Abstract
Sunflower (Helianthus annuus L.) is still considered as a recalcitrant species to in vitro culture and transformation in spite of the publication of different protocols. Here we describe a routine transformation system of this crop which requires mature HA89 genotype seeds and Agrobacterium tumefaciens EHA105 strain for gene delivery, being both easily available. Selection of transformed shoots depends on root development in kanamycin-selective media, instead of shoot color, avoiding selection of escapes. The establishment of this protocol proved successful for the incorporation of both reporter and agronomic important genes and also for the evaluation of the specific expression patterns of different promoters in transgenic sunflower plants. Stable expression of the incorporated transgenes was confirmed by RT-PCR and GUS reporter gene visualization. Stable inheritance of transgenes was successfully followed until T2 generation in several independent lines.
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Acknowledgments
This study was developed at the Instituto de Biotecnología, Instituto Nacional de Tecnología Agropecuaria (INTA), Argentina, and was supported by grants of ANPCyT (PICTO ASAGIR Nº08-13164, PICTO INTA Nº08-12925) and INTA (PE AEGR3425, PE AEGR3426).
The authors would like to thank Valeria Peralta and Agustín Montenegro for greenhouse technical support.
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Radonic, L.M., Lewi, D.M., López, N.E., Hopp, H.E., Escandón, A.S., Bilbao, M.L. (2015). Sunflower (Helianthus annuus L.). In: Wang, K. (eds) Agrobacterium Protocols. Methods in Molecular Biology, vol 1224. Springer, New York, NY. https://doi.org/10.1007/978-1-4939-1658-0_5
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DOI: https://doi.org/10.1007/978-1-4939-1658-0_5
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