Planta

, Volume 245, Issue 3, pp 611–622 | Cite as

Naturally occurring high oleic acid cottonseed oil: identification and functional analysis of a mutant allele of Gossypium barbadense fatty acid desaturase-2

  • Jay Shockey
  • Michael Dowd
  • Brian Mack
  • Matthew Gilbert
  • Brian Scheffler
  • Linda Ballard
  • James Frelichowski
  • Catherine Mason
Original Article

Abstract

Main conclusion

Some naturally occurring cotton accessions contain commercially attractive seed oil fatty acid profiles. The likely causal factor for a high-oleate trait in pima cotton (Gossypium barbadense) accession GB-713 is described here.

Vegetable oils are broadly used in the manufacture of many human and animal nutritional products, and in various industrial applications. Along with other well-known edible plant oils from soybean, corn, and canola, cottonseed oil is a valuable commodity. Cottonseed oil is a co-product derived from the processing of cottonseed fiber. In the past, it was used extensively in a variety of food applications. However, cottonseed oil has lost market share in recent years due to less than optimal ratios of the constituent fatty acids found in either traditional or partially hydrogenated oil. Increased awareness of the negative health consequences of dietary trans-fats, along with the public wariness associated with genetically modified organisms has created high demand for naturally occurring oil with high monounsaturate/polyunsaturate ratios. Here, we report the discovery of multiple exotic accessions of pima cotton that contain elevated seed oil oleate content. The genome of one such accession was sequenced, and a mutant candidate fatty acid desaturase-2 (FAD2-1D) gene was identified. The mutant protein produced significantly less linoleic acid in infiltrated Arabidopsis leaf assays, compared to a repaired version of the same enzyme. Identification of this gene provides a valuable resource. Development of markers associated with this mutant locus will be very useful in efforts to breed the high-oleate trait into agronomic fiber accessions of upland cotton.

Keywords

Cottonseed Fatty acid desaturase Oleic acid Linoleic acid 

Abbreviations

CTAB

Cetyltrimethylammonium bromide

DNA

Deoxyribonucleic acid

FAD

Fatty acid desaturase

FAME

Fatty acid methyl ester

FID

Flame ionization detection

GC

Gas chromatography

GRAS

Generally recognized as safe

GRIN

Germplasm Resources Information Network

NCGC

U.S. National Cotton Germplasm Collection

PCR

Polymerase chain reaction

PHO

Partially hydrogenated oil

RT

Reverse transcription

Supplementary material

425_2016_2633_MOESM1_ESM.docx (44 kb)
Supplementary material 1 (DOCX 44 kb)
425_2016_2633_MOESM2_ESM.xlsx (25 kb)
Supplementary material 2 (XLSX 24 kb)

