Generation of Camelina lines overexpressing CsGGCT2;1
To overexpress CsGGCT2;1 in Camelina, the gene constructCaMV35Sp::CsGGCT2;1::Nos-t (Figure 1a) was introduced into the Camelina nuclear genome under the control of the constitutive 35Spromoter via flower dip method. Resulting seeds from the transgenic lines were identified using DS Red fluorescence (Figure 1b) and grown up to T3 generation to obtain homozygous lines. Two weeks old seedlings of T3 homozygous lines were analyzed for the confirmation of the overexpression of CsGGCT2;1 transcripts using qRT- PCR. The PCR assays of three elite lines indicated that the relative expression of CsGGCT2;1 transcripts was 6 to 17-fold higher in the Camelina transgenic lines compared to WT seedlings (Figure 1c). The qRT-PCR findings confirmed that the CsGGCT2;1 construct resulted in GGCT2;1 transcriptional overexpression in Camelina.