Global Gene Expression Analysis during Germination in the Chytridiomycete
            <i>Blastocladiella emersonii</i>

Salem-Izacc, Sílvia Maria; Koide, Tie; Vêncio, Ricardo Z. N.; Gomes, Suely L.

doi:10.1128/ec.00330-08

Cited by 13 publications

(17 citation statements)

References 49 publications

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“…Transcriptome studies with N. crassa, Fusarium graminearum, and Ustilago maydis have been performed using spores germinated in minimal (N. crassa and U. maydis) or full (F. graminearum) media in (partly shaken) suspensions, with no permanent surface contact of the spores (17,18,19). Blumeria microarray-based studies with germinated conidia forming infection structures, including haustoria, on the surface and within host epidermal cells have focused mainly on transcriptomic changes related to metabolic pathways, such as lipid degradation, glycolysis, pentose phosphate pathway, and amino acid biosynthesis (20,21).…”

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Transcriptome Profiling of Botrytis cinerea Conidial Germination Reveals Upregulation of Infection-Related Genes during the Prepenetration Stage

et al. 2013

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Botrytis cinerea causes gray mold on a great number of host plants. Infection is initiated by airborne conidia that invade the host tissue, often by penetration of intact epidermal cells. To mimic the surface properties of natural plant surfaces, conidia were incubated on apple wax-coated surfaces, resulting in rapid germination and appressorium formation. Global changes in gene expression were analyzed by microarray hybridization between conidia incubated for 0 h (dormant), 1 h (pregermination), 2.5 h (postgermination), 4 h (appressoria), and 15 h (early mycelium). Considerable changes were observed, in particular between 0 h and 1 h. Genes induced during germination were enriched in those genes encoding secreted proteins, including lytic enzymes. Comparison of wild-type and a nonpathogenic MAP kinase mutant (bmp1) revealed marked differences in germination-related gene expression, in particular related to secretory proteins. Using promoter-GFP reporter strains, we detected a strictly germination-specific expression pattern of a putative chitin deacetylase gene (cda1). In contrast, a cutinase gene (cutB) was found to be expressed only in the presence of plant lipids, in a developmentally less stringent pattern. We also identified a coregulated gene cluster possibly involved in secondary metabolite synthesis which was found to be controlled by a transcription factor also encoded in this cluster. Our data demonstrate that early conidial development in B. cinerea is accompanied by rapid shifts in gene expression that prepare the fungus for germ tube outgrowth and host cell invasion.

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confidence: 99%

“…Transcriptome profiling analyses during germination have been published for N. crassa (17), the saprotrophic chytridiomycete Blastocladiella emersonii (18), and the wheat pathogen F. graminearum (19). These studies revealed major changes of gene expression patterns starting already during the pregermination swelling stage.…”

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confidence: 99%

Transcriptome Profiling of Botrytis cinerea Conidial Germination Reveals Upregulation of Infection-Related Genes during the Prepenetration Stage

et al. 2013

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“…In contrast, data revealed that most genes involved in signal transduction showed the same expression profile during the initial stages of germination with all the inducers investigated. These results indicate that the same signaling pathways are activated irrespective of the initial stimulus triggering germination (29).…”

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“…The expression profiles of 530 genes differentially expressed during (29) and sporulation (this work) of B. emersonii were compared through clustering analysis using the algorithm K-means. Data revealed that approximately 95% of the genes analyzed are inversely regulated between these two differentiation stages (see Fig.…”

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confidence: 99%

“…Recently, global gene expression changes occurring during B. emersonii germination were analyzed in cells germinating both in nutrient medium and in inorganic solution containing either potassium or adenine as inducers of this differentiation process (29). Data revealed that more than 900 genes out of 3,563 distinct genes spotted in the microarray chips were differentially expressed during germination in nutrient medium, over 500 of them being upregulated.…”

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Global Gene Expression Analysis during Sporulation of the Aquatic Fungus Blastocladiella emersonii

