Conserved defense responses between maize and sorghum to Exserohilum turcicum.

BACKGROUND:Exserohilum turcicum is an important pathogen of both sorghum and maize, causing sorghum leaf blight and northern corn leaf blight. Because the same pathogen can infect and cause major losses for two of the most important grain crops, it is an ideal pathosystem to study plant-pathogen evolution and investigate shared resistance mechanisms between the two plant species. To identify sorghum genes involved in the E. turcicum response, we conducted a genome-wide association study (GWAS). RESULTS:Using the sorghum conversion panel evaluated across three environments, we identified a total of 216 significant markers. Based on physical linkage with the significant markers, we detected a total of 113 unique candidate genes, some with known roles in plant defense. Also, we compared maize genes known to play a role in resistance to E. turcicum with the association mapping results and found evidence of genes conferring resistance in both crops, providing evidence of shared resistance between maize and sorghum. CONCLUSIONS:Using a genetics approach, we identified shared genetic regions conferring resistance to E. turcicum in both maize and sorghum. We identified several promising candidate genes for resistance to leaf blight in sorghum, including genes related to R-gene mediated resistance. We present significant advancements in the understanding of host resistance to E. turcicum, which is crucial to reduce losses due to this important pathogen

Inhibition of ethylene involved in resistance to E. turcicum in an exotic-derived double haploid maize population

Northern corn leaf blight (NCLB) is an economically important disease of maize. While the genetic architecture of NCLB has been well characterized, the pathogen is known to overcome currently deployed resistance genes, and the role of hormones in resistance to NCLB is an area of active research. The objectives of the study were (i) to identify significant markers associated with resistance to NCLB, (ii) to identify metabolic pathways associated with NCLB resistance, and (iii) to examine role of ethylene in resistance to NCLB. We screened 252 lines from the exotic-derived double haploid BGEM maize population for resistance to NCLB in both field and greenhouse environments. We used a genome wide association study (GWAS) and stepwise regression to identify four markers associated with resistance, followed by a pathway association study tool (PAST) to identify important metabolic pathways associated with disease severity and incubation period. The ethylene synthesis pathway was significant for disease severity and incubation period. We conducted a greenhouse assay in which we inhibited ethylene to examine the role of ethylene in resistance to NCLB. We observed a significant increase in incubation period and a significant decrease in disease severity between plants treated with the ethylene inhibitor and mock-treated plants. Our study confirms the potential of the BGEM population as a source of novel alleles for resistance. We also confirm the role of ethylene in resistance to NCLB and contribute to the growing body of literature on ethylene and disease resistance in monocots

Insights Into Disease Resistance: Genetic Architecture, Genes, And Pleiotropy In Maize

The genes and mechanisms underlying quantitative disease resistance remain largely elusive. The objective of this dissertation was to resolve the structure of multiple disease resistance loci, explore the dynamics that shape the genome at those loci, and identify genes associated with plant defense. In order to do this, both locus-specific and genome-wide approaches were taken, as each resistance locus has a unique resistance profile and mechanism(s) of resistance. Bins 1.02 and 1.06 of the maize genome carry loci of interest conditioning multiple disease resistance. The two loci differ in allelic diversity, pathogen specificity, and mechanism of resistance. The locus in bin 1.06 is particularly interesting, as it has been characterized as yield-stabilizing and exhibits signs of genome plasticity. I have used fine-mapping, association mapping, expression evidence, and mutant analysis to dissect these loci, identify candidate genes, and demonstrate the role of candidate genes in plant defense. Each locus was unique, although common themes arose. Both loci may have multiple underlying genes, demonstrating that the genetic architecture of disease resistance is complex. Resistance to multiple diseases appears to be due to linkage, although there may be a role for pleiotropy at both loci. Fine-mapping narrowed the intervals, and was complemented by association mapping and expression analysis to evaluate candidate genes. A putative remorin was implicated by fine-mapping and expression analysis; roughsheath2-interacting KH domain protein (rik) and pangloss1 (pan1) were identified through fine-mapping and association mapping. rik was later eliminated as a candidate for the QTL of interest through fine-mapping and association mapping. Mutants were used to confirm the role of candidate genes in plant defense, including for pan1 and the putative remorin. Based on these results, pan1 was inferred to be a susceptibility gene for NLB and Stewart's wilt, and increased resistance was correlated with decreased expression. Susceptibility conditioned by wild-type pan1 could be due to a passive mechanism, such as altered anatomical structures, or an active process, such as actin re-organization during pathogen attack. To test genome-wide association mapping candidate genes, mutants were identified and evaluated for NLB phenotype. Approximately 37% of the 123 families tested differed in disease phenotype from the background line. One of these was the putative remorin gene, which was inferred to contribute to resistance. Overall, I have examined candidate genes, explored genomic structure at these loci, and demonstrated a role for pan1 in resistance to multiple diseases

