Abstract
Cecropin B is a peptide of approximately 4 kDA which shows antimicrobial activityin vitro against Gram positive and Gram negative bacteria. Potato cvs Agria, Bintje, Karnico, Kondor and Producent were transformed with a gene construct encoding a cecropin B precursor polypeptide. In total, 49 independent transgenic potato clones were obtained. Northern blot analysis of these plants revealed that the introduced gene was transcribed to detectable levels in almost all plants, the highest transcription level being approximately 0.6% of total mRNA. No cecropin B peptide could be detected in transgenic plants, probably as a result of rapid proteolytic degradation of newly synthesized cecropin B by potato endogenous proteases. Neither small tubers of a group of 11 clones with moderate to high transcription levels nor slices from field grown tubers of any of the transgenic clones obtained, showed significantly (P<0.01) less rot after inoculation withErwinia carotovora subspatroseptica orE. chrysanthemi, the plant pathogenic bacteria that cause potato soft rot.
Compendio
Cecropin B es un peptido de aproximadamente 4 kDA que muestra actividad antimicrobianain vitro contra las bacterias Gram positivas y Gram negativas. Los cultivares de papa Agria, Bintje, Karnico, Kondor y Producent fueron transformados con un prototipo de gene que codifica un polipeptido precursor de cecropin B. En total, se obtuvieron 49 clones independientes de papa transgénica. El análisis de estas plantas, mediante la transferencia del RNA a placas de nitrocelulosa (Northern Blot Analysis), reveló que el gene introducido fue transcrito a niveles detectables en casi todas las plantas, siendo el nivel más alto de transcription de aproximadamente 0.6% del mRNA total. No se pudo detectar al peptido de cecropin B en las plantas transgénicas, probablemente debido a una degradación proteolítica rápida del cecropin B nuevamente sintetizado, por las proteasas endogenas de la papa. Ni los pequenos tubérculos de un grupo de 11 clones con niveles de transcriptión moderados a altos ni las rebanadas de tubérculos obtenidos del campo, de cualesquiera de los clones transgénicos obtenidos, mostraron una pudrición significativamente menor (P<0.01 ) después de la inoculation conErwinia carotovora subsp.atroseptica oE. chrysanthemi, las bacterias patógenas causantes de la pudrición blanda de la papa.
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Allefs, S.J.H.M., Florack, D.E.A., Hoogendoorn, C. et al. Erwinia soft rot resistance of potato cultivars transformed with a gene construct coding for antimicrobial peptide cecropin B is not altered. American Potato Journal 72, 437–445 (1995). https://doi.org/10.1007/BF02851677
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DOI: https://doi.org/10.1007/BF02851677