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Capsicum (pepper) species are economically important crops that are recalcitrant to genetic transformation by Agrobacterium ( Agrobacterium tumefaciens ). A number of protocols for pepper transformation have been described but are not routinely applicable. The main bottleneck in pepper transformation is the low frequency of cells that are both susceptible for Agrobacterium infection and have the ability to regenerate. Here, we describe a protocol for the effi cient regeneration of transgenic sweet pepper ( C. annuum ) through inducible activation of the BABY BOOM (BBM) AP2/ERF transcription factor. Using this approach, we can routinely achieve a transformation effi ciency of at least 0.6 %. The main improvements in this protocol are the reproducibility in transforming different genotypes and the ability to produce fertile shoots. An added advantage of this protocol is that BBM activity can be induced subsequently in stable transgenic lines, providing a novel regeneration system for clonal propagation through somatic embryogenesis.
|Title of host publication||Agrobacterium Protocols: Volume 1 (third edition)|
|Place of Publication||New York|
|Number of pages||368|
|Publication status||Published - 2015|
|Name||Methods in Molecular Biology|
|Publisher||Springer Science+Business Media|