High-efficiency transformation by biolistics of soybean, common bean and cotton transgenic plants.
This protocol describes a method for high-frequency recovery of transgenic soybean, bean and cotton plants, by combining resistance to the herbicide imazapyr as a selectable marker, multiple shoot induction from embryonic axes of mature seeds and biolistics techniques. This protocol involves the following stages: plasmid design, preparation of soybean, common bean and cotton apical meristems for bombardment, microparticle-coated DNA bombardment of apical meristems and in vitro culture and selection of transgenic plants. The average frequencies (the total number of fertile transgenic plants divided by the total number of bombarded embryonic axes) of producing germline transgenic soybean and bean and cotton plants using this protocol are 9, 2.7 and 0.55%, respectively. This protocol is suitable for studies of gene function as well as the production of transgenic cultivars carrying different traits for breeding programs. This protocol can be completed in 7-10 months.
Rech
Elibio L
EL
Embrapa Genetic Resources and Biotechnology, Laboratory of Gene Transfer, Biotechnology Unit, Parque Estação Biológica PqEB W5 Norte, Brasília 70770-900, DF, Brazil. rech@cenargen.embrapa.br
Vianna
Giovanni R
GR
Aragão
Francisco J L
FJ
D019470:Biolistics / Q000379:methods*; D005819:Genetic Markers; D053209:Herbicide Resistance / Q000235:genetics; D019684:Magnoliopsida / Q000235:genetics*; D030821:Plants, Genetically Modified; D014170:Transformation, Genetic
D005819:Genetic Markers