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.