RNAi knockdown of fatty acid elongase1 alters fatty acid composition in Brassica napus

Biochem Biophys Res Commun. 2015 Oct 23;466(3):518-22. doi: 10.1016/j.bbrc.2015.09.062. Epub 2015 Sep 14.

Abstract

The quality and end-use of oil from oilseed crops is determined by its fatty acid composition. In particular, the relative proportions of erucic and oleic acids are key selection traits for breeders. The goal of our research is to genetically improve the nutritional quality of Brassica napus cultivar CY2, the oil of which is high in erucic acid (about 40%) and low in oleic acid (about 20%). Here, we report the use of a seed-specific napin A promoter to drive the knockdown of BnFAE1 in transgenic CY2. Southern blotting results confirmed the presence of the transgene. RT-PCR analysis showed that the levels of BnFAE1 were greatly decreased in BnFAE1-Ri lines compared with the CY2 cultivar. Knockdown of BnFAE1 sharply decreased the levels of erucic acid (less than 3%), largely increased the contents of oleic acid (more than 60%) and slightly increased the polyunsaturated chain fatty acids. Compared with high erucic acid parents, expression of BnFAE1 was dramatically decreased in developing F1 seeds derived from reciprocally crossed BnFAE1-Ri lines and high erucic acid cultivars. In addition, F1 seeds derived from reciprocal crosses between BnFAE1-Ri lines and high erucic acid cultivars showed significantly increased oleic acid (more than 52%) and sharply decreased erucic acid (less than 4%), demonstrating that the RNAi construct of BnFAE1 can effectively interfere with the target gene in F1 seeds. Taken together, our results demonstrate that BnFAE1 is a reliable target for genetic improvement of rapeseed in seed oil quality promotion.

Keywords: BnFAE1; Erucic acid; Oleic acid; RNAi knockdown.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetyltransferases / antagonists & inhibitors*
  • Acetyltransferases / genetics*
  • Acetyltransferases / metabolism
  • Brassica napus / genetics*
  • Brassica napus / metabolism*
  • Erucic Acids / metabolism
  • Fatty Acid Elongases
  • Fatty Acids / metabolism*
  • Fatty Acids, Monounsaturated
  • Fatty Acids, Unsaturated / metabolism
  • Gene Expression
  • Gene Knockdown Techniques
  • Genes, Plant
  • Hybridization, Genetic
  • Oleic Acid / metabolism
  • Plant Oils / metabolism
  • Plant Proteins / antagonists & inhibitors*
  • Plant Proteins / genetics*
  • Plant Proteins / metabolism
  • Plants, Genetically Modified
  • Promoter Regions, Genetic
  • RNA Interference
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • RNA, Plant / genetics
  • RNA, Plant / metabolism
  • Rapeseed Oil

Substances

  • Erucic Acids
  • Fatty Acids
  • Fatty Acids, Monounsaturated
  • Fatty Acids, Unsaturated
  • Plant Oils
  • Plant Proteins
  • RNA, Messenger
  • RNA, Plant
  • Rapeseed Oil
  • erucic acid
  • Oleic Acid
  • Acetyltransferases
  • Fatty Acid Elongases