American chestnut (Castanea dentata) is one of the United States' most valuable resources for its nuts and timber. Many scientists are exploring genetic transformation techniques to improve chestnut blight resistance in addition to conventional breeding. In vitro shoot production must be first obtained and optimized in order to establish an efficient transformation system. Although shoot proliferation has been achieved, chestnut is still considered difficult for tissue culture with poor rooting. Therefore, this research has focused on improving rooting ability of micropropagated chestnut shoots. In vitro shoot production was established and maintained in WPM supplemented with 0.1 mg/l BA, 3% sucrose, and 0.7% agar with the pH adjusted to 5.8. The shoots were then transferred to rooting medium containing the same components as for shoot proliferation plus an auxin at various concentrations. Right after placing shoots onto rooting medium, a very thin layer (5 ml) of the same auxin (diluted) was added to provide a quick stimulation of rooting. Detailed discussion will be presented.
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