Effect of Overexpression of γ-Tocopherol Methyltransferase on α-Tocopherol and Fatty Acid Accumulation and Tolerance to Salt Stress during Seed Germination in Brassica napus L.

Rapeseed (Brassica napus L.) is an important oil crop and a major source of tocopherols, also known as vitamin E, in human nutrition. Enhancing the quality and composition of fatty acids (FAs) and tocopherols in seeds has long been a target for rapeseed breeding. The gene gamma-Tocopherol methyltransferase (gamma-TMT) encodes an enzyme catalysing the conversion of gamma-tocopherol to alpha-tocopherol, which has the highest biological activity. However, the genetic basis of gamma-TMT in B. napus seeds remains unclear. In the present study, BnaC02.TMT.a, one paralogue of Brassica napus gamma-TMT, was isolated from the B. napus cultivar Zhongshuang11 by nested PCR, and two homozygous transgenic overexpression lines were further characterised. Our results demonstrated that the overexpression of BnaC02.TMT.a mediated an increase in the alpha- and total tocopherol content in transgenic B. napus seeds. Interestingly, the FA composition was also altered in the transgenic plants; a reduction in the levels of oleic acid and an increase in the levels of linoleic acid and linolenic acid were observed. Consistently, BnaC02.TMT.a promoted the expression of BnFAD2 and BnFAD3, which are involved in the biosynthesis of polyunsaturated fatty acids during seed development. In addition, BnaC02.TMT.a enhanced the tolerance to salt stress by scavenging reactive oxygen species (ROS) during seed germination in B. napus. Our results suggest that BnaC02.TMT.a could affect the tocopherol content and FA composition and play a positive role in regulating the rapeseed response to salt stress by modulating the ROS scavenging system. This study broadens our understanding of the function of the Bn gamma-TMT gene and provides a novel strategy for genetic engineering in rapeseed breeding.

Automated summary

In this study, the overexpression of the BnaC02.TMT.a gene in transgenic rapeseed increased the content of α-tocopherol and total tocopherol in seeds and altered the fatty acid composition. The overexpression also enhanced the tolerance of rapeseed seeds to salt stress. These findings expand our knowledge of the function of the Bnγ-TMT gene and provide a new genetic engineering strategy for rapeseed breeding.