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Transgene expression in strawberries driven by a heterologous phloem-specific promoter

  • Genetic Transformation and Hybridization
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Abstract

Strawberry is susceptible to diseases caused by phytoplasmas, mycoplasma-like prokaryotes restricted to sieve elements in the phloem tissue of infected plants. One strategy to improve strawberry resistance to phytoplasmas involves transgenic expression of anti-microbial peptide genes in phloem. For targeted phloem-specific expression, we constructed a binary vector with an expression cassette bearing the β-glucuronidase (GUS) reporter gene (uidA) under control of the Arabidopsis sucrose-H+ symporter gene (AtSUC2) promoter. Transgenic strawberry lines were generated with high efficiencies by a modified transformation protocol, which combines the adoption of a 3-day pre-selection period following transformation, and the addition of 10-μM thidiazuron to the regeneration medium. Histological GUS activity indicated that the reporter gene was expressed specifically in phloem of leaves, petioles, and roots of transgenic plants. The results suggest that the transformation protocol and the AtSUC2 promoter may be useful for engineering phytoplasma-resistant transgenic strawberries.

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References

  • Alsheikh MK, Suso HP, Robson M, Battey NH, Wetten A (2002) Appropriate choice of antibiotic and Agrobacterium strain improves transformation of anti biotic-sensitive Fragaria vesca and F.v. semperflorens. Plant Cell Rep 20:1173–1180

    Article  CAS  Google Scholar 

  • Batjer LP, Schneider H (1960) Relation of pear decline to rootstocks and sieve-tube necrosis. Proc Am Soc Hortic Sci 76:85–97

    Google Scholar 

  • Belbahri L, Boucher C, Candresse T, Nicole M, Ricci P, Keller H (2001) A local accumulation of the Ralstonia solanacearum PopA protein in transgenic tobacco renders a compatible plant-pathogen interaction incompatible. Plant J 28:419–430

    Article  CAS  PubMed  Google Scholar 

  • Bertamini M, Grando MS, Muthuchelian K, Nedunchezhian N (2002a) Effect of phytoplasmal infection on photosystem II efficiency and thylakoid membrane protein changes in field grown apple (Malus pumila) leaves. Physiol Mol Plant Pathol 61:349–356

    Article  CAS  Google Scholar 

  • Bertamini M, Nedunchezhian N, Tomasi F, Grando MS (2002b) Phytoplasma [Stolbur-subgroup (Bois Noir-BN)] infection inhibits photosynthetic pigments, ribulose-1,5-bisphosphate carboxylase and photosynthetic activities in field grown grapevine (Vitis vinifera L. cv. Chardonnay) leaves. Physiol Mol Plant Pathol 61:357–366

    Article  CAS  Google Scholar 

  • Braun EJ, Sinclair WA (1976) Histopathology of phloem necrosis in Ulmus americana. Phytopathology 66:598–607

    Google Scholar 

  • Dickman MB, Park YK, Oltersdorf T, Li W, Clemente T, French R (2001) Abrogation of disease development in plants expressing animal antiapoptotic genes. Proc Natl Acad Sci USA 98:6957–6962

    Article  CAS  PubMed  Google Scholar 

  • Gamborg OL, Miller RA, Ojima K (1968) Nutrient requirements of suspension cultures of soybean root cells. Exp Cell Res 50:151–158

    CAS  PubMed  Google Scholar 

  • Gottwald JR, Krysan PJ, Young JC, Evert RF, Sussman MR (2000) Genetic evidence for the in planta role of phloem-specific plasma membrane sucrose transporters. Proc Natl Acad Sci USA 97:13979–13984

    Article  CAS  PubMed  Google Scholar 

  • Haymes KM, Davis TM (1998) Agrobacterium-mediated transformation of ‘Alpine’ Fragaria vesca, and transmission of transgenes to R1 progeny. Plant Cell Rep 17:279–283

    Article  CAS  Google Scholar 

  • Hood EE, Gelvin SB, Melchers LS, Hoekema A (1993) New Agrobacterium helper plasmids for gene transfer to plants. Transgenic Res 2:208–218

    CAS  Google Scholar 

  • Imlau A, Truernit E, Sauer N (1999) Cell-to-cell and long-distance trafficking of the green fluorescent protein in the phloem and symplastic unloading of the protein into sink tissues. Plant Cell 11:309–322

    Article  CAS  PubMed  Google Scholar 

  • James DJ, Passey AJ, Rugini E (1988) Factors affecting high frequency plant regeneration from apple leaf tissues cultured in vitro. J Plant Physiol 132:148–154

    CAS  Google Scholar 

  • Jefferson RA (1987) Assaying chimeric genes in plants: the GUS gene fusion system. Plant Mol Biol Rep 5:387–405

    CAS  Google Scholar 

  • Kartte S, Seemüller E (1991) Histopathology of apple proliferation in Malus taxa and hybrids of different susceptibility. J Phytopathol 131:149–160

