Euphytica

, Volume 202, Issue 3, pp 373–383 | Cite as

Cry1Ac insecticidal protein levels in genetically modified Coffea canephora Pierre coffee plants were negatively correlated with the growth speed in a field experiment

  • Bernard Perthuis
  • Jean-Michel Vassal
  • Catherine Fenouillet
  • Thierry Leroy
Article

Abstract

Genetically-modified coffee clones (GMCs) were presented in a previous work. They were created from a single commercial clone of Coffea canephora Pierre (clone 126). Therefore they all have the same genotype, except for the localization of the transgenic insertions. They synthesize the Bacillus thuringiensis endotoxin Cry1Ac against Leucoptera coffeella Guérin-Méneville, a secondary pest of C. canephora and one of the main pest of C. arabica in South America. The synthetic Cry1Ac gene is regulated by the EF1α-A1 promoter of Arabidopsis thaliana (L.) Heynh. They were tested in an experimental field for their resistance against the pest insect in a previous work. In the present work, levels of Cry1Ac were measured in the leaves. The insecticidal protein was evenly distributed in all the leaves. Cry1Ac levels were measured once in the coffee plants of a sensitive GMC and of 14 resistant ones grown in the experimental field and in plants grown in a greenhouse. Some resistant GMCs contained higher levels but it is not possible to confirm that it would be enough for a sustainable resistance to the pest. Growth speed was variable in the plot. The correlation with plant height and other indicators of plant growth was examined. Cry1Ac levels in the GMCs were negatively correlated with growth speed. The latter was not statistically influenced neither by Cry1Ac synthesis nor by the genetic modification in itself as seen by comparing the GMCs and the unmodified control clone 126. Hence the conclusion is that the growth conferred to field-grown plants by environmental factors and especially the soil was probably the underlying cause of the negative correlation. Other field experiments would be necessary in order to confirm this result. It would be important that the genetic construct inserted in these GMCs and mainly the EF1α-A1 promoter of the Cry1Ac gene be reconsidered since Cry1Ac levels might be too low to provide efficient and sustainable protection against L. coffeella in a highly favourable environment for coffee plants. Alternatives are discussed.

Keywords

Coffea canephora Pierre Coffee plant Cry1Ac level EF1α-A1 promoter Environmental factors Genetic modification Quantification 

Abbreviations

GMC (s)

Genetically-modified clone(s)

