Transgenic Research

, Volume 20, Issue 6, pp 1285–1292 | Cite as

Uniform accumulation of recombinant miraculin protein in transgenic tomato fruit using a fruit-ripening-specific E8 promoter

  • Tadayoshi Hirai
  • You-Wang Kim
  • Kazuhisa Kato
  • Kyoko Hiwasa-Tanase
  • Hiroshi Ezura
Original Paper

Abstract

The E8 promoter, a tomato fruit-ripening-specific promoter, and the CaMV 35S promoter, a constitutive promoter, were used to express the miraculin gene encoding the taste-modifying protein in tomato. The accumulation of miraculin protein and mRNA was compared among transgenic tomatoes expressing the miraculin gene driven by these promoters. Recombinant miraculin protein predominantly accumulated in transgenic tomato lines using the E8 promoter (E8-MIR) only at the red fruit stage. The accumulations were almost uniform among all fruit tissues. When the 35S promoter (35S-MIR) was used, miraculin accumulation in the exocarp was much higher than in other tissues, indicating that the miraculin accumulation pattern can be regulated by using different types of promoters. We also discuss the potential of the E8-MIR lines for practical use.

Keywords

E8 promoter Miraculin Taste-modifying protein Tomato 

Supplementary material

11248_2011_9495_MOESM1_ESM.pdf (3.9 mb)
Supplementary material 1 (PDF 4027 kb)