References

  1. AOCS (1998) Official method Ce 1e-91. In: Firestone D (ed) Official methods and recommended practices of the American Oil Chemists’ Society. 5th edn. AOCS Press, Champaign, p 3Google Scholar
  2. Badami RC, Patil KB (1980) Structure and occurrence of unusual fatty acids in minor seed oils. Prog Lipid Res 19:119–153CrossRefPubMedGoogle Scholar
  3. Boetzer M, Pirovano W (2012) Toward almost closed genomes with GapFiller. Genome Biol 13:R56CrossRefPubMedPubMedCentralGoogle Scholar
  4. Browse J, Kunst L, Anderson S, Hugly S, Somerville CR (1989) A mutant of Arabidopsis deficient in the chloroplast 16:1/18:1 desaturase. Plant Physiol 90:522–529CrossRefPubMedPubMedCentralGoogle Scholar
  5. Chapman KD, Austin-Brown S, Sparace SA, Kinney AJ, Ripp KG, Pirtle IL, Pirtle RM (2001) Transgenic cotton plants with increased seed oleic acid content. J Am Oil Chem Soc 78:941–947CrossRefGoogle Scholar
  6. Chen ZJ, Scheffler BE, Dennis E, Triplett BA, Zhang et al (2007) Toward sequencing cotton (Gossypium) genomes. Plant Physiol 145:1303–1310CrossRefPubMedPubMedCentralGoogle Scholar
  7. 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). Plant Cell Rep 30:1393–1404CrossRefPubMedGoogle Scholar
  8. Dowd MK (2012) Identification of the unsaturated heptadecyl fatty acids in the seed oils of Thespesia populnea and Gossypium hirsutum. J Am Oil Chem Soc 89:1599–1609CrossRefGoogle Scholar
  9. Dyer JM, Chapital DC, Kuan J-CW, 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 130:2027–2038CrossRefPubMedPubMedCentralGoogle Scholar
  10. Federal Register (2013) 78:67179–67185Google Scholar
  11. Fox BG, Shanklin J, Somerville C, Münck E (1993) Stearoyl-acyl carrier protein delta 9 desaturase from Ricinus communis is a diiron-oxo protein. Proc Natl Acad Sci USA 90:2486–2490CrossRefPubMedPubMedCentralGoogle Scholar
  12. Gleave AP (1992) A versatile binary vector system with a T-DNA organisational structure conducive to efficient integration of cloned DNA into the plant genome. Plant Mol Biol 20:1203–1207CrossRefPubMedGoogle Scholar
  13. Guo HH, Li QQ, Wang TT, Hu Q, Deng WH, Xia XL, Gao HB (2014) XsFAD2 gene encodes the enzyme responsible for the high linoleic acid content in oil accumulated in Xanthoceras sorbifolia seeds. J Sci Food Agric 94:482–488CrossRefPubMedGoogle Scholar
  14. Khadake RM, Ranjekar PK, Harsulkar AM (2009) Cloning of a novel omega-6 desaturase from flax (Linum usitatissimum L.) and its functional analysis in Saccharomyces cerevisiae. Mol Biotechnol 42:168–174CrossRefPubMedGoogle Scholar
  15. 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 509:189–194CrossRefPubMedGoogle Scholar
  16. Li J-F, Norville JE, Aach J, McCormack M, Zhang D, Bush J, Church GM, Sheen J (2013) Multiplex and homologous recombination–mediated genome editing in Arabidopsis and Nicotiana benthamiana using guide RNA and Cas9. Nature 31:688–691Google Scholar
  17. Li F, Fan G, Wang K, Sun F, Yuan Y, Song G, Li Q, Ma Z et al (2014) Genome sequence of the cultivated cotton Gossypium arboreum. Nat Genet 46:567–572CrossRefPubMedGoogle Scholar
  18. Li F, Fan G, Lu C, Xiao G, Zou C, Kohel RJ, Ma Z, Shang H et al (2015) Genome sequence of cultivated Upland cotton (Gossypium hirsutum TM-1) provides insights into genome evolution. Nat Biotechnol 33:524–530CrossRefPubMedGoogle Scholar
  19. Li X, Mei D, Liu Q, Fan J, Singh S, Green A, Zhou XR, Zhu LH (2016) Down-regulation of crambe fatty acid desaturase and elongase in Arabidopsis and crambe resulted in significantly increased oleic acid content in seed oil. Plant Biotechnol J 14:323–331CrossRefPubMedGoogle Scholar
  20. Lindqvist Y, Huang W, Schneider G, Shanklin J (1996) Crystal structure of delta9 stearoyl-acyl carrier protein desaturase from castor seed and its relationship to other di-iron proteins. EMBO J 15:4081–4092PubMedPubMedCentralGoogle Scholar
