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Functional identification of oleate 12-desaturase and ω-3 fatty acid desaturase genes from Perilla frutescens var. frutescens.
Plant Cell Rep. 2016 Dec; 35(12):2523-2537.PC

Abstract

KEY MESSAGE

We described identification, expression, subcellular localization, and functions of genes that encode fatty acid desaturase enzymes in Perilla frutescens var. frutescens. Perilla (Perilla frutescens var. frutescens) seeds contain approximately 40 % of oil, of which α-linolenic acid (18:3) comprise more than 60 % in seed oil and 56 % of total fatty acids (FAs) in leaf, respectively. In perilla, endoplasmic reticulum (ER)-localized and chloroplast-localized ω-3 FA desaturase genes (PfrFAD3 and PfrFAD7, respectively) have already been reported, however, microsomal oleate 12-desaturase gene (PfrFAD2) has not yet. Here, four perilla FA desaturase genes, PfrFAD2-1, PfrFAD2-2, PfrFAD3-2 and PfrFAD7-2, were newly identified and characterized using random amplification of complementary DNA ends and sequence data from RNAseq analysis, respectively. According to the data of transcriptome and gene cloning, perilla expresses two PfrFAD2 and PfrFAD3 genes, respectively, coding for proteins that possess three histidine boxes, transmembrane domains, and an ER retrieval motif at its C-terminal, and two chloroplast-localized ω-3 FA desaturase genes, PfrFAD7-1 and PfrFAD7-2. Arabidopsis protoplasts transformed with perilla genes fused to green fluorescence protein gene demonstrated that PfrFAD2-1 and PfrFAD3-2 were localized in the ER, and PfrFAD7-1 and PfrFAD7-2 were localized in the chloroplasts. PfrFAD2 and perilla ω-3 FA desaturases were functional in budding yeast (Saccharomyces cerevisiae) indicated by the presence of 18:2 and 16:2 in yeast harboring the PfrFAD2 gene. 18:2 supplementation of yeast harboring ω-3 FA desaturase gene led to the production of 18:3. Therefore, perilla expresses two functional FAD2 and FAD3 genes, and two chloroplast-localized ω-3 FA desaturase genes, which support an evidence that P. frutescens cultivar is allotetraploid plant.

Authors+Show Affiliations

Department of Agricultural Biotechnology, National Institute of Agricultural Science, Rural Development Administration, Jeonju, 54874, Republic of Korea.

Division of Integrative Biosciences and Biotechnology, Pohang University of Science and Technology, Pohang, 37673, Republic of Korea.

Department of Agricultural Biotechnology, National Institute of Agricultural Science, Rural Development Administration, Jeonju, 54874, Republic of Korea.

Department of Agricultural Biotechnology, National Institute of Agricultural Science, Rural Development Administration, Jeonju, 54874, Republic of Korea.

Department of Agricultural Biotechnology, National Institute of Agricultural Science, Rural Development Administration, Jeonju, 54874, Republic of Korea.

Department of Agricultural Biotechnology, National Institute of Agricultural Science, Rural Development Administration, Jeonju, 54874, Republic of Korea.

Department of Agricultural Biotechnology, National Institute of Agricultural Science, Rural Development Administration, Jeonju, 54874, Republic of Korea.

Department of Bioindustry and Bioresource Engineering, Plant Engineering Research Institute, Sejong University, Seoul, 05006, Republic of Korea. hukim64@sejong.ac.kr.

MeSH

Amino Acid SequenceChromatography, GasCloning, MolecularEstersFatty Acid DesaturasesFatty AcidsGene Expression Regulation, EnzymologicGene Expression Regulation, PlantGenes, PlantPerilla frutescensPhylogenyPlant ProteinsSaccharomyces cerevisiaeSeedsSequence AlignmentSubcellular Fractions

Pub Type(s)

Journal Article

Language

eng

PubMed ID

27637203

Citation

Lee, Kyeong-Ryeol, et al. "Functional Identification of Oleate 12-desaturase and Ω-3 Fatty Acid Desaturase Genes From Perilla Frutescens Var. Frutescens." Plant Cell Reports, vol. 35, no. 12, 2016, pp. 2523-2537.

