High throughput profiling of tocochromanols in leaves and seeds of Arabidopsis and Maize
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(2020) 16:126 Bao et al. Plant Methods https://doi.org/10.1186/s13007-020-00671-9
Open Access
METHODOLOGY
High throughput profiling of tocochromanols in leaves and seeds of Arabidopsis and Maize Yan Bao*† , Maria Magallenes‑Lundback†, Nicholas Deason and Dean DellaPenna*
Abstract Background: Tocochromanols are a group of lipid-soluble antioxidants produced by all plants and include the toco‑ pherols, which are ubiquitous in the plant kingdom, and the biosynthetically-related compounds tocotrienols, which along with tocopherols commonly occur in seed of monocots. Most tocochromanols have some level of vitamin E activity, with α-tocopherol being the highest, and as such are essential nutrients in the human diet. Tocochromanols are particularly abundant in seeds and are critical for maintaining seed longevity and proper germination and as seed oils are a major component of the human diet, they are an important source of dietary vitamin E. In vegetative tissues, tocochromanols are important components in plant responses to stressful environments and can accumulate to high levels in response to various stresses including high light, heat, and dark. Results: We report a robust, high throughput extraction and HPLC analysis method to quantify the levels of tocophe‑ rols and tocotrienols in leaves and seeds of plants, using Arabidopsis and maize tissues as examples. Conclusion: The described method provides a rapid, high-throughput, cost-effective approach to quantifying the composition and content of tocopherols, and if needed simultaneously tocotrienols, in vegetative tissues and seeds. Optimized extraction methods are described for the two tissue types and have been used to study tocochromanol (vitamin E) natural variation in seed of large Arabidopsis and maize diversity panels, to assess gene function in T-DNA and Mu-tagged populations of Arabidopsis and maize, respectfully, and study the impact of environmental stresses, including high light stress, heat stress, and dark on tocopherols content and composition of vegetative tissue. Keywords: Vitamin E, Tocopherol, Stress, Arabidopsis, Maize, Leaf, Seed Background Tocochromanols, which include tocopherols and tocotrienols, are a class of lipid-soluble, amphipathic compounds bearing a chromanol ring and polyprenyl side chain. The side chain of tocopherols is fully saturated and derived from phytyl diphosphate (phytyl-DP) while that of tocotrienols is derived from geranylgeranyl diphosphate (GGDP) and contains three additional double *Correspondence: [email protected]; [email protected] † Yan Bao and Maria Magallenes-Lundback contributed equally to this work Department of Biochemistry and Molecular Biology, Michigan State University, East Lansing, MI 28824, USA
bonds. Tocopherols are ubiquitous in plants while tocotrienols are more restricted in occurrence and are primarily produced, along with tocopherols, by monocots. Four isoforms of tocopherols and tocotrienols (α, β, δ and γ) occur and differ in their degree and location of methylations on the chromanol ring he
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