Quantitative Proteomics Reveals the Defense Response of Wheat against Puccinia striiformis f. sp. tritici.
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ABSTRACT:
SUBMITTER: Yang Y
PROVIDER: S-EPMC5039691 | biostudies-literature | 2016 Sep
REPOSITORIES: biostudies-literature
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Quantitative Proteomics Reveals the Defense Response of Wheat against Puccinia striiformis f. sp. tritici.
Yang Yuheng Y Yu Yang Y Bi Chaowei C Kang Zhensheng Z
Scientific reports 20160928
Wheat stripe rust, caused by Puccinia striiformis f. sp. tritici (Pst), is considered one of the most aggressive diseases to wheat production. In this study, we used an iTRAQ-based approach for the quantitative proteomic comparison of the incompatible Pst race CYR23 in infected and non-infected leaves of the wheat cultivar Suwon11. A total of 3,475 unique proteins were identified from three key stages of interaction (12, 24, and 48 h post-inoculation) and control groups. Quantitative analysis sh ...[more]
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Project description:MicroRNAs are small RNAs that regulate gene expression in eukaryotes. In this study, we analyzed the small RNA profiles of two cultivars that exhibit different reactions to stripe rust infection: one susceptible, the other partially resistant. Using small RNA libraries prepared from the two wheat cultivars infected with stripe rust fungus (Puccinia striiformis f. sp. tritici), we identified 182 previously known miRNAs, 91 variants of known miRNAs, and 163 candidate novel wheat miRNAs. Known miRNA loci were usually copied in all three wheat sub-genomes, whereas novel miRNA loci were often specific to a single sub-genome. DESeq2 analysis of differentially expressed microRNAs revealed 23 miRNAs that exhibit cultivar-specific differences. TA078/miR399b showed cultivar-specific differential regulation in response to infection. Using different target prediction algorithms, 145 miRNAs were predicted to target wheat genes, while 69 miRNAs were predicted to target fungal genes. We also confirmed reciprocal expression of TA078/miR399b and tae-miR9664 and their target genes in different treatments, providing evidence for miRNA-mediated regulation during infection. Both known and novel miRNAs were predicted to target fungal genes, suggesting trans-kingdom regulation of gene expression. Overall, this study contributes to the current repository of wheat miRNAs and provides novel information on the yet-uncharacterized roles for miRNAs in the wheat-stripe rust pathosystem.
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Project description:BACKGROUND: Puccinia striiformis f. sp. tritici is a fungal pathogen causing stripe rust, one of the most important wheat diseases worldwide. The fungus is strictly biotrophic and thus, completely dependent on living host cells for its reproduction, which makes it difficult to study genes of the pathogen. In spite of its economic importance, little is known about the molecular basis of compatible interaction between the pathogen and wheat host. In this study, we identified wheat and P. striiformis genes associated with the infection process by conducting a large-scale transcriptomic analysis using cDNA-AFLP. RESULTS: Of the total 54,912 transcript derived fragments (TDFs) obtained using cDNA-AFLP with 64 primer pairs, 2,306 (4.2%) displayed altered expression patterns after inoculation, of which 966 showed up-regulated and 1,340 down-regulated. 186 TDFs produced reliable sequences after sequencing of 208 TDFs selected, of which 74 (40%) had known functions through BLAST searching the GenBank database. Majority of the latter group had predicted gene products involved in energy (13%), signal transduction (5.4%), disease/defence (5.9%) and metabolism (5% of the sequenced TDFs). BLAST searching of the wheat stem rust fungus genome database identified 18 TDFs possibly from the stripe rust pathogen, of which 9 were validated of the pathogen origin using PCR-based assays followed by sequencing confirmation. Of the 186 reliable TDFs, 29 homologous to genes known to play a role in disease/defense, signal transduction or uncharacterized genes were further selected for validation of cDNA-AFLP expression patterns using qRT-PCR analyses. Results confirmed the altered expression patterns of 28 (96.5%) genes revealed by the cDNA-AFLP technique. CONCLUSION: The results show that cDNA-AFLP is a reliable technique for studying expression patterns of genes involved in the wheat-stripe rust interactions. Genes involved in compatible interactions between wheat and the stripe rust pathogen were identified and their expression patterns were determined. The present study should be helpful in elucidating the molecular basis of the infection process, and identifying genes that can be targeted for inhibiting the growth and reproduction of the pathogen. Moreover, this study can also be used to elucidate the defence responses of the genes that were of plant origin.
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Project description:BackgroundPlant fungal pathogens can rapidly evolve and adapt to new environmental conditions in response to sudden changes of host populations in agro-ecosystems. However, the genomic basis of their host adaptation, especially at the forma specialis level, remains unclear.ResultsWe sequenced two isolates each representing Puccinia striiformis f. sp. tritici (Pst) and P. striiformis f. sp. hordei (Psh), different formae speciales of the stripe rust fungus P. striiformis highly adapted to wheat and barley, respectively. The divergence of Pst and Psh, estimated to start 8.12 million years ago, has been driven by high nucleotide mutation rates. The high genomic variation within dikaryotic urediniospores of P. striiformis has provided raw genetic materials for genome evolution. No specific gene families have enriched in either isolate, but extensive gene loss events have occurred in both Pst and Psh after the divergence from their most recent common ancestor. A large number of isolate-specific genes were identified, with unique genomic features compared to the conserved genes, including 1) significantly shorter in length; 2) significantly less expressed; 3) significantly closer to transposable elements; and 4) redundant in pathways. The presence of specific genes in one isolate (or forma specialis) was resulted from the loss of the homologues in the other isolate (or forma specialis) by the replacements of transposable elements or losses of genomic fragments. In addition, different patterns and numbers of telomeric repeats were observed between the isolates.ConclusionsHost adaptation of P. striiformis at the forma specialis level is a complex pathogenic trait, involving not only virulence-related genes but also other genes. Gene loss, which might be adaptive and driven by transposable element activities, provides genomic basis for host adaptation of different formae speciales of P. striiformis.
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Project description:Stripe rust, caused by Puccinia striiformis f. sp. tritici (Pst), is one of the most serious diseases of wheat; therefore, exploring effective resistance-related genes is critical for breeding and studying resistance mechanisms. However, only a few stripe rust resistance genes and defence-related genes have been cloned. Moreover, transgenic wheat with enhanced stripe rust resistance has rarely been reported. Receptor-like proteins (RLPs) are known to be involved in defence and developmental pathways. In this research, a novel RLP gene TaRLP1.1 was characterized as an important stripe rust defence gene. TaRLP1.1 was screened by GeneChip and was found to be induced by Pst specifically in the resistant variety. Knock down of TaRLP1.1 in the stripe rust-resistant plants resulted in increased susceptibility to Pst, and phenolic autofluorogen accumulation at the pathogen-host interaction sites, usually correlated with the hypersensitive response, was decreased dramatically. However, when the TaRLP1.1 gene was transformed into the susceptible wheat variety Yangmai158, the transgenic plants showed highly increased resistance to Pst, and the hypersensitive response was enhanced at the infection sites. Meanwhile, the expression of pathogenesis-related genes decreased in the TaRLP1.1-silenced plants and increased in the TaRLP1.1-overexpressing plants. Thus, it was proposed that TaRLP1.1 greatly contributed to the hypersensitive response during the pathogen-host interaction. Along with the functional analysis, an evolutionary study of the TaRLP1 family was performed. Characterization of TaRLP1.1 may facilitate breeding for stripe rust resistance and better understanding of the evolution of the RLP genes in wheat.
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