Abstract
We have previously developed a protocol for the Agrobacterium-mediated transformation of internodal stem sections of Citrus seedlings [Moore et al., Plant Cell Rep. (1992) 11:238-242]. However, most studies were done with a single genotype, Carrizo citrange [C. sinensis (L.) Osb. × Poncirus trifoliata (L.) Raf.]; putatively transformed, β-glucuronidase positive (GUS+) shoots were obtained infrequently; and, rooting of shoots was problematic. Recent studies have led to the development of an improved protocol for Citrus transformation that is applicable to a range of citrus types. Factors influencing transformation frequencies included plant genotype, Agrobacterium strain used for inoculation, and, especially, vector plasmid used. The effect of wounding the explant tissue by particle bombardment prior to inoculation with Agrobacterium was also evaluated. Rooting frequency of regenerated shoots was found to be influenced by the concentration of benzyladenine in the regeneration medium and was increased through the use of an improved soil mixture.
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