Preliminary analyses of three kinds of promoter activity showed that a-actin gene promoter was chosen to initiate the hormone transcription for the first consideration. We have fused the cDNAs encoding the intact somatolactins in frame to a zebrafish a-actin gene promoter to generate transgenic zebrafish lines co-injected with a GFP protein driven by the same promoter. The transgenic zebrafish were selected from GFP expression and confirmed by genomic PCR and Southern blot analysis, then maintained as transgenic founders. Measurement of the transgenes' expressions and the expressions of marker genes in different pathways by using real-time PCR provided a general understanding of SLs' actions. The data obtained indicated that the over-expressing of SLalpha and SLbeta in vivo significantly enhance the transcriptions of the insulin-like growth factors, IGF1 (5.46-fold and 6.77-fold), IGF2a (4.38-fold and 4.35-fold) and IGF2b (2.83-fold and 3.94-fold), but down-regulated IGF3 (a novel member found specifically in gonad) in larvae. However, the stimulation by administration of recombinant proteins (SLalpha and SLbeta) only showed a slight induction of the mRNA levels of IGFs (IGF1, IGF2a and IGF2b) on ZFL cells in vitro. / Somatolactin (SL) is a novel member of pituitary polypeptide hormone found only in fish; it shares significant structural homology with prolactin and growth hormone. Since somatolactin receptor (SLR) was first defined as GHR1 and orthologous to the growth hormone receptor GHR2, SL and GH may share similar actions in growth and development. Recently, two SLs have been identified as SLalpha and SLbeta with similar structures, freshwater fish have these two isoforms found in the same species and only one isoform (SLalpha) is found in marine species. The two isoforms of SL may have different functions and physiological actions. To investigate the roles of SLs on vertebrate development and embryogenesis, we generated transgenic fish models with "all zebrafish" elements in origin to study the physiological functions of SLs in zebrafish. / The ectopic expression of somatolactins also results in up-regulating gene expression of insulin, leptin, sterol regulatory element binding protein 1 (SREBP1) and fatty acid synthase (FAS), as well as the expression of vitellogenin and melanocyte-stimulating hormone (MSH) levels while causing reduction of catalase (CAT) and glutathione S-transferase (GST) levels in larvae. The results here represent the similar function between SLalpha and SLbeta and reveal more details in fish of the endocrinology system involvement in growth development, glucose synthesis, lipid metabolism, reproduction, pigmentation and antioxidant defense system through the actions of SLs. / Three different gene promoters of zebrafish have been isolated to initiate the ectopic expression of somatolactins in vivo, which including a constitutional beta-actin gene promoter, a liver specific transferrin gene promoter and a zinc ion inducible metallothionein (MT) gene promoter. The promoter activities were tested in fish cell-line by using luciferase reporter assay. In MT gene promoter, two alleles of a zebrafish metallothionein II gene (zMT-II) promoter (zMT-IIA and zMT-IIB) containing 10 MREs in the 5'-flanking region (1,514 bp) were identified in zebrafish. These putative MREs were confirmed via electrophoretic mobility shift assay (EMSA) to have binding activities from the cellular and nuclear extracts of a zebrafish cell line, ZFL. Transient gene expression studies using zebrafish liver (ZFL) cell lines also confirmed that the most distal cluster of MREs contributed to the maximal induction of zMT-IIA activity by Zn2+ and the Zn 2+ induction was dose-dependent. EMSA also identified transcription factor(s) of two different sizes from the cytoplasmic and nuclear extracts of the ZFL cells that were able to bind with the MREs, but no increase in MRE binding was detected in the extracts of these cells after Zn2+ or Cd2+ treatment, compared with untreated control cells. The mechanisms of MT gene transcription induction via metal ions are discussed herein. / Wan, Guohui. / Adviser: Chan King Ming. / Source: Dissertation Abstracts International, Volume: 73-01, Section: B, page: . / Thesis (Ph.D.)--Chinese University of Hong Kong, 2009. / Includes bibliographical references (leaves 139-163). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [201-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese.

