Free Standard AU & NZ Shipping For All Book Orders Over $80!
Register      Login
Functional Plant Biology Functional Plant Biology Society
Plant function and evolutionary biology
RESEARCH ARTICLE

Production of high oleic rice grains by suppressing the expression of the OsFAD2-1 gene

Ella Simone Zaplin A B C , Qing Liu A C , Zhongyi Li A C , Vito M. Butardo Jr. A C , Christopher L. Blanchard B and Sadequr Rahman A C D E
+ Author Affiliations
- Author Affiliations

A CSIRO Food Future National Research Flagship, GPO Box 1600, Canberra, ACT 2601, Australia.

B School of Biomedical Science, Charles Sturt University, Wagga Wagga, NSW 2678, Australia.

C CSIRO Plant Industry, GPO Box 1600, Canberra, ACT 2601, Australia.

D School of Science, Monash University, 46150 Bandar Sunway, Selangor, Malaysia.

E Corresponding author. Email: sadequr.rahman@monash.edu

Functional Plant Biology 40(10) 996-1004 https://doi.org/10.1071/FP12301
Submitted: 11 October 2012  Accepted: 27 March 2013   Published: 29 May 2013

Abstract

The composition of rice (Oryza sativa L.) grain fatty acids (18% palmitic acid, 36% oleic acid and 37% linoleic acid) is suboptimal for rice storage and utilisation of rice bran oil as food grade oil or a source of biodiesel. Genetic manipulation of fatty acid composition in rice bran oil to increase oleic acid levels at the expense of linoleic acid and palmitic acid would not only add extra value to the rice, but also enhance health benefits for consumers. Four putative rice microsomal Δ12-fatty acid desaturase (OsFAD2) genes were identified as potentially important target genes to achieve this improvement. Reverse transcription–PCR analysis indicated that OsFAD21 was the most highly expressed gene in rice grains. RNA interference (RNAi) suppression of the expression of OsFAD21 resulted in an increase of oleic acid and a reduction of linoleic and palmitic acids in T3 grains. The research here showed that in the rice grains, the OsFAD2–1 enzyme was an effective target for raising oleic acid levels at the expense of the oxidatively unstable linoleic acid and the cholesterol-raising palmitic acid.

Additional keywords: FAD2, linoleic acid, oleic acid, Oryza sativa, palmitic acid, RNAi.


References

Alt JL, Fehr WR, Welke GA, Sandhu D (2005) Phenotypic and molecular analysis of oleate content in the mutant soybean line M23. Crop Science 45, 1997–2000.
Phenotypic and molecular analysis of oleate content in the mutant soybean line M23.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2MXhtVOrt7fO&md5=a31886f4dcbceac0c59689a34398ebdaCAS |

Broadwater JA, Whittle E, Shanklin J (2002) Desaturation and hydroxylation. Residues 148 and 324 of Arabidopsis FAD2, in addition to substrate chain length, exert a major influence in partitioning of catalytic specificity. The Journal of Biological Chemistry 277, 15613–15620.
Desaturation and hydroxylation. Residues 148 and 324 of Arabidopsis FAD2, in addition to substrate chain length, exert a major influence in partitioning of catalytic specificity.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD38Xjs1Wjsrc%3D&md5=27ad313bcd9e06743bd52c14d7dfac19CAS | 11864983PubMed |

Butardo VM, Fitzgerald MA, Bird AR, Gidley MJ, Flanagan BM, Larroque O, Resurreccion AP, Laidlaw HKC, Jobling SA, Morell MK, Rahman S (2011) Impact of down-regulation of starch branching enzyme IIb in rice by artificial microRNA- and hairpin RNA-mediated RNA silencing. Journal of Experimental Botany 62, 4927–4941.
Impact of down-regulation of starch branching enzyme IIb in rice by artificial microRNA- and hairpin RNA-mediated RNA silencing.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3MXhtlejs77I&md5=eaf070010e15fd573a92ebb2f32c7022CAS | 21791436PubMed |

Canakci M, Monyem A, Van Gerpen J (1999) Accelerated oxidation processes is biodiesel. Transactions of the American Society of Agricultural Engineers 42, 1565–1572.

