Identification and Characterization of Late Blight Resistance Genes in Potato

Identification and Characterization of Late Blight Resistance Genes in Potato
Author: Sripad Joshi
Publisher:
Total Pages:
Release: 2021
Genre:
ISBN:

"Potato (Solanum tuberosum) is one of the major non-cereal staple food crops grown across the world. Canada produces 4.4 million tonnes of potato. The major constraints in potato production are the biotic and abiotic stresses. Among the biotic stresses, late blight of potato is one of the destructive diseases, caused by an oomycete, Phytophthora infestans. Late blight of potato causes up to 40% yield loss worldwide. Resistance to late blight is either qualitative or quantitative. Even though quantitative resistance is durable, the genetic bed rock underlying resistance is not well deciphered, which limits its applications. The objective of this study was to identify the resistance genes and their mechanisms, in a resistant genotype (Libertas) and a commercial susceptible genotype (AG704). The first study aimed at transcriptomics and metabolomics to identify the induced resistance related metabolites and genes, including the large impact transcription factors which regulate these genes and metabolites that are in higher fold change in a resistant genotype than in a susceptible genotype. A total of 160 induced metabolites and 611 induced genes were identified. A Transcription Factor (TF) enrichment study identified a total of 134 regulatory TFs, which were highly enriched in the promoters of induced genes. A correlation study among the induced genes further mapped several primary and secondary TFs. Among these bHLH66, MYB61, NAC56, WRKY51, MYB like, ERF RAP2-3 and MADS-box AGL15 had regulating sites in more than 208 downstream genes, of which many were secondary TFs. Hence, a two-tier transcriptional regulation of defense response genes was mapped. The metabolic profiling identified Hydroxycinnamic acid amides (HCAAs). Feruloylagmatine was highly accumulated in the resistant genotype post pathogen inoculation. Two major genes StACT1 and StACT2 which might be responsible were characterized by gene sequencing, qPCR and in silico protein docking data and it revealed the StACT2 gene to be mainly responsible for feruloylagmatine accumulation in potato. We found a NAC TF regulating StACT2 promoter region and it was identified as NAC72, which was accumulated more in the resistant than in the susceptible genotype. When the NAC72 CDS and promoter regions were sequenced a mutation was found in the promoter of NAC72. This gene was edited in a late blight susceptible commercial genotype, AG704, to enhance resistance. The expressions of genes StACT2 and NAC72 were significantly increased, however, the disease severity showed no significant difference. It is possible that there may be a missing chain in the hierarchy of genes involved in the regulation and/or in the metabolic pathway network to synthesize the metabolite feruloylagmatine, which is known to enhance disease resistance through deposition of these metabolites to reinforce the secondary cell walls, thus containing the pathogen to initial infection area. Discovery and editing of other mutated gene(s) should enhance feruloylagmatine production, as in resistant genotype. The other genes identified here also can be used in future for genome editing to increase RR metabolite accumulation in potato to enhance late blight and other disease resistance in potato"--

Field assessment of resistance in potato to Phytophthora infestans

Field assessment of resistance in potato to Phytophthora infestans
Author: Forbes, G
Publisher: International Potato Center
Total Pages: 40
Release: 2014-06-27
Genre:
ISBN: 9290604409

The present guide is designed to assist professionals and technicians in charge of evaluation trials designed to screen selected potato genotypes for resistance to this disease. The evaluation of breeding families, which is carried out under greenhouse or field conditions, can use the same methodology. The guide can help to organize trials, improve data collection and analysis and introduces new criteria for resistance measurement based on epidemiological principles.

Genetic Characterization and Mapping of Late Blight Resistance Genes in the Wild Tomato Accessions PI 163245 and PI 224710

Genetic Characterization and Mapping of Late Blight Resistance Genes in the Wild Tomato Accessions PI 163245 and PI 224710
Author: Erik Ohlson
Publisher:
Total Pages:
Release: 2015
Genre:
ISBN:

Late blight (LB), caused by the oomycete Phytophthora infestans (Mont.) de Bary is one of the most destructive diseases of tomato and potato worldwide. Development of fungicide resistant and more aggressive P. infestans clonal lineages has emphasized the importance of discovering and incorporating new genetic resistance in tomato cultivars. Although the cultivated tomato, Solanum lycopersicum L., contains limited genetic diversity, several related wild species of tomato are suitable for identification of new desirable traits. Previously, 67 S. pimpinellifolium accessions were screened for LB resistance in field, greenhouse and detached leaflet trials and 12 accessions with strong resistance to LB were identified. In this dissertation, two resistant accessions, PI 163245 and PI 224710, were selected for further genetic characterization. PI 163245 and PI 224710 were each hybridized with a LB susceptible tomato breeding line, Fla. 8059, and F1 progeny were self-fertilized to develop F2 populations. Large F2 populations were grown and screened for LB resistance under greenhouse conditions, and the most resistant and most susceptible individuals in each F2 population were retained for conducting heritability studies as well as identifying and mapping of resistance loci.To characterize the genetic basis of resistance in the two accessions, estimates of heritability (h2) were obtained based on F2:F3 parent-offspring (P:O) correlation analyses. An additional estimate of h2 was obtained based on F3:F4 generations when using the accession PI 163245. Estimates of h2 were moderately-high for both PI 163245 (h2F2:F3 = 0.78, h2F3:F4 = 0.94) and PI 224710 (h2F2:F3 = 0.87). The heritable nature of the resistance suggested that PI 163245 and PI 224710 were potentially viable for breeding LB resistance in tomato, and that mapping of LB resistance loci was warranted. To discover SNP markers for genetic mapping studies, reduced representation libraries (RRLs) for each of PI 163245, PI 224710 and Fla. 8059 were constructed and sequenced. Comparisons of accessions PI 163245 and PI 224710 with breeding line Fla. 8059 resulted in the identification of 33,385 and 20,894 single nucleotide polymorphisms (SNPs), respectively. The most resistant (n = 39) and susceptible (n = 35) F2 individuals in the PI 163245 mapping population were genotyped with 233 SNP markers, which were distributed throughout the genome. This selective genotyping approach identified four genomic intervals (quantitative trait loci, QTLs) on chromosomes 2, 3, 10, and 11 associated with LB resistance. Similarly, the most resistant (n = 40) and susceptible (n = 40) F2 individuals in the PI 224710 mapping population were genotyped with 144 SNPs, and LB resistance QTLs were identified on chromosomes 1, 2, 10, and 12. Resistance QTLs on chromosomes 2 and 10 co-localized with two previously identified LB resistance genes, qPh2.1 and Ph-2 respectively, however fine mapping and cloning is necessary to determine how these QTLs correspond to previously identified LB resistance genes. The remaining resistance QTLs did not appear to correspond to known LB resistance genes or QTLs in tomato, and thus are likely unique to this study. Efforts to fine map these resistance QTLs and incorporate them into elite tomato breeding lines are currently in progress.