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Gene Rpi-bt1 from Solanum bulbocastanum Confers Resistance to Late Blight in Transgenic Potatoes

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Abstract

Phytophthora infestans, the causal agent of late blight is the most devastating pathogen of cultivated potato worldwide. Utilizing map based cloning; a genomic region containing a cluster of six nucleotide binding site-leucine-rich repeat resistance gene analogs was isolated from the wild potato species Solanum bulbocastanum. Four genes were pseudogenes, with coding sequences interrupted by either frame shift mutations or premature stop codons. However, neither of the two uninterrupted genes conferred resistance to P. infestans when introduced into susceptible potatoes. Specific primers for one of the pseudogenes were used to amplify an uninterrupted cDNA from P. infestans-infected S. bulbocastanum leaves. A corresponding gDNA was amplified from a late blight-resistant bulbocastanum–tuberosum introgression line (Rpi-bt1). The Rpi-bt1 gene under transcriptional control of the constitutive potato Ubi3 promoter was found to confer resistance to P. infestans in several transgenic potato lines in a whole plant greenhouse assay.

Resumen

Phytophthora infestans, el agente causal del tizón tardío, es el patógeno mas devastador de la papa cultivada en el mundo. Mediante la clonación basada en mapas, se aisló de la especie silvestre de papa Solanum bulbocastanum una región genómica que incluía un grupo de repetición con seis análogos de genes de resistencia de sitios de unión de nucleótidos ricos en leucina. Cuatro genes fueron pseudogenes, con secuencias de codificación interrumpidas ya fuera por mutaciones por cambio de marcos o por codones de terminación prematura. No obstante, ninguno de los dos genes sin interrupción confirió resistencia a P. infestans cuando se introdujeron a papas susceptibles. Se utilizaron iniciadores específicos para uno de los pseudogenes para amplificar un ADNc ininterrumpido de hojas de S. bulbocastanum infectadas con P. infestans. También se amplificó un ADNg correspondiente de una línea de introgresión (Rpi-bt1) de bulbocastanum-tuberosum resistente al tizón tardío. Se encontró que el gen Rpi-bt1 bajo control transcripcional del promotor de papa Ubi3 confiere resistencia a P. infestans en varias líneas de papa transgénica en un ensayo con planta completa bajo invernadero.

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Abbreviations

NBS-LRR:

Nucleotide Binding Site-Leucine-Rich Repeat

RACE:

Rapid Amplification of cDNA Ends

BAC:

Bacterial Artificial Chromosome

-ps:

pseudogene

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Acknowledgements

Funding is acknowledged from the Agricultural Research Service National Programs (CRIS Project Number 5325-21420-003-00D) and Dry Creek Laboratories (CRADA Number 58-3 k95-M-434).

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Author notes

  1. T. Oosumi

    Present address: J.R. Simplot Company, Simplot Plant Sciences, Boise, ID, 83706, USA

  2. M. M. Maccree

    Present address: USDA, Agricultural Research Service, Crops Pathology and Genetics Research Unit, UC Davis Plant Pathology Department, Davis, CA, 95616, USA

Authors and Affiliations

  1. Dry Creek Laboratories, Hughson, CA, 95326, USA

    T. Oosumi

  2. USDA, Agricultural Research Service, Western Regional Research Center, Crop Improvement and Utilization Research Unit, Albany, CA, 94710, USA

    D. R. Rockhold, M. M. Maccree, K. F. McCue & W. R. Belknap

  3. USDA, Agricultural Research Service, Genetic Improvement of Fruits and Vegetables Laboratory, Beltsville, MD, 20705, USA

    K. L. Deahl

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  1. T. Oosumi
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  4. K. L. Deahl
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Correspondence to K. F. McCue.

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Oosumi, T., Rockhold, D.R., Maccree, M.M. et al. Gene Rpi-bt1 from Solanum bulbocastanum Confers Resistance to Late Blight in Transgenic Potatoes. Am. J. Pot Res 86, 456–465 (2009). https://doi.org/10.1007/s12230-009-9100-4

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  • DOI: https://doi.org/10.1007/s12230-009-9100-4

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