The Point Mutation G461S in the <i>MfCYP51</i> Gene is Associated with Tebuconazole Resistance in <i>Monilinia fructicola</i> Populations in Brazil

Lichtemberg, Paulo S. F.; Luo, Yong; Morales, Rafael Gustavo Ferreira; Muehlmann-Fischer, Juliana M.; Michailides, Themis J.; Mio, Louise Larissa May De

doi:10.1094/phyto-02-17-0050-r

Cited by 49 publications

(35 citation statements)

References 55 publications

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“…Many point mutations have been reported in CYP51 of various plant pathogens, among which the mutation at AA136 conferring the change from tyrosine to phenylalanine (Y136F) is the most common. [24][25][26][27][28] However, only a mutation at AA511 conferring the change of arginine to tryptophan (R511W) was found in one of our resistant isolates, indicating other mechanisms were responsible for the development of DMI resistance. It is surprising that no Alternaria spp.…”

Section: Discussionmentioning

confidence: 99%

Induced expression of CYP51 associated with difenoconazole resistance in the pathogenic Alternaria sect. on potato in China

Zhang

Zhou

Tian

et al. 2019

Pest Management Science

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BACKGROUND Early blight caused by Alternaria spp. is amongst the most important diseases in potato. Demethylation inhibitor (DMI) fungicides are widely used to control the disease but long‐term use may decrease its control efficacy due to fungicide resistance. This study investigated the occurrence of difenoconazole resistance in Alternaria spp. and molecular resistant mechanisms. RESULTS EC50 values of 160 isolates to difenoconazole ranged from 0.026 μg mL−1 to 15.506 μg mL−1 and the frequency of difenoconazole sensitivity formed a non‐normal distribution curve with a major and a minor peak. Isolates with EC50 values of 4.121 and 5.461 μg mL−1 were not controlled effectively at fungicide doses of 50 and 100 μg mL−1. Cross‐resistance was observed between DMI fungicides difenoconazole and propiconazole, but not between difenoconazole and other fungicide groups, including boscalid, iprodione, or carbendazim. The CYP51gene was 1673 bp encoding 525 amino acids in length and contained two introns. All sensitive and resistant isolates had the identical amino acid sequence of CYP51, with the exception of one resistant isolate carrying a mutation of R511W. A 6 bp insertion in the upstream region was observed in half of the resistant isolates. In the absence of propiconazole, the relative expression of CYP51 was not significantly different in sensitive and resistant isolates. In the presence of difenoconazole, expression of CYP51 gene was induced significantly in the DMI‐resistant isolates but not in the sensitive ones. CONCLUSION Induced expression of CYP51 in resistant isolates exposed to difenoconazole is an important determinant for DMI resistance in potato pathogens Alternaria sect. © 2019 Society of Chemical Industry

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Section: Discussionmentioning

confidence: 99%

Induced expression of CYP51 associated with difenoconazole resistance in the pathogenic Alternaria sect. on potato in China

Zhang

Zhou

Tian

et al. 2019

Pest Management Science

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“…PCR was conducted using DNA from M. fructicola isolates. The primers MfgaF1/MfgaR1 (Lichtemberg et al , ) were used to identify the occurrence of the G461S mutation in the MfCYP51 gene. PCR conditions were the same as those described by Lichtemberg et al ().…”

Section: Methodsmentioning

confidence: 99%

“…The primers MfgaF1/MfgaR1 (Lichtemberg et al , ) were used to identify the occurrence of the G461S mutation in the MfCYP51 gene. PCR conditions were the same as those described by Lichtemberg et al (). After PCR, the products were applied to 1.5% agarose gel in Tris‐borate‐EDTA (TBE) buffer, stained with GelRed, and the DNA fragments visualized on a UV transilluminator and photographed.…”

Section: Methodsmentioning

confidence: 99%

“…Reduction in the sensitivity of M. fructicola isolates has been associated with a mutation at codon position 136 (Y136F) that changes the amino acid from tyrosine to phenylalanine in the MfCYP51 protein (Chen et al , ); overexpression of the MfCYP51 gene encoding 14α‐demethylase (Luo & Schnabel, ); and overexpression of the MfABC1 gene (Schnabel et al , ). In Brazil, the sensitivity reduction of M. fructicola isolates was associated with a mutation at codon 461 in the MfCYP51 gene (G461S), which causes glycine to be replaced by serine (Lichtemberg et al , ). Although the G461S mutation has been identified, its frequency in the M. fructicola population in Brazil is still unknown.…”

Section: Introductionmentioning

confidence: 99%

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Discontinuance of tebuconazole in the field restores sensitivity of Monilinia fructicola in stone fruit orchards

et al. 2019

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Demethylation inhibitor (DMI) fungicides are used to control brown rot in stone fruit worldwide. However, their specific mode of action can select resistant isolates of Monilinia fructicola. Monilinia fructicola resistant to DMI fungicides are associated with a fitness cost in the absence of selective pressure, indicating that the sensitive population can be re‐established when discontinuing the fungicide in the field. This work aimed to build up the sensitive population of M. fructicola after discontinuing the use of tebuconazole for successive crop seasons. The sensitivity of M. fructicola to tebuconazole was assessed in four commercial peach orchards in Paraná and São Paulo States from 2012/13 to 2015/16. Different fungicide programmes were used and DMI fungicides were discontinued from 2013/14. The sensitivity of M. fructicola to tebuconazole was assessed by a mycelial growth assay in vitro and by determining the frequency of the G461S mutation in the MfCYP51 gene. The isolates from Paraná had high sensitivity to the fungicide across all seasons and the frequency of the G461S mutation remained below 5%. The isolates from São Paulo were highly resistant in the 2012/13 season; however, there was a gradual decline until 2015/16. In addition, the G461S mutation frequency in Sao Paulo State was about 80% in the 2012/13 season, but reduced until it was completely undetectable in 2015/16. These results provide evidence that resistance can be managed in orchards with high selective pressure to tebuconazole after discontinuing the use of the fungicide for at least 3 years.

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“…‘Mona’‐associated resistance to DMIs in M. fructicola was first found in isolates from the southeastern USA, then from the northeast, including Ohio, New York, New Jersey, Pennsylvania and Maryland . However, some isolates with reduced sensitivity to propiconazole from New York and Brazil lacked ‘Mona’, indicating that other mechanisms may occur …”

Section: Introductionmentioning

confidence: 99%

LAMP detection of the genetic element ‘Mona’ associated with DMI resistance in Monilinia fructicola

Chen

Schnabel

Yuan

et al. 2018

Pest Management Science

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The Point Mutation G461S in the MfCYP51 Gene is Associated with Tebuconazole Resistance in Monilinia fructicola Populations in Brazil

Cited by 49 publications

References 55 publications

Induced expression of CYP51 associated with difenoconazole resistance in the pathogenic Alternaria sect. on potato in China

Induced expression of CYP51 associated with difenoconazole resistance in the pathogenic Alternaria sect. on potato in China

Discontinuance of tebuconazole in the field restores sensitivity of Monilinia fructicola in stone fruit orchards

LAMP detection of the genetic element ‘Mona’ associated with DMI resistance in Monilinia fructicola

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