Abstract

Tilletia indica (Ti) - a quarantined fungal pathogen of wheat and its pathogenesis is chiefly governed by pathogen effectors secreted inside the host plant. The de novo genome sequencing of several field isolates and stages available could be used for understanding the molecular pathogenesis. The presence of gaps and low coverage of assembled genomes poses a problem in accurate functional annotation of such functions. In the present study attempts were made to improve the Ti draft genome through reconciliation of globally available datasets of three highly virulent monoteliospore cultures of Ti field isolates. It has sequence depth of 107x and N50 scaffold size of 80,772 (more than 26 times as large as achieved in the draft assembly) with highest sequence contiguity, more accurate and nearly complete. Functional annotation revealed that Ti genome contains 9209 genes evolved with many expanded gene families and arranged mostly in a cluster. About 79% of Ti genes were orthologous to other basidiomycetes fungi, Around 7.93% proteins were having secretary signals and 6.66% were identified as highly virulent pathogenicity genes. Using improved Ti genome as a reference, the genomic variation was assessed with respect to repeats, SNPs/InDel, gene families and correct set of virulence associated genes during its life cycle. The comparative intra-species, inter-stage and inter-species genomic variation will have broader implications to understand the gene regulatory networks involved in growth, mating and virulence behaviour of Tilletia f. spp. and also for better appreciation of fungal biology and disease management.

Highlights

  • Karnal bunt (KB) of wheat crops was first reported in Karnal in India in 19311 and caused by the smut fungus Tilletia indica (Ti) basidiomycetes belonging to the subphylum Ustilaginomycotina

  • Genome assemblies using hybridSPAdes performed with multiple k-mer combination ranging from 21 to 101 suggested that the best assemblies was built with k-mers 63, 65, 67 and 69

  • Pathogen interactions are dynamic and involve high rates of crossing resulting in increased pathogenicity and enhanced capability to resist the host plant immune system

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Summary

Introduction

Karnal bunt (KB) of wheat crops was first reported in Karnal in India in 19311 and caused by the smut fungus Tilletia indica (Ti) basidiomycetes belonging to the subphylum Ustilaginomycotina. The improved Ti genome sequence with decreased redundancy and genomes described by different Ti projects was employed to examine inter-stage genomic diversity in teliospores (mycelium), monosporidial lines (+ and − mating types) and dikaryon level during different developmental stages in Ti lifecycle. Such comparative genomic analysis of monosporidial and monoteliosporic cultures will help in unravelling the complexity of molecular pathogenesis of Ti pathogen of wheat and understanding the mechanisms associated with intra-species and inter-species genetic variability at molecular level. Identified SSR, SNP, Indel and repeat elements among different field isolates analyzed apart from establishing accurate phylogenic relationships that may help in better understanding the plasticity and dynamic behaviour of Ti genomes to overcome the host plant immune systems

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