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The beta-ketoacyl-ACP synthase II (KASII) is an enzyme in fatty acid biosynthesis, catalyzing the elongation of 16:0-acyl carrier protein (ACP) to 18:0-ACP in plastids. Mutations in  KASII  genes in higher plants can lead to lethality, which makes it difficult to utilize the gene for lipid metabolic engineering. We demonstrated previously that transient expression of plastid-directed fatty acyl reductases and wax ester synthases could result in different compositions of wax esters. We hypothesized that changing the ratio between C16 (palmitoyl-compounds) and C18 (stearoyl-compounds) in the plastidic acyl-ACP pool by inhibition of  KASII  expression would change the yield and composition of wax esters via substrate preference of the introduced enzymes. Here, we report that transient inhibition of  KASII  expression by three different RNAi constructs in leaves of  N. benthamiana  results in almost complete inhibition of  KASII  expression. The transient RNAi approach led to a shift of carbon flux from a pool of C18 fatty acids to C16, which significantly increased wax ester production in  AtFAR6 -containing combinations. The results demonstrate that transient inhibition of  KASII  in vegetative tissues of higher plants enables metabolic studies towards industrial production of lipids such as wax esters with specific quality and composition.

Transient silencing of the KASII genes is feasible in Nicotiana benthamiana for metabolic engineering of wax ester composition