Comparative proteomic analysis reveals molecular differences between incompatible and compatible interaction of Erysiphe pisi in garden pea.

Comparative proteome analysis of Erysiphe pisi-infected pea genotypes; JI-2480 carrying er2 resistant gene and Arkel, the susceptible genotype by liquid chromatography- mass spectrometry (LCMS/MS QTOF) at 72 h post inoculation (hpi) revealed several differentially abundant proteins (DAPs) of both the host and the pathogen. The functional annotation of proteins through gene enrichment and KEGG pathway analyses revealed strong up-regulation of pathogenesis related protein NPR1, proteins related to defense, transportation and signal transduction, hypersensitive response, cell wall modifications, phenylpropanoid and metabolic pathways in J-72. Significant abundance of membrane-related polypeptides, kinase domains and small GTPase signal transduction-related proteins suggested their major role in plant defense. The abundance of cellular antioxidant protein, catalase and its isozyme along with calreticulin-1 and 2 in J-72 confirmed their intervention in maintaining a redox balance in powdery mildew defense. High abundance levels of Glycolysis-related proteins indicated it as a major pathway for energy source during fungal growth. The majority of pathogenicity and virulence genes were downregulated in J-72 compared to A-72, while four EKA (Effectors homologues to Avk1 and Avra10) like avirulence proteins were significantly upregulated in incompatible interaction suggesting their role in eliciting hypersensitive response in pea against E. pisi.

Comparative transcriptome of compatible and incompatible interaction of Erysiphe pisi and garden pea reveals putative defense and pathogenicity factors.

Comparative transcriptome analysis of Erysiphe pisi-infected pea (Pisum sativum) genotypes JI-2480 (resistant) and Arkel (susceptible) at 72 hours post-inoculation (hpi) was carried to detect molecular components involved in compatible and incompatible interactions. Differential gene expression was observed in Arkel and JI-2480 genotype at 72 hpi with E. pisi isolate (Ep01) using EdgeR software. Out of 32 217 transcripts, 2755 transcripts showed significantly altered gene expression in case of plants while 530 were related to E. pisi (P < 0.05). The higher transcript number of differentially expressed genes demonstrated peak activity of pathogenicity genes in plants at 72 hpi. Glycolysis was observed to be the major pathway for energy source during fungal growth. Differential gene expression of plant transcripts revealed significant expression of putative receptor and regulatory sequences involved in defense in the resistant, JI-2480 compared to susceptible, Arkel genotype. Expression of genes involved in defense and hormonal signaling, genes related to hypersensitive response, reactive oxygen species and phenylpropanoid pathway in JI-2480 indicated their crucial role in disease resistance against E. pisi. Down-regulation of transcription factors like-WRKY-28 and up-regulation of several putative pattern recognition receptors in JI-2480 compared to Arkel also suggested activation of host-mediated defense responses against E. pisi in pea.

Detection of putative pathogenicity and virulence genes of Erysiphe pisi using genome-wide in-silico search and their suppression by er2 mediated resistance in garden pea.

The biotrophic fungus, Erysiphe pisi is the chief causal agent of powdery mildew disease of garden pea. A genome-wide search using in-silico approach was carried to detect putative pathogenicity and virulence genes of E. pisi, since information about these genes and their interaction with pea is limited. Nineteen putative pathogenicity gene sequences were detected through genome-wide pathogenicity gene-search and confirmed them to be conserved in E. pisi through genomic PCRs. Fifteen of these genes expressed through reverse transcriptase-polymerase chain reaction (RT-PCR) amplifying expected band size along with fungal and plant specific internal controls. Gene sequencing and annotation revealed them to be Erysiphe-specific. A time course study was carried to monitor expression of nine of these genes through real-time quantitative (qRT)-PCR in Erysiphe-challenged plants of powdery mildew resistant pea genotype, JI-2480 carrying er2 gene and susceptible pea cultivar, Arkel. Expression of these genes was differentially and temporally regulated. They were found mostly related to signaling; cAMP-PKA (cPKA, CRP and AC) and MAPK (MST7) pathways along with MFP, TRE and PEX which are reported pathogenicity factors in other ascomycete members indicating that similar conserved pathways function in E. pisi also. These genes expressed at higher level at initial hours post inoculation (hpi) as early as 6 hpi in Arkel compared to JI-2480 implying them as pathogenicity factors. The elevated level of expression of MFP, TRE, CRP and cPKA gene sequences in E. pisi-challenged JI-2480 genotype at 12 hpi alone suggests these genes to possess a role in avirulence in JI-2480, conferring er2 mediated resistance.