Browse > Article
http://dx.doi.org/10.5010/JPB.2014.41.3.146

Isolation and functional analysis of three microsomal delta-12 fatty acid desaturase genes from Camelina sativa (L.) cv. CAME  

Kim, Hyojin (Department of Bioenergy Science and Technology, Chonnam National University)
Go, Young Sam (Department of Bioenergy Science and Technology, Chonnam National University)
Kim, Augustine Yonghwi (Department of Food Science & Technology, Sejong University)
Lee, Sanghyeob (Department of Bioresource engineering, Sejong University)
Kim, Kyung-Nam (Department of Molecular Biology, Sejong University)
Lee, Geung-Joo (Department of Horticultural Science, Chungnam National University)
Kim, Gi-Jun (Breeding Institute, Asia Seed Co. Ltd.)
Suh, Mi Chung (Department of Bioenergy Science and Technology, Chonnam National University)
Publication Information
Journal of Plant Biotechnology / v.41, no.3, 2014 , pp. 146-158 More about this Journal
Abstract
Camelina sativa that belongs to Brassicaceae family is an emerging oilseed crop. Camelina seeds contain approximately 40% storage oils per seed dry weight, which are useful for human and animal diets and industrial applications. Microsomal delta-12 fatty acid desaturase2 (FAD2) catalyzes the conversion of oleic acid to linoleic acid. The polymorphisms of FAD2 genes are correlated with the levels of oleic acids in seed oils. Microsomal delta-12 fatty acid desaturase2 (FAD2) catalyzes the conversion of oleic acid to linoleic acid. The polymorphisms of FAD2 genes are correlated with the levels of oleic acids in seed oils. In this study, three CsFAD2 genes (CsFAD2-1, CsFAD2-2 and CsFAD2-3.1) were isolated from developing seeds of Camelina sativa (L.) cv. CAME. The nucleotide and deduced amino acid sequences of three CsFAD2 genes were compared with those from dicotyledon and monocotyledon plants including Camelina cultivars Sunesone and SRS933. Three histidine motifs (HECGH, HRRHH, and HVAHH) required for FAD activity and a hydrophobic valine or isoleucine residue, which is a SNP (single nucleotide polymorphism) marker related with enzyme activity are well conserved in three CsFAD2s. The expressions of CsFAD2-1 and CsFAD2-3.1 were ubiquitously detected in various Camelina organs, whereas the CsFAD2-2 transcripts were predominantly detected in flowers and developing seeds. The contents of oleic acids decreased, whereas the amounts of linoleic acid increased in dry seeds of transgenic fad2-2 lines expressing each CsFAD2 gene compared with fad2-2 mutant, indicating that three CsFAD2 genes are functionally active. The isolated CsFAD2 genes might be applicable in metabolic engineering of storage oils with high oleic acids in oilseed crops.
Keywords
Citations & Related Records
연도 인용수 순위
  • Reference
1 Cheng J, Zhu L-H, Salentijn EMJ, Huang B, Gruber J, Dechesne AC, Krens FA, Qi W, Visser RG, and van Loo EN (2013) Functional analysis of the omega-6 fatty acid desaturase (CaFAD2) gene family of the oil seed crop Crambe abyssinica. BMC Plant Biol 13:146   DOI
2 Bechtold N, Pelletier G (1998) In planta Agrobacterium-mediated transformation of adult Arabidopsis thaliana plants by vacuum infiltration. Methods Mol Biol 82:259-266
3 Belo A, Zheng P, Luck S, Shen B, Meyer DJ, Li B, Tingey S, Rafalski A (2008) Whole genome scan detects an allelic variant of fad2 associated with increased oleic acid levels in maize. Mol Genet Genomics 279:1-10   DOI
4 Budin J, Breene W, Putnam D (1995) Some compositional properties of camelina (Camelina sativa L. Crantz) seeds and oils. J Am Oil Chem Soc 72:309-315   DOI
5 Chi X, Yang Q, Pan L, Chen M, He Y, Yang Z, Yu S (2011) Isolation and characterization of fatty acid desaturase genes from peanut (Arachis hypogaea L.). Plant Cell Rep 30:1393-1404   DOI