A planta daninha Amaranthus palmeri é nativa dos Estados Unidos, porém foi pela primeira vez relatada no Brasil no ano de 2015. Embora comprovadamente com resistência múltipla aos herbicidas inibidores da ALS e da EPSPS, até o momento não foram investigadas as bases moleculares da resistência. Além disso, por causa da recente introdução da planta daninha no país, alternativas de manejo com culturas tolerantes a herbicidas necessitam ser estudadas. Sendo assim, os objetivos desse trabalho são de caracterizar a espécie de planta daninha introduzida no país, identificar os mecanismos de resistência aos herbicidas inibidores da ALS e da EPSPS presentes no biótipo, e propor abordagens de manejo em ambientes dos novos eventos transgênicos resistentes a herbicidas. Um bioensaio utilizando marcadores genéticos foi desenvolvido para confirmar que a população coletada no estado do Mato Grosso (BR-R) é A. palmeri, e não A. tuberculatus, outra espécie dióica do gênero Amaranthus. Os resultados de experimentos de curvas de dose-resposta e acúmulo de chiquimato indicaram que a BR-R possui alto nível de resistência, com DL50 de 4.426 e 3.400 g glyphosate ha-1 no primeiro e segundo experimento, respectivamente, mais que o dobro da dose típicamente recomendada para o controle da espécie e, adicionalmente, observou se acúmulo mínimo de chiquimato a concentração de 1 mM nos tecidos das plantas tratadas com o herbicida. BR-R também foi resistente a herbicidas dos grupos químicos das sulfoniluréias e imidazolinonas. O mecanismo de resistência ao glyphosate encontrado nesta população foi a super expressão gência, através do aumento no número de cópias do gene da EPSPS no genoma da planta BR-R, entre 50 e 179 cópias adicionais. Além disso, duas substituições de aminoácidos foram observadas na sequência da ALS, W574L e S653N, conferindo resistência tanto a sulfoniluréias quanto a imidazolinonas. No experimento utilizandos os herbicidas correspondentes às culturas geneticamente modificadas com novos traits de tolerância a herbicidas observou se, de uma forma geral, que as associações de herbicidas apresentaram níveis de controle mais satisfatórios. Assim, esta pesquisa confirma a introdução de da espécie A. palmeri no Brasil, assim como a resistência múltipla aos herbicidas inibidores da EPSPS e da ALS. Seu manejo é mais eficaz através da associação de herbicidas, garantindo assim o uso racional das novas tecnologias de culturas geneticamente modificadas com tolerância a herbicidas. / Palmer Amaranth (Amaranthus palmeri) is a weed species native to the United States, but it was reported in Brazil for the first time in 2015. Despite this population being resistant to EPSPS and ALS inhibitors, the molecular basis of its multiple resistance is unknown up to date. Because of this species introduction to Brazil, alternatives of management with the new herbicide-tolerant crops technologies need to be studied. The objectives of this research are to characterize the weed species introduced to Brazil, identify the mechanisms conferring resistance to ALS and EPSPS inhibitors herbicides, and to propose management approaches in environments with the new genetically modified herbicide-tolerant crops. A genotyping bioassay using genetic markers was developed to confirm that the species collected in the state of Mato Grosso (BR-R) is indeed A. palmeri and not A. tuberculatus, another dioceous species in the Amaranthus genus. Dose-response experiments and shikimate accumulation bioassay data indicate high level of resistance, with LD50 of 4,426 and 3,400 g glyphosate ha-1 in the first and second experiments, respectively, higher than the double rate tipically recommended to control it, and minimal accumulation in BR-R with 1 mM of glyphosate in treated plants in the leaf disks assay. BR-R also was resistanto to sulfonilurea and imidazolinone herbicides. The mechanism conferring resistance to glyphosate identified in this population was gene amplification, with increased EPSPS copy number - between 50 and 179 more copies in BR-R. Besides, two target-site mutations were identified in the ALS gene sequencing, W574L and S653N, conferring resistance to sulfonilureas and imidazolinones. The weed control experiment, overal, herbicide tank mixtures achieved higher levels of control. Therefore, this research confirms the introduction of A. palmeri to Brazil, as well as its multiple resistance to EPSPS and ALS inhibitor herbicides. Its control is more efficient with herbicide mixtures, which guarantees more susteinable use of the new herbicide-tolerant crop technologies.

