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Current approaches to identification of Fusarium fungi infecting wheat

Karelov A.V., Borzykh O.I., Kozub N.O., Sozinov I.O., Yanse L.A., Sozinova O.I., Tkalenko H.M., Mishchenko L.T., Blume Ya.B.


SUMMARY. Fungi of the genus Fusarium are especially dangerous phytopathogens affecting common wheat (Triticum aestivum L.) among other crops, as they may cause not only crop losses but poisoning of humans and livestock. The review highlights current approaches to identify fungi of the genus Fusarium infecting common wheat. Microbiological techniques for identification of Fusa-rium species are still among laboratory protocols and recommendations, therefore some of the most popular genus- and species-specific media are mentioned in the review. However, in the modern literature, much more attention is paid to identification of Fusarium fungi with the use of the polymerase chain reaction (PCR). Therefore conventional PCR assays for identification of representatives of the genus Fusarium in general or only species producing especially dangerous metabolites (nivalenol, deoxynivalenol, 3-acetyldeoxynivalenol, 4-acetyl-nivalenol, enniatin) are highlighted in the review. The primer pairs to identify the presence of certain Fusarium species or their combinations in samples are described. For real-time PCR assays, which may be used for more precise quantitative and qualitative genus- and species-specific identification of Fusarium fungi, protocol details, primer and probe sequences are described, as well as recommended dyes are mentioned for the probes. For some primer pairs, additional details regarding their validation and the assay sensitivity are mentioned. Thus, the techniques described in the review are precise and comprehensive enough and may be used in combinations and separately for genus- and species-specific quantitative or qualitative identification of the fungi of the genus Fusarium.

Key words: Fusarium, wheat, Fusarium head blight, pink snow mold, molecular markers

Tsitologiya i Genetika 2021, vol. 55, no. 5, pp. 36-52

  1. Institute of Plant Protection of the National Academy of Agrarian Sciences of Ukraine, 03022, 33 Vasykivska St., Kyiv
  2. Institute of Food Biotechnology and Genomics of the National Academy of Sciences of Ukraine, 04123, 2a Osypovskogo St., Kyiv
  3. Education and Scientific Center Institute of Biology and Medicine of Taras Shevchenko National University of Kyiv, 01601, 64/13 Volodymyrska St., Kyiv

E-mail: hromogenblack, natalkozub, blume.yaroslav, lmishchenko

Karelov A.V., Borzykh O.I., Kozub N.O., Sozinov I.O., Yanse L.A., Sozinova O.I., Tkalenko H.M., Mishchenko L.T., Blume Ya.B. Current approaches to identification of Fusarium fungi infecting wheat, Tsitol Genet., 2021, vol. 55, no. 5, pp. 36-52.

In "Cytology and Genetics":
A. V. Karelov, O. I. Borzykh, N. O. Kozub, I. O. Sozinov, L. A. Yanse, O. I. Sozinova, H. M. Tkalenko, L. T. Mishchenko & Ya. B. Blume Current Approaches to Identification of Fusarium Fungi Infecting Wheat, Cytol Genet., 2021, vol. 55, no. 5, pp. 433446
DOI: 10.3103/S0095452721050030


1. Abass, M.H., Madhi, Q.H., and Matrood, A.A.A., Identity and prevalence of wheat damping-off fungal pathogens in different fields of Basrah and Maysan provinces, Bull. Natl. Res. Cent., 2021, vol. 45, p. 51.

2. Amato, B., Pfohl, K., Tonti, S., et al., Fusarium proliferatum and fumonisin B1 co-occur with Fusarium species causing Fusarium Head Blight in durum wheat in Italy. JABFQ 88:288233.

3. Anderson, M.G. and Atkinson, R.G., Comparison of media for the isolation of Fusarium oxysporum f. sp lycopersici from sawdust used for growing tomatoes, Can. J. Plant Sci., 1974, vol. 54, no. 2, pp. 373374.

