Assessing and monitoring impacts of genetically modified plants on agro-ecosystems: the approach of AMIGA project

The environmental impacts of genetically modified crops is still a controversial issue in Europe. The overall risk assessment framework has recently been reinforced by the European Food Safety Authority(EFSA) and its implementation requires harmonized and efficient methodologies. The EU-funded research project AMIGA − Assessing and monitoring Impacts of Genetically modified plants on Agro-ecosystems − aims to address this issue, by providing a framework that establishes protection goals and baselines for European agro-ecosystems, improves knowledge on the potential long term environmental effects of genetically modified (GM) plants, tests the efficacy of the EFSA Guidance Document for the Environmental Risk Assessment, explores new strategies for post market monitoring, and provides a systematic analysis of economic aspects of Genetically Modified crops cultivation in the EU. Research focuses on ecological studies in different EU regions, the sustainability of GM crops is estimated by analysing the functional components of the agro-ecosystems and specific experimental protocols are being developed for this scope.

A proposed impact assessment method for genetically modified plants (AS-GMP Method).

An essential step in the development of products based on biotechnology is an assessment of their potential economic impacts and safety, including an evaluation of the potential impact of transgenic crops and practices related to their cultivation on the environment and human or animal health. The purpose of this paper is to provide an assessment method to evaluate the impact of biotechnologies that uses quantifiable parameters and allows a comparative analysis between conventional technology and technologies using GMOs. This paper introduces amethod to performan impact analysis associatedwith the commercial release and use of genetically modified plants, the Assessment SystemGMPMethod. The assessment is performed through indicators that are arranged according to their dimension criterion likewise: environmental, economic, social, capability and institutional approach. To perform an accurate evaluation of the GMP specific indicators related to genetic modification are grouped in common fields: genetic insert features, GMplant features, gene flow, food/feed field, introduction of the GMP, unexpected occurrences and specific indicators. The novelty is the possibility to include specific parameters to the biotechnology under assessment. In this case by case analysis the factors ofmoderation and the indexes are parameterized to perform an available assessment.

No adverse effect of genetically modified antifungal wheat on decomposition dynamics and the soil fauna community - a field study

The cultivation of genetically modified (GM) plants has raised several environmental concerns. One of these concerns regards non-target soil fauna organisms, which play an important role in the decomposition of organic matter and hence are largely exposed to GM plant residues. Soil fauna may be directly affected by transgene products or indirectly by pleiotropic effects such as a modified plant metabolism. Thus, ecosystem services and functioning might be affected negatively. In a litterbag experiment in the field we analysed the decomposition process and the soil fauna community involved. Therefore, we used four experimental GM wheat varieties, two with a race-specific antifungal resistance against powdery mildew (Pm3b) and two with an unspecific antifungal resistance based on the expression of chitinase and glucanase. We compared them with two non-GM isolines and six conventional cereal varieties. To elucidate the mechanisms that cause differences in plant decomposition, structural plant components (i.e. C:N ratio, lignin, cellulose, hemicellulose) were examined and soil properties, temperature and precipitation were monitored. The most frequent taxa extracted from decaying plant material were mites (Cryptostigmata, Gamasina and Uropodina), springtails (Isotomidae), annelids (Enchytraeidae) and Diptera (Cecidomyiidae larvae). Despite a single significant transgenic/month interaction for Cecidomyiidae larvae, which is probably random, we detected no impact of the GM wheat on the soil fauna community. However, soil fauna differences among conventional cereal varieties were more pronounced than between GM and non-GM wheat. While leaf residue decomposition in GM and non-GM wheat was similar, differences among conventional cereals were evident. Furthermore, sampling date and location were found to greatly influence soil fauna community and decomposition processes. The results give no indication of ecologically relevant adverse effects of antifungal GM wheat on the composition and the activity of the soil fauna community.

Biological characteristics of black armyworm Spodoptera cosmioides on genetically modified soybean and corn crops that express insecticide Cry proteins.

This study aimed to evaluate the development and reproduction of the black armyworm, Spodoptera cosmioides when larvae fed on leaves of Bt-corn hybrids, expressing a single Cry1F and also Cry1F, Cry1A.105 and Cry2Ab2 in pyramided corn and their non-Bt-isoline (hybrid 2B688), as well as on leaves of two soybean isolines expressing the Cry1Ac protein and its non-Bt isoline (A5547-227). We also assessed the effect of these Bt and non-Bt plants on the leaf consumption rate of S. cosmioides larvae. This pest was unable to develop when fed on any of the corn isolines (Bt and non-Bt). When both 1st and 3rd instar larvae were fed on corn leaf, mortality was 100% in both Bt and non-Bt corn. In contrast, when corn leaves were offered to 5th instar larvae, there were survivors. Defoliation and leaf consumption was higher with non-Bt corn than with both of the Bt corn isolines. There was no negative effect of Bt soybean leaves on the development and reproduction of S. cosmioides with respect to all evaluated parameters. Our study indicates that both Bt and non-Bt corn adversely affect the development of S. cosmioides while Bt soybean did not affect its biology, suggesting that this lepidopteran has major potential to become an important pest in Bt soybean crops.

