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Analysis of gene transfer between microorganisms and plants

Obiettivo

We propose to investigate the potential for transfer and expression of model genes under conditions that employ different sets of interactions between microbes and plants. We expect to develop new extraction and detection methods appropriate to these studies. Environmental factors (eg, moisture and bacterial motility) will be investigated for potential roles in survival and transfer.
The possibility that genetic information can transfer horizontally between distantly related organisms is being explored. A better understanding of this is necessary in order to establish rules for biological containment of genetically manipulated organism which are consistent with potential hazards to man and the environment. Genetic and physiochemical factors which affect gene transfer both under well controlled conditions in the laboratory and in the natural environment are to be established. Studies are designed to understand more about possible mechanisms which allow deoxyribonucleic acid (DNA) transfer including conjugation, transformation, or unknown processes which might allow gene transfer between plants and microbes.

The study is being conducted in the following areas:
effects of environmental conditions on conjugative gene transfer;
bacterial conjugative mechanisms;
plant bacteria gene transfer;
characterization of DNA soil interactions and its influence on transformation of plants and bacteria in the soil.
The research is based essentially on the use of a group of specific marker genes (gentamicin, phosphinothricin and bromoxynil) that are excellent selective markers for a variety of microbes and plants to monitor the transfer of genetic information between species in nature and under natural conditions.

Several aspects of plasmid transfer in soil will be considered; transfer between gram negatives such as Rhizobium leguminosarum and Agrobacterium tumefaciens into Pseudomonas syringae in soil, using the marker genes and detection methods developed, and also transfer between gram positives. In the latter case we have shown transfer of broad host range gram negative plasmids to streptomycetes by conjugation, the plasmids can, surprisingly, replicate and express marker genes. We have designed a system of entrapment by which we will add a selectable, marked streptomycetes to soil and then screen for transfer to this recipient, after growth and selection.

We will also investigate the effects of motility, moisture, soil composition, and other chemical and physical factors. Experiments have been planned to try to estimate how often mating partners make contact, and what factors may influence the efficiency of this contact.

Transfer between microbes in nodules and the possibility of transfer from plant to microbe will be investigated. For these experiments, three different model systems will be used. The first will investigate transfer of a marker gene from transgenic plants into Rhizobium leguminosarum. The second will examine the possibility of transfer between transgenic tobacco plants and indigenous soil microbes surrounding the roots as well as microbes associated with the tobacco plants during the fermentative processes that take place during curing. Studies investigating gene transfer in plant/microbial interaction will consider transfer from Rhizobium meliloti to plant in alfalfa nodules when the marker genes are carried by a promiscuous plasmid. The marker gene will be used in several types of constructions involving both bacterial and plant promoter sequences. Concomitantly, transfer of the same genes from plant to bacteria will be screened in nodules, tumours (Agrobacterium), following infections (Xanthomonas), and uring the normal curing process of tobacco.

Campo scientifico (EuroSciVoc)

CORDIS classifica i progetti con EuroSciVoc, una tassonomia multilingue dei campi scientifici, attraverso un processo semi-automatico basato su tecniche NLP. Cfr.: Il Vocabolario Scientifico Europeo.

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Coordinatore

INSTITUT PASTEUR
Contributo UE
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Indirizzo
Rue du Docteur Roux 25
75724 PARIS
Francia

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