Abstract
Tocopherols (Tocs) are a kind of lipid-soluble substance required for the normal physiological function of mammals, particularly their antioxidant capacity. Rapeseed (Brassica napus) oil is an important source of exogenous Tocs. However, the genotypic differences in the total Toc contents, the Toc composition in the seeds, and the molecular markers associated with the seed Toc remain largely unknown. Here, we selected 290 rapeseed accessions based on the resequencing of 991 genomes in a worldwide collection of rapeseed germplasm. The contents of the four Toc isoforms, namely, α-, β-, γ-, and δ-Tocs, were also measured. Results show that the total Toc content and the γ-/α-Toc ratio varied greatly across the accessions from 85.34 to 387.00 mg/mg and 0.65 to 5.03, respectively. Furthermore, we conducted genome-wide association studies on the Tocs, which identified 28 and 73 single nucleotide polymorphisms significantly associated with the variation of total Toc content and γ-/α-Toc ratio, respectively. Bna.C02.VTE4, a putative orthologue of Arabidopsis VITAMIN E DEFICIENT 4, was tightly associated with the γ-/α-Toc ratio. This study recommends specific genetic materials with particularly high total Toc and/or low γ-/α-Toc ratio and the molecular markers and haplotypes associated with these quality traits for rapeseed breeding.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Data availability
The supporting data of figures and tables are available in Supplemental Tables S1–S3. The raw reads of the rapeseed accessions have been deposited in the public database of the National Center of Biotechnology Information under SRP155312 (https://www.ncbi.nlm.nih.gov/sra/SRP155312) and China National Center for Bioinformation (NGDC) (https://ngdc.cncb.ac.cn/gsa/browse/CRA001854).
Abbreviations
- Tocs:
-
Tocopherols
- GWAS:
-
Genome-wide association studies
- SNPs:
-
Single nucleotide polymorphisms
- VTE4 :
-
VITAMIN E DEFICIENT 4
- HPT:
-
Homogentisate phytyltransferase
- HGA:
-
Homogentisate
- PDP:
-
Phytyldiphosphate
- MPBQ:
-
2-Methyl-6-phytylbenzoquinol
- ROS:
-
Reactive oxygen species
- PUFA:
-
Polyunsaturated fatty acids
- LD:
-
Linkage disequilibrium
- CAPS:
-
Cleaved amplified polymorphic sequences
- γ- TMT :
-
GAMMA-TOCOPHEROL METHYLTRANSFERASE
- HTTC:
-
High total Toc content
- LTTC:
-
Low total Toc content
- PAV:
-
Presence-and-absence variations
- CNV:
-
Copy number variations
References
Alexander DH, Novembre J, Lange K (2009) Fast model-based estimation of ancestry in unrelated individuals. Genome Res 19(9):1655–1664. https://doi.org/10.1101/gr.094052.109
Ali E, Hussain S, Hussain N et al (2022) Tocopherol as plant protector: an overview of tocopherol biosynthesis enzymes and their role as antioxidant and signaling molecules. Acta Physiol Plant 44:1–11. https://doi.org/10.1007/s11738-021-03350-x
Alqudah AM, Sallam A, Baenziger PS, Börner A (2020) GWAS : Fast-forwarding gene identification and characterization in temperate Cereals : lessons from Barley – a review. J Adv Res 22:119–135. https://doi.org/10.1016/j.jare.2019.10.013
Arifuzzaman M, Oladzadabbasabadi A, McClean P, Rahman M (2019) Shovelomics for phenotyping root architectural traits of rapeseed/canola (Brassica napus L.) and genome-wide association mapping. Mol Genet Genomics 294:985–1000. https://doi.org/10.1007/s00438-019-01563-x
Berman K, Brodaty H (2004) Tocopherol (vitamin E) in Alzheimer’s disease and other neurodegenerative disorders. CNS Drugs 18:807–825. https://doi.org/10.2165/00023210-200418120-00005
Boonnoy P, Karttunen M, Wong-ekkabut J (2017) Alpha-tocopherol inhibits pore formation in oxidized bilayers. Phys Chem Chem Phys 5699–5704. https://doi.org/10.1039/c6cp08051k
Chin K, Pang K, Soelaiman I (2016) Tocotrienol and its role in chronic diseases. In: Gupta S, Prasad S, Aggarwal B (eds) Anti-inflammatory nutraceuticals and chronic diseases. Springer, Cham, pp 97–130
Collakova E, Dellapenna D (2003) The role of homogentisate phytyltransferase and other tocopherol pathway enzymes in the regulation of tocopherol synthesis during abiotic stress. Plant Physiol 133:930–940. https://doi.org/10.1104/pp.103.026138.930
Demurin Y, Skoric D, Karlovic D (1996) Genetic variability of tocopherol composition in sunflower seeds as a basis of breeding for improved oil quality. Plant Breed 115:33–36. https://doi.org/10.1111/j.1439-0523.1996.tb00867.x
Evans HM, Bishop KS (1922) On the existence of a hitherto unrecognized dietary factor essential for reproduction. Science 56:650–651. https://doi.org/10.1126/science.56.1458.650
