Optimization of molecular diagnostics of fusarium pathogens in winter triticale (×Triticosecale Wittm. & A. Camus)

Phytopathogenic fungi are among the most common and dangerous plant pathogens, causing significant crop losses and considerable economic costs. The greatest damage to agricultural crops is caused by fungi of the genus Fusarium, making the study of their distribution, species diversity and diagnostic methods more in demand every year. This is particularly important for breeding purposes. The determination of the species composition of this pathogen allows not only to evaluate the level of infestation, but also to perform preventive selection for resistance to Fusarium. The aim of the study was to test the developed primers and probes for the identification of the most common representatives of the genus Fusarium such as F. culmorum, F. graminearum and F. poae. Pure cultures of fungi obtained from different collections of microorganisms were used as research material. We studied 35 varieties of winter triticale from the RSAU-MTAA collection. In order to differentiate the studied species, oligonucleotides were designed for a fragment of the Galactose oxidase (GalOx) gene and its homologous genes from the NCBI database. Diagnosis was performed by real-time polymerase chain reaction (qPCR). The main analytical characteristics of the developed primers such as specificity and sensitivity were determined. Screening of 35 varieties of winter triticale showed their infestation with fungi of the genus Fusarium. It was revealed that the predominant species was F. poae, which was found in 95.2% of the varieties. For F. graminearum, the highest prevalence was found between 2020 and 2021, when the infestation of triticale samples was 31.4% and 91.4%, respectively. F. culmorum had the lowest percentage of infestation, being found in only 20% of all samples. In addition, the winter triticale varieties of the 2022 harvest K-1–19#14 K-3851 PRAG502 (Dagestan) and K-1–19#42 K3727 SNT-5/92 (Omsk region) were identified as the least susceptible to infestation by the studied species of fungi.

1. Golikov N.N. Castor, resistant to Fusarium. Plant Protection and Quarantine. 2003;3:44. (In Russ.)

2. Ivashchenko V.G. Fusarium graminearum Schwabe geographical distribution and features of bioecology. Mycology and Phytopathology. 1998;32(5):1–10. (In Russ.)

3. Kosterina N.A. Analysis of the current state of the problem of fusarium ear and grain of wheat in the Russian Federation. Agrarian Bulletin of the Urals. 2023;05(234):49–59. (In Russ.)

4. Kurilova D.A. Harmfulness of Fusarium species on soybean depending on a degree of plants affection. Oil Crops. 2010;2:144–145. (In Russ.)

5. Phytopathology. Diseases of tree and shrub species: a textbook. Ed. by I.I. Minkevich. St. Petersburg, Russia: Izdatel’stvo Lan’, 2011:160. (In Russ.)

6. Monastyrskii O.A. Mycotoxins – a global problem of food and feed safety. Agrohimia. 2016;6:67–71. (In Russ.)

7. Platonova Yu.V., Surin N.A. Geography of fungi of the genus Fusarium (literary review). Fundamental’nye issledovaniya. 2004; 4: 95–97. (In Russ.)

8. Popov V.S., Samburov N.V., Vorob’eva N.V. The problem of mycotoxicoses in modern conditions and principles of preventive solutions: monografiya. Kursk, Russia: Planeta+, 2018:158. (In Russ.)

9. Popova S.A., Skoptsova T.I., Losyakova E.V. Mycotoxins in feed: causes, consequences, prevention. Izvestiya Velikolukskoy gosudarstvennoy sel’skokhozyaystvennoy akademii. 2017;1:16–23. (In Russ.)

10. Sokolova G.D. Intraspecific diversity of phytopathogenic fungus Fusarium graminearum. Mycology and Phytopathology. 2015;2(49):71–79. (In Russ.)

11. Toropova E.Yu., Selyuk M.P., Kazakova O.A. Dominance factors of fungi genus fusarium in grain crops root rot pathocomplex. Agrohimia. 2018;5:73–82. (In Russ.)

12. Brennan J., Fagan B., Van Maanen A. et al. Studies on in vitro growth and pathogenicity of European Fusarium fungi. European Journal of Plant Pathology. 2003;109(6): 577–587.

13. Cirlini M., Generotti S., Dall’Erta A. et al. Durum Wheat (Triticum Durum Desf.) Lines Show Different Abilities to Form Masked Mycotoxins under Greenhouse Conditions. Toxins. 2014;6(1):81–95.

14. Ekwomadu T.I., Mwanza M. Fusarium Fungi Pathogens, Identification, Adverse Effects, Disease Management, and Global Food Security: A Review of the Latest Research. Agriculture. 2023;13(9):1810.

15. Gil–Serna J., Vázquez C., González–Jaén M.T. et al. Mycotoxins: Toxicology. Encyclopedia of Food Microbiology. 2014:887–892.

16. Matny O., Bates S., Song Z. Geographic distribution of Fusarium culmorum chemotypes associated with wheat crown rot in Iraq. Journal of Plant Protection Research. 2017;57(1).

17. Parry D., Jenkinson P., Mcleod L. Fusarium ear blight (scab) in small grain cereals: a review. Plant Pathology. 1995;44:207–238.

18. Rassat A. Fusarium poae and Fusarium langsethiae in an oat field–time point of infection and possible inoculum sources. Biology, Environmental Science, Agricultural and Food Sciences. 2019.

19. Töth A., Barna–Vetrö I., Gyöngyösi Ä. et al. Prevalence and mycotoxin production of Fusarium species isolated from wheat grains in Hungary. Acta Phytopath. Entomol. Hung. 1993;28: 3–12.

20. Wagacha J., Muthomi J. Fusarium culmorum: Infection process, mechanisms of mycotoxin production and their role in pathogenesis in wheat. Crop Protection. 2007;26(7):877–885.

21. Wilcoxson R., Kommedahl T., Ozmon E. et al. Occurrence of Fusarium species in scabby wheat from Minnesota and their pathogenicity to wheat. Phytopathology. 1988;78:586–589.

22. Xu X., Nicholson P., Thomsett M. et al. Relationship between the fungal complex causing Fusarium head blight of wheat and environmental conditions. Phytopathology. 2008;98(1):69–78.

23. Yli–Mattila T., Opoku J., Ward T.J. Population structure and genetic diversity of Fusarium graminearum from southwestern Russia and the Russian Far East as compared with northern Europe and North America. Mycologia. 2023:1–11.