / Chapter 4.1.1 --- General guidelines --- p.96 / Chapter 4.1.2 --- UV irradiation --- p.97 / Chapter 4.1.3 --- Inactivation protocols --- p.93 / Chapter 4.1.4 --- Positive and negative controls --- p.99 / Chapter 4.1.5 --- PCR verification --- p.99 / Chapter 4.1.6 --- Equipment decontamination --- p.100 / Chapter 4.2 --- Materials & Methods --- p.101 / Chapter 4.2.1 --- Selection of primers for external control --- p.101 / Chapter 4.2.2 --- Development of the external control --- p.101 / Chapter 4.2.3 --- Selection of primers for internal control --- p.103 / Chapter 4.3 --- Results --- p.104 / Chapter 4.4 --- Discussion --- p.107 / Chapter Chapter 5 --- DNA extraction from food samples --- p.110 / Chapter 5.1 --- Introduction --- p.111 / Chapter 5.2 --- Reagents and Buffers for DNA extraction from food samples --- p.112 / Chapter 5.2.1 --- Cetyltrimethylammonium bromide (CTAB) extraction method --- p.112 / Chapter 5.2.2 --- Organic-based extraction method --- p.113 / Chapter 5.2.3 --- Potassium acetate/sodium dodecyl sulphate precipitation method --- p.113 / Chapter 5.2.4 --- Hexane-based extraction method --- p.114 / Chapter 5.3 --- Weight and names of samples --- p.115 / Chapter 5.4 --- DNA extraction methods --- p.115 / Chapter 5.4.1 --- CTAB extraction method --- p.115 / Chapter 5.4.2 --- Qiagen DNeasy´ёØ plant mini kit --- p.116 / Chapter 5.4.3 --- Promega Wizard® genomic DNA purification --- p.116 / Chapter 5.4.4 --- Promega Wizard® Magnetic DNA purification system --- p.117 / Chapter 5.4.5 --- Promega Wizard® DNA Clean-Up system --- p.118 / Chapter 5.4.6 --- Qiagen QIAshreddrer´ёØ and QIAamp spin column --- p.119 / Chapter 5.4.7 --- Chelex-based extraction method --- p.119 / Chapter 5.4.8 --- Organic-based extraction method --- p.120 / Chapter 5.4.9 --- Nucleon PhytoPure extraction and purification method --- p.120 / Chapter 5.4.10 --- Potassium acetate/SDS precipitation method --- p.121 / Chapter 5.4.11 --- Hexane-based extraction method --- p.122 / Chapter 5.5 --- Results --- p.123 / Chapter 5.5.1 --- Comparison of eleven extraction methods --- p.123 / Chapter 5.5.2 --- Comparison of DNA extraction on selected methods --- p.125 / Chapter 5.6 --- Discussion --- p.132 / Chapter Chapter 6 --- Quantitative Analysis --- p.136 / Chapter 6.1 --- Introduction --- p.137 / Chapter 6.1.1 --- Chemistry of quantitative PCR --- p.138 / Chapter 6.1.2 --- PCR system --- p.140 / Chapter 6.2 --- Materials & Methods --- p.142 / Chapter 6.2.1 --- Design of primers and probes --- p.142 / Chapter 6.2.2 --- Methods --- p.145 / Chapter 6.3 --- Results --- p.146 / Chapter 6.3.1 --- Selection of primer/probe --- p.146 / Chapter 6.3.2 --- Primer optimization --- p.149 / Chapter 6.3.3 --- Quantitative analysis of real samples --- p.158 / Chapter 6.4 --- Discussion --- p.152 / Chapter Chapter 7 --- Conclusion --- p.168 / References --- p.175 / Appendix --- p.193

Thesis (M.Tech.: Biotechnology)-Dept. of Biotechnology and Food Technology, Durban University of Technology, 2007 iv, 75 leaves / The development of transgenic plants resulted in the need to utilize the various molecular methods (e.g., ELISA, real - time PCR etc.) for the detection or analysis of the presence or absence of a specific trait in a particular plant (Bt in this study). The overall aim of this study was to optimize a half – seed extraction technique as part of a laboratory protocol for transgenic maize plants and to explore the possibility of using the following molecular techniques: horizontal isoelectric focusing, real - time PCR and ELISA, as methods for detection of the Bt trait for incorporation into the half – seed extraction protocol.