References

  1. Benfey PN, Ellis J, Pelletier G (1990) Tissue-specific expression from CaMV 35S enhancer subdomains in early stages of plant development. EMBO J 9:1677–1684PubMedGoogle Scholar
  2. De La Garza RD, Quinlivan EP, Klaus SMJ et al (2004) Folate biofortification in tomatoes by engineering the pteridine branch of folate synthesis. Proc Natl Acad Sci USA 101:13720–13725CrossRefGoogle Scholar
  3. Deblaere R, Bytebier B, De greve H, Deboeck FSJ, Van Montagu M, Leemans J (1985) Efficient octopine Ti plasmid-derived vectors for Agrobacterium-mediated gene transfer to plants. Nucl Acid Res 13:4777–4788CrossRefGoogle Scholar
  4. Deikman J, Fischer RL (1988) Interaction of a DNA binding factor with the 5¢-flanking region of an ethylene-responsive fruit ripening gene from tomato. EMBO J 7:3315–3320PubMedGoogle Scholar
  5. Deikman J, Kline R, Fischer RL (1992) Organization of ripening and ethylene regulatory regions in a fruit-specific promoter from tomato (Lycopersicon esculentum). Plant Physiol 100:2013–2017PubMedCrossRefGoogle Scholar
  6. Deikman J, Xu R, Kneissl ML, Ciardi JA, Kim KN, Pelah D (1998) Separation of cis elements responsive to elements responsive to ethylene, ethylene, fruit development, and ripening in the 5′-flanking region of the ripening-related E8 gene. Plant Mol Biol 37:1001–1011PubMedCrossRefGoogle Scholar
  7. Dellapenna D, Lincoln JE, Fischer RL, Bennett AB (1989) Transcriptional analysis of polygalacturonase and other ripening associated genes in rutgers, rin, nor, and Nr tomato fruit. Plant Physiol 90:1372–1377PubMedCrossRefGoogle Scholar
  8. Estornell LH, Orzaez D, Lopez-Pena L, Pineda B, Anton MT, Moreno V, Granell A (2009) A multisite gateway-based toolkit for targeted gene expression and hairpin RNA silencing in tomato fruits. Plant Biotechnol J 7:298–309PubMedCrossRefGoogle Scholar
  9. Fluhr R, Kuhlemeier C, Nagy F, Chua NH (1986) Organ-specific and light-induced expression of plant genes. Science 232:1106–1112PubMedCrossRefGoogle Scholar
  10. Giovannoni JJ, DellaPenna D, Bennett AB, Fischer RL (1989) Expression of a chimeric polygalacturonase gene in transgenic rin (ripening inhibitor) tomato fruit results in polyuronide degradation but not fruit softening. Plant Cell 1:53–63PubMedCrossRefGoogle Scholar
  11. Good X, Kellogg JA, Wagoner W, Langhoff D, Matsumura W, Bestwick RK (1994) Reduced ethylene synthesis by transgenic tomatoes expressing S-adenosylmethionine hydrolase. Plant Mol Biol 26:781–790PubMedCrossRefGoogle Scholar
  12. He ZM, Jiang XL, Qi Y, Luo DQ (2008) Assessment of the utility of the tomato fruit-specific E8 promoter for driving vaccine antigen expression. Genetica 133:207–214PubMedCrossRefGoogle Scholar
  13. Hirai T, Sato M, Toyooka K, Sun HJ, Yano M, Ezura H (2010) Miraculin, a taste-modifying protein is secreted into intercellular spaces in plant cells. J Plant Physiol 167:209–215PubMedCrossRefGoogle Scholar
  14. Hiwasa-Tanase K, Nyarubona M, Hirai T, Kato K, Ichikawa T, Ezura H (2011) High-level accumulation of recombinant miraculin protein in transgenic tomatoes expressing a synthetic miraculin gene with optimized codon usage terminated by the native miraculin terminator. Plant Cell Rep 30:113–124Google Scholar
  15. Jiang XL, He ZM, Peng ZQ, Qi Y, Chen Q, Yu SY (2007) Cholera toxin B protein in transgenic tomato fruit induces systemic immune response in mice. Transgenic Res 16:169–175PubMedCrossRefGoogle Scholar
  16. Kato K, Yoshida R, Kikuzaki A, Hirai T, Kuroda H, Hiwasa-Tanase K, Takane K, Ezura H, Mizoguchi T (2010) Molecular breeding of tomato lines for mass production of miraculin in a plant factory. J Agric Food Chem 58:9505–9510PubMedCrossRefGoogle Scholar
  17. Kaulen H, Schell J, Kreuzaler F (1986) Light-induced expression of the chimeric chalcone synthase-NPTII gene in tobacco cells. EMBO J 5:1–8PubMedGoogle Scholar
  18. Kim YW, Kato K, Hirai T, Hiwasa-Tanase K, Ezura H (2010) Spatial and developmental profiling of miraculin accumulation in transgenic tomato fruits expressing the miraculin gene constitutively. J Agric Food Chem 58:282–286CrossRefGoogle Scholar
  19. Lewinsohn E, Schalechet F, Wilkinson J, Matsui K, Tadmor Y, Nam KH, Amar O, Lastochkin E, Larkov O, Ravid U, Hiatt W, Gepstein S, Pichersky E (2001) Enhanced levels of the aroma and flavor compound S-linalool by metabolic engineering of the terpenoid pathway in tomato fruits. Plant Physiol 127:1256–1265PubMedCrossRefGoogle Scholar
  20. Lincoln JE, Fischer RL (1988a) Diverse mechanisms for the regulation of ethylene-inducible gene expression. Mol Gen Genet 212:71–75PubMedCrossRefGoogle Scholar