Yang, Fan. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2010. / Includes bibliographical references (leaves 111-132). / Abstracts in English and Chinese. / ACKNOWLEDGEMENTS --- p.iii / ABSTRACT --- p.iv / TABLE OF CONTENTS --- p.viii / LIST OF FIGURES --- p.xii / LIST OF TABLES --- p.xv / LIST OF ABBREVIATIONS --- p.xvi / Chapter Chatper 1. --- General Introduction --- p.1 / Chapter Chapter 2. --- Literature Review --- p.5 / Chapter 2.1 --- Facts on food allergy --- p.5 / Chapter 2.1.1 --- Food allergy and its prevalence --- p.5 / Chapter 2.1.2 --- Pathogenesis of food allergy --- p.6 / Chapter 2.1.3 --- Clineal disorders caused and diagnosis of food allergy --- p.8 / Chapter 2.2 --- Allergenicity assessment of genetically engineered food --- p.13 / Chapter 2.2.1 --- The structural and sequence homology of proteins as a criterion for food allergenicity assessment --- p.14 / Chapter 2.2.2 --- Digestion stability as a criterion for food allergenicity assessment --- p.15 / Chapter 2.2.3 --- Animal models for Food Allergenicity Assessment --- p.21 / Chapter 2.3 --- The importance of rice and its nutritional facts --- p.27 / Chapter 2.3.1 --- The importance of rice --- p.27 / Chapter 2.3.2 --- Rice nutritional facts and its relationship with malnutrition --- p.28 / Chapter 2.4 --- Food allergenicity research in rice --- p.30 / Chapter 2.5 --- Glutelin overexpression transgenic rice GT3 --- p.33 / Chapter 2.6 --- Recent and future perspectives for treatment of food allergy --- p.36 / Chapter Chapter 3. --- Materials and Methods --- p.39 / Chapter 3.1 --- Rice Seed Protein Extraction --- p.39 / Chapter 3.1.1 --- Rice varieties for protein extraction --- p.39 / Chapter 3.1.2 --- Protein extraction from rice seeds --- p.39 / Chapter 3.1.3 --- Fractionation of major rice seed storage proteins --- p.40 / Chapter 3.1.4. --- Protein quantification --- p.41 / Chapter 3.1.5 --- Tricine SDS-PAGE --- p.42 / Chapter 3.2 --- Simulated Gastric Digestibility Assay --- p.43 / Chapter 3.2.1 --- Assay System --- p.43 / Chapter 3.2.2 --- Preparation of Simulated Gastric Fluid --- p.43 / Chapter 3.2.3 --- Assay Procedures --- p.44 / Chapter 3.2.4 --- Results Interpretation --- p.44 / Chapter 3.3 --- Construction of Mouse Models --- p.45 / Chapter 3.3.1 --- Mouse strain and reagents used --- p.45 / Chapter 3.3.2 --- Mouse Model I --- p.46 / Chapter 3.3.3 --- Mouse Model II --- p.50 / Chapter 3.3.4 --- Mouse Model III --- p.51 / Chapter 3.4 --- Bioinformatic Analysis of Glutelin Sequence --- p.52 / Chapter 3.5 --- Epitope Mapping of Glutelin --- p.55 / Chapter 3.5.1 --- Bioinformatic Analysis --- p.55 / Chapter 3.5.2 --- Direct and Competitive ELISA --- p.56 / Chapter 3.5.3 --- Western Blot Analysis --- p.57 / Chapter 3.5.4 --- IgE-binding assay --- p.58 / Chapter Chapter 4. --- Results and Discussion --- p.60 / Chapter 4.1 --- Rice Seed Protein Extraction --- p.60 / Chapter 4.1.1 --- Rice Protein Extraction --- p.60 / Chapter 4.1.2 --- Extraction of rice major seed storage protein fractions --- p.62 / Chapter 4.2 --- Simulated Gastric Digestibility Assay --- p.64 / Chapter 4.2.1 --- Pepsin Digestibility of total protein from GT3 and WT rice seeds --- p.64 / Chapter 4.2.2 --- Pepsin Digestibility of major storage protein fractions in GT3 and WT rice --- p.68 / Chapter 4.2.3 --- Summary of Pepsin Digestibility Assay --- p.74 / Chapter 4.3 --- Mouse Model I --- p.75 / Chapter 4.3.1 --- Protein-specific IgE levels --- p.75 / Chapter 4.3.2 --- Protein-specific IgG1 and IgG2a levels --- p.77 / Chapter 4.3.3 --- Allergic Response Test --- p.79 / Chapter 4.3.4 --- Summary from Mouse Model I --- p.81 / Chapter 4.4 --- Mouse Model II --- p.83 / Chapter 4.4.1 --- Proteins specific IgE levels --- p.84 / Chapter 4.4.2 --- Proteins specific IgG1 and IgG2a levels --- p.85 / Chapter 4.4.3 --- Allergic Response Test --- p.87 / Chapter 4.4.4 --- Summary from Mouse Model II --- p.88 / Chapter 4.5 --- Mouse Model III --- p.90 / Chapter 4.5.1 --- Protein-specific IgE levels --- p.90 / Chapter 4.5.2 --- Proteins specific IgG1 and IgG2a levels --- p.91 / Chapter 4.5.3 --- Allergic Response Test --- p.93 / Chapter 4.5.4 --- Summary from Mouse Model III --- p.93 / Chapter 4.6 --- Potential allergenicity of rice glutelin by bioinformatics and epitope mapping --- p.94 / Chapter 4.6.1 --- Bioinformatic analysis --- p.94 / Chapter 4.6.2 --- ELISA analysis of synthesized epitopes --- p.97 / Chapter 4.6.3 --- Western Blot Analysis --- p.99 / Chapter 4.6.4 --- IgE-binding assay --- p.103 / Chapter Chapter 5. --- Conclusion and Future Perspectives --- p.109 / References --- p.111

Genetically modified (GM) foods technology is a novel idea for improving food and crop production, but the supposed health risk of GM foods, such as possible negative long-term health effects to humans, animals and the environment, have provoked the European Union to create assessment protocols to monitor and regulate the safety of GM foods and crops. This research investigates the perception and attitudes of unlabeled GM foods of the WKU faculty and staff. A survey was administered via WKU Qualtrics, and chi-square tests were performed to see how the benefits and disadvantages of GM foods may affect the purchasing decisions of the educated consumer, and to see if the WKU faculty and staff wants GM foods to be labeled or not. The research confirms that the benefits and disadvantages of GM foods do affect the purchasing decisions of the educated consumer. The survey revealed that about 60% of the WKU faculty and staff buys GM foods, and 40% do not buy GM foods, and approximately 92% of the WKU faculty and staff wants GM foods to have proper labeling and information. The research provides information about how the educated consumer of Kentucky may feel about unlabeled GM foods. The research also recommends some trade-off benefits of GM foods, including that approximately 35% of the WKU faculty and staff reported that they would buy GM foods if it helps to lower cholesterol and fight diabetes, and 20% say they would buy GM foods if it is cheaper than other foods. Some of the disadvantages of GM foods are that the pesticide chemicals used in the production methods of GM foods are toxic to humans, animals, and the environment. Approximately 54% of the participants say they would not buy GM foods because they are concerned about how it may affect their long-term health, and about 35% reported that they don’t buy GM foods because of improper labeling and information.

With the adoption of biotechnology in many agricultural products with first-generation biotechnology traits such as increased pest resistance, greater herbicide resistance, and increased yields the growers have accepted them. The next wave of biotech crops have second-generation traits, such as improved nutrient content, extended shelf life, reduced pesticide and herbicide application (a consumer demanded trait), and better taste. Will these consumer benefits offset any concern that the consumer has regarding biotechnology? What are those benefits and how should the information be communicated to the consumer? Three focus groups give insight to the proposed questions. The focus groups were done in three California cities, with participants screened to be: 18-65 years of age, the primary shopper for the household, and with an education level up to a bachelor’s degree. We found that the consumer has little knowledge of biotechnology, but that they assume any concern over these products is reduced if the grocery store or point of purchase is a reputable location. The consumer does look for added utility in products, but they are not willing to pay more unless they understand the production of biotechnology developed products. The consumer feels that there should be labeling of these products, but will likely purchase the least expensive option.

