European Journal of Plant Pathology

, Volume 149, Issue 4, pp 895–910 | Cite as

Molecular and experimental evidence of multi-resistance of Cercospora beticola field populations to MBC, DMI and QoI fungicides

  • Nenad R. TrkuljaEmail author
  • Anja G. Milosavljević
  • Milana S. Mitrović
  • Jelena B. Jović
  • Ivo T. Toševski
  • Mohamed F. R. Khan
  • Gary A. Secor


Cercospora leaf spot (CLS) caused by Cercospora beticola occurs annually in Serbia causing severe yield losses of sugar beet, which requires intensive use of fungicides. In recent years we have observed unsatisfactory control of CLS originating from northwestern Serbia. Frequency of C. beticola populations resistant to Quinone outside inhibitors (QoI) was 81% (51/63 isolates), 98% (62/63) to sterol-demethylation inbibitors (DMI) and 54% (34/63) to methyl-2-benzimidazole carbamate fungicides (MBC). The genetic basis underlying the resistance was tested by characterizing the cob, CYP51 and ß-tubulin genes, associated with resistance to QoI, DMI and MBC fungicides, respectively. Isolates that were resistant to QoI fungicides had the G143A mutation in the cob gene. Characterization of the CYP51 gene revealed seven diverse haplotypes; however, no correlation with sensitivity or resistance to DMI fungicides could be identified. Resistance to MBC fungicides was associated with the presence of the E198A mutation in the ß-tubulin gene of all resistant isolates. From a total of 63 isolates originating from sugar beet fields of northwestern Serbia, 62 isolates showed resistance to multiple modes of action. Three multi-resistant phenotypes were identified: MR1 (N = 29) - resistant to QoI and DMI fungicides (QoI-R and DMI-R) but sensitive to MBC fungicides (MBC-S); MR2 (N = 11, QoI-S, DMI-R and MBC-R); and MR3 (N = 22), resistant to all three groups of fungicides (QoI-R, DMI-R and MBC-R). This is the first report of C. beticola resistance to QoI fungicides in Serbia. This study revealed development of multi-resistance of C. beticola isolates to MBC, DMI and QoI fungicides, which represents the first record of this phenomenon in C. beticola populations.


Multi-resistance Cercospora beticola Molecular characterization CYP51 ß-tubulin Cob 



This research has been funded by the national projects III43001 and TR31018 financed by the Ministry of Education, Science and Technological development of Serbia.


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Copyright information

© Koninklijke Nederlandse Planteziektenkundige Vereniging 2017

Authors and Affiliations

  • Nenad R. Trkulja
    • 1
    Email author
  • Anja G. Milosavljević
    • 1
  • Milana S. Mitrović
    • 2
  • Jelena B. Jović
    • 2
  • Ivo T. Toševski
    • 2
    • 3
  • Mohamed F. R. Khan
    • 4
    • 5
  • Gary A. Secor
    • 4
  1. 1.Department of Plant DiseaseInstitute for Plant Protection and EnvironmentBelgradeSerbia
  2. 2.Department of Plant PestsInstitute for Plant Protection and EnvironmentZemunSerbia
  3. 3.CABIDelémontSwitzerland
  4. 4.Department of Plant PathologyNorth Dakota State UniversityFargoUSA
  5. 5.University of MinnesotaSt. PaulUSA

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