6 Dyer JM, Chapital DC, Kuan JC, Mullen RT, Turner C, McKeon TA, Pepperman AB (2002) Molecular analysis of a bifunctional fatty acid conjugase/desaturase from tung. Implications for the evolution of plant fatty acid diversity. Plant Physiol 130:2027-2038   DOI   ScienceOn
7 Jung JH, Kim H, Go YS, Lee SB, Hur CG, Kim HU, Suh MC (2011) Identification of functional BrFAD2-1 gene encoding microsomal delta-12 fatty acid desaturase from Brassica rapa and development of Brassica napus containing high oleic acid contents. Plant Cell Rep 30:1881-1892   DOI
8 Gimeno RE, Cao J (2008) Thematic review series: glycerolipids. Mammalian glycerol-3-phosphate acyltransferases: new genes for an old activity. J Lipid Res 49:2079-2088   DOI   ScienceOn
9 Guo HH, Li QQ, Wang TT, Hu Q, Deng WH, Xia XL, Gao HB (2013) XsFAD2 gene encodes the enzyme responsible for the high linoleic acid content in oil accumulated in Xanthoceras sorbifolia seeds. J Sci Food Agric 94:482-488
10 Hutcheon C, Ditt RF, Beilstein M, Comai L, Schroeder J, Goldstein E, Shewmaker CK, Nguyen T, De Rocher J, Kiser J (2010) Polyploid genome of Camelina sativa revealed by isolation of fatty acid synthesis genes. BMC Plant Biol 10:233   DOI
11 Kagale S, Koh C, Nixon J, Bollina V, Clarke WE, Tuteja R, Spillane C, Robinson SJ, Links MG, Clarke C, Higgins EE, Huebert T, Sharpe AG, Parkin IA (2014) The emerging biofuel crop Camelina sativa retains a highly undifferentiated hexaploid genome structure. Nat Commun 5:3706
12 Lee KR, Sohn IS, Jung JH, Kim SH, Roh KH, Kim JB, Suh MC, Kim HU (2013) Functional analysis and tissue-differential expression of four FAD2 genes in amphidiploid Brassica napus derived from Brassica rapa and Brassica oleracea. Gene 531:253-262   DOI
13 Kang J, Snapp AR, Lu C (2011) Identification of three genes encoding microsomal oleate desaturases (FAD2) from the oilseed crop Camelina sativa. Plant Physiol Biochem 49:223-229   DOI
14 Kennedy EP (1961) Biosynthesis of complex lipids. Fed Proc Fed Am Soc Exp Biol 20:934-940
15 Kim MJ, Kim H, Shin JS, Chung CH, Ohlrogge JB, Suh MC (2006) Seed-specific expression of sesame microsomal oleic acid desaturase is controlled by combinatorial properties between negative cis-regulatory elements in the SeFAD2 promoter and enhancers in the 5'-UTR intron. Mol Genet Genomics 276:351-368   DOI   ScienceOn
16 Kim HU, Li Y, Huang AH (2005) Ubiquitous and endoplasmic reticulum-located lysophosphatidyl acyltransferase, LPAT2, is essential for female but not male gametophyte development in Arabidopsis. Plant Cell 17:1073-1089   DOI   ScienceOn
17 Le Corre V, Roux F, Reboud X (2002) DNA polymorphism at FRIGIDA gene in Arabidopsis thaliana: extensive nonsynonymous variation is consistent with local selection for flowering time. Mol Biol Evol 19(8):1261-1271   DOI
18 Lee KR, Kim SH, Go YS, Jung SM, Roh KH, Kim JB, Suh MC, Lee S, Kim HU (2012) Molecular cloning and functional analysis of two FAD2 genes from American grape (Vitis labrusca L.). Gene 509:189-194   DOI
19 Lemieux B, Miquel M, Somerville C, Browse J. 1990. Mutants of Arabidopsis with alterations in seed lipid fatty acid composition. Theor Appl Genet 80:234-240
20 Liu Q, Cao S, Zhou XR, Wood C, Green A, Singh S (2013) Nonsense-mediated mRNA degradation of CtFAD2-1 and development of a perfect molecular marker for olol mutation in high oleic safflower (Carthamus tinctorius L.). Theor Appl Genet 126:2219-2231   DOI
21 Lou Y, Schwender J, Shanklin J (2014) FAD2 and FAD3 desaturases form heterodimers that facilitate metabolic channeling in vivo. J Biol Chem 289(26):17996-8007   DOI
22 Lozinsky S, Yang H, Forseille L, Cook GR, Ramirez-Erosa I, Smith MA (2014) Characterization of an oleate 12-desaturase from Physaria fendleri and identification of 5'UTR introns in divergent FAD2 family genes. Plant Physiol Biochem 75:114-122   DOI
23 Moser BR (2012) Biodiesel from alternative oilseed feedstocks: camelina and field pennycress. Biofuels 3:193-209   DOI