Vermont is the first and only state in the US to establish mandatory labels for food containing genetically modified organisms (GMOs). This thesis investigates the impact of the mandatory labeling law as it relates to changes in prices, quantities sold, and opinions of GMOs. First, grocery store scanner data from Vermont and Oregon are compared using triple difference (difference-in-difference-in-difference) models. Next, Vermont, Oregon, and Colorado survey response data are compared using difference-in-difference models. The findings reveal that there is a general price premium for non-GMO goods of $0.05/oz across all states and times, that mandatory labeling laws do not result in a short-term change in quantities sold or prices of GMO products, and that both mandatory labeling laws and failed mandatory labeling referendums cause an increase in support for GMOs in the food supply. The implications of this research are that mandatory GMO labels did not impact short-term prices or sales and increased the level of support for GMOs.

As economies develop, novel products are created and markets for these products arise. Genetically modified food (GMF) is an example of such a novel product and provides economists with the opportunity to investigate an infant market. Of particular interest with GMF is the impact of consumer reactions on the market. The response of consumers to GMF and their willingness to pay for it has emerged as an important factor in the development of this technology. This research investigates these consumer responses. Prior research suggests that two aspects of consumer behaviour may be relevant for the GMF market. First, consumers may react differently to different types of GMF, so that some products are potentially more economically viable. Secondly, some consumers appear to prefer not having GMF at all. Consumer behaviour is often framed according to neoclassical economic theory. Consumer preferences over goods and the attributes of those goods are generally held to have certain properties. The aspects of consumers’ reactions to GMF noted above, however, may be in conflict with two properties of preferences in neoclassical theory. First, preferences over food attributes are not separable, but may interact with each other. Secondly, some consumers may have preferences regarding GMF that are not continuous. As a result, aggregate impacts of introducing GMF may be difficult to measure, which raises a third issue for investigation, aggregation. Finally, an alternative model of consumer behaviour is bounded rationality, which theorises that choices may be discontinuous as a result of specific protocols. It also suggests that consumers seek to make good-enough choices, rather than attempting to maximise their satisfaction. Thus, optimisation or maximisation is the fourth issue considered in this thesis. In order to investigate these properties of consumers’ preferences, a choice experiment survey was developed. The strength of a choice experiment for examining these issues is its focus on the impact of each product attribute on a respondent’s choices. Thus, it may be possible to identify potentially discontinuous choice patterns and to identify choices affected by interactions between GM technology and other food attributes. Results from a neoclassical analysis of the survey data suggest that some consumers consider the type of benefit created with GM technology in making their choices. In addition, one-quarter to one-half of respondents may have had discontinuous preferences with respect to GMF. Reactions to GMF appear related to respondents’ attitudes, but not to socio-economic or demographic descriptors. As a result, aggregate measures of the impact of GMF may not fully account for consumers’ responses. A boundedly rational model also has reasonable goodness of fit, and may provide a different perspective on consumer behaviour. It is hoped that the results of this research provide a better understanding of consumer behaviour regarding GMF and, by extension, of the process of consumer adoption of novel products. It is further hoped that this attempt to incorporate choice protocols into discrete choice analysis will provide a useful example for further research.

This dissertation traces the historical transformation of the Canada-UK commodity chain for wheat-bread as a lens on processes of local and global change in agrofood relations. During the 1990s, the Canadian Wheat Board (Canada’s monopoly wheat seller) and Warburtons, a British bakery, pioneered an innovative identity-preserved sourcing relationship that ties contracted prairie farmers to consumers of premium bread in the UK. Emblematic of the increasing importance of quality claims, traceability, and private standards in the reorganization of agrifood supply chains, I argue that the changes of the 1990s cannot be understood outside of historical legacies giving shape to unique institutions for regulating agrofood relations on the Canadian prairies and in the UK food sector. I trace the rise, fall, and re-invention of the Canada-UK commodity chain across successive food regimes, examining the changing significance of wheat- bread, inter-state relations between Canada, the UK, and the US, and public and private forms of agrofood regulation over time. In particular, I focus on the way in which changing food regime relations transformed the CWB, understood as the nexus of institutions tying prairie farmers into global circuits of accumulation. When in the 1990s, the CWB and Warburtons responded to structural crises in their respective industries by re-inventing the Canada-UK wheat trade, the result was significant organizational and industry change. On the prairies, the CWB has shown how – contrary to expectations -- centralized marketing and quality control may help prairie farmers adapt to the demands of end-users in the emerging ‘economy of qualities’. In the UK, Warburtons has led the ‘premiumisation’ of the bread sector, traditionally defined by consumer taste for cheap bread, over the last 15 years. The significance of the shift towards quality chains in the wheat-bread sector is analyzed in light of conflicts over the proposed introduction of genetically engineered (GE) wheat to the Canadian prairies.

