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High-oleate peanut mutants result from a MITE insertion into the FAD2 gene


A high-oleate trait in the cultivated peanut (Arachis hypogaea L.) was reported to rely on the allelic composition of the two homeologous, microsomal oleoyl-PC desaturase genes (ahFAD2A or ahFAD2B). The enzyme activity of either ahFAD2A or ahFAD2B is sufficient for a normal oleate phenotype, and a significant reduction in the levels of ahFAD2B and a mutation in ahFAD2A were reported to be responsible for the high-oleate phenotype in one chemically induced mutant (M2-225) and one derived from a naturally occurring (8-2122) mutant. Here, we report an insertion of the same miniature inverted-repeat transposable element (MITE) in the ahFAD2B gene in another chemically induced mutant (Mycogen-Flavo) and the previously characterized M2-225 mutant. In both cases, this MITE insertion in ahFAD2B causes a frameshift, resulting in a putatively truncated protein sequence in both mutants. The insertion of this MITE in ahFAD2B, in addition to the point mutation in ahFAD2A, appears to be the cause of the high-oleate phenotype in Mycogen-Flavo and M2-225 mutants. Utilizing sequences of the MITE, we developed a DNA marker strategy to differentiate the two insertion-containing mutants from the normal oleate peanut variety (AT-108) and the naturally occurring, high-oleate mutant 8-2122. Reverse transcript-PCR/differential digestion results reveal the expression of both ahFAD2A and ahFAD2B genes in Mycogen-Flavo mutant. This result is in contrast to the observation that ahFAD2B transcripts are greatly reduced in the M2-225 mutant having the MITE insertion further 3′ in ahFAD2B gene.

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  1. Arnoff R, Baran R, Hodgkin J (2001) Molecular identification of smg-4, required for mRNA surveillance in C. elegans. Gene 268:153–164

  2. Ashri A (1988) Mutagenic studies and breeding of peanut (Arachis hypogaea) and (Sesame indicum). In: Improvement of grain legume production using induced mutations. International Atomic Energy Agency, Paris, pp 509–511

  3. Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K (1994) Current protocols in molecular biology. Wiley, New York

  4. Braquart C, Royer C, Bouhin H (1999) DEC: a new miniature inverted-repeat transposable element from the genome of the beetle Tenebrio molitor. Insect Mol Biol 8:571–574

  5. Brown P, Gettner S, Somerville C (1999) Genetic engineering of plant lipids. Ann Rev Nutr 19:197–216

  6. Bureau TE, Wessler S (1992) Tourist: a large family of small inverted-repeat elements frequently associated with maize genes. Plant Cell 4:1283–1294

  7. Bureau TE, Wessler S (1994) Mobile inverted-repeat elements of the Tourist family are associated with the genes of many cereal grasses. Proc Natl Acad Sci USA 91:1411–1415

  8. Cali BM, Anderson P (1998) mRNA surveillance mitigates genetic dominance in Caenorhabditis elegans. Mol Gen Genet 260:176–184

  9. Charrier B, Foucher F, Kondorosi E, Aubenton-Carafay Y, Thermer C, Kondorosi A, Ratel P (1999) Bigfoot: a new family of MITE elements characterized from the Medicago genus. Plant J 18:431–441

  10. Dickey LF, Nguyen TT, Allen GC, Thompson WF (1994) Light modulation of ferredoxin mRNA requires an open reading frame. Plant Cell 6:1171–1176

  11. Feschotte C, Mouchè C (2000) Evidence that a family of miniature inverted repeat transposable elements (MITES) from the Arabidopsis thaliana genome has arisen from a pogo-like DNA transposon. Mol Biol Evol 17(5):730–737

  12. Garcés R, Mancha M (1989) Oleate desaturation in seeds of two genotypes of sunflower. Phytochemistry 28:2593–2596

  13. Garcés R, Mancha M (1991) In vitro oleate desaturase in developing sunflower seeds. Phytochemistry 30:2127–2130

  14. Grundy SM (1986) Comparison of monounsaturated fatty acids and carbohydrates for lowering plasma cholesterol in man. N Engl J Med 314:745–748

  15. Heiner RE, Konzak CF, Nilan RA, Legunlt RR (1960) Diverse ratios of mutations to chromosome aberrations in barley treated with diethylsulfate and gamma rays. Genetics 46:1215–1221

