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Expression of Drosera rotundifolia Chitinase in Transgenic Tobacco Plants Enhanced Their Antifungal Potential

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Abstract

In this study, a chitinase gene (DrChit) that plays a role in the carnivorous processes of Drosera rotundifolia L. was isolated from genomic DNA, linked to a double CaMV35S promoter and nos terminator in a pBinPlus plant binary vector, and used for Agrobacterium-mediated transformation of tobacco. RT-qPCR revealed that within 14 transgenic lines analysed in detail, 57% had DrChit transcript abundance comparable to or lower than level of a reference actin gene transcript. In contrast, the transgenic lines 9 and 14 exhibited 72 and 152 times higher expression level than actin. The protein extracts of these two lines exhibited five and eight times higher chitinolytic activity than non-transgenic controls when measured in a fluorimetric assay with FITC-chitin. Finally, the growth of Trichoderma viride was obviously suppressed when the pathogen was exposed to 100 μg of crude protein extract isolated from line 9 and line 14, with the area of mycelium growth reaching only 56.4% and 45.2%, of non-transgenic control, respectively. This is the first time a chitinase from a carnivorous plant with substrate specificity for long chitin polymers was tested in a transgenic plant with the aim of exploring its antifungal potential.

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Acknowledgements

The authors would like to thank Anna Fabelova for in vitro plant care. This work was co-funded by a grant from the Slovak Grant Agency VEGA 2/0075/17 and Research Centre AgroBioTech built in the framework of European Community project Building Research Centre “AgroBioTech” ITMS 26220220180.

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  1. Dominika Durechova and Martin Jopcik contributed equally to this study.

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  1. Institute of Plant Genetics and Biotechnology, Plant Science Biodiversity Center, Slovak Academy of Sciences, Akademicka 2, P. O. Box 39A, 950 07, Nitra, Slovak Republic

    Dominika Durechova, Martin Jopcik, Miroslav Rajninec, Jana Moravcikova & Jana Libantova

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  1. Dominika Durechova
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JL and MJ and designed research. DD, MJ, MR and JM conducted the experiments. MJ analysed data. JL wrote the manuscript.

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Correspondence to Jana Libantova.

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Durechova, D., Jopcik, M., Rajninec, M. et al. Expression of Drosera rotundifolia Chitinase in Transgenic Tobacco Plants Enhanced Their Antifungal Potential. Mol Biotechnol 61, 916–928 (2019). https://doi.org/10.1007/s12033-019-00214-1

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