4. Aoki, T. and ODonnell, K., Morphological and molecular characterization of Fusarium pseudograminearum recognized as the Group 1 population of F. graminearum, Mycologia, 1999, vol. 91, no. 4, pp. 597609. doi 10.1080/00275514.1999.12061058

5. Arif, M., Chawla, S., Zaidi, N.W., et al., Development of specific primers for genus Fusarium and F. solani using rDNA sub-unit and transcription elongation factor (TEF-1α) gene, Afr. J. Biotechnol., 2012, vol. 11, no. 2, pp. 444447.

6. Birr, T., Hasler, M., Verreet, J.A., et al., Composition and predominance of Fusarium species causing Fu-sarium head blight in winter wheat grain depending on cultivar susceptibility and meteorological factors, Microorganisms, 2020, vol. 8, no. 4, p. 617.

7. Bluhm, B.H., Flaherty, J.E., Cousin, M.A., et al., Multiplex polymerase chain reaction assay for the differential detection of trichothecene- and fumonisin-producing species of Fusarium in cornmeal, J. Food Prot., 2002, vol. 65, no. 12, pp. 19551961.

8. Brandfass, C. and Karlovsky, P., Simultaneous detection of Fusarium culmorum and F. graminearum in plant material by duplex PCR with melting curve analysis, BMC Microbiol., 2006, vol. 6, p. 4.

9. Casasnovas, F., Fantini, E.N., Palazzini, J.M., et al., Development of amplified fragment length polymorphism (AFLP)-derived specific primer for the detection of Fusarium solani aetiological agent of peanut brown root rot, J. Appl. Microbiol., 2013, vol. 114, no. 6, pp. 17821792.

10. Castanares, E., Albuquerque, D.R., Dinolfo, M.I., et al., Trichothecene genotypes and production profiles of Fusarium graminearum isolates obtained from barley cultivated in Argentina, Int. J. Food Microbiol., 2014, vol. 179, pp. 5763.

11. Covarelli, L., Beccari, G., and Salvi, S., Infection by mycotoxigenic fungal species and mycotoxin contamination of maize grain in Umbria, central Italy, Food Chem. Toxicol., 2011, vol. 49, pp. 236 52369.

12. Demeke, T., Clear, R.M., Patrick, S.K., et al., Species-specific PCR-based assays for the detection of Fusarium species and a comparison with the whole seed agar plate method and trichothecene analysis, Int. J. Food Microbiol., 2005, vol. 103, no. 3, pp. 271284.

13. Deng, Y.Y., Li, W., Zhang, P., et al., Fusarium pseudograminearum as an emerging pathogen of crown rot of wheat in eastern China, Plant Pathol., 2020, vol. 69, pp. 240248.

14. Desmond, O.J., Manners, J.M., Stephens, A.E., et al., The Fusarium mycotoxin deoxynivalenol elicits hydrogen peroxide production; programmed cell death and defence responses in wheat, Mol. Plant Pathol., 2008, vol. 9, no. 4, pp. 435445.

15. Doohan, F.M., Parry, D.W., Jenkinson, P., et al., The use of species-specific PCR-based assays to analyze Fusarium ear blight of wheat, Plant Pathol., 1998, vol. 47, pp. 197205.

16. Elbelt, S., Siou, D., Gelisse, S., et al., Optimized real-time qPCR assays for detecting and quantifying the Fusarium and Microdochium species responsible for wheat head blight, as defined by MIQE guidelines, bioRxiv, 2018, art. 272534.

17. Faria, C.B., Abe, C.A., da Silva, C.N., et al., New PCR assays for the identification of Fusarium verticillioides, Fusarium subglutinans, and other species of the Gibberella fujikuroi complex, Int. J. Mol. Sci., 2012, vol. 13, no. 1, pp. 115132.

18. Fisher, N.L., Burgess, L.W., Toussoun, T.A., et al., Carnation leaves as a substrate and for preserving cultures of Fusarium species, Phytopathology, 1982, vol. 72, no. 1, pp. 151155.