    Google Scholar 

  • Kiefer E, Heller W, Ernst D (2000) A simple and efficient protocol for isolation of functional RNA from plant tissues rich in secondary metabolites. Plant Mol Biol Rep 18:33–39

    CAS  Google Scholar 

  • Le Gall F, Bové JM, Garnier M (1998) Engineering of a single-chain variable-fragment (scFv) antibody specific for the stolbur phytoplasma (Mollicute) and its expression in Escherichia coli and tobacco plants. Appl Environ Microbiol 64:4566–4572

    PubMed  Google Scholar 

  • Lee I-M, Davis RE (1992) Mycoplasmas which infect plants and insects. In: Maniloff J, McElhansey RN, Finch LR, Baseman JB (eds) Mycoplasmas: molecular biology and pathogenesis. American Society for Microbiology, Washington, D.C., pp 379–390

  • Lee I-M, Davis RE, Gundersen-Rindal DE (2000) Phytoplasma: phytopathogenic mollicutes. Annu Rev Microbiol 54:221–255

    Article  CAS  PubMed  Google Scholar 

  • Lepka P, Stitt M, Moll E, Seemüller E (1999) Effect of phytoplasmal infection on concentration and translocation of carbohydrates and amino acids in periwinkle and tobacco. Physiol Mol Plant Pathol 55:59–68

    Article  CAS  Google Scholar 

  • Lincoln JE, Richael C, Overduin B, Smith K, Bostock R, Gilchrist DG (2002) Expression of the antiapoptotic baculovirus p35 gene in tomato blocks programmed cell death and provides broad-spectrum resistance to disease. Proc Natl Acad Sci USA 99:15217–15221

    Article  CAS  PubMed  Google Scholar 

  • Mass JL (1998) Compendium of strawberry diseases, 2nd edn. American Phytopathology Society, St Paul, Minn.

  • McCoy RE, Caudwell A, Chang CJ, Chen T-A, Chiykowski LN et al (1989) Plant diseases associated with mycoplasma-like organisms. In: Whitcomb RF, Tully JG (eds) The mycoplasmas, vol 5. Academic, New York, pp 545–560

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol Plant 15:473–497

    CAS  Google Scholar 

  • Murthy BNS, Murch S, Saxena PK (1998) Thidiazuron: a potent regulator of plant morphogenesis. In Vitro Cell Dev Biol 34:267–275

    CAS  Google Scholar 

  • Osusky M, Zhou G, Osuska L, Hancock RE, Kay WW, Misra S (2000) Transgenic plants expressing cationic peptide chimeras exhibit broad-spectrum resistance to phytopathogens. Nat Biotechnol 18:1162–1166

    Article  CAS  PubMed  Google Scholar 

  • Passey AJ, Barrett KJ, James DJ (2003) Adventitious shoot regeneration from seven commercial strawberry cultivars (Fragaria × ananassa Duch.) using a range of explant types. Plant Cell Rep 21:397–401

    CAS  PubMed  Google Scholar 

  • Sauer N, Stolz J (1994) SUC1 and SUC2: two sucrose transporters from Arabidopsis thaliana; expression and characterization in baker’s yeast and identification of the histidine-tagged protein. Plant J 6:67–77

    Article  CAS  PubMed  Google Scholar 

  • Stadler R, Sauer N (1996) The Arabidopsis thaliana AtSUC2 gene is specifically expressed in companion cells. Bot Acta 109:299–306

    CAS  Google Scholar 

  • Truernit E, Sauer N (1995) The promoter of the Arabidopsis thaliana SUC2 sucrose-H+ symporter gene directs expression of beta-glucuronidase to the phloem: evidence for phloem loading and unloading by SUC2. Planta 196:564–570

    CAS  PubMed  Google Scholar 

  • Wright KM, Roberts AG, Martens HJ, Sauer N, Oparka KJ (2003) Structural and functional vein maturation in developing tobacco leaves in relation to AtSUC2 promoter activity. Plant Physiol 131:1555–1565

    Article  CAS  PubMed  Google Scholar 

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Authors and Affiliations

  1. Molecular Plant Pathology Laboratory, USDA-Agriculture Research Service, Beltsville, MD, 20705, USA

    Yan Zhao, Qingzhong Liu & Robert E. Davis

  2. Shandong Institute of Pomology, Taian, 271000, People’s Republic of China

    Qingzhong Liu

Authors
  1. Yan Zhao
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  2. Qingzhong Liu
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  3. Robert E. Davis
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Correspondence to Yan Zhao.

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Communicated by K.K. Kamo

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Zhao, Y., Liu, Q. & Davis, R.E. Transgene expression in strawberries driven by a heterologous phloem-specific promoter. Plant Cell Rep 23, 224–230 (2004). https://doi.org/10.1007/s00299-004-0812-0

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  • DOI: https://doi.org/10.1007/s00299-004-0812-0

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