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Characterisation of benzimidazole resistance of Cercospora beticola in Serbia using PCR-based detection of resistance-associated mutations of the beta-tubulin gene

Само за регистроване кориснике
2013
Аутори
Trkulja, Nenad
Ivanović, Žarko
Pfaf-Dolovac, Erika
Dolovac, Nenad
Mitrović, Milana
Toševski, Ivo
Jović, Jelena
Чланак у часопису (Објављена верзија)
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Апстракт
A survey to detect and characterise benzimidazole resistance within populations of Cercospora beticola in Serbia was performed. From 52 field isolates collected from sugar beet and beet root, only eight were found to be benzimidazole-sensitive based on the inhibition of mycelial growth by discriminatory concentrations of carbendazim and thiophanate-methyl. Sensitivity tests revealed the presence of three resistant phenotypes among the tested isolates: high-resistance (HR), low-resistance (LR) and moderate-resistance (MR). The benzimidazole resistant isolates were characterised based on the DNA sequence of the beta-tubulin gene and temperature sensitivity. The HR isolates showed no temperature sensitivity regardless of carbendazim concentration, whereas the LR and MR isolates were sensitive at lower temperatures. Analysis of the beta-tubulin gene sequence revealed two amino acid replacements in the benzimidazole-resistant isolates of C. beticola. One was a glutamic acid to alanine chang...e at position 198 (codon GAG to GCG) that was identified in HR isolates; this mutation has previously been reported to be associated with the development of benzimidazole resistance in C. beticola. The second replacement was a novel point mutation of phenylalanine (TTC) to tyrosine (TAC) at position 167, identified in low and moderate benzimidazole-resistant isolates, sharing a single LR/MR beta-tubulin genotype. A diagnostic PCR-RFLP assay utilising a BsaI restriction site present in the benzimidazole sensitive and LR/MR genotypes but absent in the HR genotype was developed for the routine detection of high resistance. A mutation-specific PCR assay was developed for the diagnosis of LR/MR genotype based on a mutation from T to A at codon 167, which is unique to this genotype.

Кључне речи:
Cercospora leaf spot / Novel point mutation Phe167Tyr / PCR-RFLP diagnostics / PCR mutation-specific diagnostic assay / Sugar beet / Temperature sensitivity
Извор:
European Journal of Plant Pathology, 2013, 135, 4, 889-902
Издавач:
  • Springer, Dordrecht
Пројекти:
  • Разрада интегрисаног управљања и примене савремених принципа сузбијања штетних организама у заштити биља (RS-31018)
  • Агробиодиверзитет и коришћење земљишта у Србији: интегрисана процена биодиверзитета кључних група артропода и биљних патогена (RS-43001)

DOI: 10.1007/s10658-012-0135-x

ISSN: 0929-1873

WoS: 000315601500025

Scopus: 2-s2.0-84874569380
[ Google Scholar ]
15
13
URI
http://plantarum.izbis.bg.ac.rs/handle/123456789/272
Колекције
  • Radovi istraživača / Researchers' publications
Институција
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