Abstract
Tilletia indica incites Karnal bunt (KB) disease in wheat. To date, no KB resistant wheat cultivar could be developed due to non-availability of potential biomarkers related to pathogenicity/virulence for screening of resistant wheat genotypes. The present study was carried out to compare the proteomes of T. indica highly (TiK) and low (TiP) virulent isolates. Twenty one protein spots consistently observed as up-regulated/differential in the TiK proteome were selected for identification by MALDI-TOF/TOF. Identified sequences showed homology with fungal proteins playing essential role in plant infection and pathogen survival, including stress response, adhesion, fungal penetration, invasion, colonization, degradation of host cell wall, signal transduction pathway. These results were integrated with T. indica genome sequence for identification of homologs of candidate pathogenicity/virulence related proteins. Protein identified in TiK isolate as malate dehydrogenase that converts malate to oxaloacetate which is precursor of oxalic acid. Oxalic acid is key pathogenicity factor in phytopathogenic fungi. These results were validated by GC-MS based metabolic profiling of T. indica isolates indicating that oxalic acid was exclusively identified in TiK isolate. Thus, integrated omics approaches leads to identification of pathogenicity/virulence factor(s) that would provide insights into pathogenic mechanisms of fungi and aid in devising effective disease management strategies.
Highlights
Tilletia indica is a smut fungus that incites Karnal bunt (KB), an economically important disease of wheat
The pathogenicity/virulence related proteins may serve as potential biomarkers for screening of resistant wheat genotypes and diagnosis of KB pathogen
The T. indica isolates can be categorized into three distinct aggressive types, namely Karnal Bunt- High Aggressive type (KB-HAg), Karnal Bunt- Moderate Aggressive type (KB-MAg) and Karnal Bunt- Low Aggressive type (KB-LAg)
Summary
Tilletia indica is a smut fungus that incites Karnal bunt (KB), an economically important disease of wheat. The pathogenicity/virulence related proteins may serve as potential biomarkers for screening of resistant wheat genotypes and diagnosis of KB pathogen In this context, it is pertinent to differentiate the T. indica isolates based on the degree of virulence/aggressiveness which in turn can be employed for the plant breeding programme. In the field of fungal proteomics, 2-DE coupled to tandem mass spectrometry has been extensively used for studying fungal plant pathogens with respect to their proteome maps, pathogenicity proteins and virulence factors[6,7,8] With this view, the present study was aimed to carry out the comparative proteomic analysis of mycelial proteins from T. indica isolates exhibiting varied virulence behaviour. To monitor and identify the difference between the T. indica isolates exhibiting varying virulence levels, metabolomes were carried out that would further complement our results obtained by proteomics and genomics studies
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