[Obtaining new germplast of Brassica napus with high oleic acid content by RNA interference and marker-free transformation of Fad2 gene].

Abstract

A seed-specific Fad2 gene expression cassette, which is free-marker gene and with sense and antisense structure, was constructed by using the promoter and terminator of rape seed storage protein cruciferin gene. Transgenic rape plants without selection marker genes were obtained by Agrobacterium-mediated transformation. The oleic acid content of transgenic plant seeds is 83.9%, which is nearly the same as that of Brassica napus with double Fad2 mutation (85%). The result of RT-PCR analysis shows that the raising of oleic acid content may be due to the degradation of Fad2 mRNA induced by co-transformation of sense-antisense Fad2 gene. These transgenic plants with high oleic acid trait grew normally and without the disadvantageous agronomic traits such as weak cold resistance, tardy development, death of buds and low rate of seed setting caused by Fad2 inactivation in mutant Brassica napus plants. This work would serve as a good base for breeding of more lines with high oleic acid content.