19. Glazebrook, J., Contrasting mechanisms of defense against biotrophic and necrotrophic pathogens, Annu. Rev. Phytopathol., 2005, vol. 43, pp. 205227.

20. Góral, T., Wisniewska, H., Ochodzki, P., et al., Relationship between Fusarium head blight, kernel damage, concentration of Fusarium biomass, and Fusarium toxins in grain of winter wheat inoculated with Fusarium culmorum, Toxins, 2018, vol. 11, no. 1, p. 2. doi 10.3390/toxins11010002

21. Halstensen, A.S., Nordby, K.C., Eduard, W., et al., Real-time PCR detection of toxigenic Fusarium in airborne and settled grain dust and associations with trichothecene mycotoxins, J. Environ. Monit., 2006, vol. 8, no. 12, pp. 12351241.

22. Hayashi, Y., Kozawa, T., Aiuchi, D., et al., A selective medium to isolate airborne spores of Microdochium nivale, causing winter wheat scab, Eur. J. Plant Pathol., 2014, vol. 138, pp. 247256.

23. Hrytsev, O.A., Zozulya, O.L., Vorobiova, N.G., et al., Monitoring of species composition of fungi of the genus Fusarium in seed materials of winter wheat on Ukrainian territory, Micribiol. Biotechnol., 2018, vol. 2, pp. 8189.

24. Johnson, D.D., Flaskerud, G.K., Taylor, R.D., et al., Fusarium head blight of wheat and barley, in Quantifying Economic Impacts of Fusarium Head Blight in Wheat, Leonard, K.J. and Bushnell, W.R., Eds., St. Paul, USA: APS Press, 2003, pp. 461483.

25. Jung, B., Lee, S., Ha, J., et al., Development of a selective medium for the fungal pathogen Fusarium graminearum using toxoflavin produced by the bacterial pathogen Burkholderia glumae, Plant Pathol. J., 2013, vol. 29, no. 4, pp. 446450.

26. Jurado, M., Vazquez, C., Patico, B., et al., PCR detection assays for the trichothecene-producing species Fusarium graminearum, Fusarium culmorum, Fusarium poae, Fusarium equiseti and Fusarium sporotrichioides, Syst. Appl. Microbiol., 2005, vol. 28, pp. 562568.

27. Jurado, M., Vázquez, C., Marín, S., et al., PCR-based strategy to detect contamination with mycotoxigenic Fusarium species in maize, Syst. Appl. Microbiol., 2006, vol. 29, no. 8, pp. 681689.

28. Kazan, K. and Gardiner, D.M., Fusarium crown rot caused by Fusarium pseudograminearum in cereal crops: recent progress and future prospects, Mol. Plant Pathol., 2018, vol. 19, no. 7, pp. 15471562.

29. Kerenyi, Z., Moretti, A., Waalwijk, C., et al., Mating type sequences in asexually reproducing Fusarium species, Appl. Environ. Microbiol., 2004, vol. 70, pp. 44194423.

30. Khaledi, N., Taheri, P., and Falahati, R.M., Identification, virulence factors characterization, pathogenicity and aggressiveness analysis of Fusarium spp., causing wheat head blight in Iran, Eur. J. Plant Pathol., 2017, vol. 147, pp. 897918.

31. Komada, H., A new selective medium for isolating Fusarium from natural soil, Proc. Am. Phytopathol. Soc., 1976, vol. 3, p. 221.

32. Konstantinova, P. and Yli-Mattila, T., IGS-RFLP analysis and development of molecular markers for identification of Fusarium poae, Fusarium langsethiae, Fusarium sporotrichioides and Fusarium kyushuense, Int. J. Food Microbiol., 2004, vol. 95, pp. 321331.

33. Kovalyshyna, H.M., Murashko, L.A., and Kovalyshyn, A.B., Spike diseases of winter wheat from the Forest-Steppe of Ukraine, Bull. Ukr. Soc. Genet. Breed., 2008, vol. 6, no. 2, pp. 223239.