  21. Liu Q, Singh SP, Brubaker CL, Sharp PJ, Green AG, Marshall DR (1999) Molecular cloning and expression of a cDNA encoding a microsomal u-6 fatty acid desaturase from cotton (Gossypium hirsutum). Plant Physiol 26:101–106Google Scholar
  22. Liu QL, Singh SP, Green AG (2002) High-stearic and high-oleic cottonseed oils produced by hairpin RNA-mediated post-transcriptional gene silencing. Plant Physiol 129:1732–1743CrossRefPubMedPubMedCentralGoogle Scholar
  23. Liu X, Zhao B, Zheng H-J, Hu Y, Lu G, Yang C-Q, Chen J-D, Chen J-J et al (2015) Gossypium barbadense genome sequence provides insight into the evolution of extra-long staple fiber and specialized metabolites. Sci Reports 5:14139CrossRefGoogle Scholar
  24. Luo R, Liu B, Xie Y, Li Z, Huang W, Yuan J, He G, Chen Y et al (2012) SOAPdenovo2: an empirically improved memory-efficient short-read de novo assembler. Giga Sci 1:18CrossRefGoogle Scholar
  25. Mangano S, Gonzalez CD, Petruccelli S (2014) Chapter 8: Agrobacterium tumefaciens -mediated transient transformation of Arabidopsis thaliana leaves. In: Sanchez-Serrano Jose J, Salinas Julio (eds) Arabidopsis Protocols, Methods in Molecular Biology, vol 1062. Springer, New York, p 8Google Scholar
  26. Mauney JR, Phillips LL (1963) Influence of daylength and night temperature on flowering of Gossypium. Bot Gaz 124:278–283CrossRefGoogle Scholar
  27. 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 37:156–173CrossRefPubMedGoogle Scholar
  28. McGarry RC, Ayre BG (2012) Geminivirus-mediated delivery of florigen promotes determinate growth in aerial organs and uncouples flowering from photoperiod in cotton. PLoS One 7:e36746CrossRefPubMedPubMedCentralGoogle Scholar
  29. Milcamps A, Tumaney AW, Paddock T, Pan DA, Ohlrogge J, Pollard M (2005) Isolation of a gene encoding a 1,2-diacylglycerol-sn-acetyl-CoA acetyltransferase from developing seeds of Euonymus alatus. J Biol Chem 280:5370–5377CrossRefPubMedGoogle Scholar
  30. Miquel M, Browse J (1992) Arabidopsis mutants deficient in polyunsaturated fatty acid synthesis. Biochemical and genetic characterization of a plant oleoyl-phosphatidylcholine desaturase. J Biol Chem 267:1502–1509PubMedGoogle Scholar
  31. Mozaffarian D, Katan MB, Ascheri A, Stampfe MD, Willet WC (2006) Trans fatty acids and cardiovascular disease. N Engl J Med 354:1601–1613CrossRefPubMedGoogle Scholar
  32. Naim F, Shrestha P, Singh SP, Waterhouse PM, Wood CC (2016) Stable expression of silencing-suppressor protein enhances the performance and longevity of an engineered metabolic pathway. Plant Biotechnol J 14:1418–1426CrossRefPubMedGoogle Scholar
  33. Napier JA, Haslam RP, Beaudoin F, Cahoon EB (2014) Understanding and manipulating plant lipid composition: metabolic engineering leads the way. Current Opin Plant Biol 19:68–75CrossRefGoogle Scholar
  34. 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 6:147–158CrossRefPubMedPubMedCentralGoogle Scholar
  35. Page RD (1996) TreeView: an application to display phylogenetic trees on personal computers. Comput Appl Biosci 12:357–358PubMedGoogle Scholar
  36. Paterson AH, Wendel JF, Gundlach H, Guo H, Jenkins J et al (2012) Repeated polyploidization of Gossypium genomes and the evolution of spinnable cotton fibres. Nature 492:423–427CrossRefPubMedGoogle Scholar
  37. Pirtle IL, Kongcharoensuntorn W, Nampaisansuk M, Knesek JE, Chapman KD, Pirtle RM (2001) Molecular cloning and functional expression of the gene for a cotton Delta-12 fatty acid desaturase (FAD2). Biochim Biophys Acta 1522:122–129CrossRefPubMedGoogle Scholar
  38. Priyam A, Woodcroft BJ, Rai V, Munagala A, Moghul I, Ter F, Gibbins MA, Moon H, Leonard G, Rumpf W, Wurm Y (2015) Sequenceserver: a modern graphical user interface for custom BLAST databases. Biorxiv: 033142Google Scholar
