Biotechnology Letters

, Volume 39, Issue 7, pp 1049–1058 | Cite as

A comparison between constitutive and inducible transgenic expression of the PhRIP I gene for broad-spectrum resistance against phytopathogens in potato

  • Romel Gonzales-Salazar
  • Bianca Cecere
  • Michelina Ruocco
  • Rosa Rao
  • Giandomenico Corrado
Original Research Paper



To engineer broad spectrum resistance in potato using different expression strategies.


The previously identified Ribosome-Inactivating Protein from Phytolacca heterotepala was expressed in potato under a constitutive or a wound-inducible promoter. Leaves and tubers of the plants constitutively expressing the transgene were resistant to Botrytis cinerea and Rhizoctonia solani, respectively. The wound-inducible promoter was useful in driving the expression upon wounding and fungal damage, and conferred resistance to B. cinerea. The observed differences between the expression strategies are discussed considering the benefits and features offered by the two systems.


Evidence is provided of the possible impact of promoter sequences to engineer BSR in plants, highlighting that the selection of a suitable expression strategy has to balance specific needs and target species.


Fungal resistance Potato Ribosome-inactivating protein Solanum tuberosum Transgenic plants 


Supporting information

Supplementary Table 1—Primers employed and their main features.

Supplementary Table 2—Analysis of variance of the measures of the lesion area produced by B. cinerea as a function of the genotype (Desirèe, COST and IND) and time (2, 4 and 7 days following inoculations). Post-hoc test on lesions indicated that the COST and IND genotypes are not different (p = 0.125).

Supplementary Fig. 1—Statistical analysis of the severity of the symptoms 7 days following inoculation. The graph reports mean values and its standard error of the lesion area. Different letters represent statistically different groups (Tukey; p < 0.05).

Supplementary material

10529_2017_2335_MOESM1_ESM.docx (85 kb)
Supplementary material 1 (DOCX 84 kb)