34. Krnjaja, V., Stanković, S., Obradović, A., et al., Trichothecene genotypes of Fusarium graminearum populations isolated from winter wheat crops in Serbia, Toxins, 2018, vol. 10, no. 11, p. 460.

35. Kulik, T., Detection of Fusarium tricinctum from cereal grain using PCR assay, J. Appl. Genet., 2008, vol. 49, pp. 305311.

36. Kulik, T., Fordonski, G., Pszczylkowska, A., et al., Development of PCR assay based on ITS2 rDNA polymorphism for the detection and differentiation of Fusarium sporotrichioides, FEMS Microbiol. Lett., 2004, vol. 239, pp. 181186.

37. Kuzdraliński, A., Kot, A., Szczerba, H., et al., A review of conventional PCR assays for the detection of selected phytopathogens of wheat, J. Mol. Microbiol. Biotechnol., 2017a, vol. 27, pp. 175189.

38. Kuzdraliński, A., Nowak, M., Szczerba, H., et al., The composition of Fusarium species in wheat husks and grains in south-eastern Poland, J. Integr. Agric., 2017b, vol. 16, no. 7, pp. 15301536. 52-6

39. Kyslukh, T.M. and Shevchuk, O.V., Harmfulness of the main pathogens of Fusarium head blight of winter wheat in the Forest-Steppe zone of Ukraine, Bull. Agricult. Sci., 2006, vol. 1, pp. 1618.

40. Leslie, J.F., Summerel, B.A., and Bullock, S., The Fusarium Laboratory Manual, Wiley, 2006. ISBN 0813819199, 9780813819198

41. Ma, H., Zhang, X., Yao, J., et al., Breeding for the resistance to Fusarium head blight of wheat in China, Front. Agr. Sci. Eng., 2019, vol. 6, no. 3, pp. 251264. doi 10.15302/J-FASE-2019262

42. Martinez, M., Castanares, E., Dinolfo, M.I., et al., Presencia de Fusarium graminearum en muestras de trigo destinado al consumo humano, Rev. Argent. Microbiol., 2014, vol. 46, no. 1, pp. 4144.

43. Miller, J.D., Greenhalgh, R., Wang, Y., et al., Trichothecene chemotypes of three Fusarium species, Mycologia, 1991, vol. 83, pp. 121130.

44. Minati, M.H. and Mohammed-Ameen, M.K., Novel report on six Fusarium species associated with head blight and crown rot of wheat in Basra province, Iraq. Bull. Natl. Res. Cent., 2019, vol. 43, p. 139.

45. Mishra, P.K., Fox, R.T., and Culham, A., Development of a PCR-based assay for rapid and reliable identification of pathogenic Fusaria, FEMS Microbiol. Lett., 2003, vol. 218, no. 2, pp. 329332. tb11537.x

46. Möller, E.M., Chelkowski, J., and Geiger, H.H., Species-specific PCR assays for the fungal pathogens Fusarium moniliforme and Fusarium subglutinans and their application to diagnose maize ear rot disease, J. Phytopathol., 1999, vol. 147, pp. 497508.

47. Mulè, G., Susca, A., Stea, G., et al., A species-specific PCR assay based on the calmodulin partial gene for identification of Fusarium verticillioides, F. proliferatum and F. subglutinans, Eur. J. Plant Pathol., 2004, vol. 110, pp. 495 502.

48. Nicholson, P. and Parry, D.W., Development of a PCR assay to detect Fusarium poae in wheat, Plant Pathol., 1996, vol. 45, pp. 872883.

49. Nicholson, P., Simpson, D.R., Weston, G., et al., Detection and quantification of Fusarium culmorum and Fusarium graminearum in cereals using PCR assays, Physiol. Mol. Plant Pathol., 1998, vol. 53, pp. 1737.

