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Regulation of FATTY ACID ELONGATION1 expression and production in Brassica oleracea and Capsella rubella

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Abstract

Main conclusion

The contribution of variations in coding regions or promoters to the changes in FAE1 expression levels have be quantified and compared in parallel by specifically designed swapping constructs.

FATTY ACID ELONGATION1 (FAE1) is a key gene in control of erucic acid synthesis in plant seeds. The expression of FAE1 genes in Brassica oleracea and Capsella rubella, representatives of high and low erucic acid species, respectively, was characterized to provide insight into the regulation of very long-chain fatty-acid biosynthesis in seeds. Virtually, no methylation was detected either in B. oleracea or in C. rubella, suggesting that modification of promoter methylation might not be a predominant mechanism. Swapping constructs were specifically designed to quantify and compare the contribution of variations in coding regions or promoters to the changes in FAE1 expression levels in parallel. A significantly higher fold change in erucic acid content was observed when swapping coding regions rather than when swapping promoters, indicating that the coding region is a major determinant of the catalytic power of β-ketoacyl-CoA synthase proteins. Common motifs have been proposed as essential for the preservation of basic gene expression patterns, such as seed-specific expression. However, the occurrence of variation in common cis-elements or the presence of species-specific cis-elements might be plausible mechanisms for changes in the expression levels in different organisms. In addition, conflicting observations in previous reports associated with FAE1 expression are discussed, and we suggest that caution should be taken when selecting a plant transformation vector and in interpreting the results obtained from vectors carrying the CaMV 35S promoter.

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Abbreviations

DAF:

Days after flowering

FAE1:

FATTY ACID ELONGATION1

KCS:

β-Ketoacyl-CoA synthase

HEA:

High erucic acid

LEA:

Low erucic acid

VLCFA:

Very long-chain fatty acid

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Acknowledgements

This work was supported by grants from the National Natural Science Foundation of China (31470448), Jiangsu Province Key Laboratory for Plant Ex Situ Conservation (QIAN201202), Project for Ability Improvement of Provincial Public Institutes (BM2015019) granted to X. Q. Sun, and the Jiangsu Key Laboratory for the Research and Uti1ization of Plant Resources (JSPKLB201510) granted to G. C. Zhou.

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Authors and Affiliations

  1. Institute of Botany, Jiangsu Province and Chinese Academy of Sciences, No. 1 Qianhu Houcun, Nanjing, 210014, Jiangsu, China

    Dinghong Li, Zhao Lei, Jiayu Xue, Guangcan Zhou, Yueyu Hang & Xiaoqin Sun

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  1. Dinghong Li
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  2. Zhao Lei
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  3. Jiayu Xue
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Correspondence to Xiaoqin Sun.

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This work does not contain any study with human participants and/or animals, and fully complies with ethical standards.

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425_2017_2731_MOESM1_ESM.tif

Fig. S1 Sliding window analysis of the TA content in the promoters of eight Brassicaceae FAE1 orthologs. TA content was calculated in the sliding window analysis (window width: 50, step size: 10) in the promoter region starting from the site at position −1 relative to the start codon (TIFF 645 kb)

425_2017_2731_MOESM2_ESM.tif

Fig. S2 Bioinformatic analysis of the promoters of eight Brassicaceae FAE1 orthologs. Promoter sequences upstream of the ATG start codon of the FAE1 genes were retrieved from the plant genome database, and the promoter cis-elements (using the presented color code) were analyzed using PLANTCARE (http://bioinformatics.psb.ugent.be/webtools/plantcare/html/) and PLACE (http://www.dna.affrc.go.jp/PLACE/) (TIFF 1282 kb)

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Li, D., Lei, Z., Xue, J. et al. Regulation of FATTY ACID ELONGATION1 expression and production in Brassica oleracea and Capsella rubella . Planta 246, 763–778 (2017). https://doi.org/10.1007/s00425-017-2731-7

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