Perturbation of lignin biosynthesis pathway in Brassica napus (canola) plants using RNAi

Abstract

Brassica napus meal contains high levels of lignin, which is one of the most important compositional factors affecting feed utilization by ruminants. We attempted to modify the concentration and composition of lignin in B. napus plants using the RNAi approach. Four genes were targeted for silencing by this approach either independently or in combination; caffeic acid O-methyltransferase (COMT), cinnamate 4-hydroxylase (C4H); coumarate 3-hydroxylase (C3H); ferulic acid 5-hydroxylase (F5H). We successfully developed transgenic B. napus lines expressing CaMV35S:C3H-C4H RNAi, CaMV35S:F5H-COMT RNAi, and Cruciferin:COMT RNAi that contained up to 40% less seed lignin in the transgenic seeds compared to the control. Despite successfully achieving suppression of these lignin biosynthesis genes and reduction in lignin content in B. napus seeds, we observed minor phenotypic effects on the transgenic plants. In lines carrying the cruciferin:COMT RNAi construct we observed a decrease in lignin content (40%) in the seed and anatomical variations when stem sections were examined. While our silencing had no major negative effect on plant growth it resulted in deformation of vessel elements, and minor changes in S-units. Taken together, these results clearly show that by employing RNAi strategy, it is possible to alter seed lignin content and composition in a manner non-detrimental to B. napus plants.Key words: Brassica napus, cruciferin, lignin, COMT, RNAi