50. Nicholson, P., Lees, A.K., Maurin, N., et al., Development of a PCR assay to identify and quantify Microdochium nivale var. nivale and Microdochium nivale var. majus in wheat, Physiol. Mol. Plant Pathol., 1996, vol. 48, pp. 257271.

51. Nicholson, P., Simpson, D., Wilson, A.H., et al., Detection and differentiation of trichothecene and enniatin-producing Fusarium species on small-grain cereals, Eur. J. Plant Pathol., 2004, vol. 110, pp. 503514.

52. Nicolaisen, M., Suproniene, S., Nielsen, L.K., et al., Real-time PCR for quantification of eleven individual Fusarium species in cereals, J. Microbiol. Methods, 2009, vol. 76, no. 3, pp. 234240.

53. Niessen, L., Schmidt, H., and Vogel, R.F., The use of tri5 gene-sequences for PCR detection and taxonomy of trichothecene-producing species in the Fusarium section Sporotrichiella, Int. J. Food Microbiol., 2004, vol. 95, pp. 305319.

54. Nishimura, N., Selective media for Fusarium oxysporum, J. Gen. Plant Pathol., 2007, vol. 73, pp. 342348.

55. Papavizas, G.C., Evaluation of various media and antimicrobial agents for isolation of Fusarium from soil, Phytopathology, 1967, vol. 57, pp. 848852.

56. Patino, B., Mirete, S., González-Jaén, M.T., et al., PCR detection assay of fumonisin-producing Fusarium verticillioides strains, J. Food Prot., 2004, vol. 67, no. 6, pp. 12781283.

57. Pollard, A.T. and Okubara, P.A., Real-time PCR quantification of Fusarium avenaceum in soil and seeds, J. Microbiol. Methods, 2019, vol. 157, pp. 2130. 2018.12.009

58. Puhall, J., Classification of strains of Fusarium oxysporum on the basis of vegetative compatibility, Can. J. Bot., 1985, vol. 63, pp. 179183.

59. Quarta, A., Mita, G., Haidukowski, M., et al., Assessment of trichothecene chemotypes of Fusarium culmorum occurring in Europe, Food Additives Contam., 2005, vol. 22, no. 4, pp. 309315.

60. Quarta, A., Mita, G., Haidukowski, M., et al., Multiplex PCR assay for the identification of nivalenol, 3-and 15-acetyl-deoxynivalenol chemotypes in Fusarium, FEMS Microbiol. Lett., 2006, vol. 259, no. 1, pp. 713.

61. Ramdass, A.C., Villafana, R.T., and Rampersad, S.N., TRI genotyping and chemotyping: a balance of power, Toxins, 2020, vol. 12, p. 64.

62. Reischer, G.H., Lemmens, M., Farnleitner, A., et al., Quantification of Fusarium graminearum in infected wheat by species specific real-time PCR applying a TaqMan Probe, J. Microbiol. Methods, 2004, vol. 59, no. 1, pp. 141146.

63. Rossi, V., Terzi, V., Moggi, F., et al., Assessment of Fusarium infection in wheat heads using a quantitative PCR assay, Food Addit. Contam., 2007, vol. 24, no. 10, pp. 11211130.

64. Sadhasivam, S., Britzi, M., Zakin, V., et al., Rapid detection and identification of mycotoxigenic fungi and mycotoxins in stored wheat grain, Toxins, 2017, vol. 9, no. 10, p. 302.

65. Salgado, J.D., Madden, L.V., and Paul, P.A., Quantifying the effects of Fusarium head blight on grain yield and test weight in soft red winter wheat, Phytopathology, 2015, vol. 105, no. 3, pp. 295306.

66. Sanoubar, R., Bauer, A., and Seigner, L., Detection, identification and quantification of Fusarium graminearum and Fusarium culmorum in wheat kernels by PCR techniques, J. Plant Pathol. Microbiol., 2015, vol. 6, p. 287.

67. Schilling, A.G., Möller, E.M., and Geiger, H.H., Polymerase chain reaction-based assays for species-specific detection of Fusarium culmorum, F. graminearum and F. avenaceum, Mol. Plant Pathol., 1996, vol. 86, pp. 515522.

