Transgenic,transplastomic and other genetically modified plants: a Canadian perspective

Since the mid 1990s, genetically modified (GM) crops have been grown commercially in Canada on a scale that has increased steadily over the years. An intense debate ensued, as elsewhere, and many fears were expressed regarding not only the technology itself but some of the main GM crops being grown. It would seem appropriate at this time to examine how these novel crops compare to crops bred by more traditional means and what impacts these GM crops have had based on experience and not merely on conjecture. To begin, we will put things in a historical perspective and recall how domestication and conventional plant breeding have shaped the crops of today. Then, we will describe briefly the distinctive features of GM plants (obtained so far mainly by nuclear transgenesis) and how these novel crops are regulated in Canada. We will then give two examples of widely grown GM crops in Canada (insect-resistant corn and herbicide-tolerant canola) and examine the main questions that were raised as well as the actual impacts these crops have had on the farm. These examples will help us outline some of the limitations of the current generation of GM plants and, finally, we will try to get a glimpse of the future by examining some recent technical developments in the field of recombinant DNA technologies applied to plant breeding.     

转基因植物的食品安全性问题及评价策略

转基因植物经历了20年的快速发展,已经成为21世纪农业的重要支柱之一。转基因植物从诞生以来,其食用安全性一直是科学界、政府和广大消费者关心的焦点问题。各类转基因安全事件层出不穷,严重影响着转基因产业的发展。本文从营养、毒性、过敏性、抗生素抗性、非期望效应等五个方面系统总结了转基因植物食用安全的问题,详细介绍了转基因植物食用安全评价的六大原则以及国际与国内对转基因植物食用安全性评价的内容,最后介绍了我国的转基因植物安全管理体系。以期使读者对转基因植物食用安全问题有一个系统全面的了解。     

Problem formulation in the environmental risk assessment for genetically modified plants

Problem formulation is the first step in environmental risk assessment (ERA) where policy goals, scope, assessment endpoints, and methodology are distilled to an explicitly stated problem and approach for analysis. The consistency and utility of ERAs for genetically modified (GM) plants can be improved through rigorous problem formulation (PF), producing an analysis plan that describes relevant exposure scenarios and the potential consequences of these scenarios. A properly executed PF assures the relevance of ERA outcomes for decision-making. Adopting a harmonized approach to problem formulation should bring about greater uniformity in the ERA process for GM plants among regulatory regimes globally. This paper is the product of an international expert group convened by the International Life Sciences Institute (ILSI) Research Foundation.     

DNA barcoding simplifies environmental risk assessment of genetically modified crops in biodiverse regions

Transgenes encoding for insecticidal crystal (Cry) proteins from the soil-dwelling bacterium Bacillus Thuringiensis have been widely introduced into Genetically Modified (GM) crops to confer protection against insect pests. Concern that these transgenes may also harm beneficial or otherwise valued insects (so-called Non Target Organisms, NTOs) represents a major element of the Environmental Risk Assessments (ERAs) used by all countries prior to commercial release. Compiling a comprehensive list of potentially susceptible NTOs is therefore a necessary part of an ERA for any Cry toxin-containing GM crop. In partly-characterised and biodiverse countries, NTO identification is slowed by the need for taxonomic expertise and time to enable morphological identifications. This limitation represents a potentially serious barrier to timely adoption of GM technology in some developing countries. We consider Bt Cry1A cowpea (Vigna unguiculata) in Nigeria as an exemplar to demonstrate how COI barcoding can provide a simple and cost-effective means of addressing this problem. Over a period of eight weeks, we collected 163 insects from cowpea flowers across the agroecological and geographic range of the crop in Nigeria. These individuals included 32 Operational Taxonomic Units (OTUs) spanning four Orders and that could mostly be assigned to genus or species level. They included 12 Lepidopterans and two Coleopterans (both potentially sensitive to different groups of Cry proteins). Thus, barcode-assisted diagnoses were highly harmonised across groups (typically to genus or species level) and so were insensitive to expertise or knowledge gaps. Decisively, the entire study was completed within four months at a cost of less than 10,000 US$. The broader implications of the findings for food security and the capacity for safe adoption of GM technology are briefly explored.     

Ecological risk assessment of genetically modified crops based on cellular automata modeling

The assessment of ecological risk in genetically modified (GM) biological systems is critically important for decision-making and public acceptance. Cellular automata (CA) provide a potential modeling and simulation framework for representing relationships and interspecies interactions both temporally and spatially. In this paper, a simple subsystem contains only four species: crop, target pest, non-target pest and enemy insect, and a three layer arrangement of L × L stochastic cellular automata with a periodic boundary were established. The simulation of this simplified system showed abundant and sufficient complexity in population assembly and densities, suggesting a prospective application in ecological risk assessment of GM crops.     

