Tomato lines expressing human lactoferrin gene are characterized by enhanced resistance to late blight

TitleTomato lines expressing human lactoferrin gene are characterized by enhanced resistance to late blight
Publication TypeJournal Article
Year of Publication2020
AuthorsBuziashvili, АY, Cherednichenko, LМ, Kropyvko, SV, Yemets, АІ
Abbreviated Key TitleDopov. Nac. akad. nauk Ukr.
DOI10.15407/dopovidi2020.05.095
Issue5
SectionBiology
Pagination95-102
Date Published5/2020
LanguageUkrainian
Abstract

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.
Keywordshuman lactoferrin gene, Lycopersicon esculentum Mill., Phytophthora infestans, resistance
References: 

1. 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
2. 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
3. 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).
4. 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
5. 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
6. 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
7. 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
8. 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
9. 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
10. 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
11. 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
12. 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
13. Tkachyk, S. O. (Ed.) (2014). Methods of phytopathological researches with artificial infection of plants. Kyiv: Nylan-LTD (in Ukrainian).
14. 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...
15. 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