  16. Hongtrakul V, Slabaugh MB, Knapp SJ (1998) A seed specific δ12-oleate desaturase gene is duplicated, rearranged, and weakly expressed in high-oleic acid sunflower lines. Crop Sci 38:1245–1249

  17. Hoof A van, Green PJ (1996) Premature nonsense codon decreases the stability of phytohemagglutnin mRNA in a position-dependent manner. Plant J 10:415–424

  18. Izsvák Z, Ivics Z, Shimoda N, Mohn D, Okamoto H, Hackett PB (1999) Short inverted-repeat transposable elements in teleost fish and implications for a mechanism for their amplification. J Mol Evol 48:13–21

  19. Jacobson A, Peltz S (1996) Interrelationships of the pathways of mRNA decay and translation in eukaryotic cells. Annu Rev Biochem 65:693–739

  20. Jarret RL, Austin D (1994) Genetic diversity and systemic relationships in sweet potato (Ipomoea batatas (L.) Lam.) and related species as revealed by RAPD analysis. Genet Res Crop Evol 31:165–173

  21. Jokufu KD, Schipper RD, Goldberg RB (1989) A frameshift mutation prevents Kunitz trypsin inhibitor mRNA accumulation in soybean embryos. Plant Cell 1:427–435

  22. Jung S, Powell G, Moore K, Abbott A (2000a) The high-oleate trait in cultivated peanut [Arachis hypogaea L.]. II. Molecular basis and genetics of the trait. Mol Gen Genet 263:806–811

  23. Jung S, Swift D, Sengoku E, Patel M, Teulé F, Powell G, Moore K, Abbott A (2000b) The high-oleate trait in the cultivated peanut [Arachis hypogaea L.]. I. Isolation and characterization of two genes encoding microsomal oleoyl-PC desaturases. Mol Gen Genet 263:796–805

  24. Kabbaj A, Vervoort V, Abbott AG, Tersac M, Bervillé A (1996) Polymorphism in Helianthus and expression of stearate, oleate and linoleate desaturase genes in sunflower with normal and high-oleic contents. Helia 19:1–18

  25. Kidwell MG, Lisch D (1997) Transposable elements as sources of variation in animals and plants. Proc Natl Acad Sci USA 94:7704–7711

  26. Kinney AJ (1998a) Manipulating flux through plant metabolic pathways. Curr Opin Plant Biol 1:173–178

  27. Kinney AJ (1998b) Plants as industrial chemical factories—new oils from genetically engineered soybeans. Fett/Lipid 100:173–176

  28. Lee MS, Guerra DJ (1994) Biochemical characterization of temperature-induced changes in lipid metabolism in a high-oleic acid mutant of Brassica rapa. Arch Biochem Biophys 315:203–211

  29. Lohuis MR ten, Meyer A, Meyer P (1993) An improved method of high molecular weight >300 kb plant DNA in liquid phase. Biotechniques 14:6 890–892

  30. Lopéz Y, Nadaf HL, Smith OD, Connell JP, Reddy AS, Fritz AK (2000) Isolation and characterization of the δ12-fatty acid desaturase in peanut (Arachis hypogaea L.) and search for polymorphisms for the high-oleate trait in Spanish market-type lines. Theor Appl Genet 101:1131–1138

  31. Lopéz Y, Nadaf HL, Smith OD, Simpson CE, Fritz AK (2002) Expressed variants of the δ12-fatty acid desaturase for the high-oleate trait in Spanish market-type peanut lines. Mol Breed 9:183–190

  32. Martin BA, Rinne RW (1986) A comparison of oleic acid metabolism in the soybean (Glycine max [L.] Merr.) genotypes Williams and A5, a mutant with decreased linoleic acid in the seed. Plant Physiol 81:41–44

  33. Martínez-Rivas JM, Sperling P, Lühs W, Heinz E (1998) Isolation of three different microsomal oleate desaturase cDNA clones from sunflower. Expression studies in normal type and high-oleic mutant. In: Sánchez J, Cerdá-Olmedo E, Martínez-Force E (eds) Advances in plant lipid research. Secretariado de Publicaciones de la Universidad de Sevilla, Seville, Spain, pp 137–139

