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Evaluation of Neomycin Phosphotransferase II and Cry3A as Targets for the Serological Detection of Transgenic Potato Tubers

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Abstract

Neomycin phosphotransferase II (NptII) and Cry3A were evaluated as potential targets in ELISA for the detection of transgenic potato tubers. The highest concentrations of NptII (0.23 – 2.95 μg /g) were measured in cultivar NL 10-ATL tubers, while very low (<0.007 μg /g) or undetectable levels of NptII were found in tuber tissue of the other cultivars. Polymerase chain reaction (PCR) analysis revealed that the nptII transgene could not be detected in cultivars NL 10-SUP or in NL-30-RBK-082. It was concluded that a serological method targeting NptII would not provide an effective approach for detecting transgenic potato tubers. Cry3A could be detected in all the cultivars, ranging between 0.02 μg /g in NL 20-SHE and NL 10-ATL to 1.98 μg /g in NL 10-SUP. Environmental conditions appeared to influence Cry3A protein accumulation in tubers of cultivar NL 30-RBK-350 but no such effect on cultivars NL 10-ATL or NL 20-SHE could be detected. It was concluded that ELISA methods for Cry3A detection could be used for confirming the presence of cry3A in individual tubers. With improved sensitivity for Cry3A, serological methods may be useful in screening larger lots for the presence of transgenic potato tubers.

Resumen

Se evaluaron neomicina fosfotransferasa II (NptII) y Cry 3A como objetivos potenciales en ELISA para la detección de tubérculos de papa transgénica. Las concentraciones más altas de NptII (0.23–2.95 µg/g) se midieron en tubérculos del cultivar NL-10-ATL, mientras que se encontró niveles muy bajos (<0.007 µg/g) o indetectables de NptII en la pulpa del tejido de tubérculo de otros cultivares. El análisis de la reacción en cadena de la polimerasa (PCR), reveló que el transgen nptII no pudo ser detectado en los cultivares NL10-SUP o en NL-30-RBK-082. Se concluyó que el método serológico dirigido a NptII no proporciona un enfoque efectivo para detectar tubérculos de papa transgénica. El Cry3A puede ser detectado en todos los cultivares en el rango entre 0.02 µg/g en NL 20-SHE y NL 10-ATL a 1.98 µg/g en NL 10-SUP. Las condiciones ambientales parecen influenciar la acumulación de la proteína Cry3A en tubérculos del cultivar NL 30 RBK 350 pero no se pudo detectar tal efecto sobre los cultivares NL10-ATL o NL 20-SHE. Se concluyó que el método ELISA para la detección de Cry3A podría usarse para confirmar la presencia de cry3A en tubérculos individuales. Con sensibilidad mejorada para Cry3A, los métodos serológicos pueden ser útiles en el tamizado de lotes grandes para presencia de tubérculos de papa transgénica.

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Acknowledgements

The authors thank Bob Jenkins of the Canadian Food Inspection Agency for maintaining the transgenic potato tissue culture lines. The authors are also grateful to Christine Noronha and Mary Smith of Agriculture and Agri-Food Canada for providing the transgenic field tubers.

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Authors and Affiliations

  1. Canadian Food Inspection Agency, Charlottetown Laboratory, 93 Mount Edward Road, Charlottetown, Prince Edward Island, C1A 5T1, Canada

    Donna S. Smith, Philip W. Maxwell & Solke H. De Boer

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  1. Donna S. Smith
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  2. Philip W. Maxwell
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  3. Solke H. De Boer
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Correspondence to Donna S. Smith.

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Smith, D.S., Maxwell, P.W. & De Boer, S.H. Evaluation of Neomycin Phosphotransferase II and Cry3A as Targets for the Serological Detection of Transgenic Potato Tubers. Am. J. Pot Res 85, 60–68 (2008). https://doi.org/10.1007/s12230-008-9007-5

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  • DOI: https://doi.org/10.1007/s12230-008-9007-5

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