Isolation and Functional Characterisation of the Genes Encoding Δ8-Sphingolipid Desaturase from Brassica rapa

a.
Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China
b.
Graduate University of Chinese Academy of Sciences, Beijing 100049, China
c.
Biological College, Harbin Normal University, Harbin 150080, China
d.
USDA-ARS, FRRL, Utah State University, Logan, UT 84322-6300, USA

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Corresponding author: E-mail address: zmhu@genetics.ac.cn (Zan-Min Hu)

摘要

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Abstract: Δ⁸-Sphingolipid desaturase is the key enzyme that catalyses desaturation at the C8 position of the long-chain base of sphingolipids in higher plants. There have been no previous studies on the genes encoding Δ⁸-sphingolipid desaturases in Brassica rapa. In this study, four genes encoding Δ⁸-sphingolipid desaturases from B. rapa were isolated and characterised. Phylogenetic analyses indicated that these genes could be divided into two groups: BrD8A, BrD8C and BrD8D in group I, and BrD8B in group II. The two groups of genes diverged before the separation of Arabidopsis and Brassica. Though the four genes shared a high sequence similarity, and their coding desaturases all located in endoplasmic reticulum, they exhibited distinct expression patterns. Heterologous expression in Saccharomyces cerevisiae revealed that BrD8A/B/C/D were functionally diverse Δ⁸-sphingolipid desaturases that catalyse different ratios of the two products 8(Z)- and 8(E)-C18-phytosphingenine. The aluminium tolerance of transgenic yeasts expressing BrD8A/B/C/D was enhanced compared with that of control cells. Expression of BrD8A in Arabidopsis changed the ratio of 8(Z):8(E)-C18-phytosphingenine in transgenic plants. The information reported here provides new insights into the biochemical functional diversity and evolutionary relationship of Δ⁸-sphingolipid desaturase in plants and lays a foundation for further investigation of the mechanism of 8(Z)- and 8(E)-C18-phytosphingenine biosynthesis.
- Biochemical functional diversity /
- Expression pattern /
- Long-chain base (LCB)

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