RESUMO
Interactions between genetic perturbations and segregating loci can cause perturbations to show different phenotypic effects across genetically distinct individuals. To study these interactions on a genome scale in many individuals, we used combinatorial DNA barcode sequencing to measure the fitness effects of 8046 CRISPRi perturbations targeting 1721 distinct genes in 169 yeast cross progeny (or segregants). We identified 460 genes whose perturbation has different effects across segregants. Several factors caused perturbations to show variable effects, including baseline segregant fitness, the mean effect of a perturbation across segregants, and interacting loci. We mapped 234 interacting loci and found four hub loci that interact with many different perturbations. Perturbations that interact with a given hub exhibit similar epistatic relationships with the hub and show enrichment for cellular processes that may mediate these interactions. These results suggest that an individual's response to perturbations is shaped by a network of perturbation-locus interactions that cannot be measured by approaches that examine perturbations or natural variation alone.
Assuntos
Epistasia Genética , Genoma Fúngico , Saccharomyces cerevisiae , Saccharomyces cerevisiae/genética , Variação Genética , Aptidão Genética , Sistemas CRISPR-Cas , Fenótipo , Código de Barras de DNA TaxonômicoRESUMO
Interactions between genetic perturbations and segregating loci can cause perturbations to show different phenotypic effects across genetically distinct individuals. To study these interactions on a genome scale in many individuals, we used combinatorial DNA barcode sequencing to measure the fitness effects of 7,700 CRISPRi perturbations targeting 1,712 distinct genes in 169 yeast cross progeny (or segregants). We identified 460 genes whose perturbation has different effects across segregants. Several factors caused perturbations to show variable effects, including baseline segregant fitness, the mean effect of a perturbation across segregants, and interacting loci. We mapped 234 interacting loci and found four hub loci that interact with many different perturbations. Perturbations that interact with a given hub exhibit similar epistatic relationships with the hub and show enrichment for cellular processes that may mediate these interactions. These results suggest that an individual's response to perturbations is shaped by a network of perturbation-locus interactions that cannot be measured by approaches that examine perturbations or natural variation alone.
RESUMO
The Solanaceae family includes commercially important vegetable crops characterized by their relative sensitivity to salinity. Evaluation of 8 eggplant (Solanum melongena), 7 tomato (Solanum lycopersicum), and 8 pepper (Capsicum spp.) heirloom cultivars from different geographic regions revealed significant variation in salt tolerance. Relative fruit yield under salt treatment varied from 52 to 114% for eggplant, 56 to 84% for tomato, and 52 to 99% for pepper. Cultivars from all three crops, except Habanero peppers, restricted Na transport from roots to shoots under salinity. The high salt tolerance level showed a strong association with low leaf Na concentration. Additionally, the leaf K-salinity/K-control ratio was critical in determining the salinity tolerance of a genotype. Differences in relative yield under salinity were regulated by several component traits, which was consistent with the gene expression of relevant genes. Gene expression analyses using 12 genes associated with salt tolerance showed that, for eggplant and pepper, Na+ exclusion was a vital component trait, while sequestration of Na+ into vacuoles was critical for tomato plants. The high variability for salt tolerance found in heirloom cultivars helped characterize genotypes based on component traits of salt tolerance and will enable breeders to increase the salt tolerance of Solanaceae cultivars.