Thesis (MSc)--University of Stellenbosch, 2002. / ENGLISH ABSTRACT: Soil salinity is a major limiting factor of plant and crop growth, because the absorption of water and nutrients is such a complex process while low and moderate salinity are omnipresent. Plant growth is affected negatively if a specific ion concentration exceeds its threshold and becomes toxic. The detrimental effect of soil affected by salt on crop production is increasing worldwide (Tanji, 1990). The level to which plants can tolerate high salinity levels is genetically controlled with several physiological and genetic mechanisms contributing to salt tolerance (Epstein & Rains, 1987). The most effective way of addressing the limitations of crop productivity in saline areas, is the development of salt tolerant varieties. Understanding the genetics of salt tolerance is, therefore, necessary for the development of an effective breeding strategy for salt tolerance. The department of Genetics (US) conducts a wide crosses research programme aiming to transfer genes for salt tolerance to wheat and triticale. The donor species, Thinopyrum disticum, an indigenous coastal wheat grass, adapted to high concentrations of salt, was crossed with cultivated rye (Secale cereale) in an attempt to study the genetics of salt tolerance (Marais et al., 1998). The primary goal of this study was to find molecular markers (RAPD and AFLP) which associate with chromosomes promoting salt tolerance for later attempts to transfer the genes to triticale. Seventy clones of secondary hybrids (Th disticum /4x-rye 1/2x-rye) were tested for salt tolerance and showed different levels of salt tolerance. RAPD-marker analyses were used to identify polymorphisms between salt tolerant and salt sensitive plants. Twelve RAPD primers produced clear, analyzable and repetitive polymorphic . fragments that can be used as useful markers. Different AFLP-primer combinations were tested against the genotypes of 15 clones (Marais & Marais 2001, unpublished data) and produced approximately 2000 clearly distinguishable AFLP fragments, of which 54 (3%) were polymorphic fragments. Two RAPD fragments and 4 AFLP fragments that can be used as possible markers for the presence of chromosomes that contribute to salt tolerance were identified. The interpretation of the markers was complicated by heterogeneity among plants with regard to the origin of their chromosomes and the genetic diversity of the rye genome. It is also possible that chromosome re-arrangement took place during backcrossing, which could have complicated the data. / AFRIKAANSE OPSOMMING: Versouting is een van die groot beperkende faktore op plant- en gewasgroei, omdat die opname van water en voedingstowwe so In ingewikkelde proses is en die effek van lae of matige versouting so alomteenwoordig is. Plantgroei word nadelig geaffekteer as 'n spesifieke ioonkonsentrasie sy drempelwaarde oorskry en toksies word. Die nadelige effek van soutgeaffekteerde grond op gewasproduksie, is wêreldwyd aan die toeneem (Tanji, 1990). Die vlak waartoe plante hoë konsentrasies sout kan hanteer is onder genetiese beheer met verskeie fisiologiese en genetiese meganismes wat 'n bydrae maak tot soutverdraagsaamheid (Epstein & Rains, 1987). Die mees effektiewe manier om die beperkinge op gewas produktiwiteit in versoute gebiede te oorkom, is die ontwikkeling van soutverdraagsame variëteite. Begrip van die genetika van soutverdraagsaamheid is dus noodsaaklik vir die ontwikkeling van In effektiewe telingsstrategie. Die departement Genetika (US) bedryf tans 'n wye-kruisings navorsingsprogram waarmee gepoog word om gene vir soutverdraagsaamheid na korog en koring oor te dra. Die skenkerspesie, Thinopyrum disticum, In inheemse strandkoringgras wat aangepas is by hoë konsentrasies sout, is gekruis met verboude rog (Secale cereale) in 'n poging om die oorerwing van soutverdraagsaamheid te bestudeer (Marais et al., 1998). Die hoofdoel van hierdie studie was om molekulêre merkers (RAPD en AFLP) te vind, wat assosieer met chromosome wat soutverdraagsaamheid bevorder en om nuttige merkers daar te stel vir latere pogings om die gene na korog en koring oor te dra. Ongeveer 70 klone van sekondêre hibriede (Th distichum I 4x-rog /I 2x-rog) is onderwerp aan souttoetse en het verskillende grade van soutverdraagsaamheid getoon. RAPDmerker analise is gebruik om polimorfismes te identifiseer tussen soutverdraagsame en soutsensitiewe plante. Twaalf RAPD inleiers het duidelike, ontleedbare en herhalende polimorfiese fragmente opgelewer en moontlike nuttige merkers uitgewys. Verskillende AFLP-inleier kombinasies, wat getoets is teen die genotipes van 15 klone (Marais & Marais, 2001 ongepubliseerde data) het ongeveer 2000 duidelik onderskeibare AFLP fragmente geproduseer, waarvan 54 (3%) polimorfiese fragmente was. Twee RAPD fragmente en 4 AFLP fragmente is geïdentifiseer wat as moontlike kandidaat merkers gebruik kan word vir die identifisering van chromosome wat bydra tot soutverdraagsaamheid . Die interpretasie van die merkers is bemoeilik deur heterogeniteit tussen die plante wat betref die agtergrond van chromosome wat hulle besit en die genetiese diversiteit van die rog genoom. Dit is ook moontlik dat chromosoom herrangskikking plaasgevind het tydens terugkruising, wat die data verder kon kompliseer.

