Sunflower (Helianthus annuus L.)

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 tumefaci...

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Published inMethods in molecular biology (Clifton, N.J.) Vol. 1224; p. 47
Main Authors Radonic, Laura M, Lewi, Dalia M, López, Nilda E, Hopp, H Esteban, Escandón, Alejandro S, Bilbao, Marisa López
Format Journal Article
LanguageEnglish
Published United States 01.01.2015
Subjects
Agrobacterium tumefaciens - genetics
Agrobacterium tumefaciens - growth & development
Coculture Techniques
Disinfection
Flowers - growth & development
Gene Transfer Techniques
Genetic Engineering - methods
Helianthus - genetics
Helianthus - growth & development
Helianthus - physiology
Plants, Genetically Modified
Regeneration
Seeds - growth & development
Soil
Transformation, Genetic
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Summary: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.
ISSN:1940-6029
DOI:10.1007/978-1-4939-1658-0_5