Methods to enhance sweetgum (Liquidambar styraciflua L.) in vitro axillary shoot formation and shoot establishment into soil are presented. Sweetgum shoots grown in an automated plant culture system (APCS) produced 400 to 500 shoots via axillary branching compared to only 40 shoots produced within Magenta vessels containing agar medium after 8 weeks of incubation. Vitrification was observed in as many as 80% of the axillary shoots produced in the APCS. A continuous carbon dioxide (CO2)-flow-through system was tested on both vitrified and non-vitrified sweetgum shoots transferred from the APCS to soil. One- and two-cm-long vitrified shoots were grown within CO2-flow-through system chambers and subjected to 350, 1500, 3000, 10,000, or 30,000 μL·L–1 (ppm) CO2 for 4 weeks. Administering 10,000 μL·L–1 CO2 improved culture survival and enhanced overall shoot and root growth compared to shoots grown under ambient atmosphere (i.e., 350 μL·L–1 CO2).
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