High-Throughput Resequencing of Maize Landraces at Genomic Regions Associated with Flowering Time

Publisher's PDFDespite the reduction in the price of sequencing, it remains expensive to sequence and assemble whole, complex genomes of multiple samples for population studies, particularly for large genomes like those of many crop species. Enrichment of target genome regions coupled with next generation sequencing is a cost-effective strategy to obtain sequence information for loci of interest across many individuals, providing a less expensive approach to evaluating sequence variation at the population scale. Here we evaluate amplicon-based enrichment coupled with semiconductor sequencing on a validation set consisting of three maize inbred lines, two hybrids and 19 landrace accessions. We report the use of a multiplexed panel of 319 PCR assays that target 20 candidate loci associated with photoperiod sensitivity in maize while requiring 25 ng or less of starting DNA per sample. Enriched regions had an average on-target sequence read depth of 105 with 98% of the sequence data mapping to the maize ‘B73’ reference and 80% of the reads mapping to the target interval. Sequence reads were aligned to B73 and 1,486 and 1,244 variants were called using SAMtools and GATK, respectively. Of the variants called by both SAMtools and GATK, 30% were not previously reported in maize. Due to the high sequence read depth, heterozygote genotypes could be called with at least 92.5% accuracy in hybrid materials using GATK. The genetic data are congruent with previous reports of high total genetic diversity and substantial population differentiation among maize landraces. In conclusion, semiconductor sequencing of highly multiplexed PCR reactions is a cost-effective strategy for resequencing targeted genomic loci in diverse maize materials.Department of Plant and Soil Science

DataSheet_1_Inhibition of ethylene involved in resistance to E. turcicum in an exotic-derived double haploid maize population.csv

Northern corn leaf blight (NCLB) is an economically important disease of maize. While the genetic architecture of NCLB has been well characterized, the pathogen is known to overcome currently deployed resistance genes, and the role of hormones in resistance to NCLB is an area of active research. The objectives of the study were (i) to identify significant markers associated with resistance to NCLB, (ii) to identify metabolic pathways associated with NCLB resistance, and (iii) to examine role of ethylene in resistance to NCLB. We screened 252 lines from the exotic-derived double haploid BGEM maize population for resistance to NCLB in both field and greenhouse environments. We used a genome wide association study (GWAS) and stepwise regression to identify four markers associated with resistance, followed by a pathway association study tool (PAST) to identify important metabolic pathways associated with disease severity and incubation period. The ethylene synthesis pathway was significant for disease severity and incubation period. We conducted a greenhouse assay in which we inhibited ethylene to examine the role of ethylene in resistance to NCLB. We observed a significant increase in incubation period and a significant decrease in disease severity between plants treated with the ethylene inhibitor and mock-treated plants. Our study confirms the potential of the BGEM population as a source of novel alleles for resistance. We also confirm the role of ethylene in resistance to NCLB and contribute to the growing body of literature on ethylene and disease resistance in monocots.</p

Image_1_Inhibition of ethylene involved in resistance to E. turcicum in an exotic-derived double haploid maize population.pdf

Northern corn leaf blight (NCLB) is an economically important disease of maize. While the genetic architecture of NCLB has been well characterized, the pathogen is known to overcome currently deployed resistance genes, and the role of hormones in resistance to NCLB is an area of active research. The objectives of the study were (i) to identify significant markers associated with resistance to NCLB, (ii) to identify metabolic pathways associated with NCLB resistance, and (iii) to examine role of ethylene in resistance to NCLB. We screened 252 lines from the exotic-derived double haploid BGEM maize population for resistance to NCLB in both field and greenhouse environments. We used a genome wide association study (GWAS) and stepwise regression to identify four markers associated with resistance, followed by a pathway association study tool (PAST) to identify important metabolic pathways associated with disease severity and incubation period. The ethylene synthesis pathway was significant for disease severity and incubation period. We conducted a greenhouse assay in which we inhibited ethylene to examine the role of ethylene in resistance to NCLB. We observed a significant increase in incubation period and a significant decrease in disease severity between plants treated with the ethylene inhibitor and mock-treated plants. Our study confirms the potential of the BGEM population as a source of novel alleles for resistance. We also confirm the role of ethylene in resistance to NCLB and contribute to the growing body of literature on ethylene and disease resistance in monocots.</p

DataSheet_2_Inhibition of ethylene involved in resistance to E. turcicum in an exotic-derived double haploid maize population.csv