Biosynthesis of 5-hydroxy-L-tryptophan by a genetically modified strain ofEscherichia coli in presence of 5-hydroxyindole

Summary A derivative ofEscherichia coli strain W3110 with increased tryptophan synthas (TS) activity was studied in the biosynthesis of L-tryptophan and 5-hydroxy-L-tryptophan, respectively, in presence of precursors. Indole or 5-hydroxyindole was added to growing cells in minimal medium supplemented with tetracycline. The specific activity of TS for 5-hydroxyindole was about 5-fold lower compared with indole. However, this difference in enzyme activity was not observed when the specific productivity (q_p) of L-tryptophan or 5-hydroxy-L-tryptophan, which was 0.14–0.15 g (g dry wt cells)^–1 · h^–1 was determined. In minimal medium L-serine was shown to limit the production of both tryptophan and its hydroxylated derivative. In presence of L-serine, q_p, for L-tryptophan and 5-hydroxy-L-tryptophan were increased by a factor of about 3 and 2, respectively.     

Farm Scale Evaluations of spring-sown genetically modified herbicide-tolerant crops: a statistical assessment

Primary results from the Farm Scale Evaluations (FSEs) of spring-sown genetically modified herbicide-tolerant crops were published in 2003. We provide a statistical assessment of the results for count data, addressing issues of sample size (n), efficiency, power, statistical significance, variability and model selection. Treatment effects were consistent between rare and abundant species. Coefficients of variation averaged 73% but varied widely. High variability in vegetation indicators was usually offset by large n and treatment effects, whilst invertebrate indicators often had smaller n and lower variability; overall, achieved power was broadly consistent across indicators. Inferences about treatment effects were robust to model misspecification, justifying the statistical model adopted. As expected, increases in n would improve detectability of effects whilst, for example, halving n would have resulted in a loss of significant results of about the same order. 40% of the 531 published analyses had greater than 80% power to detect a 1.5-fold effect; reducing n by one-third would most likely halve the number of analyses meeting this criterion. Overall, the data collected vindicated the initial statistical power analysis and the planned replication. The FSEs provide a valuable database of variability and estimates of power under various sample size scenarios to aid planning of more efficient future studies.     

Monitoring the presence of genetically modified food on the market of the Republic of Croatia

From the beginning of the human race people have been applying different methods to change the genetic material of either plants or animals in order to increase their yield as well as to improve the quality and quantity of food. Genetically modified organism (GMO) means an organism in which the genetic material has been altered in a way that does not occur naturally by mating and/or natural recombination. Analysing the presence of GMO in food is done by detecting the presence of either specific DNA sequences inserted in the genome of transgenic organism, or detecting proteins as a result of the expression of the inserted DNA. In this work food testing for the presence of genetically modified organisms was conducted during the period from 2004 to 2007 in the GMO laboratory of the Croatian National Institute of Public Health. According to the regulations, among the samples in which the presence of GMO was detected, all those which had more than 0.9% of GMO content were either rejected from the border or removed from the market, because such GM food has to be appropriately labelled. Among the food samples which were analysed in 2004: 127 (2.37%) of a total of 1226 samples contained more than 0.9% of GMOs; in 2005 there was only one in 512 (0.20%) samples in total; in 2006 there were 4 out of 404 samples (0.99%), and in 2007: 7 of a total of 655 samples (1.07%) had GMO content above the allowed threshold of 0.9%.     

Methods of identification and assessment of safety of genetically modified microorganisms in manufacture food production

Methods of identification of genetically modified microorganisms (GMM), used in manufacture food on control probes are presented. Results of microbiological and molecular and genetic analyses of food products and their components important in microbiological and genetic expert examination of GMM in foods are considered. Examination of biosafety of GMM are indicated.     

Cutaneous melanoma in genetically modified animals

Cutaneous melanomas are tumors originating from skin melanocytes which are present in hair follicles, and interfollicular epidermal and dermal layers. Experimental work in model systems involves in silico, in vitro and in vivo analyses. Such models allow to mimick melanocytic aberrations characteristic of melanoma, and to potentially exploit novel therapies. Transgenic technologies can be used to modify specifically the genome of the model organism and thereby generate transgenic strains, and combinations of such strains, which may develop metastasizing melanoma. In such strains, metastasizing melanoma either arises spontaneously after a period of latency or requires additional physical or chemical induction. In this review, we summarize the work of currently available transgenic melanoma models and discuss the most recent progress in creating improved and/or inducible models reflecting the human disease.     