FAOSTAT (2009) Section D Consumption table D9 Consumption of 10 major vegetal foods (2003–2005). Available at http://www.fao.org/docrep/014/am079m/PDF/am079m09d.pdf

Jeon JS, Lee S, Jung KH, Jun SH, Kim C, An G (2000) Tissue-preferential expression of a rice alpha-tubulin gene, OsTubA1, mediated by the first intron. Plant Physiology 123, 1005–1014.
Tissue-preferential expression of a rice alpha-tubulin gene, OsTubA1, mediated by the first intron.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3cXlt1Sltrs%3D&md5=3c533b95de6901de1abc0a00bf4b5dd2CAS | 10889249PubMed |

Juliano BO (1983) Lipids in rice and rice processing. In ‘Lipids in cereal technology’. (Ed. PJ Barnes) pp. 403–438. (Academic Press: London)

Kato T, Yamaguchi Y, Uyehara T, Yokoyama T, Namai T, Yamanaka S (1983) Defense mechanism of the rice plant against rice blast disease. Naturwissenschaften 70, 200–201.
Defense mechanism of the rice plant against rice blast disease.Crossref | GoogleScholarGoogle Scholar |

Kris-Etherton PM, Derr J, Mitchell DC, Mustard VA, Russell ME, McDonnell ET, Salabsky D, Pearson TA (1993) The role of fatty acid saturation on plasma lipids, lipoproteins and apolipoproteins: I. Effects of whole food diets high in cocoa butter, olive oil, soybean oil, dairy butter and milk chocolate on plasma lipids of young men. Metabolism: Clinical and Experimental 42, 121–129.
The role of fatty acid saturation on plasma lipids, lipoproteins and apolipoproteins: I. Effects of whole food diets high in cocoa butter, olive oil, soybean oil, dairy butter and milk chocolate on plasma lipids of young men.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK38XmsFSntbw%3D&md5=2a1a9aeac61100886238cb30c595934aCAS |

Liu Q, Singh SP, Green AG (2002) High-stearic and high-oleic cottonseed oils produced by hairpin RNA-mediated post-transcriptional gene silencing. Plant Physiology 129, 1732–1743.
High-stearic and high-oleic cottonseed oils produced by hairpin RNA-mediated post-transcriptional gene silencing.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD38Xmtl2gt7o%3D&md5=c6df5a10644ec0d899e3e6205e9d4192CAS | 12177486PubMed |

Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-delta delta C(T)) method. Methods (San Diego, Calif.) 25, 402–408.
Analysis of relative gene expression data using real-time quantitative PCR and the 2(-delta delta C(T)) method.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD38XhtFelt7s%3D&md5=cda22da73d6034ad2e4c74827f633baaCAS |

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. The Plant Journal 37, 156–173.
Membrane-bound fatty acid desaturases are inserted co-translationally into the ER and contain different ER retrieval motifs at their carboxy termini.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2cXhsVGltb0%3D&md5=66a7d61a151a8b6dccb62f166ee9833aCAS | 14690501PubMed |

Möllers C, Schierholt A (2002) Genetic variation of palmitate and oil content in a winter oilseed rape doubled haploid population segregating for oleate content. Crop Science 42, 379–384.
Genetic variation of palmitate and oil content in a winter oilseed rape doubled haploid population segregating for oleate content.Crossref | GoogleScholarGoogle Scholar |

Mozaffarian D, Katan MB, Ascherio A, Stampfer MJ, Willett WC (2006) Trans fatty acids and cardiovascular disease. The New England Journal of Medicine 354, 1601–1613.
Trans fatty acids and cardiovascular disease.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD28XjsVKmsLo%3D&md5=1c7fab8e93e553b4c0eae27fd907fda2CAS | 16611951PubMed |

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. The Plant Cell 6, 147–158.

Orthoefer FT (1996a) Rice bran oil. In ‘Bailey’s industrial oils and fat products. Vol. 2.’ 5th edn. (Ed. YH Hui) pp. 393–410. (Wiley: New York)

Orthoefer FT (1996b) Rice bran oil: healthy lipid source. Food Technology 50, 62–64.

