Project description:According to the key words, the gene set, including oxidation-reduction, RNA silence, disease resistance, phytohormone, phosphorylation, dephosphorylation, transcription factor, receptor, kinase, ubiquitination and RNA binding,etc. from sugarcane and the whole CDS sequence from smut genome, was achieved and used as targets in the present microarray assay. Based on smut infection samples from smut-susceptible sugarcane genotype YC71-374 and smut-resistant sugarcane genotype NCo376, the hybridization was conducted and validated by real-time fluorescent quantitative PCR. It would provide a basic data for the study on sugarcane-smut interaction mechanism, which referred to sugarcane smut resistance and smut pathogenesis.
Project description:Dimorphic fungi have the ability to change morphology during their lifecycle, a crucial feature for the establishment of infection and fungal growth and development in planta. Life cycle of the dimorphic sugarcane smut fungi, Sporisorium scitamineum, involves recognition and mating of compatible saprophytic yeast-like haploid sporidia (MAT-1 and MAT-2) that upon fusion, develop into infective dikaryotic mycelia. Although the dimorphic transition is intrinsically linked with the pathogenicity and virulence of S. scitamineum, it has never been studied using a proteomics approach. In the present study, an iTRAQ-based comparative proteomic analysis of three distinct stages covering the dimorphic transition period - haploid sporidial stage (MAT-1 and MAT-2) to the transition phase (24 hours post co-culturing (hpc)) and dikaryotic mycelial stage (48 hpc) was carried out. Functional categorization showed that the most altered biological processes were energy production, primary metabolism especially carbohydrate, amino acid, fatty acid, followed by translation, post-translation and protein turnover. The identified proteins could be grouped into 8 distinct clusters with different trends in abundance. Enrichment analysis of the clusters showed that biological processes related to energy production through oxidative phosphorylation, citrate cycle, and β-oxidation, transcription, translation and redox homeostasis were specifically altered. In addition, an overall downregulation of carbohydrate metabolism and reprogrammed amino acid metabolism were observed. Several differentially abundant proteins (DAPs), especially in the dikaryotic mycelial stage were predicted as effectors. Taken together, key molecular mechanisms underpinning the dimorphic transition in S. scitamineum at the proteome level were highlighted. A catalogue of stage-specific and dimorphic transition-associated -proteins and potential effectors identified herein are potential candidates for defective mutant screening to elucidate their functional role in the dimorphic transition and pathogenicity in S. scitamineum.
Project description:Sugarcane smut caused by the basidiomycetes fungus Sporisorium scitamineum is a devastating disease for the sugarcane industry worldwide. As the initial step, the smut teliospores germinate on sugarcane buds, and subsequently, the mycelium infects the bud tissues. However, chemical signals that induce spore germination are still unknown. By comparison of the behavior of the teliospores on the buds of both resistant and susceptible varieties, we found that spore germination rates were significantly lower on the buds of resistant cultivars ZZ1, ZZ6, and ZZ9 than on the susceptible varieties GT42 and ROC22. It was found that the levels of hexacosanol and octacosanol were higher on the buds of smut-susceptible varieties than on the smut-resistant varieties. These observations were extended to the smut-resistant and smut-susceptible sub-genetic populations derived from the cross of ROC25 and YZ89-7. In artificial surface assays, we found that hexacosanol and octacosanol promoted smut teliospore germination. Transcriptome analysis of smut teliospores under the induction by octacosanol revealed that genes in the MAPK signaling pathway and fatty acid metabolism were significantly differentially expressed. Overall, our results provide evidence that alkanol plays important roles in smut teliospore germination and thus could be used as a potential marker for smut resistance in sugarcane breeding programs.
Project description:Head smut of maize, which is caused by the Sporisorium reilianum f. sp. Zeae (Kühn), has been a serious disease in maize. In order to find head smut resistant candidate genes, microarrays were used to monitor the gene expression profiles between disease resistant near isogenic lines (NIL) L282 and L43, highly resistant inbred line Q319 and highly susceptible inbred line Huangzao4 after 0 to7 days post inoculation of S.reiliana by artificial inoculation method.
Project description:The plant cell wall is known to be the first barrier against plant pathogens. Detailed information about sugarcane cell wall-associated defense responses to infection by the causal agent of smut, Sporisorium scitamineum, is scarce. Herein, (immuno)histochemical analysis of two smut resistant and two susceptible sugarcane cultivars was conducted to understand host cell wall structural and compositional modifications in response to fungal infection. Results showed that the fungus grew on the surface and infected the outermost bud scale of both susceptible and resistant cultivars. The present findings also supported the existence of early (24 h after inoculation) and later (72-96 h after inoculation) inducible histopathological responses related to the cell wall modification in resistant cultivars. Lignin and phenolic compounds accumulated during early stages of infection. Later infection response was characterized by the formation of a protective barrier layer with lignin, cellulose and arabinoxylan in the cell walls. Overall, the results suggest possible induction of cell wall-modified responses in smut resistant cultivars to prevent initial entry of the fungus into the meristematic tissues.
Project description:Sporisorium scitamineum is the fungal pathogen causing severe sugarcane smut disease that leads to massive economic losses globally. S. scitamineum invades host cane by dikaryotic hyphae, formed after sexual mating of two haploid sporidia of opposite mating type. Therefore, mating/filamentation is critical for S. scitamineum pathogenicity, while its molecular mechanisms remain largely unknown. The AGC (cyclic AMP [cAMP]-dependent protein kinase 1 [protein kinase A {PKA}], cGMP-dependent protein kinase [PKG], and protein kinase C [PKC]) kinase family is a group of serine/threonine (Ser/Thr) protein kinases conserved among eukaryotic genomes, serving a variety of physiological functions, including cell growth, metabolism, differentiation, and cell death. In this study, we identified an AGC kinase, named SsAgc1 (for S. scitamineum Agc1), and characterized its function by reverse genetics. Our results showed that SsAgc1 is critical for S. scitamineum mating/filamentation and pathogenicity, and oxidative stress tolerance under some circumstances. Transcriptional profiling revealed that the SsAgc1 signaling pathway may control expression of the genes governing fungal mating/filamentation and tryptophan metabolism, especially for tryptophol production. We showed that tryptophan and tryptophol could at least partially restore ssagc1Δ mating/filamentation. Overall, our work revealed a signaling pathway mediated by AGC protein kinases to regulate fungal mating/filamentation, possibly through sensing and responding to tryptophol as signal molecules.IMPORTANCE The AGC signaling pathway represents a conserved distinct signaling pathway in regulation of fungal differentiation and virulence, while it has not been identified or characterized in the sugarcane smut fungus Sporisorium scitamineum In this study, we identified a PAS domain-containing AGC kinase, SsAgc1, in S. scitamineum Functional analysis revealed that SsAgc1 plays a regulatory role on the fungal dimorphic switch.
Project description:Head smut of maize, which is caused by the Sporisorium reilianum f. sp. Zeae (Kühn), has been a serious disease in maize. In order to find head smut resistant candidate genes, microarrays were used to monitor the gene expression profiles between disease resistant near isogenic lines (NIL) L282 and L43, highly resistant inbred line Q319 and highly susceptible inbred line Huangzao4 after 0 to7 days post inoculation of S.reiliana by artificial inoculation method. Maize leaves were selected at 0d, 1d, 2d, 4d, 7d post inoculation for RNA extraction and hybridization on Affymetrix microarrays. We sought to obtain different expression genes of different varieties at each inoculation stage in order to find head smut resistant candidate genes.