Banana (Musa sp.).
Cultivated bananas are vegetatively propagating herbs, which are difficult to breed because of widespread male and female sterility. As a complementary gene transfer method in banana, the described Agrobacterium protocol relies on highly regenerable embryogenic cell cultures. Embryogenic cells are infected and co-cultivated in the presence of acetosyringone with Agrobacterium tumefaciens harboring a binary plasmid vector to obtain a mixed population of transformed and untransformed plant cells. Transformed plant cells are promoted to grow for 2 to 3 mo on a cell colony induction medium containing the antibiotics geneticin or hygromycin as selective agents, while agrobacteria are counterselected by timentin. The whole procedure, including plant regeneration, takes approx 6 mo and results in an average frequency of 25 to 50 independent transgenic plants per plate, which equals 50 mg of embryogenic cells. This method has been applied to a wide range of cultivars and to generate large populations of transgenic colonies and plants for tagging genes and promoters in banana.
Pérez Hernández
Juan B
JB
Departamento de Fruticultura Tropical, Instituto Canario de Investigaciones Agrarias, Apartado 60, 38200 La Laguna, Santa Cruz de Tenerife, Spain.
Remy
Serge
S
Swennen
Rony
R
Sági
László
L
D016960:Agrobacterium tumefaciens / Q000166:cytology / Q000235:genetics*; D018929:Cell Culture Techniques; D018920:Coculture Techniques; D003470:Culture Media; D018014:Gene Transfer Techniques; D005819:Genetic Markers; D005822:Genetic Vectors; D028521:Musa / Q000166:cytology / Q000196:embryology / Q000235:genetics*; D012639:Seeds / Q000166:cytology / Q000235:genetics; D012987:Soil; D014170:Transformation, Genetic
D003470:Culture Media; D005819:Genetic Markers; D012987:Soil