24 McCartney AW, Dyer JM, Dhanoa PK, Kim PK, Andrews DW, McNew JA, Mullen RT (2004) Membrane-bound fatty acid desaturases are inserted co-translationally into the ER and contain different ER retrieval motifs at their carboxy termini. Plant J 37:156-173   DOI
25 Miquel M, James Jr D, Dooner H, Browse J (1993) Arabidopsis requires polyunsaturated lipids for low-temperature survival. Proc Natl Acad Sci USA 90:6208-6212   DOI   ScienceOn
26 Moser BR (2010) Camelina (Camelina sativa L.) oil as a biofuels feedstock: Golden opportunity or false hope? Lipid Technol 22:270-273   DOI
27 Nam JW, Kappock TJ (2007) Cloning and transcriptional analysis of Crepis alpina fatty acid desaturases affecting the biosynthesis of crepenynic acid. J Exp Bot 58:1421-1432   DOI
28 Odell JT, Nagy F, Chua NH (1985) Identification of DNA sequences required for activity of the cauliflower mosaic virus 35S promoter. Nature 313:810-812   DOI   ScienceOn
29 Onate-Sanchez L, Vicente-Carbajosa J (2008) DNA-free RNA isolation protocols for Arabidopsis thaliana, including seeds and siliques. BMC Res Notes 1:93   DOI   ScienceOn
30 Okuley J, Lightner J, Feldmann K, Yadav N, Lark E, Browse J (1994) Arabidopsis FAD2 gene encodes the enzyme that is essential for polyunsaturated lipid synthesis. Plant Cell 6:147-158   DOI
31 Pham AT, Lee JD, Shannon JG, Bilyeu KD (2010) Mutant alleles of FAD2-1A and FAD2-1B combine to produce soybeans with the high oleic acid seed oil trait. BMC Plant Biol 10:195   DOI
32 Putnam DH, Budin JT, Field LA, Breene WM (1993) Camelina: A promising low-input oilseed. In: Janick J, Simon JE, (eds.), New crops, Wiley, New York, pp 314-322
33 Shanklin J, Cahoon EB (1998) Desaturation and related modifications of fatty acids1. Annu Rev Plant Physiol Plant Mol Biol 49:611-641   DOI
34 Qu J, Mao HZ, Chen W, Gao SQ, Bai YN, Sun YW, Geng YF, Ye J (2012) Development of marker-free transgenic Jatropha plants with increased levels of seed oleic acid. Biotechnol Biofuels 5:10   DOI
35 Rodriguez-Rodriguez MF, Sanchez-Garcia A, Salas JJ, Garces R, Martinez-Force E (2013) Characterization of the morphological changes and fatty acid profile of developing Camelina sativa seeds. Ind Crops Prod 50:673-679   DOI
36 Rounsley SD, Last RL (2010). Shotguns and SNPs: how fast and cheap sequencing is revolutionizing plant biology. The Plant J 61:922-927   DOI
37 Shanklin J, Whittle E, Fox BG (1994). Eight histidine residues are catalytically essential in a membrane-associated iron enzyme, stearoyl-CoA desaturase, and are conserved in alkane hydroxylase and xylene monooxygenase. Biochem 33:12787-12794   DOI   ScienceOn
38 Waraich EA, Ahmed Z, Ahmad R, Ashraf MY, Naeem MS, Rengel Z (2013) Camelina sativa, a climate proof crop, has high nutritive value and multiple-uses: a review. Aust J Crop Sci 7:1551-1559
39 Shi J, Cao Y, Fan X, Li M, Wang Y, Ming F (2012) A rice microsomal delta-12 fatty acid desaturase can enhance resistance to cold stress in yeast and Oryza sativa. Mol Breed 29(3):743-757   DOI
40 Vander Zwan C, Brodie SA, Campanella JJ (2000) The Intraspecific phylogenetics of Arabidopsis thaliana in worldwide populations. Systematic Botany 25:47-59   DOI
41 Zheng P, Allen WB, Roesler K, Williams ME, Zhang S, Li J, Glassman K, Ranch J, Nubel D, Solawetz W, Bhattramakki D, Llaca V, Deschamps S, Zhong G-Y, Tarczynski MC, Shen B (2008) A phenylalanine in DGAT is a key determinant of oil content and composition in maize. Nat Genet 40(3):367-372   DOI   ScienceOn
42 Zou J, Wei Y, Jako C, Kumar A, Selvaraj G, Taylor DC (1999) The Arabidopsis thaliana TAG1 mutant has a mutation in a diacylglycerol acyltransferase gene. The Plant J 19:645-653   DOI   ScienceOn
43 Zhang M, Fan J, Taylor DC, Ohlrogge JB (2009) DGAT1 and PDAT1 acyltransferases have overlapping functions in Arabidopsis triacylglycerol biosynthesis and are essential for normal pollen and seed development. Plant Cell 21:3885-3901   DOI