Abstract
Maize genetic transformation is a critical tool for functional genomics and crop improvement. Many laboratories, however, continue to face multiple challenges in attempting to achieve routine genetic transformation of maize inbred genotypes. Here, we describe a rapid and robust maize B104 transformation method using immature embryos as explants. This method uses anAgrobacteriumternary vector system, which includes a conventional T-DNA binary vector (pCBL101-RUBY) and a compatible ternary helper plasmid (pKL2299) that carries extra copies of essential virulence genes. The T-DNA binary vector carries theneomycin phosphotransferase II(NptII) gene for selection and a betalain biosynthesis marker,RUBY,for visual screening. We provide step-by-step instructions for immature embryo explant preparation,Agrobacteriuminfection, tissue culture procedures, and greenhouse care for acclimatization of regenerated plantlets.
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