Colletotrichum lentis causes anthracnose, which is a serious disease on lentil and can account for up to 70% crop loss. Two pathogenic races, 0 and 1, have been described in the C. lentis population from lentil. To unravel the genetic control of virulence, an isolate of the virulent race 0 was sequenced at 1481-fold genomic coverage. The 56.10-Mb genome assembly consists of 50 scaffolds with N scaffold length of 4.89 Mb. A total of 11 436 protein-coding gene models was predicted in the genome with 237 coding candidate effectors, 43 secondary metabolite biosynthetic enzymes and 229 carbohydrate-active enzymes (CAZymes), suggesting a contraction of the virulence gene repertoire in C. lentis. Scaffolds were assigned to 10 core and two minichromosomes using a population (race 0 × race 1, n = 94 progeny isolates) sequencing-based, high-density (14 312 single nucleotide polymorphisms) genetic map. Composite interval mapping revealed a single quantitative trait locus (QTL), qClVIR-11, located on minichromosome 11, explaining 85% of the variability in virulence of the C. lentis population. The QTL covers a physical distance of 0.84 Mb with 98 genes, including seven candidate effector and two secondary metabolite genes. Taken together, the study provides genetic and physical evidence for the existence of a minichromosome controlling the C. lentis virulence on lentil.
Resistance to anthracnose caused by the fungal pathogen Colletotrichum lentis was explored through transcriptome sequencing over a period of 24 to 96 h post-inoculation (hpi) of the partially resistant recombinant inbred lines (RIL) LR-66-528 and susceptible LR-66-524 of the crop wild relative Lens ervoides population LR-66. The development of infection vesicles and primary hyphae by C. lentis were significantly higher on susceptible RIL LR-66-524 compared to partially resistant LR-66-528 at 24 and 48 hpi, but exponential trends in fungal growth were observed between 24 to 96 hpi in both RILs. Comparison of inoculated with mock-inoculated samples revealed 3091 disease responsive genes, among which 477 were differentially expressed between the two RILs. These were clustered into six expression clusters with genes that had either high or low expression in one of the RILs. Differentially expressed genes (DEGs) were functionally annotated and included genes coding LRR and NB-ARC domain disease resistance proteins, protein detoxification, LRR receptor-like kinase family proteins, and wall-associated Ser/Thr Kinases. DEGs were compared to genes in previously published anthracnose resistance QTLs mapped in LR-66 and revealed 22 DEGs located in 3 QTLs. Expression of 21 DEGs was validated using RT-qPCR confirming expression trends in RNA-seq.
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