Sánchez-García A, Mancha M, Heinz E, Martínez-Rivas JM (2004) Differential temperature regulation of three sunflower microsomal oleate desaturase (FAD2) isoforms overexpressed in Saccharomyces cerevisiae. European Journal of Lipid Science and Technology 106, 583–590.
Differential temperature regulation of three sunflower microsomal oleate desaturase (FAD2) isoforms overexpressed in Saccharomyces cerevisiae.Crossref | GoogleScholarGoogle Scholar |

Shanklin J, Cahoon EB (1998) Desaturation and related modifications of fatty acids. Annual Review of Plant Physiology and Plant Molecular Biology 49, 611–641.
Desaturation and related modifications of fatty acids.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK1cXjvVSgs7s%3D&md5=0807f4b247a911f21f125eb7dd090fddCAS | 15012248PubMed |

Sheng ZW, Fen LY, Wu ZS, Bao XX (2006) Obtaining new germplast of Brassica napus with high oleic acid content by RNA interference and marker-free transformation of Fad2 gene. Journal of Plant Physiology and Molecular Biology 32, 665–671.

Shu O, Hamilton J, Lin H, Campbell M, Childs K, Thibaud-Nissen F, Malek RL, Lee Y, Zheng L, Orvis J, Haas B, Wortman J, Buell CR (2007) The TIGR Rice Genome Annotation Resource: improvements and new features. Nucleic Acids Research 35, D883–D887.
The TIGR Rice Genome Annotation Resource: improvements and new features.Crossref | GoogleScholarGoogle Scholar |

Spiteller G (1998) Linoleic acid peroxidation – the dominant lipid peroxidation process in low density lipoprotein – and its relationship to chronic diseases. Chemistry and Physics of Lipids 95, 105–162.
Linoleic acid peroxidation – the dominant lipid peroxidation process in low density lipoprotein – and its relationship to chronic diseases.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK1cXmvFWgtL4%3D&md5=a918884dbb1caf9be2ac35fc2facd89dCAS | 9853364PubMed |

Stoutjesdijk PA, Singh SP, Liu Q, Hurlstone CJ, Waterhouse PA, Green AG (2002) hpRNA-mediated targeting of the Arabidopsis FAD2 gene gives highly efficient and stable silencing. Plant Physiology 129, 1723–1731.
hpRNA-mediated targeting of the Arabidopsis FAD2 gene gives highly efficient and stable silencing.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD38Xmtl2gt70%3D&md5=909d3f176b961947228365fa9195d15fCAS | 12177485PubMed |

Suzuki Y, Ise K, Li C, Honda I, Iwai Y, Matsukura U (1999) Volatile components in stored rice [Oryza sativa (L.)] of varieties with and without lipoxygenase-3 in seeds. Journal of Agricultural and Food Chemistry 47, 1119–1124.
Volatile components in stored rice [Oryza sativa (L.)] of varieties with and without lipoxygenase-3 in seeds.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK1MXosFShtA%3D%3D&md5=0f1c88b988556c4020b891b96895d05cCAS | 10552425PubMed |

Taira H, Nakagahra M, Nagamine T (1988) Fatty acid composition of Indica, Sinica, Javanica, Japonica groups of nonglutinous brown rice. Journal of Agricultural and Food Chemistry 36, 45–47.
Fatty acid composition of Indica, Sinica, Javanica, Japonica groups of nonglutinous brown rice.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaL1cXlsVKltQ%3D%3D&md5=3e5d4fe5bc6e31a54127bc09efa6283aCAS |

Trapnell C, Williams BA, Pertea G, Mortazavi A, Kwan G, van Baren MJ, Salzberg SL, Wold BJ, Pachter L (2010) Transcript assembly and abundance estimation from RNA-Seq reveals thousands of new transcripts and switching among isoforms. Nature Biotechnology 28, 511–515.
Transcript assembly and abundance estimation from RNA-Seq reveals thousands of new transcripts and switching among isoforms.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3cXlsVyitLY%3D&md5=b43471a0d193e0a105b6f6b843313f08CAS | 20436464PubMed |

Upadhyaya NM, Surin B, Ramm K, Gaudron J, Schunmann PHD, Taylor W, Waterhouse PM, Wang MB (2000) Agrobacterium-mediated transformation of Australian rice cultivars Jarrah and Amaroo using modified promoters and selectable markers. Australian Journal of Plant Physiology 27, 201–210.

Zhou Z, Robards K, Helliwell S, Blanchard C (2002) Ageing of stored rice: changes in chemical and physical attributes. Journal of Cereal Science 35, 65–78.
Ageing of stored rice: changes in chemical and physical attributes.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD38Xhtlylsbo%3D&md5=1cc59f0c3aea59ef0a569a027d756040CAS |