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Assessment of Risks induced by virus-derived transgene products in plants, using luteoviruses carrying the green flourescent protein as a visible reporter

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The large number of genetically-engineered virus resistant plants which is expected to be put forward for approval for marketing in the EU prompts the need for risk assessment studies on the potential interactions between viral transgene products and infecting viruses or viroids. Major concern centres on three main classes of hazards: heterologous-encapsidation, RNA recombination and synergism which may result in viruses and viroids with altered genetic traits, novel combinations of properties, or novel' diseases. Little information is available concerning the probability of the occurrence of the undesired interactions in transgenic plants, making it impossible to reach generally acceptable conclusions about the environmental safety related to the introduction of these plants. The major bottleneck in obtaining sufficiently large data sets on recombination heterologous encapsidation and synergism is to detect these events.
The current proposal aims to tackle this problem by using a visible reporter, the jellyfish green fluorescent protein (GFP), which will markedly simplify and speed up the detection of the aforesaid events, thus allowing large numbers of virus-infected transgenic plants to be screened. The GFP-encoding gene will be incorporated into the genome of beet western yellows luteovirus (BWYV) and potato leafroll luteovirus (PLRV). The GFP-carrying viruses will then be used to challenge transgenic plants expressing luteoviral and potyviral sequences which are commonly used to achieve pathogen-derived resistance in a large number of important arable and vegetable crops. Luteoviruses are most appropriate for this type of risk assessment studies because (i) they are known to readily undergo heterologous-encapsidation during co-infections in nature, and encapsidate viroid RNA, (ii) their genetic maps suggest that they arose by RNA recombination between different ancestor viruses and several areas on their RNA are potential 'hot spots' for recombination, and (iii) synergism is a common phenomenon in mixed infection of a luteovirus and a potyvirus, leading to enhanced symptom expression and higher virus litres.
A consortium of research partners will combine their complementary expertise in order to address the following objectives:
1. To identify heterologous-encapsidation of PLRV and BWYV in transgenic plants expressing the viral capsid-associated proteins of these viruses, and to assess whether potato spindle tuber viroids (PSTVd) will be encapsidated by these transgenic proteins.
2. To identify RNA recombination between luteovirus-derived transgenes and infecting BvVYV or PLRV, and between PVY-derived transgenes and infecting luteoviruses.
3. To identify the occurrence of synergism between potyviral transgene products involved in virus movement and infecting luteoviruses.
4. To evaluate the probability of the aforementioned events, particularly in relation to naturally occurring mixed infections of the viruses and viroids involved.
The project is organized in work packages in which each of these points will be addressed. The project starts off with generating the required transgenic plants and the full-length infectious luteoviral clones carrying the GFP gene. Measurable deliverables are (i) well described methods to detect possible interactions between viral transgenic products and co-infecting viruses and viroids, and (ii) procedures to assess the probability of these interactions. These methods and procedures are pivotal for regulatory authorities carrying out risk assessment under Community legislation.

Wissenschaftliches Gebiet (EuroSciVoc)

CORDIS klassifiziert Projekte mit EuroSciVoc, einer mehrsprachigen Taxonomie der Wissenschaftsbereiche, durch einen halbautomatischen Prozess, der auf Verfahren der Verarbeitung natürlicher Sprache beruht. Siehe: Das European Science Vocabulary.

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