  21. Lincoln JE, Fischer RL (1988b) Regulation of gene expression by ethylene in wild-type and rin tomato (Lycopersicon esculentum) fruit. Plant Physiol 88:370–374PubMedCrossRefGoogle Scholar
  22. Lincoln JE, Cordes S, Read E, Fischer RL (1987) Regulation of gene expression by ethylene during Lycopersicon esculentum (tomato) fruit development. Proc Natl Acad Sci USA 84:2793–2797PubMedCrossRefGoogle Scholar
  23. Mehta RA, Cassol T, Li N, Ali N, Handa AK, Mattoo AK (2002) Engineered polyamine accumulation in tomato enhances phytonutrient content, juice quality, and vine life. Nat Biotechnol 20:613–618PubMedCrossRefGoogle Scholar
  24. Nagaya S, Kawamura K, Shinmyo A, Kato K (2010) The HSP terminator of Arabidopsis thaliana increase gene expression in plant cells. Plant Cell Physiol 51:328–332PubMedCrossRefGoogle Scholar
  25. Ramirez YJP, Tasciotti E, Gutierrez-Ortega A, Donayre AJ, Flores MTO, Giacca M, Lim MAG (2007) Fruit-specific expression of the human immunodeficiency virus type 1 Tat gene in tomato plants and its immunogenic potential in mice. Clin Vaccine Immunol 14:685–692PubMedCrossRefGoogle Scholar
  26. Sandhu JS, Rasnyanski SF, Domier LL, Korban SS, Osadjan MD, Buetow DE (2000) Oral immunisation of mice with transgenic tomato fruit expressing respiratory syncytial virus-F protein induces a systemic immune response. Trans Res 9:127–135CrossRefGoogle Scholar
  27. Satoh J, Kato K, Shinmyo A (2004) The 5′-untranslated region of tobacco alcohol dehydrogenase gene functions as an effective translational enhancer in plant. J Biosci Bioeng 1:1–8Google Scholar
  28. Stougaard J, Sandal NN, Gron A, Kuhle A, Marcker KA (1987) 5′ Analysis of the soybean leghaemoglobin lbc3 gene: regulatory elements required for promoter activity and organ specificity. EMBO J 6:3565–3569PubMedGoogle Scholar
  29. Sugaya T, Yano M, Sun HJ, Hirai T, Ezura H (2008) Transgenic strawberry expressing a taste-modifying protein, miraculin. Plant Biotechnol 25:329–333CrossRefGoogle Scholar
  30. Sun HJ, Cui ML, Ma B, Ezura H (2006a) Functional expression of the taste-modifying protein, miraculin, in transgenic lettuce. FEBS Lett 580:620–626PubMedCrossRefGoogle Scholar
  31. Sun HJ, Uchii S, Watanabe S, Ezura H (2006b) A highly efficient transformation protocol for Micro-Tom, a model cultivar of tomato functional genomics. Plant Cell Physiol 47:426–431PubMedCrossRefGoogle Scholar
  32. Sun HJ, Kataoka H, Yano M, Ezura H (2007) Genetically stable expression of functional miraculin, a new type of alternative sweetener, in transgenic tomato plants. Plant Biotechnol J 5:768–777PubMedCrossRefGoogle Scholar
  33. Sunilkumar G, Mohr L, Lopata-Finch E, Emani C, Kathore KS (2002) Developmental and tissue-specific expres sion of a CaMV 35S promoter in cotton as revealed by GFP. Plant Mol Biol 50:463–474PubMedCrossRefGoogle Scholar
  34. Szwacka M, Siedlecka E, Zawirska-Wojtasiak R, Wisniewski L, Malepszy S (2009) Expression pattern of the pre-prothaumatin II gene under the control of the CaMV 35S promoter in transgenic cucumber (Cucumis sativus L.) flower buds and fruits. J Appl Genet 50:9–16PubMedCrossRefGoogle Scholar
  35. Theerasilp S, Kurihara Y (1988) Complete purification and characterization of the taste-modifying protein, miraculin, from miracle fruit. J Biol Chem 263:11536–11539PubMedGoogle Scholar
  36. Theologis A, Oeller PW, Wong LM, Rottmann WH, Gantz DM (1993) Use of a tomato mutant constructed with reverse genetics to study fruit ripening, a complex developmental process. Dev Genet 14:282–295PubMedCrossRefGoogle Scholar
  37. Van Haaren MJJ, Houck CM (1993) A functional map of the fruit specific promoter of the tomato 2A11 gene. Plant Mol Biol 21:625–640PubMedCrossRefGoogle Scholar
  38. Williamson JD, Hirsch-Wyncott ME, Larkins BA, Gelvin SB (1989) Differential accumulation of a transcript driven by the CaMV 35S promoter in transgenic tobacco. Plant Physio 90:1570–1576CrossRefGoogle Scholar
  39. Xu R, Goldman S, Coupe S, Deikman J (1996) Ethylene control of E4 transcription during tomato fruit ripening involves two cooperativecis elements. Plant Mol Biol 31:1117–1127PubMedCrossRefGoogle Scholar
  40. Yano M, Hirai T, Kato K, Hiwasa-Tanase K, Fukuda N, Ezura H (2010) Tomato is a suitable material for producing recombinant miraculin protein in genetically stable manner. Plant Sci 178:469–473CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2011

Authors and Affiliations

  • Tadayoshi Hirai
    • 1
  • You-Wang Kim
    • 1
  • Kazuhisa Kato
    • 1
  • Kyoko Hiwasa-Tanase
    • 1
  • Hiroshi Ezura
    • 1
  1. 1.Graduate School of Life and Environmental SciencesUniversity of TsukubaTsukubaJapan

Personalised recommendations