using a dual luciferase-based system. The dual luciferase reporter
plasmid harbored the MaACS1- and MaACO1- promoter fused to
LUC, and the REN driven by the CaMV35S promoter (CaMV35S-
REN/MaACS1/MaACO1 pro-LUC), while an effector plasmid carried
MaC2H2-1/2 expressed under the control of the CaMV 35S
promoter (Fig. 6a). As exhibited in Fig. 6b, compared with empty
control, the LUC/REN ratio was significantly reduced when either
the MaACS1 pro-LUC or the MaACO1 pro-LUC reporter construct
was cotransfected with the effector of CaMV35S-MaC2H2-1 or
CaMV35S-MaC2H2-2. It has been reported that many TFs regulate
ethylene biosynthetic genes (ACS and ACO) via binding to their
promoters during fruit ripening. For example, LeHB-1, an HD-Zip
homeobox protein, can bind to the homeo box cis-elements of the
tomato LeACO1 promoter and modulate the gene's expression
during fruit ripening (Lin et al., 2008). RIN regulates the expression
of two ethylene biosynthetic genes (ACS2 and ACS4) by binding to
CArG box elements in their promoters (Ito et al., 2008; Fujisawa
et al., 2011). In addition, overexpression of ERF022 led to the
inhibition of two ACC synthases (ACS7 and ACS8), suggesting that
ERF022 negatively regulates ethylene biosynthesis (Nowak et al.,
2015). Our previous study showed that banana ERFs differently
regulated the ethylene biosynthesis, in which MaERF9 activated
MaACO1 promoter activity, while the potential repressor
MaERF11 suppressed MaACS1 and MaACO1 activities
using a dual luciferase-based system. The dual luciferase reporterplasmid harbored the MaACS1- and MaACO1- promoter fused toLUC, and the REN driven by the CaMV35S promoter (CaMV35S-REN/MaACS1/MaACO1 pro-LUC), while an effector plasmid carriedMaC2H2-1/2 expressed under the control of the CaMV 35Spromoter (Fig. 6a). As exhibited in Fig. 6b, compared with emptycontrol, the LUC/REN ratio was significantly reduced when eitherthe MaACS1 pro-LUC or the MaACO1 pro-LUC reporter constructwas cotransfected with the effector of CaMV35S-MaC2H2-1 orCaMV35S-MaC2H2-2. It has been reported that many TFs regulateethylene biosynthetic genes (ACS and ACO) via binding to theirpromoters during fruit ripening. For example, LeHB-1, an HD-Ziphomeobox protein, can bind to the homeo box cis-elements of thetomato LeACO1 promoter and modulate the gene's expressionduring fruit ripening (Lin et al., 2008). RIN regulates the expressionof two ethylene biosynthetic genes (ACS2 and ACS4) by binding toCArG box elements in their promoters (Ito et al., 2008; Fujisawaet al., 2011). In addition, overexpression of ERF022 led to theinhibition of two ACC synthases (ACS7 and ACS8), suggesting thatERF022 negatively regulates ethylene biosynthesis (Nowak et al.,2015). Our previous study showed that banana ERFs differentlyregulated the ethylene biosynthesis, in which MaERF9 activatedMaACO1 promoter activity, while the potential repressorMaERF11 ระงับกิจกรรม MaACS1 และ MaACO1
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