Functional characterization of TPSs and CYP in vivo
To further identify the products of target proteins in plants,LaTPS7and LaTPS8 were separately transiently transfected into tobacco leaves. Similarly,LaCYP71D582was co-expressed with either LaTPS7 or LaTPS8 to investigate the function of LaCYP71D582 . Our results showed thatLa TPS7 produced only limonene within tobacco, whereas La TPS8 producedα- pinene and sylvestrene; these results are consistent with those of the in-vitro enzymatic assay (Figure 5a). Notably, carveol levels were assessed in tobacco plants co-expressing LaTPS7 andLaCYP71D582 , but no changes in caveol levels were observed in plants co-expressing LaTPS8 -LaCYP71D582 (Figure 5b). In other words, limonene could be hydroxylated at C6 byLa CYP71D582 to form carveol (Figure 5c).