Project description:Maize histone ZmH2B belongs to the histone H2B family. In the current field of molecular biology, the majority of histone-related studies focus on their post-translational modification functions. However, studies specifically addressing histone H2B itself are relatively scarce, particularly concerning its response mechanism against Bipolaris maydis.In this study, a nucleus-localized ZmH2B was identified. To characterize the disease resistance phenotype of ZmH2B, we employed virus-induced gene silencing (VIGS) and transient overexpression (VOX) techniques to obtain ZmH2B-silenced material FoMV:ZmH2B and overexpressed material FoMV:ZmH2B-VOX. It was discovered that FoMV:ZmH2B promoted the infestation of B. maydis and inhibit the reactive oxygen species (ROS) burst induced by chitin. Conversely, FoMV:ZmH2B-VOX exhibited the opposite effects. Furthermore, ZmH2B was verified to positively regulate the infestation of B. maydis through pathogenesis-related (PR) genes. Subsequently, transcriptome analysis was carried out on the ZmH2B-silenced material and the differentially expressed genes were predominantly enriched in photosynthesis-related pathways.Collectively, these results suggested that ZmH2B plays a positive role in regulating maize resistance to B. maydis.
Project description:In maize crops Southern Leaf Blight (SLB) is a severe disease caused by the fungus Bipolaris maydis (Y. Nisik & C. Miyake) leading to several losses in crop production. Unlike in maize, popcorn resistant varieties are not yet commercially available and this trait have been the subject of many popcorn breeding programs. The present study aimed to identify differentially accumulated proteins (DAPs) associated with resistance to B. maydis in two contrasting popcorn inbred lines using the comparative proteomic analysis. Forty-day old popcorn plants from resistant and susceptible inbred lines at the V4 growth stage were inoculated with B. maydis by spraying the conidium suspension. At four and ten days after inoculation, the morphological aspects of lesions in leaves were checked and the proteomic analysis was performed. Our results showed that the resistant inbred line exhibited minor foliar lesions in comparison with the SLB-susceptible genotype. In the genotype’s comparison, 644 DAPs were identified at 4 days and 613 at 10 days after B. maydis inoculation. Besides that, in resistant plants were identified DAPs upaccumulated related to response to stress, response to stimulus, photosynthesis, cellular growth, maintenance and detoxification process, which might be involved in plant response against the pathogen. It was identified proteins as salicylic acid-related and nucleotide-binding leucine-rich repeat that may be responsible to initiate a response to fungus in SBL-resistant inbred line. A differential response was identified in chloroplast proteins that is the main organelle involved in perception of the fungus to initiate a signalizing response. The changes in the proteomic profile in resistant inbred line may be effective in the response under B. maydis infection. This is the first work presenting the proteomic profile alteration under B. maydis inoculation and these findings may be useful for identifying candidate biomarkers in popcorn resistance to B. maydis being relevant for further researches on genetic breeding to develop resistant genotypes.
Project description:Hxt1 is a high affinity hexose transporter important for virulence of the phytopathogenic basidiomycete Ustilago maydis on its host plant maize. Hxt1 shows the highest similarities to the glucose sensors Rgt2 and Snf3 from S. cereviciae (42% and 39% identity on amino acid level, respectively). In these sensors, the substitution of a highly conserved arginine to lysine leads to a constitutive active signal, resulting in expression of several glucose induced genes. Introduction of an analogous mutation in Hxt1 leads to loss of transport activity in the resulting Hxt1(R164K) protein. Expression of Hxt1(R164K) in hxt1 deletion strains results in a completely avirulent phenotype. Pathogenic developement of M-bM-^HM-^Fhxt1 hxt1(R164K) strains is blocked immediately after plant penetration. Expression analysis of M-bM-^HM-^Fhxt1 hxt1(R164K) cells 24 hours post inoculation revealed downregulation of a set of genes involved in carbohydrate metabolism indicating a role of Hxt1 in carbohydrate signaling during initiation of the pathogenic stage. To analyze expression changes of SG200 and SG200M-bM-^HM-^Fhxt1 and SG200M-bM-^HM-^Fhxt1 hxt1(R164K) cells were harvested from the surface of maize leaves 24 hour post inoculation. For each strain three independent replicates were conducted.
Project description:The rep1 gene of the maize pathogen Ustilago maydis encodes a pre-pro-protein that is processed in the secretory pathway into 11 peptides. These so-called repellents form amphipathic amyloid fibrils at the surface of aerial hyphae. Strains in which the rep1 gene is inactivated (M-bM-^HM-^Frep1 strain) are affected in aerial hyphae formation. This makes these strains instrumental to assess changes in global gene expression as a consequence of aerial growth. Microarray analysis revealed that only 31 genes in the M-bM-^HM-^Frep1 SG200 strain had a fold change in expression of >= 2. Twenty-two of these genes are up-regulated and half of them encode small secreted proteins (SSPM-bM-^@M-^Ys) with unknown functions. Seven of the SSP genes and two other genes that are over-expressed in the M-bM-^HM-^Frep1 SG200 strain encode secreted cysteine-rich proteins (SCRPM-bM-^@M-^Ys). Interestingly, most of the SCRPM-bM-^@M-^Ys are predicted to form amyloids. The SCRP gene um00792 showed the highest up-regulation in the M-bM-^HM-^Frep1 strain. Using GFP as a reporter, it was shown that this gene is over-expressed in the layer of hyphae at the medium-air interface. Taken together, it is concluded that only minor changes occur in the expression profile when U. maydis forms aerial structures. Key words: aerial hypha, repellent, hydrophobin-like protein, Ustilago maydis, SSP, SCRP, fungal pathogenicity. To analyze expression changes in aerial hyphae upon deletion of the rep1 gene, strains SG200 and SG200M-bM-^HM-^Frep1 were grown on charcoal nitrate minimal array media supplemented with vitamins and 1% glucose at 22 C for 48 h. For each experiment three biological replicates were performed.
Project description:Goals: Comparing the infection between Ustilago maydis SG200 with the wild-type strain FB1xFB2 previously published Methods: Comparative RNASeq analysis between U. maydis SG200 and U. maydis FB1xFB2 at three timepoints (axenic, 2dpi, 12dpi) Results: The RNASeq analysis in SG200 identifies differences in gene expression with FB1xFB2. These differences could be the result of a unequal contribution of each nuclei to transcription. Further analysis identified a set of differentially transcribed genes.