Analysis of genetically modified oils

Genetically modified oils with altered functional or nutritional characteristics are being introduced into the marketplace. A wide array of analytical techniques has been utilized to facilitate developing these oils. This article attempts to review the utilization of these analytical procedures for characterizing both the chemistry and some functionality of these oils. Although techniques to assess oxidative stability in frying and food applications are covered, measurement of nutritional characteristics are not.     

Gene flow in genetically modified wheat

Understanding gene flow in genetically modified (GM) crops is critical to answering questions regarding risk-assessment and the coexistence of GM and non-GM crops. In two field experiments, we tested whether rates of cross-pollination differed between GM and non-GM lines of the predominantly self-pollinating wheat Triticum aestivum. In the first experiment, outcrossing was studied within the field by planting "phytometers" of one line into stands of another line. In the second experiment, outcrossing was studied over distances of 0.5-2.5 m from a central patch of pollen donors to adjacent patches of pollen recipients. Cross-pollination and outcrossing was detected when offspring of a pollen recipient without a particular transgene contained this transgene in heterozygous condition. The GM lines had been produced from the varieties Bobwhite or Frisal and contained Pm3b or chitinase/glucanase transgenes, respectively, in homozygous condition. These transgenes increase plant resistance against pathogenic fungi. Although the overall outcrossing rate in the first experiment was only 3.4%, Bobwhite GM lines containing the Pm3b transgene were six times more likely than non-GM control lines to produce outcrossed offspring. There was additional variation in outcrossing rate among the four GM-lines, presumably due to the different transgene insertion events. Among the pollen donors, the Frisal GM line expressing a chitinase transgene caused more outcrossing than the GM line expressing both a chitinase and a glucanase transgene. In the second experiment, outcrossing after cross-pollination declined from 0.7-0.03% over the test distances of 0.5-2.5 m. Our results suggest that pollen-mediated gene flow between GM and non-GM wheat might only be a concern if it occurs within fields, e.g. due to seed contamination. Methodologically our study demonstrates that outcrossing rates between transgenic and other lines within crops can be assessed using a phytometer approach and that gene-flow distances can be efficiently estimated with population-level PCR analyses.     

Lifespan extension in genetically modified mice

Major advances in aging research have been made by studying the effect of genetic modifications on the lifespan of organisms, such as yeast, invertebrates (worms and flies) and mice. Data from yeast and invertebrates have been the most plentiful because of the ease in which genetic manipulations can be made and the rapidity by which lifespan experiments can be performed. With the ultimate focus on advancing human health, testing genetic interventions in mammals is crucial, and the mouse has proven to be the mammal most amenable to this task. Lifespan studies in mice are resource intensive, requiring up to 4 years to complete. Therefore, it is critical that a set of scientifically-based criteria be followed to assure reliable results and establish statistically significant findings so other laboratories can replicate and build on the data. Only then will it be possible to confidently determine that the genetic modification extends lifespan and alters aging.     

Safety assessment of rice genetically modified with soybean glycinin by feeding studies on rats

Feeding studies on rice genetically modified with soybean glycinin were performed on rats for four weeks. The rats were divided into three groups, each being fed on (I) only a commercial diet, (II) this diet plus control rice and (III) this diet plus rice genetically modified with glycinin. The rats were fed with 10 g/kg-weight of rice every day by oral administration. During the test period, the rats in every group grew well without marked differences in appearance, food intake, body weight, or cumulative body weight gain. There were also no significant differences in the blood count, blood composition or internal organ weights among the rats. Necropsy at the end of the experiment indicated neither pathological symptoms nor histopathological abnormalities in the liver and kidney. Judging from these results, the rice genetically modified with glycinin is considered to have been essentially the same in nutritional and biochemical characteristics as the control rice.     

A statistical assessment of differences and equivalences between genetically modified and reference plant varieties

Background  

Safety assessment of genetically modified organisms is currently often performed by comparative evaluation. However, natural variation of plant characteristics between commercial varieties is usually not considered explicitly in the statistical computations underlying the assessment.     

Detection of genetically modified organisms in foods

Legislation enacted worldwide to regulate the presence of genetically modified organisms (GMOs) in crops, foods and ingredients, necessitated the development of reliable and sensitive methods for GMO detection. In this article, protein- and DNA-based methods employing western blots, enzyme-linked immunosorbant assay, lateral flow strips, Southern blots, qualitative-, quantitative-, real-time- and limiting dilution-PCR methods, are discussed. Where information on modified gene sequences is not available, new approaches, such as near-infrared spectrometry, might tackle the problem of detection of non-approved genetically modified (GM) foods. The efficiency of screening, identification and confirmation strategies should be examined with respect to false-positive rates, disappearance of marker genes, increased use of specific regulator sequences and the increasing number of GM foods.