Fritsche S, Wang X, Li J et al (2012) A candidate gene-based association study of tocopherol content and composition in rapeseed (Brassica napus). Front Plant Sci 3:1–24. https://doi.org/10.3389/fpls.2012.00129
Fritsche S, Wang X, Jung C (2017) Recent advances in our understanding of tocopherol biosynthesis in plants : an overview of key genes, functions, and breeding of vitamin E improved crops. Antioxidants 6:99. https://doi.org/10.3390/antiox6040099
Gugliandolo A, Bramanti P, Mazzon E (2017) Role of vitamin E in the treatment of Alzheimer’s disease: evidence from animal models. Int J Mol Sci 18:2504. https://doi.org/10.3390/ijms18122504
Havlickova L, He ZS, Wang LH et al (2018) Validation of an updated associative transcriptomics platform for the polyploid crop species Brassica napus by dissection of the genetic architecture of erucic acid and tocopherol isoform variation in seeds. Plant J 93:181–192. https://doi.org/10.1111/tpj.13767
He Y, Wu D, Wei D et al (2017) GWAS, QTL mapping and gene expression analyses in Brassica napus reveal genetic control of branching morphogenesis. Sci Rep 7:1–9. https://doi.org/10.1038/s41598-017-15976-4
Hunter SC, Cahoon EB (2007) Enhancing vitamin E in oilseeds : unraveling tocopherol and tocotrienol biosynthesis. Lipids 42:97–108. https://doi.org/10.1007/s11745-007-3028-6
Hussain N, Irshad F, Jabeen Z et al (2013a) Biosynthesis, structural, and functional attributes of tocopherols in planta; past, present, and future perspectives. J Agric Food Chem 61:6137–6149. https://doi.org/10.1021/jf4010302
Hussain N, Jabeen Z, Yuan-long LI et al (2013b) Detection of tocopherol in oilseed rape ( Brassica napus L.) using gas chromatography with flame ionization detector. J Integr Agric 12:803–814. https://doi.org/10.1016/S2095-3119(13)60301-9
Jiang J, Jia H, Feng G et al (2016) Overexpression of Medicago sativa TMT elevates the α-tocopherol content in Arabidopsis seeds, alfalfa leaves, and delays dark-induced leaf senescence. Plant Sci 249:93–104. https://doi.org/10.1016/j.plantsci.2016.05.004
Kang HM, Jae HS, Susan KS et al (2010) Variance component model to account for sample structure in genome-wide association studies. Nat Genet 42:348–354. https://doi.org/10.1038/ng.548.Variance
Kitts DD, Singh A, Fathordoobady F et al (2019) Plant extracts inhibit the formation of hydroperoxides and help maintain vitamin E levels and omega-3 fatty acids during high temperature processing and storage of hempseed and soybean oils. J Food Sci 84:3147–3155. https://doi.org/10.1111/1750-3841.14817
Kulas E, Ackman RG (2001) Properties of α -, γ -, and δ -tocopherol in purified fish oil triacylglycerols 1. J Amer Oil Chem Soc 78:361–367
Lai GY, Weinstein SJ, Albanes D et al (2014) Association of serum α-tocopherol, β-carotene, and retinol with liver cancer incidence and chronic liver disease mortality. Br J Cancer 111:2163–2171. https://doi.org/10.1038/bjc.2014.365
Li Y, Cao K, Zhu G et al (2019) Genomic analyses of an extensive collection of wild and cultivated accessions provide new insights into peach breeding history. Genome Biol 20:36. https://doi.org/10.1186/s13059-019-1648-9
Long Z, Tu M, Xu Y et al (2023) Genome-wide-association study and transcriptome analysis reveal the genetic basis controlling the formation of leaf wax in Brassica napus. J Exp Bot erad047. https://doi.org/10.1093/jxb/erad047
Lu K, Wei L, Li X et al (2019) Whole-genome resequencing reveals Brassica napus origin and genetic loci involved in its improvement. Nat Commun 10:1–12. https://doi.org/10.1038/s41467-019-09134-9
Ma J, Qiu D, Gao H et al (2020) Over-expression of a γ-tocopherol methyltransferase gene in vitamin e pathway confers PEG-simulated drought tolerance in alfalfa. BMC Plant Biol 20:1–16. https://doi.org/10.1186/s12870-020-02424-1
Ohkatsu Y, Kajiyama T, Arai Y (2001) Antioxidant activities of tocopherols. Polym Degrad Stab 72:303–311. https://doi.org/10.1016/S0141-3910(01)00022-2
Olofsson P, Hultqvist M, Hellgren LI et al (2014) Phytol: a chlorophyll component with anti-inflammatory and metabolic properties. In: Jacob C, Kirsch G, Slusarenko A et al (eds) Recent advances in redox active plant and microbial products. Springer, Dordrecht, pp 345–359
Quadrana L, Almeida J, Asís R et al (2014) Natural occurring epialleles determine vitamin E accumulation in tomato fruits. Nat Commun 5:. https://doi.org/10.1038/ncomms5027
Sattler SE, Gilliland LU, Magallanes-Lundback M et al (2004) Vitamin E is essential for seed longevity and for preventing lipid peroxidation during germination. Plant Cell 16:1419–1432. https://doi.org/10.1105/tpc.021360
