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

Shu-Fen Li; Li-Ying Song; Wei-Bo Yin; Yu-Hong Chen; Liang Chen; Ji-Lin Li; Richard R.-C. Wang; Zan-Min Hu

doi:10.1016/j.jgg.2011.12.002

Volume 39 Issue 1

Jan. 2012

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Article Navigation > Journal of Genetics and Genomics > 2012 > 39(1): 47-59

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

doi: 10.1016/j.jgg.2011.12.002

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)
Received Date: 2011-12-06
Accepted Date: 2011-12-23
Rev Recd Date: 2011-12-16

Available Online: 2011-12-30

Publish Date: 2012-01-20

Abstract

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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References(50)

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