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Overexpression of Arabidopsis WRI1 enhanced seed mass and storage oil content in Camelina sativa

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Abstract

Vegetable oils are essential for human and animal diets and have been used for industrial applications such as cosmetics and lubricants. Camelina sativa, which contains 35–45 % storage oils of seed dry weight, is an emerging oilseed crop. In addition, Arabidopsis WRINKLED1 (AtWRI1) is known to be an AP2/EREBP-type transcription factor that regulates the expression of genes that encode enzymes involved in the glycolytic pathway and fatty acid synthesis. In this study, AtWRI1 was expressed in C. sativa under the control of the seed-specific SiW6 promoter. The introduction of the AtWRI1 gene was identified from polymerase chain reaction (PCR) analysis using genomic DNA, and its mRNA expression was analyzed by reverse transcription-PCR (RT-PCR) using gene-specific primers in transgenic C. sativa plants. The expression of AtWRI1 caused an increase of seed mass through an increase in size, but not through an increase in number of cells. Moreover, the total seed oil contents increased by approximately 14 % in the T3 transgenic C. sativa lines compared with the non-transgenic plants. It was revealed that the elevation in storage oil contents is caused by the upregulation of three isoforms encoding a pyruvate dehydrogenase E1α subunit and three isoforms encoding a biotin carboxyl carrier protein of acetyl-CoA carboxylase complex, which are involved in fatty acid biosynthesis. Finally, the increased expression levels of C. sativa expansin1, which may be involved in cell-wall loosening during cell expansion, was observed in transgenic C. sativa developing seeds. Transgenic C. sativa with enhanced seed oil contents will be useful for the production of non-petroleum-based biomaterials and biofuels.

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References

  • An G (1987) Binary Ti vector for plant transformation and promoter analysis. Method Enzymol 153:292–305

    Article  CAS  Google Scholar 

  • Angelini LG, Moscheni E, Colonna G, Belloni P, Bonari E (1997) Variation in agronomic characteristics and seed oil composition of new oilseed crops in central Italy. Ind Crops Prod 6:313–323

    Article  Google Scholar 

  • Bae JM, Kwak MS, Noh SA, Oh MJ, Kim YS, Shin JS (2014) Overexpression of sweetpotato expansin cDNA (IbEXP1) increases seed yield in Arabidopsis. Transgenic Res 23:657–667

    Article  CAS  PubMed  Google Scholar 

  • Baud S, Wuillème S, Lemoine R, Kronenberger J, Caboche M, Lepiniec L, Rochat C (2005) The AtSUC5 sucrose transporter specifically expressed in the endosperm is involved in early seed development in Arabidopsis. Plant J 43:824–836

    Article  CAS  PubMed  Google Scholar 

  • Baud S, Mendoza MS, To A, Harscoët E, Lepiniec L, Dubreucq B (2007) WRINKLED1 specifies the regulatory action of leafy cotyledon2 towards fatty acid metabolism during seed maturation in Arabidopsis. Plant J 50:825–838

    Article  CAS  PubMed  Google Scholar 

  • Baud S, Wuillème S, To A, Rochat C, Lepiniec L (2009) Role of WRINKLED1 in the transcriptional regulation of glycolytic and fatty acid biosynthetic genes in Arabidopsis. Plant J 60:933–947

    Article  CAS  PubMed  Google Scholar 

  • Cernac A, Benning C (2004) WRINKLED1 encodes an AP2/EREB domain protein involved in the control of storage compound biosynthesis in Arabidopsis. Plant J 40:575–585

    Article  CAS  PubMed  Google Scholar 

  • Clough RC, Matthis AL, Barnum SR, Jaworski JG (1992) Purification and characterization of 3-ketoacyl-acyl carrier protein synthase I11 from Spinach. J Bio Chem 267:20992–20998

    CAS  Google Scholar 

  • Cosgrove DJ, Li LC, Cho HT, Hoffmann-Benning S, Moore RC, Blecker D (2002) The growing world of expansins. Plant Cell Physiol 160:2064–2082

