Acquisition of Wax Biosynthesis Related Gene OsCER4 Antisense-RNA Transgenic Plants by the Cognate Promoter

OsCER4 gene is a member of fatty aldehyde decarbonylase gene family and is highly homologous to CER1 gene in Arabidopsis, however, it is still not known whether OsCER4 participates in the synthesis of cuticular wax. A 1946-bp genomic fragment located on the upstream of the annotated ATG start codon was amplified and used as cognate promoter for antisense RNA vector construction, and a 624-bp fragment of the OsCER4 cDNA was amplified from total RNAs isolated from Zhonghua 11, and then the cognate promoter and OsCER4 cDNA fragments were cloned into the pCAMBIA1380 which was transformed into rice by Agrobacterium-mediated method. The results showed that the antisense transgene driven by the cognate promoter was constructed and transformed into rice successfully, and most of the OsCER4 antisense RNA transgenic plants were positive and segregated as single T-DNA insertion by PCR and segregation analysis of Hyg-resistant in T₁ progeny, and the expression of OsCER4 significantly decreased in the OsCER4 antisense RNA transgenic plants by Western blotting.