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Methods Mol Biol ; 1223: 255-64, 2015.
Article in English | MEDLINE | ID: mdl-25300846

ABSTRACT

Agrobacterium tumefaciens-mediated transformation is an efficient method for incorporating genes and recovering stable transgenic plants in cowpea because this method offers several advantages such as the defined integration of transgenes, potentially low copy number, and preferential integration into transcriptional active regions of the chromosome. Cotyledonary node explants of cowpea present an attractive target for T-DNA delivery followed by regeneration of shoots via axillary proliferation without involvement of a de novo regeneration pathway. In this chapter, we describe a detailed protocol for Agrobacterium-mediated transformation of the cowpea variety Pusa Komal. The seedling cotyledonary node explants are used for cocultivation with an Agrobacterium strain EHA105 harboring standard binary vector, pCAMBIA2301 or pNOV2819, and putative transformed plants are selected using aminoglycoside antibiotic or mannose as sole carbon source, respectively. The entire process includes explant infection to transgenic seed generation in greenhouse.


Subject(s)
Fabaceae/genetics , Genetic Techniques , Plants, Genetically Modified , Agriculture/methods , Agrobacterium tumefaciens/genetics , Cotyledon/genetics , DNA, Bacterial , Fabaceae/growth & development , Genetic Vectors , Germination , Plant Roots/genetics , Seeds/genetics , Selection, Genetic , Sterilization/methods , Transformation, Bacterial
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