    Google Scholar 

  • Durrett TP, Benning C, Ohlrogge J (2008) Plant triacylglycerols as feedstocks for the production of biofuels. Plant J 54:593–607

    Article  CAS  PubMed  Google Scholar 

  • Dyer JM, Stymne S, Green AG, Carlsson AS (2008) High-value oils from plants. Plant J 54:640–655

    Article  CAS  PubMed  Google Scholar 

  • Eastmond PJ, Quettier AL, Kroon JT, Craddock C, Adams N, Slabas AR (2010) Phosphatidic acid phosphohydrolase1 and 2 regulate phospholipid synthesis at the endoplasmic reticulum in Arabidopsis. Plant Cell 22:2796–2811

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Enjalbert JN, Zheng S, Johnson JJ, Mullen JL, Byrne PF, McKay JK (2013) Brassicaceae germplasm diversity for agronomic and seed quality traits under drought stress. Ind Crops Prod 47:176–185

    Article  CAS  Google Scholar 

  • Focks N, Benning C (1998) Wrinkled1: a novel, low-seed-oil mutant of Arabidopsis with a deficiency in the seed-specific regulation of carbohydrate metabolism. Plant Physiol 118:91–101

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Graham IA (2008) Seed storage oil mobilization. Annu Rev Plant Biol 59:115–142

    Article  CAS  PubMed  Google Scholar 

  • Hayashi K, Hashimoto N, Daigen M, Ashikawa I (2004) Development of PCR-based SNP markers for rice blast resistance genes at the Piz locus. Theor Appl Genet 108:1212–1220

    Article  CAS  PubMed  Google Scholar 

  • Hutcheon C, Ditt RF, Beilstein M, Comai L, Schroeder J, Goldstein E, Shewmaker CK, Nguyen T, Rocher JD, Kiser J (2010) Polyploid genome of Camelina sativa revealed by isolation of fatty acid synthesis genes. BMC Plant Biol 10:233

    Article  PubMed Central  PubMed  Google Scholar 

  • Johnston ML, Luethy MH, Miernyk JA, Randall DD (1997) Cloning and molecular analyses of the Arabidopsis thaliana plastid pyruvate dehydrogenase subunits. Biochim Biophys Acta 1321:200–206

    Article  CAS  PubMed  Google Scholar 

  • Jones A, Davies HM, Voelker TA (1995) Palmitoyl-acyl carrier protein (ACP) thioesterase and the evolutionary origin of plant acyl-ACP thioesterases. Plant Cell 7:359–371

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Kachroo A, Shanklin J, Whittle E, Lapchyk L, Hildebrand D, Kachroo P (2007) The Arabidopsis stearoyl-acyl carrier protein-desaturase family and the contribution of leaf isoforms to oleic acid synthesis. Plant Mol Biol 63:257–271

    Article  CAS  PubMed  Google Scholar 

  • Karvonen HM, Aro A, Tapola NS, Salminen I, Uusitupa MI, Sarkkinen ES (2002) Effect of α-linolenic acid-rich Camelina sativa oil on serum fatty acid composition and serum lipids in hypercholesterolemic subjects. Metabolism 51:1253–1260

    Article  CAS  PubMed  Google Scholar 

  • Kelly AA, Shaw E, Powers SJ, Kurup S, Eastmond PJ (2013) Suppression of the SUGAR-DEPENDENT1 triacylglycerol lipase family during seed development enhances oil yield in oilseed rape (Brassica napus L.). Plant Biotechnol J 11:355–361

    Article  CAS  PubMed  Google Scholar 

  • Kennedy EP (1961) Biosynthesis of complex lipids. Fed Proc 20:934–940

    CAS  PubMed  Google Scholar 

  • 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

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • 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 Gen Genomics 276:351–368

    Article  CAS  Google Scholar 

  • Kim MJ, Yang SW, Mao HZ, Veena SP, Yin JL, Chua NH (2014) Gene silencing of Sugar-dependent 1 (JcSDP1), encoding a patatin-domain triacylglycerol lipase, enhances seed oil accumulation in Jatropha curcas. Biotechnol Biofuels 7:36

