Overexpression of ShCYP51B and ShatrD in Sclerotinia homoeocarpa isolates exhibiting practical field resistance to a demethylation inhibitor fungicide.
We investigated genetic factors that govern the reduced propiconazole sensitivity of Sclerotinia homoeocarpa field isolates collected during a 2-year field efficacy study on dollar spot disease of turf in five New England sites. These isolates displayed a >50-fold range of in vitro sensitivity to a sterol demethylation inhibitor (DMI) fungicide, propiconazole, making them ideal for investigations of genetic mechanisms of reduced DMI sensitivity. The CYP51 gene homolog in S. homoeocarpa (ShCYP51B), encoding the enzyme target of DMIs, is likely a minor genetic factor for reduced propiconazole sensitivity, since there were no differences in constitutive relative expression (RE) values and only 2-fold-higher induced RE values for insensitive than for sensitive isolate groups. Next, we mined RNA-Seq transcriptome data for additional genetic factors and found evidence for the overexpression of a homolog of Botrytis cinerea atrD (BcatrD), ShatrD, a known efflux transporter of DMI fungicides. The ShatrD gene showed much higher constitutive and induced RE values for insensitive isolates. Several polymorphisms were found upstream of ShatrD but were not definitively linked to overexpression. The screening of constitutive RE values of ShCYP51B and ShatrD in isolates from two golf courses that exhibited practical field resistance to propiconazole uncovered evidence for significant population-specific overexpression of both genes. However, linear regression demonstrated that the RE of ShatrD displays a more significant relationship with propiconazole sensitivity than that of ShCYP51B. In summary, our results suggest that efflux is a major determinant of the reduced DMI sensitivity of S. homoeocarpa genotypes in New England, which may have implications for the emergence of practical field resistance in this important turfgrass pathogen.
Department of Plant, Soil, and Insect Sciences, University of Massachusetts, Amherst, Massachusetts, USA.
Jon
J
Hulvey
James T
JT
Popko
Hyunkyu
H
Sang
Andrew
A
Berg
Geunhwa
G
Jung
D018528:ATP-Binding Cassette Transporters / Q000096:biosynthesis* / Q000235:genetics; D001203:Ascomycota / Q000187:drug effects* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics; D025141:Drug Resistance, Fungal; D005659:Fungicides, Industrial / Q000378:metabolism / Q000494:pharmacology*; D018628:Gene Dosage; D008969:Molecular Sequence Data; D009511:New England; D010935:Plant Diseases / Q000382:microbiology; D006109:Poaceae / Q000382:microbiology; D017422:Sequence Analysis, DNA; D058886:Sterol 14-Demethylase / Q000096:biosynthesis* / Q000235:genetics; D014230:Triazoles / Q000378:metabolism / Q000494:pharmacology*
D018528:ATP-Binding Cassette Transporters; D004271:DNA, Fungal; D005659:Fungicides, Industrial; D014230:Triazoles; C045950:propiconazole; D058886:Sterol 14-Demethylase