Vieira

Gomes

2010

Eukaryot Cell

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The Blastocladiella emersonii life cycle presents a number of drastic biochemical and morphological changes, mainly during two cell differentiation stages: germination and sporulation. To investigate the transcriptional changes taking place during the sporulation phase, which culminates with the production of the zoospores, motile cells responsible for the dispersal of the fungus, microarray experiments were performed. Among the 3,773 distinct genes investigated, a total of 1,207 were classified as differentially expressed, relative to time zero of sporulation, at at least one of the time points analyzed. These results indicate that accurate transcriptional control takes place during sporulation, as well as indicating the necessity for distinct molecular functions throughout this differentiation process. The main functional categories overrepresented among upregulated genes were those involving the microtubule, the cytoskeleton, signal transduction involving Ca 2؉ , and chromosome organization. On the other hand, protein biosynthesis, central carbon metabolism, and protein degradation were the most represented functional categories among downregulated genes. Gene expression changes were also analyzed in cells sporulating in the presence of subinhibitory concentrations of glucose or tryptophan. Data obtained revealed overexpression of microtubule and cytoskeleton transcripts in the presence of glucose, probably causing the shape and motility problems observed in the zoospores produced under this condition. In contrast, the presence of tryptophan during sporulation led to upregulation of genes involved in oxidative stress, proteolysis, and protein folding. These results indicate that distinct physiological pathways are involved in the inhibition of sporulation due to these two classes of nutrient sources.The life cycle of Blastocladiella emersonii, an aquatic fungus of the class Blastocladiomycetes, involves a series of complex morphological and biochemical events, which involve transcriptional, posttranscriptional, and posttranslational mechanisms of control, especially during two stages of cell differentiation, the germination and the sporulation of the fungus (21). Germination starts with the zoospore, a motile uninucleated wall-less nongrowing cell, which is responsible for the dispersal of the fungus. In the presence of appropriate stimuli, the zoospore germinates, undergoing a number of biochemical and morphological changes. The early events of germination, which include retraction of the single polar flagellum, construction of a cell wall rich in chitin, and cleavage of a giant mitochondrion into normal-size ones, do not require concomitant RNA and protein synthesis and involve posttranslational regulatory events (17,20,30,31,35,36). Major changes in the pattern of RNA and protein synthesis are observed only at the late events of this stage, when the germ tube begins to branch, giving rise to a rhizoidal system through which nutrients are absorbed, and when the cells enter the vegetative growth phase (17,20...

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Agrobacterium tumefasciens-mediated transformation of the aquatic fungus Blastocladiella emersonii

Vieira

Camilo

2011

Fungal Genetics and Biology

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Agrobacterium tumefaciens is widely used for plant DNA transformation and more recently, has also been used to transform yeast, filamentous fungi and even human cells. Using this technique, we developed the first transformation protocol for the saprobic aquatic fungus Blastocladiella emersonii, a Blastocladiomycete localized at the base of fungal phylogenetic tree, which has been shown as a promising and interesting model of study of cellular function and differentiation. We constructed binary T-DNA vectors containing hygromycin phosphotransferase (hph) or enhanced green fluorescent protein (egfp) genes, under the control of Aspergillus nidulans trpC promoter and terminator sequences. 24 h of co-cultivation in induction medium (IM) agar plates, followed by transfer to PYG-agar plates containing cefotaxim to kill Agrobacterium tumefsciens and hygromycin to select transformants, resulted in growth and sporulation of resistant transformants. Genomic DNA from the pool o resistant zoospores were shown to contain T-DNA insertion as evidenced by PCR amplification of hph gene. Using a similar protocol we could also evidence the expression of enhanced green fluorescent protein (EGFP) in zoospores derived from transformed cells. This protocol can also open new perspectives for other non-transformable closely related fungi, like the Chytridiomycete class.

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Global Gene Expression Analysis during Germination in the Chytridiomycete Blastocladiella emersonii

Cited by 13 publications

References 49 publications

Transcriptome Profiling of Botrytis cinerea Conidial Germination Reveals Upregulation of Infection-Related Genes during the Prepenetration Stage

Transcriptome Profiling of Botrytis cinerea Conidial Germination Reveals Upregulation of Infection-Related Genes during the Prepenetration Stage

Global Gene Expression Analysis during Sporulation of the Aquatic Fungus Blastocladiella emersonii

Agrobacterium tumefasciens-mediated transformation of the aquatic fungus Blastocladiella emersonii

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