Skip to main content
SpringerLink
Log in

Developmental and photosynthetic regulation of δ-endotoxin reveals that engineered sugarcane conferring resistance to ‘dead heart’ contains no toxins in cane juice

  • Published:
Molecular Biology Reports Aims and scope Submit manuscript

Abstract

The phosphoenolpyruvate (PEP) carboxylase is regulated at the levels of transcription and post-translation in C4 plants in light and abundantly accumulates in leaf mesophyll cells. We report here developmental and photosynthetic regulation of stably accumulated Bacillus thuringiensis δ-endotoxin under the control of PEP-C promoter in transgenic sugarcane. In young leaves of plants, the transprotein is accumulated to 39% of the levels in mature leaves (135 ng mg−1), and is induced with the cell development, from base to tip. Nevertheless, these levels are decreased up to 99.98% in non-photosynthetic cells as cane matures, from top to bottom, suggesting the photosynthesis regulation of δ-endotoxin in cane cells. Further, transgenic plants are highly resistant to ‘dead heart’. In these studies, Scirpophaga nivela larvae causing ‘dead heart’ were killed within one hour of release to the transgenic plants. Therefore, this report may be regarded as the first report that provides a better strategy for developing transgenic sugarcane lines with absolute protection against invading larvae and no toxin residues in cane juice.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  1. Hatch MD (1987) C4 photosynthesis: a unique blend of modified biochemistry, anatomy and ultrastructure. Biochem Biophys Acta 895:81–106

    CAS  Google Scholar 

  2. Matsuoka M, Furbank R, Fukayama H, Miyao M (2001) Molecular engineering of C4 photosynthesis. Annu Rev Plant Physiol Plant Mol Biol 52:297–314

    Article  PubMed  CAS  Google Scholar 

  3. Jeanneau M, Vidal J, Gousset-Dupont A, Lebouteller B, Hodges M, Gerentes D, Perez P (2002) Manipulating PEPC levels in plants. J Exp Bot 53:1837–1845

    Article  PubMed  CAS  Google Scholar 

  4. Ku MSB, Agarie S, Nomura M, Fukayama H, Tsuchida H, Ono K, Hirose S, Toki S, Miyao M, Matsuoka M (1999) High level expression of maize phosphoenolpyruvate carboxylase in transgenic rice plants. Nat Biotechnol 17:76–80

    Article  PubMed  CAS  Google Scholar 

  5. Goatly MB, Smith H (1974) Differential properties of phosphoenolpyruvate carboxylase from etiolated and green sugar cane. Planta 117:67–73

    Article  CAS  Google Scholar 

  6. Stockhaus J, Schlue U, Koczor M, Chitty JA, Taylor WC, Westhoff P (1997) The promoter of the gene encoding the C4 form of phosphoenolpyruvate carboxylase directs mesophyll specific expression in transgenic C4 Flaveria. Plant Cell 9:479–489

    Article  PubMed  Google Scholar 

  7. Matsuoka M, Tada Y, Fujimura T, Kano-Murakami Y (1993) Tissue-specific light-regulated expression directed by the promoter of a C4 gene, maize pyruvate, orthophosphate dikinase, in a C3 plant, rice. Proc Natl Acad Sci USA 90:9586–9590

    Article  PubMed  CAS  Google Scholar 

  8. Datta K, Vasquez Z, Tu J, Torrizo L, Alam MF, Oliva N, Abrigo E, Khush GS, Datta SK (1998) Constitutive and tissue-specific differential expression of the cryI(A)b gene in transgenic rice plants conferring resistance to rice insect pests. Theor Appl Genet 97:20–30

    Article  CAS  Google Scholar 

  9. Offermann S, Danker T, Dreymüller D, Kalamajka R, Töpsch S, Weyand K, Peterhänsel C (2006) Illumination is necessary and sufficient to induce histone acetylation independent of transcriptional activity at the C4-specific phosphoenolpyruvate carboxylase promoter in maize. Plant Physiol 141:1078–1088

    Article  PubMed  CAS  Google Scholar 

  10. Höfte H, Whiteley HR (1989) Insecticidal crystal proteins of Bacillus thuringiensis. Microbiol Rev 53:242–255

    PubMed  Google Scholar 

  11. Greenplate J (1999) Quantification of Bacillus thuringiensis insect control protein Cry1Ac over time in Bollgard cotton fruit and terminals. J Econ Entomol 92:1377–1383

    CAS  Google Scholar 

  12. Enriquez-Obregon GA, Vazquez-Padron RI, Prieto-Samsonov DL, De-la-Riva GA, Selman-Housein G (1998) Herbicide-resistant sugarcane (Saccharum officinarum L.) plants by Agrobacterium-mediated transformation. Planta 206:20–27

    Article  CAS  Google Scholar 

  13. Gallo-Meagher M, Irvine JE (1996) Herbicide resistant transgenic sugarcane plants containing the bar gene. Crop Sci 36:1367–1374

    Article  CAS  Google Scholar 

  14. Chowdhury MKU, Vasil KI (1992) Stably transformed herbicide resistant callus of sugarcane via microprojectile bombardment of cell suspension cultures and electroporation of protoplasts. Plant Cell Rep 11:494–498

    Article  Google Scholar 

  15. Bower R, Elliot AR, Potier BAM, Birch RG (1996) High efficiency, microprojectile-mediated cotransformation of sugarcane, using visible or selectable markers. Mol Breed 2:239–249

    Article  CAS  Google Scholar 

  16. Liu D, Oard SV, Oard JH (2003) High transgene expression levels in sugarcane (Saccharum officinarum L.) driven by the rice ubiquitin promoter RUBQ2. Plant Sci 165:743–750

