Open AccessA pleiotropic drug resistance transporter is involved in reduced sensitivity to multiple fungicide classes in S clerotinia homoeocarpa ( F . T . B ennett)
Author(s) -
Sang Hyunkyu,
Hulvey Jon,
Popko James T.,
Lopes John,
Swaminathan Aishwarya,
Chang Taehyun,
Jung Geunhwa
Publication year - 2015
Publication title -
molecular plant pathology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.945
H-Index - 103
eISSN - 1364-3703
pISSN - 1464-6722
DOI - 10.1111/mpp.12174
Subject(s) - biology , fungicide , transporter , drug resistance , genetics , gene , botany
Summary Dollar spot, caused by S clerotinia homoeocarpa , is a prevalent turfgrass disease, and the fungus exhibits widespread fungicide resistance in N orth A merica. In a previous study, an ABC ‐ G transporter, ShatrD , was associated with practical field resistance to demethylation inhibitor ( DMI ) fungicides. Mining of ABC ‐ G transporters, also known as pleiotropic drug resistance ( PDR ) transporters, from RNA ‐ S eq data gave an assortment of transcripts, several with high sequence similarity to functionally characterized transporters from B otrytis cinerea , and others with closest blastx hits from Aspergillus and Monilinia . In addition to ShatrD , another PDR transporter showed significant over‐expression in replicated RNA ‐ S eq data, and in a collection of field‐resistant isolates, as measured by quantitative polymerase chain reaction. These isolates also showed reduced sensitivity to unrelated fungicide classes. Using a yeast complementation system, we sought to test the hypothesis that this PDR transporter effluxes DMI as well as chemically unrelated fungicides. The transporter ( ShPDR1 ) was cloned into the Gal1 expression vector and transformed into a yeast PDR transporter deletion mutant, AD12345678 . Complementation assays indicated that ShPDR1 complemented the mutant in the presence of propiconazole ( DMI ), iprodione (dicarboximide) and boscalid ( SDHI , succinate dehydrogenase inhibitor). Our results indicate that the over‐expression of ShPDR1 is correlated with practical field resistance to DMI fungicides and reduced sensitivity to dicarboximide and SDHI fungicides. These findings highlight the potential for the eventual development of a multidrug resistance phenotype in this pathogen. In addition, this study presents a pipeline for the discovery and validation of fungicide resistance genes using de novo next‐generation sequencing and molecular biology techniques in an unsequenced plant pathogenic fungus.