Transgenic sequences are frequently lost in Phytophthora parasitica transformants without reversion of the transgene-induced silenced state

Authors: Gaulin, Elodie; Haget, Nathalie; Khatib, Moustafa; Herbert, Corentin; Rickauer, Martina; Bottin, Arnaud

Source: Canadian Journal of Microbiology, Volume 53, Number 1, January 2007 , pp. 152-157(6)

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Abstract:

Little data exist on the mechanism and stability of transformation in Phytophthora parasitica, a major oomycete parasite of plants. Here, we studied the stability of drug-resistant protoplast transformants by analyzing single-zoospore derivatives. We show that the transgenic sequences are not stably integrated into the chromosomes, resulting in the loss of drug resistance in single-zoospore derivatives. However, in strains where the P. parasitica gene encoding the CBEL elicitor was silenced by transformation with sense or antisense constructs, silencing is not reversed when the transgenic sequences are lost. This suggests that instability of P. parasitica transformants is not an obstacle for loss-of-function studies in this organism.Key words: Phytophthora, transformation, sporulation, silencing, stability.

Il existe peu de données sur les mécanismes et la stabilité de la transformation chez Phytophthora parasitica, un important parasite oomycète chez les plantes. Nous étudions ici la stabilité de transformants obtenus par la méthodedes protoplastes et résistant à une drogue en analysant leurs dérivés issus de zoospores uniques. Nous démontrons que les séquences transgéniques ne sont pas intégrées de façon stable dans les chromosomes, ce qui résulte en une perte de la résistance àx la drogue chez les dérivés de zoospores uniques. Cependant, chez les souches dans lesquelles le gène de P. parasitica codant l'éliciteur CBEL a été rendu silencieux par la transformation à l'aide de constructions sens ou anti-sens, le silençage n'est pas aboli lorsque les séquences transgéniques sont perdues. Ceci suggère que l'instabilité des transformants de P. parasitica n'est pas un obstacle aux études de perte de fonction chez cet organisme.Mots-clés : Phytophthora, transformation, sporulation, silençage, stabilité.

Document Type: Research article

Publication date: 2007-01-01

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Published since 1954, this monthly journal contains new research in the field of microbiology including applied microbiology and biotechnology; microbial structure and function; fungi and other eucaryotic protists; infection and immunity; microbial ecology; physiology, metabolism and enzymology; and virology, genetics, and molecular biology. It also publishes review articles and notes on an occasional basis, contributed by recognized scientists worldwide.
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Transgenic sequences are frequently lost in Phytophthora parasitica transformants without reversion of the transgene-induced silenced state

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