GLS1 of C. graminicola Encodes a Functional GLS.
(A) Phylogenetic tree indicates close relatedness of GLS1 of C. graminicola with other GLSs of filamentous fungi: A. capsulatus, Ajellomyces capsulatus; A. dermattidis, Ajellomyces dermatitidis; A. flavus, Aspergillus flavus; A. oryzae, Aspergillus oryzae; A. fumigatus, Aspergillus fumigatus; A. nidulans, Aspergillus nidulans; C. albicans, Candida albicans; C. glabrata, Candida glabrata; C. paraplosis, Candida parapsilosis; C. gattii, Cryptococcus gattii; C. immitis, Coccidioides immitis; C. posadasii, Coccidioides posadasii; C. graminicola, Colletotrichum graminicola; C. cinerea, Coprinopsis cinerea; C. neoformans, Cryptococcus neoformans; E. dermatitis, Exophiala dermatitidis; F. solani (FKS1), Fusarium solani (ABC59463); F. solani (FKS2), Fusarium solani (XP003040299.1); L. bicolor, Laccaria bicolor; M. oryzae, Magnaporthe oryzae; M. acridum, Metarhizium acridum; N. crassa, Neurospora crassa; P. brasiliensis, Paracoccidioides brasiliensis; P. marneffei, Penicillium marneffei; P. pastoris, Pichia pastoris; P. graminis, Puccinia graminis f. sp tritici; S. cerevisiae (FKS1), Saccharomyces cerevisiae; S. cerevisiae (FKS2), Saccharomyces cerevisiae; S. cerevisiae (FKS3), Saccharomyces cerevisiae; S. pombe (Bgs1), Schizosaccharomyces pombe; S. pombe, Schizosaccharomyces pombe (Bgs2); S. pombae (Bgs3), Schizosaccharomyces pombe (NP594766.1); S. pombae (Bgs4), Schizosaccharomyces pombe; S. sclerotiorum, Sclerotinia sclerotiorum; T. stipitatus, Talaromyces stipitatus; U. maydis, Ustilago maydis; and Z. rouxii, Zygosaccharomyces rouxii. Red, filamentous Ascomycota; blue, unicellular Ascomycota (yeasts); green, Basidiomycota.
(B) Complementation of S. cerevisiae Δfks1 by GLS1 cDNA of C. graminicola suggests that GLS1 encodes an active GLS. WT, S. cerevisiae reference strain Y00000; Δfks1, FKS1-deficient S. cerevisiae strain Y05251; TpAG300, Δfks1 transformant harboring the empty binary vector pAG300; TCgGLS11 - 4, independent Δfks1 transformants expressing the GLS1 cDNA of C. graminicola; YPD, YPD medium; YPDS, YPD medium supplemented with 1 M sorbitol. Number of yeast cells inoculated were (left to right) 5 × 104, 5 × 103, 5 × 102, and 5 × 10.