68. Segalin, M. and Reis, E.M., Semi-selective medium for Fusarium graminearum detection in seed samples, Summa Phytopathol., 2010, vol. 36, no. 4, pp. 338341.

69. Shikur Gebremariam, E., Sharma-Poudyal, D., Paulitz, T.C., et al., Identity and pathogenicity of Fusarium species associated with crown rot on wheat (Triticum spp.) in Turkey, Eur. J. Plant Pathol., 2018, vol. 150, pp. 387399.

70. Snijders, C.H.A. and Perkowski, J., Effects of head blight caused by Fusarium culmorum on toxin content and weight of wheat kernels, Phytopathology, 1990, vol. 79, pp. 455469.

71. Steenkamp, E.T., Wingfield, B.D., Coutinho, T.A., et al., PCR-based identification of MAT-1 and MAT-2 in the Gibberella fujikuroi species complex, Appl. Environ. Microbiol., 2000, vol. 66, pp. 43784382.

72. Terzi, V., Morcia, C., Faccioli, P., et al., Fusarium DNA traceability along the bread production chain, Int. J. Food Sci. Technol., 2007, vol. 42, pp. 13901396.

73. Thrane, U., Comparison of three selective media for detecting Fusarium species in foods: a collaborative study, Int. J. Food Microbiol., 1996, vol. 29, no. 23, pp. 149156.

74. Turner, A.S., Lees, A.K., Rezanoor, H.N., et al., Refinement of PCR-detection of Fusarium avenaceum and evidence from DNA marker studies for phonetic relatedness to Fusarium tricinctum, Plant Pathol., 1998, vol. 47, pp. 278288.

75. Waalwijk, C., Kastelein, P., de Vries, Ph.M., et al., Major changes in Fusarium spp. in wheat in the Netherlands, Eur. J. Plant Pathol., 2003, vol. 109, pp. 743754.

76. Waalwijk, C., van der Heide, R., de Vries, I., et al., Quantitative detection of Fusarium species in wheat using TaqMan, Eur. J. Plant Pathol., 2004, vol. 110, pp. 481494.

77. Wang, C.L. and Cheng, Y.H., Identification and trichothecene genotypes of Fusarium graminearum species complex from wheat in Taiwan, Bot. Stud., 2017, vol. 58, p. 4.

78. Ward, T.J., Clear, R.M., Rooney, A.P., et al., An adaptive evolutionary shift in Fusarium head blight pathogen populations is driving the rapid spread of more toxigenic Fusarium graminearum in North America, Fungal Genet. Biol., 2008, vol. 45, no. 4, pp. 473484.

79. Wilson, A., Simpson, D., Chandler, E., et al., Development of PCR assays for the detection and differentiation of Fusarium sporotrichioides and Fusarium langsethiae, FEMS Microbiol. Lett., 2004, vol. 233, no. 1, pp. 6976.

80. Yli-Mattila, T., Mach, R., Alekhina, I.A., et al., Phylogenetic relationship of Fusarium langsethiae to Fusarium poae and F. sporotrichioides as inferred by IGS, ITS, β-tubulin sequence and UP-PCR hybridization analysis, Int. J. Food Microbiol., 2004, vol. 95, pp. 267285.

81. Yli-Mattila, T., Paavanen-Huhtala, S., Jestoi, M., et al., Real-time PCR detection and quantification of Fusarium poae, F. graminearum, F. sporotrichioides and F. langsethiae in cereal grains in Finland and Russia, Arch. Phytopathol. Pflanzenschutz., 2008, vol. 41, no. 4, pp. 243260.

82. Yoder, W.T. and Christianson, L.M., Species-specific primers resolve members of Fusarium section Fusarium. Taxonomic status of the edible Quorn fungus reevaluated, Fungal Genet. Biol., 1998, vol. 23, no. 1, pp. 6880.

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