  1. Bevan M (1984) Binary Agrobacterium vectors for plant transformation. Nucl Acids Res 12:8711–8721CrossRefPubMedPubMedCentralGoogle Scholar
  2. Cillo F, Palukaitis P (2014) Transgenic resistance. Adv Virus Res 90:35–146CrossRefPubMedGoogle Scholar
  3. Collinge D (2016) Plant pathogen resistance biotechnology. Wiley, HobokenCrossRefGoogle Scholar
  4. Collinge DB, Jørgensen HJ, Lund OS, Lyngkjær MF (2010) Engineering pathogen resistance in crop plants: current trends and future prospects. Ann Rev Phytopathol 48:269–291CrossRefGoogle Scholar
  5. Coppola V et al (2013) Transcriptomic and proteomic analysis of a compatible tomato-aphid interaction reveals a predominant salicylic acid-dependent plant response. BMC Genom 14:1CrossRefGoogle Scholar
  6. Coppola M et al (2015) Prosystemin overexpression in tomato enhances resistance to different biotic stresses by activating genes of multiple signaling pathways. Plant Mol Biol Report 33:1270–1285CrossRefPubMedGoogle Scholar
  7. Corrado G, Karali M (2009) Inducible gene expression systems and plant biotechnology. Biotechnol Adv 27:733–743CrossRefPubMedGoogle Scholar
  8. Corrado G et al (2005) Inducible expression of a Phytolacca heterotepala ribosome-inactivating protein leads to enhanced resistance against major fungal pathogens in tobacco. Phytopathol 95:206–215CrossRefGoogle Scholar
  9. Corrado G, Scarpetta M, Alioto D, Di Maro A, Polito L, Parente A, Rao R (2008) Inducible antiviral activity and rapid production of the Ribosome-inactivating protein I from Phytolacca heterotepala in tobacco. Plant Sci 174:467–474CrossRefGoogle Scholar
  10. Dai W, Bonos S, Guo Z, Meyer W, Day P, Belanger F (2003) Expression of pokeweed antiviral proteins in creeping bentgrass. Plant Cell Rep 21:497–502CrossRefPubMedGoogle Scholar
  11. Dang L, Van Damme EJ (2015) Toxic proteins in plants. Phytochem 117:51–64CrossRefGoogle Scholar
  12. Dangl JL, Horvath DM, Staskawicz BJ (2013) Pivoting the plant immune system from dissection to deployment. Science 341:746–751CrossRefPubMedGoogle Scholar
  13. Deblaere R, Bytebier B, De Greve H, Deboeck F, Schell J, 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
  14. Desmyter S, Vandenbussche F, Hao Q, Proost P, Peumans WJ, Van Damme EJ (2003) Type-1 ribosome-inactivating protein from iris bulbs: a useful agronomic tool to engineer virus resistance? Plant Mol Biol 51:567–576CrossRefPubMedGoogle Scholar
  15. Devoto A, Leckie F, Lupotto E, Cervone F, De Lorenzo G (1998) The promoter of a gene encoding a polygalacturonase-inhibiting protein of Phaseolus vulgaris L. is activated by wounding but not by elicitors or pathogen infection. Planta 205:165–174CrossRefPubMedGoogle Scholar
  16. Di Maro A, Chambery A, Daniele A, Casoria P, Parente A (2007) Isolation and characterization of heterotepalins, type 1 ribosome-inactivating proteins from Phytolacca heterotepala leaves. Phytochem 68:767–776CrossRefGoogle Scholar
  17. Durrant WE, Dong X (2004) Systemic acquired resistance. Ann Rev Phytopathol 42:185–209CrossRefGoogle Scholar
  18. Dutt M, Dhekney SA, Soriano L, Kandel R, Grosser JW (2014) Temporal and spatial control of gene expression in horticultural crops. Hortic Res 1:14047CrossRefPubMedPubMedCentralGoogle Scholar
  19. Görschen E, Dunaeva M, Hause B, Reeh I, Wasternack C, Parthier B (1997) Expression of the ribosome-inactivating protein JIP60 from barley in transgenic tobacco leads to an abnormal phenotype and alterations on the level of translation. Planta 202:470–478CrossRefPubMedGoogle Scholar
  20. Iglesias R, Citores L, Ragucci S, Russo R, Di Maro A, Ferreras JM (2016) Biological and antipathogenic activities of ribosome-inactivating proteins from Phytolacca dioica L. Biochim Biophys Acta 1860:1256–1264CrossRefPubMedGoogle Scholar
  21. Keller H et al (1999) Pathogen-induced elicitin production in transgenic tobacco generates a hypersensitive response and nonspecific disease resistance. Plant Cell 11:223–235CrossRefPubMedPubMedCentralGoogle Scholar
  22. Kou Y, Wang S (2010) Broad-spectrum and durability: understanding of quantitative disease resistance. Curr Opin Plant Biol 13:181–185CrossRefPubMedGoogle Scholar
  23. Lodge JK, Kaniewski WK, Tumer NE (1993) Broad-spectrum virus resistance in transgenic plants expressing pokeweed antiviral protein. Proc Natl Acad Sci USA 90:7089–7093CrossRefPubMedPubMedCentralGoogle Scholar
  24. Maddaloni M, Forlani F, Balmas V, Donini G, Stasse L, Corazza L, Motto M (1997) Tolerance to the fungal pathogen Rhizoctonia solani AG4 of transgenic tobacco expressing the maize ribosome-inactivating protein b-32. Transgenic Res 6:393–402CrossRefGoogle Scholar
  25. Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15:473–497CrossRefGoogle Scholar
  26. Nielsen K, Boston RS (2001) Ribosome-inactivating proteins: a plant perspective. Ann Rev Plant Biol 52:785–816CrossRefGoogle Scholar
  27. Punja ZK (2006) Recent developments toward achieving fungal disease resistance in transgenic plants. Can J Plant Pathol 28:S298–S308CrossRefGoogle Scholar
  28. Rizhsky L, Mittler R (2001) Inducible expression of bacterio-opsin in transgenic tobacco and tomato plants. Plant Mol Biol 46:313–323CrossRefPubMedGoogle Scholar
  29. Rushton PJ, Reinstädler A, Lipka V, Lippok B, Somssich IE (2002) Synthetic plant promoters containing defined regulatory elements provide novel insights into pathogen-and wound-induced signaling. Plant Cell 14:749–762CrossRefPubMedPubMedCentralGoogle Scholar
  30. Sarma BK, Singh HB, Fernando D, Silva RN, Gupta VK (2016) Enhancing Plant disease resistance without r genes. Trends Biotechnol 34:523–525CrossRefPubMedGoogle Scholar
  31. Singh KB, Foley RC, Oñate-Sánchez L (2002) Transcription factors in plant defense and stress responses. Curr Opin Plant Biol 5:430–436CrossRefPubMedGoogle Scholar
  32. Stirpe F, Gilabert-Oriol R (2015) Ribosome-inactivating proteins: an overview. In: Gopalakrishnakone P, Carlini C, Ligabue-Braun R (eds) Plant toxins. Springer, Dordrecht, pp 1–29CrossRefGoogle Scholar
  33. Van Damme EJ, Hao Q, Chen Y, Barre A, Vandenbussche F, Desmyter F, Rougé P, Peumans WJ (2001) Ribosome-inactivating proteins: a family of plant proteins that do more than inactivate ribosomes. Crit Rev Plant Sci 20:395–465CrossRefGoogle Scholar
  34. Wang P, Zoubenko O, Tumer NE (1998) Reduced toxicity and broad spectrum resistance to viral and fungal infection in transgenic plants expressing pokeweed antiviral protein II. Plant Mol Biol 38:957–964CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2017

Authors and Affiliations

  • Romel Gonzales-Salazar
    • 1
  • Bianca Cecere
    • 1
  • Michelina Ruocco
    • 2
  • Rosa Rao
    • 1
  • Giandomenico Corrado
    • 1
  1. 1.Dipartimento di AgrariaUniversità degli Studi di Napoli “Federico II”PorticiItaly
  2. 2.Istituto per la Protezione Sostenibile delle Piante, CNRPorticiItaly

Personalised recommendations