Improved Agrobacterium-mediated transformation of three maize inbred lines using MS salts.
Transformation technology as a research or breeding tool to improve maize is routinely used in most industrial and some specialized public laboratories. However, transformation of many inbred lines remains a challenging task, especially when using Agrobacterium tumefaciens as the delivery method. Here we report success in generating transgenic plants and progeny from three maize inbred lines using an Agrobacterium-mediated standard binary vector system to target maize immature embryos. Eleven maize inbred lines were pre-screened for transformation frequency using N6 salts. A subset of three maize inbred lines was then systematically evaluated for frequency of post-infection embryogenic callus induction and transformation on four media regimes: N6 or MS salts in each of two distinct media backgrounds. Transgenic plants recovered from inbred lines B104, B114, and Ky21 were analyzed for transgene integration, expression, and transmission. Average transformation frequencies of 6.4% (for B104), 2.8% (for B114), and 8% (for Ky21) were achieved using MS salts. Availability of Agrobacterium-mediated maize inbred line transformation will improve future opportunities for maize genetic and functional genomic studies.
Frame
Bronwyn R
BR
Department of Agronomy, Iowa State University, Ames, IA 50011-1010, USA.
McMurray
Jennifer M
JM
Fonger
Tina M
TM
Main
Marcy L
ML
Taylor
Kyle W
KW
Torney
François J
FJ
Paz
Margie M
MM
Wang
Kan
K
D015139:Blotting, Southern; D020090:Chromosome Segregation / Q000235:genetics; D032461:Chromosomes, Plant / Q000235:genetics; D047108:Embryonic Development; D018506:Gene Expression Regulation, Plant; D005966:Glucuronidase / Q000378:metabolism; D010641:Phenotype; D051479:Plant Infertility / Q000502:physiology; D030821:Plants, Genetically Modified; D012038:Regeneration; D012231:Rhizobium / Q000378:metabolism*; D012492:Salts / Q000378:metabolism*; D012639:Seeds / Q000378:metabolism; D046509:Tissue Culture Techniques; D014170:Transformation, Genetic; D003313:Zea mays / Q000196:embryology / Q000235:genetics* / Q000502:physiology
D012492:Salts; D005966:Glucuronidase