Bcmfs1 , a Novel Major Facilitator Superfamily Transporter from Botrytis cinerea , Provides Tolerance towards the Natural Toxic Compounds Camptothecin and Cercosporin and towards Fungicides

Abstract

Bcmfs1, a novel major facilitator superfamily gene from Botrytis cinerea, was cloned, and replacement and overexpression mutants were constructed to study its function. Replacement mutants showed increased sensitivity to the natural toxic compounds camptothecin and cercosporin, produced by the plant Camptotheca acuminata and the plant pathogenic fungus Cercospora kikuchii, respectively. Overexpression mutants displayed decreased sensitivity to these compounds and to structurally unrelated fungicides, such as sterol demethylation inhibitors (DMIs). A double-replacement mutant of Bcmfs1 and the ATP-binding cassette (ABC) transporter gene BcatrD was more sensitive to DMI fungicides than a single-replacement mutant of BcatrD, known to encode an important ABC transporter of DMIs. The sensitivity of the wild-type strain and mutants to DMI fungicides correlated with Bcmfs1 expression levels and with the initial accumulation of oxpoconazole by germlings of these isolates. The results indicate that Bcmfs1 is a major facilitator superfamily multidrug transporter involved in protection against natural toxins and fungicides and has a substrate specificity that overlaps with the ABC transporter BcatrD. Bcmfs1 may be involved in protection of B. cinerea against plant defense compounds during the pathogenic phase of growth on host plants and against fungitoxic antimicrobial metabolites during its saprophytic phase of growth.

Trehalose metabolism is important for heat stress tolerance and spore germination of Botrytis cinerea

“…In fact, its ability to produce and to withstand high concentrations of reactive oxygen species during pathogenesis has been well documented (Prins et al, 2000;Rolke et al, 2004). In addition, the grey mould fungus is equipped with a set of membrane transport proteins including several ABC and MFS transporters which confer resistance to a variety of toxic compounds (de Waard et al, 2006;Hayashi et al, 2002).…”

Molecular Tools for Detection of Plant Pathogenic Fungi and Fungicide Resistance

“…Currently, the most widespread hypothesis to explain the reduced levels of toxic products in the cell is based on active efflux of these compounds by ABC transporters (ATP binding cassette) and MFS (Major Facilitator Superfamily) (Hayashi et al, 2001(Hayashi et al, , 2002a(Hayashi et al, , 2002b(Hayashi et al, , 2003Stergiopoulos et al, 2002aStergiopoulos et al, , 2002bStergiopoulos & de Waard, 2002, Vermeulen et al, 2001. ABC and MFS transporters are the most studied so far and have been described in many fungi such as Aspergillus nidulans, Botrytis cinerea, Mycosphaerella graminicola, Magnaporthe grisea, Penicillium digitatum, etc (Andrade et al, 2000a, 2000bdel Sorbo et al, 2000;Nakaune, 2001;Schoonbeek et al, 2003;Stergiopoulos et al, 2002a;Stergiopoulos & de Waard, 2002;Vermeulen et al, 2001;Yoder and Turgeon 2001;Zwiers et al, 2003).…”

“…Botrytis cinerea shows a broad spectrum of resistance to different fungicides and their expression has been induced by many of them, particularly noteworthy Bcmfs4 induction in the presence of strobilurin (trifloxiestrobin) (Hayashi et al, 2002a(Hayashi et al, , 2002b(Hayashi et al, , 2003Schoonbeek et al, 2003;Vermeulen et al, 2001). All this means that these transporters are potential candidates for the study of factors involved in resistance based on active eflux of these toxic compounds since they have remarkably broad substrate specificity although they can also transport specific compounds.…”

Respuesta transcripcional de Populus a bifenilos policlorados: Aspectos aplicados a la fitorremediación

“…cerevisiae también están involucrados en la protección contra compuestos exógenos, tales como los inhibidores de la desmetilación de esteroles (Calabrese et al, 2000;Hayashi et al, 2002). Otros estudios en Botrytis cinerea revelaron la existencia de un transportador MFS, denominado Bcmfs1.…”

“…Son flavín monoxigenasas de la clase A dependiente de NADPH, que intervienen en la ruta de desulfuración de compuestos orgánicos, acopla la oxidación de NADH con la oxidación del sustrato en cada caso, mediante la reducción de FMN a FMNH2. Catalizan reacciones de oxigenación a partir de oxígeno molecular, que usa como sustrato para oxidar un compuesto orgánico (Hayashi et al, 2002).…”