Transgenic Research

, 15:481

Expression of the cry9Aa2 B.t. gene in tobacco chloroplasts confers resistance to potato tuber moth

  • Swarup Kumar Chakrabarti
  • Kerry Ann Lutz
  • Benjawan Lertwiriyawong
  • Zora Svab
  • Pal Maliga
Original Paper

DOI: 10.1007/s11248-006-0018-z

Cite this article as:
Chakrabarti, S.K., Lutz, K.A., Lertwiriyawong, B. et al. Transgenic Res (2006) 15: 481. doi:10.1007/s11248-006-0018-z

Abstract

We report here the control of potato tuber moth (Phthorimaea operculella) by incorporating a truncated Bacillus thuringiensis cry9Aa2 gene in the plastid genome. Plasmids pSKC84 and pSKC85 are derivatives of a new polycistronic plastid transformation vector, pPRV312L, that carries spectinomycin resistance (aadA) as a selective marker and targets insertions in the trnItrnA intergenic region. The Cry9Aa2 N-terminal region (82.1 kDa; 734 amino acids) was expressed in a cassette, which consists of 49 nucleotides of the cry9Aa2 leader and the 3′-untranslated region of the plastid rbcL gene (TrbcL), and relies on readthrough transcription from the plastid rRNA operon. In a tobacco leaf bioassay, expression of Cry9Aa2 conferred resistance to potato tuber moth. In accordance, the Cry9Aa2 insecticidal protein accumulated to high levels, ~10% of the total soluble cellular protein and ~20% in the membrane fraction. However, high-level Cry9Aa2 expression significantly delayed plant development. Thus, a practical system to control potato tuber moth by Cry9Aa2 expression calls for down-regulation of its expression.

Keywords

Bacillus thuringiensis cry9Aa2 Insect resistance Nicotiana tabacum Plastid transformation Potato tuber moth Phthorimaea operculella 

Copyright information

© Springer Science+Business Media B.V. 2006

Authors and Affiliations

  • Swarup Kumar Chakrabarti
    • 1
    • 2
  • Kerry Ann Lutz
    • 2
  • Benjawan Lertwiriyawong
    • 2
    • 3
  • Zora Svab
    • 2
  • Pal Maliga
    • 2
  1. 1.Central Potato Research InstituteShimlaIndia
  2. 2.Waksman Institute, RutgersThe State University of New JerseyPiscatawayUSA
  3. 3.Department of GeneticsKasetsart UniversityJatuchakThailand

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