  34. Morgan GT (1995) Identification in the human genome of mobile elements spread by DNA-mediated transposition. J Mol Biol 254:1–5

  35. Moore KM, Knauft DA (1989) The inheritance of high-oleic acid in peanut. J Hered 80:252–253

  36. Napier JA, Michaelson LV, Stobart AK (1999) Plant desaturases: harvesting the fat of the land. Curr Opin Plant Biol 2:123–127

  37. Norden AJ, Gorbet DW, Knauft DA, Young CT (1987) Variability in oil quality among peanut genotypes in the Florida breeding program. Peanut Sci 14:7–11

  38. Petracek ME, Nuygen T, Thompson WF, Dickey LF (2000) Premature termination codons destabilize ferredoxin-1 mRNA when ferredoxin-1 is translated. Plant J 21:563–569

  39. Ray TK, Holly SP, Knauft DA, Abbott AG, Powell GL (1993) The primary defect in developing seed from the high-oleate variety of peanut (Arachis hypogaea L.) is the absence of δ12-desaturase activity. Plant Sci 91:15–21

  40. Sambrook J, Fritsch JEF, Maniatis TE (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York

  41. Smith AF, Riggs AD (1996) Tiggers and DNA transposons fossils in the human genome. Proc Natl Acad Sci USA 93:1443–1448

  42. Southern EM (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98:503–517

  43. St. Angelo AJ, Ory RL (1973) Investigations of causes and prevention of fatty acid peroxidation in peanut butter. J Am Peanut Res Educ Soc 5:128–133

  44. Stoutjesdijk PA, Hurlestone C, Singh SP, Green AG (2000) High-oleic acid Australian Brassica napus and B. juncea varieties produced by co-suppression of endogenous δ12-desaturases. Biochem Soc Trans 28:938–940

  45. Toborek M, Lee Y W, Garrido R, Kaiser S, Hennig B (2002) Unsaturated fatty acids selectively induce an inflammatory environment in human endothelial cells. Am J Clin Nutr 75:119–125

  46. Töpfer R, Martini N, Schell J (1995) Modifications of lipid synthesis. Science 268:681–686

  47. Tu Z (1997) Three novel families of miniature inverted-repeat transposable elements are associated with gene of the yellow fever mosquito, Aedes aegypti. Proc Natl Acad Sci USA 94:7475–7480

  48. Tu Z (2001) Eight novel families of miniature inverted repeat transposable elements in the African malaria mosquito, Anopheles gambiae. Proc Natl Acad Sci USA 98:1699–1704

  49. Tudor M, Lobocka M, Goodell M, Pettitt J, O’Hare K (1992) The pogo transposable element family of Drosophila melanogaster. Mol Gen Genet 232:126–134

  50. Unsal K, Morgan GT (1995) A novel group of families of short interspersed repetitive elements (SINEs) in Xenopus: evidence of a specific target site for DNA-mediated transposition of inverted-repeat SINEs J Mol Biol 248:812–823

  51. Voelker TA, Staswick P, Willmitzer L (1986) Molecular analysis of two phytohemagglutinin genes and their expression in Phaseolus vulgaris cv. Pinto, a lectin-deficient cultivar of the bean. EMBO 5:3075–3082

  52. Voelker TA, Moreno J, Chrispeels MJ (1990) Expression of a pseudogene in transgenic tobacco: a frameshift mutation prevents mRNA accumulation. Plant Cell 2:255–261

  53. Wessler SR, Bureau TE, White SE (1995) LTR-retrotransposons and MITEs: important players in the evolution of plant genomes. Curr Opin Genet Dev 5:814–821

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We would like to thank Dr. Renate Horn for her assistance in preparation of this manuscript. This work was performed in partial fulfillment of M. Patel’s Master of Science degree in genetics. The work was supported by Agra Tech Seeds, Inc.

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Correspondence to A. Abbott.

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Communicated by C. Möllers

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Patel, M., Jung, S., Moore, K. et al. High-oleate peanut mutants result from a MITE insertion into the FAD2 gene. Theor Appl Genet 108, 1492–1502 (2004). https://doi.org/10.1007/s00122-004-1590-3

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  • Kunitz Trypsin Inhibitor
  • Homeologous Gene
  • Diethyl Sulfate
  • Peanut Variety
  • Peanut Genome