Overexpressing TOC1 in Hybrid Poplar

To create the TOC1 overexpressing construct, TOC1 transcript was amplified from hybrid poplar genome using the primers TOC1_FW 5’- ATGGAGGGAGAGGTAGATGAGC-3’ and TOC1_RV 5’-TTAAGATCCTGAAGCATCGTCCTCAG-3’. The resulting piece was cloned along with 35 S promoter into dpGreen destination vector using the MultiSite Gateway Kit (Invitrogen, MA, United States). Empty construct used as control was assembled in the same manner without TOC1 sequence. In vitro poplar plantlets were transformed following the protocol previously reported^{34 (link)}, and leaf samples were collected after 2 days at the peak of FT2 expression.