Tomato lines expressing human lactoferrin gene are characterized by enhanced resistance to late blight
DOI:
https://doi.org/10.15407/dopovidi2020.05.095Keywords:
human lactoferrin gene, Lycopersicon esculentum Mill., Phytophthora infestans, resistanceAbstract
Transgenic lines of tomatoes (Lycopersicon esculentum Mill.) of cultivars Lahidny and Money Maker expressing human lactoferrin gene (hLf) were obtained with the use of Agrobacterium-mediated transformation. For the transfer of hLf gene, the pBin35LF plasmid vector was used with the gene-of-interest under control of 35 S promoter of cauliflower mosaic virus (CaMV35S) and the gene of neomycinephosphotransferase ІІ (nptII) conferring the resistance to kanamycin. Integration of hLf into genome of transgenic lines was confirmed with the use of PCR with specific primers to hLf. Expression of lactoferrin protein was detected with the use of Western blot analysis. The resistance of transgenic lines to Phytophthora infestans was studied in vitro with the use of the disk diffusion assay, infection of aseptic plants, and detached leaves. The enhanced resistance to late blight of transgenic tomato lines was shown as compared to control lines. The obtained results demonstrate that the expression of lactoferrin improve the resistance of transgenic tomato plants to aggressive fungal pathogens such as P. infestans.
Downloads
References
Elansky, S. N., Pobedinskaya, M. A., Kokaeva, L. Y., Statsyuk, N. V. & Dyakov, Y. T. (2015). Phytophthora infestans populations from the European part of Russia: genotypic structure and metalaxyl resistance. J. Plant Pathol., 97, No. 3, pp. 449-456. https://doi.org/10.4454/JPP.V97I3.020
Blancard, D., Laterrot, H., Marchoux, G. & Candresse, Th. (Eds.). (2012). Tomato diseases identification, biology and control: A color handbook. 2 ed. London: Manson Publ. Ltd. Doi: https://doi.org/10.1201/b15145
Fedorchuk, S. V. (2019). The impact of growth regulators, chemical and biological agents on the de-velopment of Аlternaria solani and Рhytophthora infestans. Tavriiskyi naukovyi visnyk, 98, pp. 128-133 (in Ukrainian).
Jung, Y.-J., Kang K.-K. (2014). Application of antimicrobial peptides for disease control in plants. Plant Breed. Biotech., 2, No. 1, pp. 1-13. Doi: https://doi.org/10.9787/PBB.2014.2.1.001
Pribylova, R., Pavlik, I. & Bartos, M. (2006). Genetically modified potato plants in nutrition and prevention of diseases in humans and animals: a review. Vet. Med.-Czech., 51, No. 5, pp. 212-223. Doi: https://doi.org/10.17221/5540-VETMED
Yemets, A. I., Tanasienko, I. V., Krasylenko, Yu. A. & Blume, Ya. B. (2014). Plant-based biopharming of recombinant human lactoferrin. Cell Biol. Int., 38, No. 9, pp. 989-1002. Doi: https://doi.org/10.1002/cbin.10304
Muñoz, A., Marcos, J. F. (2006). Activity and mode of action against fungal phytopathogens of bovine lactoferricin-derived peptides. J. Appl. Microbiol., 101, No. 6, pp. 1199-1207. Doi: https://doi.org/10.1111/j.1365-2672.2006.03089.x
van Baarlen, P., van Belkum, A., Summerbell, R.C., Crous, P.W. & Thomma, B.P. (2007). Molecular mechanisms of pathogenicity: how do pathogenic microorganisms develop cross-kingdom host jumps? FEMS Microbiol. Rev., 31, No. 3, pp. 239-277. Doi: https://doi.org/10.1111/j.1574-6976.2007.00065.x
Adlerova, L., Bartoskova, A. & Faldyna, M. (2008). Lactoferrin: a review. Vet. Med.-Czech., 53, No. 9, pp. 457-468. Doi: https://doi.org/10.17221/1978-VETMED
Fernandes, K. E. & Carter, D. A. (2017). The antifungal activity of lactoferrin and its derived peptides: mechanisms of action and synergy with drugs against fungal pathogens. Front Microbiol., 8, Art. 2. Doi: https://doi.org/10.3389/fmicb.2017.00002
Buziashvili, А.Yu. & Yemets, А.I. (2018). The obtaining of tomato and potato plants with human lactoferrin gene hLf. Dopov. Nac. acad. nauk Ukr., No. 10, pp. 88-94 (in Ukrainian) Doi: https://doi.org/10.15407/dopovidi2018.10.088
Buziashvili, A., Cherednichenko, L., Kropyvko, S., Blume, Y. & Yemets, A. (2020). Obtaining of transgenic potato plants expressing human lactoferrin gene and analysis of their resistance to phytopathogens. Cytol. Genet., 54, No. 3, pp. 3-15. (in Russian) Doi: https://doi.org/10.3103/S0095452720030020
Tkachyk, S. O. (Ed.) (2014). Methods of phytopathological researches with artificial infection of plants. Kyiv: Nylan-LTD (in Ukrainian).
Tournas, V., Stack, M. E., Mislivec, P. B., Koch, H. A. & Bandler, R. (1998). Chapter 18. Yeasts, molds, and mycotoxins. In Bacteriological Analytical Manual. 8th ed. Revision A. AOAC International. Retrieved from https://www.fda.gov/food/laboratory-methods-food/bam-yeasts-molds-and-my.
Han, J., Lakshman, D. K., Galvez, L. C., Mitra, S., Baenziger, P. S. & Mitra, A. (2012). Transgenic expression of lactoferrin impacts enhanced resistance to head blight of wheat caused by Fusarium graminearum. BMC Plant Biol., 12, No. 1, Art. 33. Doi: https://doi.org/10.1186/1471-2229-12-33
Downloads
Published
How to Cite
Issue
Section
License
Copyright (c) 2023 Reports of the National Academy of Sciences of Ukraine
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.