Flax is one of the major cash crops in Canada. Approximately seventy percent of Canadian flaxseed was exported to European Union (EU) annually until 2009. In 2009, the EU imposed an import ban on Canadian flaxseed due to the adventitious presence of a GM flax variety - CDC Triffid was identified in Canadian flaxseed exported to the EU. The EUs decision to apply zero tolerance on CDC Triffid flax has been based on its interpretation of the precautionary principle. According to the World Trade Organisations Agreement on the Application of Sanitary and Phytosanitary Measures (SPS), however, precautionary measures are subject to a scientific risk assessment. As the EU did not base its zero tolerance for CDC Triffid flax on any scientific risk assessment, the EU is in violation of the SPS Agreement. Moreover, the EU has ignored the available scientific information regarding CDC Triffid flax. The EU did not consider the possibility of following the guidelines given by Codex Alimentarius Commission in the case of CDC Triffid flax. There are non-scientific reasons behind the EUs zero tolerance on CDC Triffid flax and they overweigh the available scientific information. The EU position would be unlikely to be supported if a complaint was brought to the World Trade Organisation Disputes Panel. A partial equilibrium model was used to provide a theoretical background to examine the changes in the flaxseed industry and the linseed oil industry due to the CDC Triffid event. A model of the supply chain of Canadian flaxseed was developed to illustrate the operationalisation of the Protocol developed by the EU and Canada to address the zero tolerance policy. Empirical estimation suggests that the operationalisation of the Protocol incurred additional cost of $7.5 million to the flax seed industry of Canada in 2009/ 2010. Out of that, cost of testing was approximately $1.2 million and cost of segregation was $4.2 million. Estimation of changes in revenue suggests that there was a loss of revenue in flaxseed trade between the EU and Canada in 2009/2010. Imports of Canadian flax by China provided an alternative market, at a considerably lower price than typically realised from the EU market. Interestingly, the EUs zero tolerance policy on CDC Triffid flax has resulted in a larger additional cost on the EU than Canada.

This thesis contains three essays on topics in agricultural economics. Essays one and two share a focus on international trade and economic development, and essays two and three apply dynamic tools to agricultural economic policy issues.<p>Essay one analyses trade-related implications of a developing country's decision to adopt genetically-modified crop technology. A fixed-proportions model is constructed that evaluates the welfare implications of a range of adoption policies and export market responses. The model in this essay illustrates the importance of the prospective adopter formulating a projection of probable export market effects before making an adoption decision and of the role that high transaction costs may play in a developing country's adoption decision. The model also considers the effects of a new policy tool; a check-off style levy on genetically-modified technology in place of a technology-use agreement. A levy could be useful tool in developing countries, which are characterised by high transaction costs. <p>Essay two models the effects of emergency food aid on a recipient country's agricultural industry. This essay formulates a definition of needed aid in the context of a food emergency and constructs an optimal control model that solves a path of aid shipments that best meets that need. The effects of a range of food aid paths on recipient-country agricultural production are illustrated through numerical simulations. There are two key results. First, a non-optimal amount of aid can hinder a recipient-country's recovery from an exogenous food shock. Second, an exogenous shock can affect farmer revenue and therefore impact planting decisions. This effect must be considered in aid allocation policies. <p>Essay three uses time-series econometric techniques to develop a demand model that assesses the effectiveness of commodity advertising. This essay describes the importance of considering long-run and dynamic effects in demand systems, especially in the case of closely substitutable commodities. A demand system that tests for and accommodates dynamic and time-series properties is developed and applied to US meat data. The results of this model are compared to a traditional static demand system. The dynamic model produces econometrically and theoretically sound results and generates some more intuitively appealing estimates.