    Article  PubMed Central  PubMed  Google Scholar 

  • Konishi T, Shinohara K, Yamada K, Sasaki (1996) Acetyl-CoA carboxylase in higher plants: most plants other than gramineae have both the prokaryotic and the eukaryotic forms of this enzyme. Plant Cell Physiol 37:117–122

    Article  CAS  PubMed  Google Scholar 

  • Koo AJ, Ohlrogge JB, Pollard M (2004) On the export of fatty acids from the chloroplast. J Biol Chem 279:16101–16110

    Article  CAS  PubMed  Google Scholar 

  • Lee SB, Kim H, Kim RJ, Suh MC (2014) Overexpression of Arabidopsis MYB96 confers drought resistance in Camelina sativa via cuticular wax accumulation. Plant Cell Rep 33:1535–1546

    Article  CAS  PubMed  Google Scholar 

  • Liu J, Hua W, Zhan G, Wei F, Wang X, Liu G, Wang H (2010) Increasing seed mass and oil content in transgenic Arabidopsis by the overexpression of wri1-like gene from Brassica napus. Plant Physiol Biochem 48:9–15

    Article  CAS  PubMed  Google Scholar 

  • Liu X, Brost J, Hutcheon C, Guilfoil R, Wilson AK, Leung S, Shewmaker CK, Rooke S, Nguyen T, Kiser J, Rocher JD (2012) Transformation of the oilseed crop Camelina sativa by Agrobacterium-mediated floral dip and simple large-scale screening of transformants. In Vitro Cell Dev Biol Plant 48:462–468

    Article  Google Scholar 

  • Lou Y, Schwender J, Shanklin JJ (2014) FAD2 and FAD3 desaturases form heterodimers that facilitate metabolic channeling in vivo. J Biol Chem 289:17996–18007

    Article  CAS  PubMed  Google Scholar 

  • Lu C, Kang J (2008) Generation of transgenic plants of a potential oilseed crop Camelina sativa by Agrobacterium-mediated transformation. Plant Cell Rep 27:273–278

    Article  CAS  PubMed  Google Scholar 

  • Lu C, Napier JA, Clemente TE, Cahoon EB (2011) New frontiers in oilseed biotechnology: meeting the global demand for vegetable oils for food, feed, biofuel, and industrial applications. Curr Opin Biotechnol 22:252–259

    Article  CAS  PubMed  Google Scholar 

  • Ma W, Kong Q, Arondel V, Kilaru A, Bates PD, Thrower NA, Benning C, Ohlogge JB (2013) WRINKLED1, a ubiquitous regulator in oil accumulating tissues from Arabidopsis embryos to oil palm mesocarp. PLoS One 8:e68887

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Maeo K, Tokuda T, Ayame A, Mitsui N, Kawai T, Tsukagoshi H, Ishiguro S, Nakamura K (2009) An AP2-type transcription factor, WRINKLED1, of Arabidopsis thaliana binds to the AW-box sequence conserved among proximal upstream regions of genes involved in fatty acid synthesis. Plant J 60:476–487

    Article  CAS  PubMed  Google Scholar 

  • Moser BR (2010) Camelina (Camelina sativa L.) oil as a biofuels feedstock: golden opportunity or false hope? Lipid Technol 22:270–273

    Article  CAS  Google Scholar 

  • Moser BR (2012) Biodiesel from alternative oilseed feedstocks: Camelina and field pennycress. Biofuels 3:193–209

    Article  CAS  Google Scholar 

  • Mou Z, He Y, Dai Y, Liu X, Li J (2000) Deficiency in fatty acid synthase leads to premature cell death and dramatic alterations in plant morphology. Plant Cell 12:405–417

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Murphy DJ (2001) The biogenesis and functions of lipid bodies in animals, plants and microorganisms. Prog Lipid Res 40:325–438