Thesis (M.Tech.: Biotechnology)-Dept. of Biotechnology and Food Technology, Durban University of Technology, 2007 iv, 75 leaves / The development of transgenic plants resulted in the need to utilize the various molecular methods (e.g., ELISA, real - time PCR etc.) for the detection or analysis of the presence or absence of a specific trait in a particular plant (Bt in this study). The overall aim of this study was to optimize a half – seed extraction technique as part of a laboratory protocol for transgenic maize plants and to explore the possibility of using the following molecular techniques: horizontal isoelectric focusing, real - time PCR and ELISA, as methods for detection of the Bt trait for incorporation into the half – seed extraction protocol.

The NRT2 (high affinity nitrate transporter 2) family is a part of the iHATS (inducible high affinity system) that studies have shown is responsible for the influx of nitrate into the plant cell after provision of nitrate. The ZmNRT2.1 from Zea mays was constitutively expressed in Nicotiana tabacum. To assess how over-expression of this foreign NRT2.1 affects nitrate influx by plants, nitrate content in leaf and root tissue, gene expression, and vegetal growth were analyzed in media with deficient or high nitrate concentrations (0.1, 1, or 10 mM). Compared to wild type plants: the transgenic lines had a significantly larger fresh weight in all nitrate conditions; primary root length was significantly longer in the 0.1 and 1 mM nitrate conditions; both the fresh weight and the primary root length were significantly higher when 50 mM NaCl was applied as a stress factor to medias containing 0.1 and 10 mM nitrate. / Includes bibliography. / Thesis (M.S.)--Florida Atlantic University, 2015. / FAU Electronic Theses and Dissertations Collection