Lee KR, Lee Y, Kim EH, et al. Functional identification of oleate 12-desaturase and ω-3 fatty acid desaturase genes from Perilla frutescens var. frutescens. Plant Cell Rep. 2016;35(12):2523-2537.

Lee, K. R., Lee, Y., Kim, E. H., Lee, S. B., Roh, K. H., Kim, J. B., Kang, H. C., & Kim, H. U. (2016). Functional identification of oleate 12-desaturase and ω-3 fatty acid desaturase genes from Perilla frutescens var. frutescens. Plant Cell Reports, 35(12), 2523-2537.

Lee KR, et al. Functional Identification of Oleate 12-desaturase and Ω-3 Fatty Acid Desaturase Genes From Perilla Frutescens Var. Frutescens. Plant Cell Rep. 2016;35(12):2523-2537. PubMed PMID: 27637203.

* Article titles in AMA citation format should be in sentence-case

TY - JOUR T1 - Functional identification of oleate 12-desaturase and ω-3 fatty acid desaturase genes from Perilla frutescens var. frutescens. AU - Lee,Kyeong-Ryeol, AU - Lee,Yongjik, AU - Kim,Eun-Ha, AU - Lee,Seul-Bee, AU - Roh,Kyung Hee, AU - Kim,Jong-Bum, AU - Kang,Han-Chul, AU - Kim,Hyun Uk, Y1 - 2016/09/16/ PY - 2016/08/18/received PY - 2016/09/05/accepted PY - 2016/9/18/pubmed PY - 2017/4/5/medline PY - 2016/9/18/entrez KW - FAD2 KW - FAD3 KW - FAD7 KW - Microsomal oleate 12-desaturase KW - Perilla frutescens var. frutescens KW - ω-3 Fatty acid desaturase SP - 2523 EP - 2537 JF - Plant cell reports JO - Plant Cell Rep VL - 35 IS - 12 N2 - KEY MESSAGE: We described identification, expression, subcellular localization, and functions of genes that encode fatty acid desaturase enzymes in Perilla frutescens var. frutescens. Perilla (Perilla frutescens var. frutescens) seeds contain approximately 40 % of oil, of which α-linolenic acid (18:3) comprise more than 60 % in seed oil and 56 % of total fatty acids (FAs) in leaf, respectively. In perilla, endoplasmic reticulum (ER)-localized and chloroplast-localized ω-3 FA desaturase genes (PfrFAD3 and PfrFAD7, respectively) have already been reported, however, microsomal oleate 12-desaturase gene (PfrFAD2) has not yet. Here, four perilla FA desaturase genes, PfrFAD2-1, PfrFAD2-2, PfrFAD3-2 and PfrFAD7-2, were newly identified and characterized using random amplification of complementary DNA ends and sequence data from RNAseq analysis, respectively. According to the data of transcriptome and gene cloning, perilla expresses two PfrFAD2 and PfrFAD3 genes, respectively, coding for proteins that possess three histidine boxes, transmembrane domains, and an ER retrieval motif at its C-terminal, and two chloroplast-localized ω-3 FA desaturase genes, PfrFAD7-1 and PfrFAD7-2. Arabidopsis protoplasts transformed with perilla genes fused to green fluorescence protein gene demonstrated that PfrFAD2-1 and PfrFAD3-2 were localized in the ER, and PfrFAD7-1 and PfrFAD7-2 were localized in the chloroplasts. PfrFAD2 and perilla ω-3 FA desaturases were functional in budding yeast (Saccharomyces cerevisiae) indicated by the presence of 18:2 and 16:2 in yeast harboring the PfrFAD2 gene. 18:2 supplementation of yeast harboring ω-3 FA desaturase gene led to the production of 18:3. Therefore, perilla expresses two functional FAD2 and FAD3 genes, and two chloroplast-localized ω-3 FA desaturase genes, which support an evidence that P. frutescens cultivar is allotetraploid plant. SN - 1432-203X UR - https://www.unboundmedicine.com/medline/citation/27637203/Functional_identification_of_oleate_12_desaturase_and_ω_3_fatty_acid_desaturase_genes_from_Perilla_frutescens_var__frutescens_ DB - PRIME DP - Unbound Medicine ER -