Previously, we reported the presence of a bifunctional gene encoding spermidine synthase (Spe) and saccharopine dehydrogenase selleck kinase inhibitor (Sdh) in the Basidiomycota fungus
Ustilago maydis, confirming previous data from Cryptococcus neoformans (Kingsbury et al., 2004). This gene contains a 5′-region encoding Spe, followed, without a termination signal or a second initiation codon, by a 3′-region encoding Sdh (Valdés-Santiago et al., 2009). Apparently, this chimeric gene is specific to Basidiomycota, because in a preliminary search, it could not be identified in several Ascomycota species. Spe catalyzes the transfer of the aminopropyl group from decarboxylated S-adenosylmethionine to putrescine during spermidine biosynthesis. Regarding lysine, it is known that fungi synthesize it via their exclusive mechanism, the α-aminoadipate pathway (see Xu et al., 2006). Sdh, also called saccharopine reductase, catalyzes the penultimate step in this pathway (Bhattacharjee, 1992). In the present work, we have performed an exhaustive analysis for the presence of a homologous gene in those Basidiomycota species whose
genome has been sequenced, in other fungal taxa, and in the rest of living organisms reported in data banks. With the results obtained and the experimental data of gene amplification by PCR in different species, we propose the use selleckchem of this gene as a molecular marker for Basidiomycota in general. Yarrowia lipolytica P01A was obtained from Claude Gaillardin (INRA), Saccharomyces cerevisiae S288C was obtained from American Type Culture
Collection (ATCC 26108), Mucor rouxii IM80 (ATCC 24905) was obtained from Salomón Bartnicki-Garcia (University of California, Riverside), Rhizopus oryzae 2672 was obtained from CECT (Colección Española de Cultivos Tipo), U. maydis FB2 was obtained from Flora Banuett (California State University, Long Beach), Coprinus cinereus UAMH4103 was obtained from University of Alberta Microfungus Collection and Herbarium, Ustilago hordei 8A was obtained Florfenicol from ATCC (90511); Ganoderma lucidum, Ganoderma sp., Schizophyllum commune, Pleurotus ostreatus, Rhizoctonia solani, Agaricus bisporus, Ustilago cynodontis, Tilletia foetida, and Bjerkandera adusta belong to the collection from Laboratorio de Micología (Universidad Autónoma de Nuevo León, Monterrey, NL, Mexico). Fungal strains were maintained in 50% glycerol at −70 °C. For propagation, strains were inoculated in liquid YPG medium [yeast extract (Difco), 2%; peptone (Difco), 1%, and glucose (Merck), 1%] and incubated at 28 °C for 18 h under shaking conditions (150 r.p.m.). Escherichia coli strain ElectroMAX™DH10B™ (Invitrogen Life Technologies) was used for transformation with the PCR-amplified products cloned previously in TOPO™4 vector (Invitrogen).