Genetically modified (GM) crops are commercially cultivated worldwide but there are concerns on their possible negative impacts on soil biodiversity. A glasshouse study was conducted to determine effects of Bt maize residues on soil microbial diversity. Residues of Bt maize (PAN 6Q-308B) and non-Bt maize (PAN 6Q-121) were incorporated into the soil and corresponding maize seeds planted. The treatments were replicated three times. Fertilizer and water application were similar for both treatments. Rhizosphere and bulk soil was destructively sampled from each treatment and analyzed for microbial community level physiological profiles using Biolog plates with 31 different carbon substrates. Absorbance in the Biolog plates was recorded after 72 h of incubation at 20oC. Arbuscular mycorrhizal fungi spore counts were also determined. Field studies were conducted at the University of Free State and University of Fort Hare Research Farms to determine the effects of growing Bt maize on soil microbial diversity. One Bt maize cultivar (PAN6Q-308B) and non-Bt maize (PAN6Q-121) were grown in a paired experiment at University of Free State farm, while two Bt maize (DKC61-25B and PAN6Q-321B) and their near-isogenic non-Bt maize lines (DKC61-24 and PAN6777) were grown in a randomized complete block design with three replicates. Fertilization, weed control and water application, were similar for both Bt maize cultivars and their non-Bt maize counterparts. Rhizosphere soil samples were collected by uprooting whole plants and collecting the soil attached to the roots. The samples were analysed for microbial diversity and for arbuscular mycorrhizae fungal spore counts. Principal component analysis showed that soil microbial diversity was affected more by sampling time whereas genetic modification had minimal effects. Presence of residues also increased the diversity of microorganisms. Mycorrhizal fungal spores were not affected by the presence of Bt maize residues. Growing Bt maize had no effect on the soil microbial diversity in the rhizosphere.

In early 2003, a persistent drought threatened about 15 million people in the Southern African region (SADC) with starvation as farmers in this region were not able to produce enough food. A similar threat was experienced in the United States of America (USA). The Americans responded by introducing GM technology, which thankfully stabilised corn production and food security. It was against this backdrop that the South African government legalised and supported GM technology in the farming industry. However, the technology became a contentious issue amongst scholars, politicians and policy makers as well as farmers. Therefore, this study analysed the perceptions of small-scale and large-scale farmers, located in Paarl, Western Cape, South Africa, on the adoption of GM technology. This qualitative study, using a case study design, collected primary data from thirty (30) farmers: fifteen (15) small-scale and fifteen (15) large-scale farmers. The findings revealed complex factors influencing farmers’ adoption decisions and that Adopter perception (AP) and Consumer perception (CP) play a key role in their adoption of GM technology. These commercially and profit-driven farmers avoid using GM technology because public opinion and the markets weigh heavily against it. It was concluded that the farmers regarded GM technology as just one of many agricultural technologies and not as an exception. It was also considered unaffordable and detrimental to the environment, the economy and their livelihoods.The study recommends that the government should fully investigate public perceptions with regard to the adoption of any new agricultural innovation prior to making policy decisions. / Development Studies / M.A. (Development Studies)

Text in English / South Africa is one of the biggest producers of genetically modified crops in the world. However, recent studies in South Africa show a low public willingness to consume genetically modified crops and accept modern agricultural biotechnology. The study analysed public perception towards consuming genetically modified crops and the acceptance of modern agricultural biotechnology in South Africa. 220 participants (N = 220) were sampled from the city of Kempton Park and the Chi-square formula was used to determine how well the sample represented the population under study. Data was collected using a 7-point Likert scale questionnaire designed following the guidelines for developing a theory of planned behaviour questionnaire in Ajzen (1991, 2001). Data analyses were carried out using the Statistical Package for Social Sciences (SPSS). The Cronbach’s alpha and Exploratory Factor Analysis were both used to determine the internal consistency and validity of the questionnaire. Correlations, independent sample t-tests, ANOVA, linear regression, and path analysis were also conducted. Findings of the study confirmed that there is low public willingness to consume genetically modified crops and to accept modern agricultural biotechnology in South Africa. / Development Studies / M.A. (Development Studies)