Green-tissue-specific, C(4)-PEPC-promoter-driven expression of Cry1Ab makes transgenic potato plants resistant to tuber moth (Phthorimaea operculella, Zeller).
An important strategy for obtaining a safer transgenic plant may be the use of a spatial- or tissue-specific promoter, instead of a constitutive one. In this study, we have used a light-inducible maize PEPC promoter to regulate the cry1Ab gene, aiming to produce transgenic potatoes that are resistant to potato tuber moth (PTM) (Phthorimaea operculella, Zeller). Out of 60 regenerated lines having normal phenotypes, 55 lines were PCR-positive for both the cry1Ab and nptII genes. Southern analysis on three selected putative transgenic lines revealed that they have only a single intact copy of the cry1Ab gene. An investigation of the Cry1Ab protein in the leaves and light-exposed (LE) tubers of the transgenic lines demonstrated the presence of the protein in the foliage and green tubers but not in the light-not exposed (LNE) tubers. A bioassay analysis of excised leaves of nine randomly selected lines showed that eight lines had 100% PTM larval mortality. Confirming results were obtained in six selected lines using the whole plant bioassay in the greenhouse. LE transgenic tubers also exhibited 100% larval mortality; however, the levels of damage to the LNE transgenic tubers were high and statistically the same as those incurred by the non-transgenic ones. Based on the results, we believe that this spatial expression of Cry1Ab using the light-inducible PEPC promoter can control PTM infestation in the field and significantly reduce pollution transmission to storage potatoes.
Ghasimi Hagh
Ziba
Z
Agricultural Biotechnology Research Institute of Iran, Karaj, Iran.
Rahnama
Hassan
H
Panahandeh
Jaber
J
Baghban Kohneh Rouz
Bahram
B
Arab Jafari
Khoda Morad
KM
Mahna
Nasser
N
Mahna
Naser
N
D000818:Animals; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000378:metabolism*; D001681:Biological Assay; D015139:Blotting, Southern; D002244:Carbon / Q000378:metabolism*; D004269:DNA, Bacterial / Q000235:genetics; D004731:Endotoxins / Q000378:metabolism*; D005434:Flow Cytometry; D005822:Genetic Vectors / Q000235:genetics; D006460:Hemolysin Proteins / Q000378:metabolism*; D007113:Immunity, Innate / Q000276:immunology; D009036:Moths; D009928:Organ Specificity / Q000235:genetics; D010935:Plant Diseases / Q000235:genetics / Q000276:immunology* / Q000469:parasitology; D018515:Plant Leaves / Q000378:metabolism / Q000469:parasitology; D035281:Plant Tubers / Q000235:genetics / Q000276:immunology / Q000382:microbiology; D030821:Plants, Genetically Modified; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic / Q000235:genetics; D017346:Protein Serine-Threonine Kinases / Q000235:genetics*; D011198:Solanum tuberosum / Q000235:genetics* / Q000276:immunology / Q000469:parasitology*; D014170:Transformation, Genetic
D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D004269:DNA, Bacterial; D004731:Endotoxins; D006460:Hemolysin Proteins; C036483:T-DNA; C030956:insecticidal crystal protein, Bacillus Thuringiensis; D002244:Carbon; C069767:phosphoenolpyruvate carboxylase kinase; D017346:Protein Serine-Threonine Kinases