Seker M, Gül MK, Ipek M et al (2008) Screening and comparing tocopherols in the rapeseed (Brassica napus L.) and olive (Olea europaea L.) varieties using high-performance liquid chromatography. Int J Food Sci Nutr 59:483–490. https://doi.org/10.1080/09637480701539484
Tanaka H, Maruta T, Tamoi M et al (2015) Plant science transcriptional control of vitamin C defective 2 and tocopherol cyclase genes by light and plastid-derived signals : the partial involvement of GENOMES UNCOUPLED 1. Plant Sci 231:20–29. https://doi.org/10.1016/j.plantsci.2014.11.007
Valentin HE, Lincoln K, Moshiri F et al (2006) The Arabidopsis vitamin E pathway gene5-1 mutant reveals a critical role for phytol kinase in seed tocopherol biosynthesis. Plant Cell 18:212–224. https://doi.org/10.1105/tpc.105.037077
Wang X, Zhang C, Li L et al (2012) Unraveling the genetic basis of seed tocopherol content and composition in rapeseed (Brassica napus L.). PLoS ONE 7:8–14. https://doi.org/10.1371/journal.pone.0050038
Wu D, Liang Z, Yan T et al (2019) Whole-genome resequencing of a worldwide collection of rapeseed accessions reveals the genetic basis of ecotype divergence. Mol Plant 12:30–43. https://doi.org/10.1016/j.molp.2018.11.007
Wu D, Li X, Tanaka R, Wood JC et al (2022) Combining GWAS and TWAS to identify candidate causal genes for tocochromanol levels in maize grain. Genetics 221:iyac091. https://doi.org/10.1093/genetics/iyac091
Xuan L, Yan T, Lu L et al (2020) Genome-wide association study reveals new genes involved in leaf trichome formation in polyploid oilseed rape (Brassica napus L.). Plant Cell Environm 43:675–691. https://doi.org/10.1111/pce.13694
Yan T, Wang Q, Maodzeka A et al (2020) BnaSNPDB: An interactive web portal for the efficient retrieval and analysis of SNPs among 1,007 rapeseed accessions. Comput Struct Biotechnol J 18:2766–2773. https://doi.org/10.1016/j.csbj.2020.09.031
Yan T, Yao Y, Wu D, Jiang L (2021) BnaGVD: A genomic variation database of rapeseed ( Brassicanapus ). Plant Cell Physiol 62:378–383. https://doi.org/10.1093/pcp/pcaa169
Zhang GY, Liu RR, Xu G et al (2013) Increased α-tocotrienol content in seeds of transgenic rice overexpressing Arabidopsis γ-tocopherol methyltransferase. Transgenic Res 22:89–99. https://doi.org/10.1007/s11248-012-9630-2
Zhang C, Zhang W, Ren G et al (2015) Chlorophyll synthase under epigenetic surveillance is critical for vitamin E synthesis, and altered expression affects tocopherol levels in Arabidopsis. Plant Physiol 168:1503–1511. https://doi.org/10.1104/pp.15.00594
Zhang H, Shi Y, Sun M et al (2022) Functional differentiation of BnVTE4 gene homologous copies in α-tocopherol biosynthesis revealed by CRISPR/Cas9 editing. Front Plant Sci 13:1–9. https://doi.org/10.3389/fpls.2022.850924
Acknowledgements
We thank Dr. Ulrike Lohwasser from Leibniz Institute of Plant Genetics and Crop Plant Research, Germany, for providing a part of the rapeseed accessions for this study and Mr. Rui Sun from Agricultural Experiment Station of Zhejiang University for the management of field experiments.
Funding
The Natural Key Research and Development Plan (Code No. 2022YFD1200400), the Key Science and Technology Project of Zhejiang Province (2021C02057), and the Jiangsu Collaborative Innovation Centre sponsored the work for Modern Crop Production.
Author information
Authors and Affiliations
Contributions
LJ conceived the experiments. LL and QH carried through the field experiments, tocopherol measurement, and data analyses. YL, JD, and YZ participated in many helpful discussions and data analysis. QH and LJ wrote the manuscript.
Corresponding authors
Ethics declarations
Competing interests
The authors declare no competing interests.
Additional information
Publisher's note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Key message.
We recommend specific genetic materials with particularly high total tocopherol and/or low γ-/α-tocopherol ratio and the SNP markers and genotypes associated with these quality traits for rapeseed breeding.
Supplementary Information
Below is the link to the electronic supplementary material.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Huang, Q., Lu, L., Xu, Y. et al. Genotypic variation of tocopherol content in a representative genetic population and genome-wide association study on tocopherol in rapeseed (Brassica napus). Mol Breeding 43, 50 (2023). https://doi.org/10.1007/s11032-023-01394-0
Received:
Accepted:
Published:
DOI: https://doi.org/10.1007/s11032-023-01394-0