  39. Shanklin J, Whittle JE, 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. Biochemistry 33:12787–12794CrossRefPubMedGoogle Scholar
  40. Shockey JM, Gidda SK, Chapital DC, Kuan JC, Dhanoa PK, Bland JM, Rothstein SJ, Mullen RT, Dyer JM (2006) Tung tree DGAT1 and DGAT2 have nonredundant functions in triacylglycerol biosynthesis and are localized to different subdomains of the endoplasmic reticulum. Plant Cell 18:2294–2313CrossRefPubMedPubMedCentralGoogle Scholar
  41. Shockey J, Mason C, Gilbert M, Cao H, Li X, Cahoon E, Dyer J (2015) Development and analysis of a highly flexible multi-gene expression system for metabolic engineering in Arabidopsis seeds and other plant tissues. Plant Mol Biol 89:113–126CrossRefPubMedGoogle Scholar
  42. Sunilkumar G, Campbell LM, Hossen M, Connell JP, Hernandez E, Reddy AS, Smith CW, Rathore KS (2005) A comprehensive study of the use of a homologous promoter in antisense cotton lines exhibiting a high seed oleic acid phenotype. Plant Biotech J 3:319–330CrossRefGoogle Scholar
  43. Teixeira MC, Coelho N, Olsson ME, Brodelius PE, Carvalho IS, Brodelius M (2009) Molecular cloning and expression analysis of three omega-6 desaturase genes from purslane (Portulaca oleracea L.). Biotechnol Lett 7:1089–1101CrossRefGoogle Scholar
  44. Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG (1997) The ClustalX windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res 24:4876–4882CrossRefGoogle Scholar
  45. Venegas-Calerón M, Sánchez R, Salas JJ, Garcés R, Martínez-Force E (2016) Molecular and biochemical characterization of the OLE-1 high-oleic castor seed (Ricinus communis L.) mutant. Planta 244:245–258CrossRefPubMedGoogle Scholar
  46. Verdaguer B, de Kochko A, Beachy RN, Fauquet C (1996) Isolation and expression in transgenic tobacco and rice plants, of the cassava vein mosaic virus (CVMV) promoter. Plant Mol Biol 31:1129–1139CrossRefPubMedGoogle Scholar
  47. Wang K, Wang Z, Li F, Ye W, Wang J, Song G, Yue Z, Cong L, Shang H, Zhu S, Zou C, Li Q et al (2012) The draft genome of a diploid cotton Gossypium raimondii. Nat Genet 44:1098–1103CrossRefPubMedGoogle Scholar
  48. Wrenn LB (1995) Chapter 5: Cottonseed-oil refining and manufacturing. In Cinderella of the New South. A History of the Cottonseed Industry, 1855–1955. The University of Tennessee Press, Knoxville, TN. p 14Google Scholar
  49. Zhang D, Pirtle IL, Park SJ, Nampaisansuk M, Neogi P, Wanjie SW, Pirtle RM, Chapman KD (2009) Identification and expression of a new delta-12 fatty acid desaturase (FAD2-4) gene in upland cotton and its functional expression in yeast and Arabidopsis thaliana plants. Plant Physiol Biochem 47:462–471CrossRefPubMedGoogle Scholar
  50. Zhang T, Hu Y, Jiang W, Fang L, Guan X, Chen J, Zhang J, Saski CA, Scheffler BE, Stelly DM, Hulse-Kemp AM, Wan Q et al (2015) Sequencing of allotetraploid cotton (Gossypium hirsutum L. acc. TM-1) provides a resource for fiber improvement. Nature Biotech 33:531–537CrossRefGoogle Scholar
  51. Zhou XR, Singh SP, Green AG (2013) Characterisation of the FAD2 gene family from Hiptage benghalensis: a ricinoleic acid accumulating plant. Phytochem 92:42–48CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg (outside the USA) 2016

Authors and Affiliations

  1. 1.Commodity Utilization Research Unit, United States Department of Agriculture-Agricultural Research ServiceSouthern Regional Research CenterNew OrleansUSA
  2. 2.Food and Feed Safety Research Unit, United States Department of Agriculture-Agricultural Research ServiceSouthern Regional Research CenterNew OrleansUSA
  3. 3.Genomics and Bioinformatics Research UnitUnited States Department of Agriculture-Agricultural Research ServiceStonevilleUSA
  4. 4.Crop Germplasm Research UnitUnited States Department of Agriculture-Agricultural Research ServiceCollege StationUSA

Personalised recommendations