Criminology and genetically modified food

Genetically modified or engineered foods are produced from rapidly expanding technologies that have sparked international debates and concerns about health and safety. These concerns focus on the potential dangers to human health, the risks of genetic pollution, and the demise of alternative farming techniques as well as biopiracy and economic exploitation by large private corporations. This article discusses the findings of the world's first Royal Commission on Genetic Modification conducted in New Zealand and reveals that there are potential social, ecological and economic risks created by genetically modified foods that require closer criminological scrutiny. As contemporary criminological discourses continue to push new boundaries in areas of crimes of the economy, environmental pollution, risk management, governance and globalization, the potential concerns posed by genetically modified foods creates fertile ground for criminological scholarship and activism.

Hairpin RNAs derived from RNA polymerase II and polymerase III promoter-directed transgenes are processed differently in plants

RNA polymerase III (Pol III) as well as Pol II (35S) promoters are able to drive hairpin RNA (hpRNA) expression and induce target gene silencing in plants. siRNAs of 21 nt are the predominant species in a 35S Pol II line, whereas 24- and/or 22-nucleotide (nt) siRNAs are produced by a Pol III line. The 35S line accumulated the loop of the hpRNA, in contrast to full-length hpRNA in the Pol III line. These suggest that Pol II and Pol III-transcribed hpRNAs are processed by different pathways. One Pol III transgene produced only 24-nt siRNAs but silenced the target gene efficiently, indicating that the 24-nt siRNAs can direct mRNA degradation; specific cleavage was confirmed by 59 rapid amplification of cDNA ends (RACE). Both Pol II- and Pol III-directed hpRNA transgenes induced cytosine methylation in the target DNA. The extent of methylation is not correlated with the level of 21-nt siRNAs, suggesting that they are not effective inducers of DNA methylation. The promoter of a U6 transgene was significantly methylated, whereas the promoter of the endogenous U6 gene was almost free of cytosine methylation, suggesting that endogenous sequences are more resistant to de novo DNA methylation than are transgene constructs. Published by Cold Spring Harbor Laboratory Press. Copyright © 2008 RNA Society.

Replicating satellite RNA induces sequence-specific DNA methylation and truncated transcripts in plants

Tobacco plants were transformed with a chimeric transgene comprising sequences encoding β-glucuronidase (GUS) and the satellite RNA (satRNA) of cereal yellow dwarf luteovirus. When transgenic plants were infected with potato leafroll luteovirus (PLRV), which replicated the transgene-derived satRNA to a high level, the satellite sequence of the GUS:Sat transgene became densely methylated. Within the satellite region, all 86 cytosines in the upper strand and 73 of the 75 cytosines in the lower strand were either partially or fully methylated. In contrast, very low levels of DNA methylation were detected in the satellite sequence of the transgene in uninfected plants and in the flanking nonsatellite sequences in both infected and uninfected plants. Substantial amounts of truncated GUS:Sat RNA accumulated in the satRNA-replicating plants, and most of the molecules terminated at nucleotides within the first 60 bp of the satellite sequence. Whereas this RNA truncation was associated with high levels of satRNA replication, it appeared to be independent of the levels of DNA methylation in the satellite sequence, suggesting that it is not caused by methylation. All the sequenced GUS:Sat DNA molecules were hypermethylated in plants with replicating satRNA despite the phloem restriction of the helper PLRV. Also, small, sense and antisense ∼22 nt RNAs, derived from the satRNA, were associated with the replicating satellite. These results suggest that the sequence-specific DNA methylation spread into cells in which no satRNA replication occurred and that this was mediated by the spread of unamplified satRNA and/or its associated 22 nt RNA molecules.

RNAi for insect-proof plants

RNA interference induced in insects after ingestion of plant-expressed hairpin RNA offers promise for managing devastating crop pests

Small RNA viruses of insects : expression in plants and RNA silencing

Interest in insect small RNA viruses (SRVs) has grown slowly but steadily. A number of new viruses have been analyzed at the sequence level, adding to our knowledge of their diversity at the level of both individual virus species and families. In particular, a number of possible new virus families have emerged. This research has largely been driven by interest in their potential for pest control, as well as in their importance as the causal agents of disease in beneficial arthropods. At the same time, research into known viruses has made valuable contributions to our understanding of an emerging new field of central importance to molecular biology-the existence of RNA-based gene silencing, developmental control, and adaptive immune systems in eukaryotes. Subject to RNA-based adaptive immune responses in their hosts, viruses have evolved a variety of genes encoding proteins capable of suppressing the immune response. Such genes were first identified in plant viruses, but the first examples known from animal viruses were identified in insect RNA viruses. This chapter will address the diversity of insect SRVs, and attempts to harness their simplicity in the engineering of transgenic plants expressing viruses for resistance to insect pests. We also describe RNA interference and antiviral pathways identified in plants and animals, how they have led viruses to evolve genes capable of suppressing such adaptive immunity, and the problems presented by these pathways for the strategy of expressing viruses in transgenic plants. Approaches for countering these problems are also discussed. © 2006 Elsevier Inc. All rights reserved.