    Article  CAS  PubMed  Google Scholar 

  • Nguyen HT, Silva JE, Podicheti R, Macrander J, Yang W, Nazarenus TJ, Nam JW, Jaworski JG, Lu C, Scheffler BE, Mockaitis K, Cahoon EB (2013) Camelina seed transcriptome: a tool for meal and oil improvement and translational research. Plant Biotechnol J 11:759–769

    Article  CAS  PubMed  Google Scholar 

  • Ohto MA, Fischer RL, Goldberg RB, Nakamura K, Harada JJ (2005) Control of seed mass by APETALA2. Proc Natl Acad Sci 102:3123–3128

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Oñate-Sánchez L, Vicente-Carbajosa J (2008) DNA-free RNA isolation protocols for Arabidopsis thaliana, including seeds and siliques. BMC Res Notes 1:93

    Article  PubMed Central  PubMed  Google Scholar 

  • Petrie JR, Shrestha P, Belide S, Kennedy Y, Lester G, Liu Q, Divi UK, Mulder RJ, Mansour MP, Nichols PD, Singh SP (2014) Metabolic engineering Camelina sativa with fish oil-like levels of DHA. PLoS One 9:e85061

    Article  PubMed Central  PubMed  Google Scholar 

  • Pidkowich MS, Nguyen HT, Heilmann I, Ischebeck T, Shanklin J (2007) Modulating seed β-ketoacyl-acyl carrier protein synthase II level converts the composition of a temperate seed oil to that of a palm-like tropical oil. Proc Natl Acad Sci 104:4742–4747

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Pouvreau B, Baud S, Vernoud V, Morin V, Py C, Gendrot G, Pichon JP, Rouster J, Paul W, Rogowsky PM (2011) Duplicate maize Wrinkled1 transcription factors activate target genes involved in seed oil biosynthesis. Plant Physiol 156:674–686

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • 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

    Google Scholar 

  • Qu J, Ye J, Geng YF, Sun YW, Gao SQ, Zhang BP, Chen W, Chua NH (2012) Dissecting functions of KATANIN and WRINKLED1 in cotton fiber development by virus-induced gene silencing. Plant Physiol 160:738–748

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Ruiz-Lopez N, Haslam RP, Napier JA, Sayanova O (2014) Successful high-level accumulation of fish oil omega-3 long-chain polyunsaturated fatty acids in a transgenic oilseed crop. Plant J 77:198–208

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Ruuska SA, Girke T, Benning C, Ohlrogge JB (2002) Contrapuntal networks of gene expression during Arabidopsis seed filling. Plant Cell 14:1191–1206

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Schwender J, Ohlrogge JB, Shachar-Hill Y (2003) A flux model of glycolysis and the oxidative pentosephosphate pathway in developing Brassica napus embryos. J Biol Chem 278:29442–29453

    Article  CAS  PubMed  Google Scholar 

  • Shcherban TY, Shi J, Durachko DM, Guiltinan MJ, McQueen-Mason SJ, Shieh M, Cosgrove DJ (1995) Molecular cloning and sequence analysis of expansins-highly conserved, multigene family of proteins that mediate cell wall extension in plants. Proc Natl Acad Sci USA 92:9245–9249

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Shen B, Allen WB, Zheng P, Li C, Glassman K, Ranch J, Nubel D, Tarczynski MC (2010) Expression of ZmLEC1 and ZmWRI1 increases seed oil production in maize. Plant Physiol 153:980–987

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Snapp AR, Kang J, Qi X, Lu C (2014) A fatty acid condensing enzyme from Physaria fendleri increases hydroxy fatty acid accumulation in transgenic oilseeds of Camelina sativa. Planta 240:599–610

    Article  CAS  PubMed  Google Scholar 

  • Tan H, Yang X, Zhang F, Zheng X, Qu C, Mu J, Fu F, Li J, Guan R, Zhang H, Wang G, Zuo J (2011) Enhanced seed oil production in canola by conditional expression of Brassica napus LEAFY COTYLEDON1 and LEC1-LIKE in developing seeds. Plant Physiol 156:1577–1588

