Containment evaluation, cold tolerance and toxicity analysis in Osmotin transgenic tomato (Solanum lycopersicum L. cv. Pusa Ruby)
The present study reports engineered cold tolerance and toxicity analysis in genetically modified tomato (Solanum lycopersicum L. cv. Pusa Ruby) developed through constitutive over expression of Nicotiana tabacum Osmotin gene. Rate of seed germination, seedling establishment and growth remained unaffected in the transgenic tomato in response to a low temperature (15 °C) treatment, but were significantly (P ≤ 0.05) reduced in the wild type. At reproductive stage, the wild type plants failed to recover at the low temperature (4.0 °C) treatment for 10 days but the transgenic plants survived successfully without any leaf senescence or other visible chilling injury symptoms. The quantitative transcript expression analysis confirmed up regulation of the transgene by 55% in the transgenic plants on cold treatment for 2 h whereas, the transcripts were not detected in the wild type. Containment evaluation under normal environmental conditions revealed similar morphology in both the transgenic and wild type tomato plants however an average fruit yield was higher in the transgenic plants (725.91 ± 39.27 g) than the wild type (679.84 ± 28.80 g). The composition of mature fruits in terms of element content was at par in both the transgenic and wild type except significantly higher Ca and Mg contents in the transgenic fruits than that of the wild type. Further, acute and sub-acute toxicity tests conducted in the adult female Wister rats revealed no mortality or significant changes in general and psychological behaviour, at par food intake and body weight and, normal biochemical, and hematological parameters for animals fed with the wild type or transgenic tomato fruits as compared to the control group, confirming its safety for animal consumption.
KeywordsGenetic engineering Cold stress Germination TaqMan® probe Toxicity
Authors are grateful to Prof. KC Bansal, National Research Centre on Plant Biotechnology, Indian Agricultural Research Institute, New Delhi, India for providing binary construct with the Osmotin gene.
VYP conceived and designed the study. VYP, AG and SMG carried out cold tolerance analysis, containment evaluation, transcript expression and statistical analysis. HSM conducted toxicity tests. First draft of the manuscript was prepared by VYP, with inputs from HSM, AG and SMG. MN arranged funds and approved final version of the manuscript.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no competing interests.
- EPA (2000) Mammalian toxicity assessment guidelines for protein plant pesticides. FIFRA Scientific Advisory Panel. Environmental Protection Agency, ArlingtonGoogle Scholar
- Goyary D (2009) Transgenic crops, and their scope for abiotic stress environment of high altitude: biochemical and physiological perspectives. DRDO Sci Spectrum 195–201Google Scholar
- Jain NC (1986) Schalm’s Veterinary Hematology. 4th ed. Lea and Febiger, 600. Washington square, Philadelphia, USAGoogle Scholar
- Ntatsi G, Savvas D (2014) Growth, yield, and metabolic responses of temperature-stressed tomato to grafting onto rootstocks differing in cold tolerance. J Amer Soc Hort Sci 139(2):230–243Google Scholar
- Patade VY (2012) Development of a rapid, reliable and sensitive real time PCR method based on probe hybridisation for validation of transgenic plants. DRDO Technol Spectrum (Supplement). 31–36Google Scholar
- Rozen S, Skaletsky HJ (2000) Primer3 on the WWW for general users and for biologists programmers. In: Krawetzs S, Misener S (eds) Bioinformatics methods and protocols: methods in molecular biology. Humana Press, Totowa, pp 365–386Google Scholar
- Sarad N, Rathore M, Singh NK, Kumar N (2004) Genetically engineered tomatoes: new vista for sustainable agriculture in high altitude regions. In: Proceedings of the Fourth international crop science congress Brisbane, AustraliaGoogle Scholar