GM

Genetically-modified

fw

Fresh weight

Bt

Bacillus thuringiensis

References

  1. Adamczyk JJ, Hardee DD, Adams LC et al (2001) Correlating differences in larval survival and development of bollworm (Lepidoptera: noctuidae) and fall armyworm (Lepidoptera: Noctuidae) to differential expression of Cry1A (c) delta-endotoxin in various plant parts among commercial cultivars of transgenic Bacillus thuringiensis cotton. J Econ Entomol 94(1):284–290CrossRefPubMedGoogle Scholar
  2. Aida R, Nagaya S, Yoshida K et al (2005) Efficient transgene expression in chrysanthemum, Chrysanthemum morifolium ramat., with the promoter of a gene for tobacco elongation factor 1 alpha protein. Jpn Agric Res Q 39(4):269–274CrossRefGoogle Scholar
  3. Brandle JE, Miki BL (1993) Agronomic performance of sulfonylurea-resistant transgenic flue-cured tobacco grown under field condition. Crop Sci 33(4):847–852Google Scholar
  4. Bruns HA, Abel CA (2007) Effects of nitrogen fertility on Bt endotoxin levels in maize. J Entomol Sci 42(1):35–43Google Scholar
  5. Callis J, Raasch JA, Vierstra RD et al (1990) Ubiquitin extension proteins of Arabidopsis thaliana. Structure, localization, and expression of their promoters in transgenic tobacco. J Biol Chem 265:12486–12493PubMedGoogle Scholar
  6. Curie C, Liboz T, Bardet C et al (1991) Cis and trans-acting elements involved in the activation of Arabidopsis thaliana A1 gene encoding the translation elongation factor EF-1α. Nucleic Acids Res 19(6):1305–1310CrossRefPubMedCentralPubMedGoogle Scholar
  7. De Guglielmo-Cróquer Z, Altosaar I, Zaidi M et al (2010) Transformation of coffee (Coffea Arabica L. cv. Catimor) with the cry1ac gene by biolistic, without the use of markers. Braz J Biol 70(2):387–393CrossRefPubMedGoogle Scholar
  8. Ducos JP, Alenton R, Reano JF et al (2003) Agronomic performance of Coffea canephora P. trees derived from large-scale somatic embryo production in liquid medium. Euphytica 131(2):215–223CrossRefGoogle Scholar
  9. Fragoso DB, Guedes RNC, Picanço MC et al (2002a) Insecticide use and organophosphate resistance in the coffee leaf miner Leucoptera coffeella (Lepidoptera: Lyonetiidae). Bull Entomol Res 92(3):203–212PubMedGoogle Scholar
  10. Fragoso DB, Jusselino Filho P, Pallini Filho A et al (2002b) Action of organophosphorate insecticides used to control Leucoptera coffeella (Guérin-Mèneville) (Lepidoptera: Lyonetiidae) on the predator mite Iphiseiodes zuluagai. Neotrop Entomol 31(3):463–467CrossRefGoogle Scholar
  11. Gould F (1998) Sustainability of transgenic insecticidal cultivars: integrating pest genetics and ecology. Annu Rev Entomol 43:701–726CrossRefPubMedGoogle Scholar
  12. Greenplate JT (1999) Quantification of Bacillus thuringiensis insect control protein Cry1Ac over time in Bollgard cotton fruit and terminals. J Econ Entomol 92(6):1377–1383CrossRefGoogle Scholar
  13. Guerreiro O, Denolf P, Peferoen M, Decazy B, Eskes AB, Frutos F (1998) Susceptibility of the coffee leaf miner (Perileucoptera spp.) to Bacillus thuringiensis δ-endotoxins: a model for transgenic perennial crops resistant to endocarpic insects. Curr Microbiol 36:175–179CrossRefGoogle Scholar
  14. Halpin C (2005) Gene stacking in transgenic plants-the challenge for 21st century plant technology. Plant Biotechnol J 3:141–155CrossRefPubMedGoogle Scholar
  15. Kay R, Chan A, Daly M et al (1987) Duplication of CaMV35S promoter sequences creates a strong enhancer for plant genes. Science 236:1299–1302CrossRefPubMedGoogle Scholar
  16. Leroy T, Henry AM, Royer M et al (2000) Genetically modified coffee plants expressing the Bacillus thuringiensis cry1Ac gene for resistance to leaf miner. Plant Cell Rep 19(4):382–389CrossRefGoogle Scholar
  17. Martins CM, Beyene G, Hofs JL et al (2008) Effect of water-deficit stress on cotton plants expressing the Bacillus thuringiensis toxin. Annals of Applied Biology 152(2):255–262CrossRefGoogle Scholar
  18. Nantes JFD, Parra JRP (1977) Biologia de Perileucoptera coffeella (Guérin-Mèneville, 1842) (Lepidoptera-Lyonetiidae), em três variedades de café (Coffea spp). Anais do Sociedade Entomológica do Brasil 6:156–163Google Scholar
  19. Perthuis B, Pradon JL, Montagnon C et al (2005) Stable resistance against the leaf miner Leucoptera coffeella expressed by genetically transformed Coffea canephora in a pluriannual field experiment in French Guiana. Euphytica 144(3):321–329CrossRefGoogle Scholar
  20. Pettigrew WT, Adamczyk JJ (2006) Nitrogen fertility and planting date effects on lint yield and Cry1Ac (Bt) endotoxin production. Agron J 98(3):691–697CrossRefGoogle Scholar
  21. Pinto FO, Maluf MP, Guerreiro-Filho O (2007) Study of simple sequence repeat markers from coffee expressed sequences associated to leaf miner resistance. Pesqui Agropecu Brasileira 42(3):377–384CrossRefGoogle Scholar
  22. Pouteau S, Huttner E, Grandbastien M et al (1991) Specific expression of the tobacco Tnt1 retrotransposon in protoplasts. EMBO J 10:1911–1918PubMedCentralPubMedGoogle Scholar
  23. Purrington CB, Bergelson J (1997) Fitness consequences of genetically engineered herbicide and antibiotic resistance in Arabidopsis. Genetics 145(3):807–814PubMedCentralPubMedGoogle Scholar
  24. Purrington CB, Bergelson J (1999) Exploring the physiological basis of costs of herbicide resistance in Arabidopsis thaliana. Am Nat 154(S1):S82–S91CrossRefGoogle Scholar
  25. Rochester IJ (2006) Effect of genotype, edaphic, environmental conditions, and agronomic practices on Cry1Ac protein expression in transgenic cotton. J Cotton Sci 10:252–262Google Scholar
  26. Smirnova OG, Ibragimova SS, Kochetov AV (2012) Simple database to select promoters for plant transgenesis. Transgenic Res 21:429–437CrossRefPubMedGoogle Scholar
  27. Van Boxtel J, Berthouly M, Carasco C, Dufour M, Eskes AB (1995) Transient expression of β-glucuronidase following biolistic delivery of foreign DNA into coffee tissues. Plant Cell Rep 14(12):748–752CrossRefPubMedGoogle Scholar
  28. Wei W, Schuler TH, Clark S et al (2005) Age-related increase in levels of insecticidal protein in the progenies of transgenic oilseed rape and its efficacy against a susceptible strain of diamondback moth. Ann Appl Biol 147(3):227–234CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2014

Authors and Affiliations

  • Bernard Perthuis
    • 1
  • Jean-Michel Vassal
    • 2
  • Catherine Fenouillet
    • 2
  • Thierry Leroy
    • 2
  1. 1.Centre International de Recherche Pour le Développement (CIRAD), UMR AGAPKourouFrance
  2. 2.Centre International de Recherche Pour le Développement (CIRAD), UMR AGAPMontpellierFrance

Personalised recommendations