7, bottom). The cgopt1-silenced mutants developed pellets with very long hyphae (hairy pellets) in CD medium and again, this morphology was not altered by IAA. Thus, the wild-type isolate developed more condensed pellets in IAA-containing media, while the morphology of the cgopt1-silenced

mutants differed from the wild type, and was unaffected by IAA. Discussion In a Quisinostat previous report, we showed that C. gloeosporioides produces auxin both in culture and in planta [16, 17]. This raised the possibility of auxin involvement in the regulation of fungal mTOR inhibitor development and pathogeniCity, and of the existence of auxin-responsive genes regulating fungal responses to IAA. As a first step towards identifying the putative IAA-responsive fungal genes, we constructed a SSH library

using mycelia from auxin-containing medium as the tester. Under culture conditions, over 95% of the IAA that is produced by C. gloeosporioides is secreted into the medium [20]. We therefore used a relatively high IAA concentration (500 μM), assuming that the endogenous concentrations would be at least 10-fold lower. We also added 500 μM IAM, the intermediate product of IAA production in C. gloeosporioides [17]. The SSH yielded limited information on putative IAA-induced genes since only three clones showed consistent induction by IAA. Thus, NSC 683864 in vivo although putative IAA-induced genes were identified, the results from the SSH approach do not support a massive change in gene transcription by IAA. However, the number of genes that could be tested by SSH was limited and more conclusive results might be obtained through robust transcript analysis using microarrays when such Levetiracetam tools become available in C. gloeosporioides. CgOPT1 exhibited consistent induction by IAA and was therefore further analyzed. Characterization of the gene as a putative OPT was strongly supported by its overall homology to other OPTs, as well as by the presence of the conserved SPYxEVRxxVxxxDDP sequence and 14 transmembrane

domains, which are common to all OPTs [18, 21, 22]. Further analyses, including complementation of yeast mutants, are needed to determine that CgOPT1 is indeed an oligopeptide transporter and to find substrate specifiCity. In S. cerevisiae, there are two genetically and physiologically distinct proton-coupled peptide transporter systems: the PTR (peptide transport) and the OPT (oligopeptide transport) protein families. Members of the PTR and OPT families differ in function and they do not share significant sequence homology (see Fig. 1C). PTR proteins are common in all organisms and transport di- or tripeptides. OPT proteins are found only in plants and fungi and transport 4- and 5-amino-acid peptides [22, 23]. Metabolically, the transport of small oligopeptides is important as an amino acid, carbon, and nitrogen source [23].