    Article  CAS  Google Scholar 

  17. Weng L-X, Deng H, Xu J-L, Li Q, Wang L-H, Jiang Z, Zhang HB, Li Q, Zhang L-H (2006) Regeneration of sugarcane elite breeding lines and engineering of stem borer resistance. Pest Manag Sci 62:178–187

    Article  PubMed  CAS  Google Scholar 

  18. Birch RG, Bower R, Elliot A, Potier B, Franks T, Cordeiro G (1995) Expression of foreign genes in sugarcane. In: Cock JH, Brekelbaum T (eds) Proceedings of the XXII Congress of the International Society of Sugarcane Technologists, vol 2. Tecnicana, Cali, pp 369–373

    Google Scholar 

  19. Arencibia A, Vasquez R, Prieto D, Tellez P, Carmona E, Coego A, Hernandez L, de la Riva G, Selman Housein G (1997) Transgenic sugarcane plants resistant to stem borer attack. Mol Breed 3:247–255

    Article  Google Scholar 

  20. Rogers SO, Bendich AJ (1988) Extraction of DNA from plant tissues. Plant Mol Biol Man 6:1–10

    Google Scholar 

  21. Kota M, Daniel H, Varma S, Garczynski F, Gould F, Moar WJ (1999) Overexpression of the Bacillus thuringiensis Cry2A protein in chloroplasts confers resistance to plants against susceptible and Bt-resistant insects. Proc Natl Acad Sci USA 96:1840–1845

    Article  PubMed  CAS  Google Scholar 

  22. Christou P, Ford TL, Kofron M (1991) Production of transgenic rice (Oryza sativa) plants from agronomically important indica and japonica varieties via electric discharge particle acceleration of exogenous DNA into immature zygotic embryos. Biotechnology 9:957–962

    Article  Google Scholar 

  23. Spencer TM, Gordon-Kamm WJ, Daines RJ, Start WG, Lemaux PG (1990) Bialaphos selection of stable transformants from maize cell culture. Theor Appl Genet 79:625–631

    Article  CAS  Google Scholar 

  24. Vasil V, Castillo A, Fromm M, Vasil IK (1992) Herbicide resistant fertile transgenic wheat plants obtained by microprojectile bombardment of regenerable embryogenic callus. Biotechnology 10:667–674

    Article  CAS  Google Scholar 

  25. Wan Y, Lemaux PG (1994) Generation of large numbers of independently transformed fertile barley plants. Plant Physiol 104:37–48

    PubMed  CAS  Google Scholar 

  26. Cao J, Duan XL, McElroy D, Wu R (1992) Regeneration of herbicide resistant transgenic rice plants following microprojectile-mediated transformation of suspension culture cells. Plant Cell Rep 11:586–591

    Article  CAS  Google Scholar 

  27. Toki S, Takamatsu S, Norjiri C, Ooba S, Anzai H, Iwata M, Christensen AH, Quail PH, Uchimiya H (1992) Expression of a maize ubiquitin gene promoter-bar chimeric gene in transgenic rice plants. Plant Physiol 100:1503–1507

    Article  PubMed  CAS  Google Scholar 

  28. D’Halluin K, Bonne E, Bossut M, De Beuckeleer M, Leemans J (1992) Transgenic maize plants by tissue electroporation. Plant Cell 4:1495–1505

    Article  PubMed  Google Scholar 

  29. Manickavasagam M, Gavapathi A, Anbazhagan VR, Sudhakar B, Selvaraj N, Vasudevan A, Kasthurirengan S (2004) Agrobacterium-mediated genetic transformation and development of herbicide-resistant sugarcane (Saccharum species hybrids) using axillary buds. Plant Cell Rep 23:134–143

    Article  PubMed  CAS  Google Scholar 

  30. Jabeen R, Khan MS, Zafar Y, Anjum T (2010) Codon optimization of cry1Ab gene for hyper expression in plant organelles. Mol Biol Rep 37:1011–1017

    Article  PubMed  CAS  Google Scholar 

  31. Ting IP, Osmond CB (1973) Multiple forms of plant phosphoenolpyruvate carboxylase associated with different metabolic pathways. Plant Physiol 51:448–453

    Article  PubMed  CAS  Google Scholar 

  32. Tachibana K, Watanabe T, Sekizawa T, Takematsu T (1986) Action mechanism of bialaphos II: accumulation of ammonia in plants treated with bialaphos. J Pest Sci 11:33–37

    CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

Download references

Acknowledgments

We thank Rob W. Briddon for critical reading of the manuscript. MSK is also thankful to Takashi Shiina for providing primers to perform transcript analysis. The research was funded by Ministry of Science and Technology (MoST), Islamabad to MSK. The idea was conceived by MSK and executed by MSK and SA at NIBGE; nevertheless student was registered at Punjab University with JI and MSK.

Author information

Author notes

  1. Muhammad Sarwar Khan

    Present address: Centre of Agricultural Biochemistry and Biotechnology (CABB), University of Agriculture, Faisalabad, Pakistan

Authors and Affiliations

  1. National Institute for Biotechnology and Genetic Engineering (NIBGE), Jhang Road, PO Box 577, Faisalabad, 38000, Pakistan

    Muhammad Sarwar Khan & Safdar Ali

  2. School of Biological Sciences, University of Punjab, Lahore, Pakistan

    Javed Iqbal

Authors
  1. Muhammad Sarwar Khan
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Safdar Ali
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Javed Iqbal
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Muhammad Sarwar Khan.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Khan, M.S., Ali, S. & Iqbal, J. Developmental and photosynthetic regulation of δ-endotoxin reveals that engineered sugarcane conferring resistance to ‘dead heart’ contains no toxins in cane juice. Mol Biol Rep 38, 2359–2369 (2011). https://doi.org/10.1007/s11033-010-0369-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11033-010-0369-7

Keywords

Search

Navigation