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Taylor DC, Katavic V, Zou J, MacKenzie SL, Keller WA, An J, Friesen W, Barton DL, Pedersen KK, Giblin EM, Ge Y, Dauk M, Sonntag C, Luciw T, Males D (2002) Field testing of transgenic rapeseed cv. Hero transformed with a yeast sn-2 acyltransferase results in increased oil content, erucic acid content and seed yield. Mol Breed 8:317–322

    Article  CAS  Google Scholar 

  • Taylor DC, Zhang Y, Kumar A, Francis T, Giblin EM, Barton DL, Ferrie JR, Laroche A, Shah S, Zhu W, Snyder CL, Hall L, Rakow G, Harwood JL, Weselake RJ (2009) Molecular modification of triacylglycerol accumulation by over-expression of DGAT1 to produce canola with increased seed oil content under field conditions. Botany 87:533–543

    Article  CAS  Google Scholar 

  • Vigeolas H, Waldeck P, Zank T, Geigenberger P (2007) Increasing seed oil content in oil-seed rape (Brassica napus L.) by over-expression of a yeast glycerol-3-phosphate dehydrogenase under the control of a seed-specific promoter. Plant Biotechnol J 5:431–441

    Article  CAS  PubMed  Google Scholar 

  • Wang Z, Huang W, Chang J, Sebastian A, Li Y, Li H, Wu X, Zhang B, Meng F, Li W (2014) Overexpression of SiDGAT1, a gene encoding acyl-CoA: diacylglycerol acyltransferase from Sesamum indicum L. increases oil content in transgenic Arabidopsis and soybean. Plant Cell Tissue Organ Cult 119:399–410

    Article  CAS  Google Scholar 

  • Waraich EA, Ahmed Z, Ahmad R, Ashraf MY, Saifullah 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

    Google Scholar 

  • Weber H, Borisjuk L, Wobus U (1996) Controlling seed development and seed size in Vicia faba: a role for seed coat-associated invertases and carbohydrate state. Plant J 10:823–834

    Article  CAS  Google Scholar 

  • Weber H, Borisjuk L, Wobus U (1997) Sugar import and metabolism during seed development. Trends Plant Sci 2:169–174

    Article  Google Scholar 

  • Wu GZ, Xue HW (2010) Arabidopsis β-ketoacyl-[acyl carrier protein] synthase I is crucial for fatty acid synthesis and plays a role in chloroplast division and embryo development. Plant Cell 22:3726–3744

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Wu XL, Liu ZH, Hu ZH, Huang RZ (2014) BnWRI1 coordinates fatty acid biosynthesis and photosynthesis pathways during oil accumulation in rapeseed. J Integr Plant Biol 56:582–593

    Article  CAS  PubMed  Google Scholar 

  • Zheng Z, Xia Q, Dauk M, Shen W, Selvaraj G, Zou J (2003) Arabidopsis AtGPAT1, a member of the membrane-bound glycerol-3-phosphate acyltransferase gene family, is essential for tapetum differentiation and male fertility. Plant Cell 15:1872–1887

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • 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. Plant J 19:645–653

    Article  CAS  PubMed  Google Scholar 

  • Zubr J (1997) Oil-seed crop: camelina sativa. Ind Crops Prod 6:113–119

    Article  Google Scholar 

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Acknowledgments

We thank Anna Sim at Chonnam National University for technical assistance. This work was supported by grants from the Korea Institute of Planning and Evaluation for Technology (No. 312033-5) and the Cooperative Research Program for Agriculture Science and Technology Development (Next-Generation BioGreen21 Program PJ0110522015), Rural Development Administration, Republic of Korea.

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  1. Department of Bioenergy Science and Technology, Chonnam National University, Gwangju, Korea

    Dahee An & Mi Chung Suh

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  1. Dahee An
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Correspondence to Mi Chung Suh.

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An, D., Suh, M.C. Overexpression of Arabidopsis WRI1 enhanced seed mass and storage oil content in Camelina sativa . Plant Biotechnol Rep 9, 137–148 (2015). https://doi.org/10.1007/s11816-015-0351-x

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