Although species within the genus Populus are, in general, easier to transform and regenerate in vitro than most other trees, many poplar species are very recalcitrant. Many protocols that previously have been reported were developed for a specific genotype or species. Thus, it has often been necessary to re-optimize a protocol each time research is initiated with a new genotype. The method presented in this chapter has been effective for a wide variety of poplar genotypes.