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Spore type-specific gene expression profiles underlying development and leaf infection processes of Colletotrichum graminicola.

Colletotrichum graminicola causes significant losses of the staple crop maize worldwide. The fungus produces two distinct asexual spore types, oval and falcate conidia, which show unique processes in development and plant interaction. Based on genome resequencing of our laboratory strain (CgM2/M1.001), we investigated the gene expression profiles of oval and falcate conidia during development and early leaf infection using RNA-seq. Our results reveal specific gene expression profiles between the two spore types, indicating fundamental differences in their developmental programs that reflect different modes of infection. We identified expression patterns discriminating both conidia types from mycelium and spore type-specific ones for genes encoding transcription factors, conserved fungal developmental genes, transporters, genes of secondary metabolite clusters, and pathogenicity-related functions, including effectors and carbohydrate-active enzymes (CAZymes). Our study shows that despite the identical genomic basis, oval and falcate conidia show unique transcriptomes across vegetative development and early plant interaction. Taking together, these results provide new insights into the molecular mechanisms determining the biology of C. graminicola and its interaction with the plant host.

Colletotrichum graminicola↗

Transformation-mediated developmental mutants of Glomerella graminicola.

Glomerella graminicola transformants were generated by insertional plasmid mutagenesis. Five transformants with developmental mutant phenotypes that segregated in crosses as single-gene mutations were selected. In four transformants, the mutant phenotype cosegregated with the inserted plasmid DNA. At least three of the mutants result from gene disruption, as demonstrated by recovery of the mutant phenotypes after transformation of wild type with "rescued" plasmid DNA. Whereas the wild type produces uninucleate, salmon-colored conidia, the tagged mutant M26 has white conidia. After exposure to either UV light or singlet oxygen, the percentage germination of M26 conidia is reduced compared to that of the wild-type conidia, indicating that the spore pigment confers protection from UV light and singlet oxygen. The tagged mutant T30 has weakened walls; falcate conidia rupture and hyphae have swollen regions unless the medium is amended with an osmoticum. The tagged mutant T29 has falcate conidia with one to four nuclei; wild-type falcate conidia are uninucleate. Two other mutants, one which grows slowly and one having conidia with increased curvature, are also described.

Ascomycota↗

The SOD2 gene, encoding a manganese-type superoxide dismutase, is up-regulated during conidiogenesis in the plant-pathogenic fungus Colletotrichum graminicola.

The SOD2 gene, encoding a manganese-type superoxide dismutase (MnSOD), was identified from Colletotrichum graminicola among a collection of cDNAs representing genes that are up-regulated during conidiogenesis. The SOD2 gene consists of a 797-bp open reading frame that is interrupted by three introns and is predicted to encode a polypeptide of 208 amino acids. All conserved residues of the MnSOD protein family, including four consensus metal binding domains, are present in the predicted SOD2 protein. However, the predicted protein does not appear to contain a signal peptide that would target it to the mitochondria. Northern hybridizations revealed that expression of the approximately 900-bp SOD2 transcript is closely associated with differentiation of both oval and falcate conidia. Southern analysis indicated that there is only a single copy of the gene. SOD2 disruption strains were morphologically and pathogenically indistinguishable from wild-type strains. The dispensability of the MnSOD enzyme may be due to the activities of two other SOD enzymes, a highly expressed iron-type superoxide dismutase and a much less abundant copper/zinc type, that were also detected in C. graminicola.

Amino Acid Sequence↗

A class Vb chitin synthase in Colletotrichum graminicola is localized in the growing tips of multiple cell types, in nascent septa, and during septum conversion to an end wall after hyphal breakage.

Previous complementation of a chitin synthase class Vb null mutant (Colletotrichum graminicola chsA) indicated that the encoded protein is responsible for approximately 30% of the conidial chitin, is essential for conidial wall strength in media with high water potential, and contributes to strength of hyphal tips. We complemented a chsA null mutant with chsA fused to the green-fluorescent protein (sgfp) gene driven by a heterologous constitutively expressed promoter. Comparisons of the strain with the ectopic chsA-sgfp to the wild type indicated that ChsA-sGFP serves the same biological functions as ChsA in that like the wild type, the chsADelta chsA::sgfp (EC) had conidia that did not explode and hyphal tips that did not swell. Confocal microscopy of ChsA-sGFP (EC) cells stained with the membrane stain FM 4-64 (N-(3-triethylammoniumpropyl)-4-(6-(4-(diethylamino)phenyl)hexatrienyl)pyridinium dibromide) indicated that ChsA is localized in the plasma membrane of the following: growing apices of hyphal branches, conidiophores, and falcate and oval conidia; in nascent septa; and in septa that are being converted to an end wall after hyphal breakage. The data support the hypothesis that chsA either directly or indirectly encodes the information for its localization, that ChsA is localized in the plasma membrane, and that the class Vb enzyme produces chitin synthase in multiple cells and after wall breakage.

Cell Membrane↗

Taxonomy and phylogeny of Synchaetomella lunatospora, a new genus and species of synnematous fungi from Southeast Asia.

The anamorphic fungus Synchaetomella lunatospora gen. et sp. nov. is described from a culture isolated from leaf litter in Singapore. The genus is characterized by the production of synnematous conidiomata with brown stipes, phialidic conidiogenesis with terminal and acropleurogenous conidiogenous cells, and falcate, 1-septate conidia, aggregating in white, mucilaginous heads. Its phylogenetic relationships, inferred from the partial nuclear small ribosomal subunit (18S) sequence, are with the coelomycetous genera Sphaerographium, Chaetomella, and Hainesia. However, the order and family affinities of this group are unclear.

Ascomycota↗

A new species and three new combinations in Cyphellophora, with a note on the taxonomic affinities of the genus, and its relation to Kumbhamaya and Pseudomicrodochium.

Cyphellophora guyanensis sp. nov., isolated from leaf litter in French Guyana, is described and illustrated. This fungus is characterized by ampulliform to flask-shaped phialides, with a conspicuous, funnel-shaped collarette, and nearly straight to falcate or slightly sigmoid, (2-)3-6 septate conidia. Cyphellophora guyanensis is compared to the other Cyphellophora species, and the latter genus is compared to Kumbhamaya and Pseudomicrodochium. Kumbhamaya indica, P. fusarioides, and P. suttonii are transferred to Cyphellophora. The phylogenetic relationships of Cyphellophora with other genera of ascomycetes and related mitosporic fungi were inferred from the analysis of the partial nuclear ribosomal DNA SSU sequences data. The analysis supports a placement of Cyphellophora in Chaetothyriales, and probably in the Herpotrichiellaceae

Ascomycota↗