Northern corn leaf blight (NCLB) is an economically important disease of maize. While the genetic architecture of NCLB has been well characterized, the pathogen is known to overcome currently deployed resistance genes, and the role of hormones in resistance to NCLB is an area of active research. The objectives of the study were (i) to identify significant markers associated with resistance to NCLB, (ii) to identify metabolic pathways associated with NCLB resistance, and (iii) to examine role of ethylene in resistance to NCLB. We screened 252 lines from the exotic-derived double haploid BGEM maize population for resistance to NCLB in both field and greenhouse environments. We used a genome wide association study (GWAS) and stepwise regression to identify four markers associated with resistance, followed by a pathway association study tool (PAST) to identify important metabolic pathways associated with disease severity and incubation period. The ethylene synthesis pathway was significant for disease severity and incubation period. We conducted a greenhouse assay in which we inhibited ethylene to examine the role of ethylene in resistance to NCLB. We observed a significant increase in incubation period and a significant decrease in disease severity between plants treated with the ethylene inhibitor and mock-treated plants. Our study confirms the potential of the BGEM population as a source of novel alleles for resistance. We also confirm the role of ethylene in resistance to NCLB and contribute to the growing body of literature on ethylene and disease resistance in monocots.</p

DataSheet_3_Inhibition of ethylene involved in resistance to E. turcicum in an exotic-derived double haploid maize population.csv

Northern corn leaf blight (NCLB) is an economically important disease of maize. While the genetic architecture of NCLB has been well characterized, the pathogen is known to overcome currently deployed resistance genes, and the role of hormones in resistance to NCLB is an area of active research. The objectives of the study were (i) to identify significant markers associated with resistance to NCLB, (ii) to identify metabolic pathways associated with NCLB resistance, and (iii) to examine role of ethylene in resistance to NCLB. We screened 252 lines from the exotic-derived double haploid BGEM maize population for resistance to NCLB in both field and greenhouse environments. We used a genome wide association study (GWAS) and stepwise regression to identify four markers associated with resistance, followed by a pathway association study tool (PAST) to identify important metabolic pathways associated with disease severity and incubation period. The ethylene synthesis pathway was significant for disease severity and incubation period. We conducted a greenhouse assay in which we inhibited ethylene to examine the role of ethylene in resistance to NCLB. We observed a significant increase in incubation period and a significant decrease in disease severity between plants treated with the ethylene inhibitor and mock-treated plants. Our study confirms the potential of the BGEM population as a source of novel alleles for resistance. We also confirm the role of ethylene in resistance to NCLB and contribute to the growing body of literature on ethylene and disease resistance in monocots.</p

An assessment of the performance of the logistic mixed model for analyzing binary traits in maize and sorghum diversity panels.

The logistic mixed model (LMM) is well-suited for the genome-wide association study (GWAS) of binary agronomic traits because it can include fixed and random effects that account for spurious associations. The recent implementation of a computationally efficient model fitting and testing approach now makes it practical to use the LMM to search for markers associated with such binary traits on a genome-wide scale. Therefore, the purpose of this work was to assess the applicability of the LMM for GWAS in crop diversity panels. We dichotomized three publicly available quantitative traits in a maize diversity panel and two quantitative traits in a sorghum diversity panel, and them performed a GWAS using both the LMM and the unified mixed linear model (MLM) on these dichotomized traits. Our results suggest that the LMM is capable of identifying statistically significant marker-trait associations in the same genomic regions highlighted in previous studies, and this ability is consistent across both diversity panels. We also show how subpopulation structure in the maize diversity panel can underscore the LMM's superior control for spurious associations compared to the unified MLM. These results suggest that the LMM is a viable model to use for the GWAS of binary traits in crop diversity panels and we therefore encourage its broader implementation in the agronomic research community

Identification of quantitative trait loci associated with maize resistance to bacterial leaf streak

Bacterial leaf streak (BLS), a foliar disease of maize (Zea mays L.) caused by Xanthomonas vasicola pv. vasculorum, recently emerged in the Americas as a disease of major importance. Little is known about the disease cycle, and consequently, management is difficult. No chemical control is available. Host resistance will likely play a major role in controlling the disease, but to date, no data regarding the resistance of maize germplasm to X. vasicola pv. vasculorum have been published. The objective of this study was to examine the genetic architecture of resistance to BLS. We conducted quantitative trait locus (QTL) mapping for BLS resistance in three maize populations: the Z022 (B73 × Oh43 recombinant inbred line) nested association mapping (NAM) population, the Z023 (B73 × Oh7B recombinant inbred line) NAM population, and the DRIL78 (NC344 × Oh7B chromosome segment substitution line) population. A total of five QTL were detected across two of the mapping populations. Of the QTL detected, one conferred a moderate effect, whereas the others conferred small effects. We also examined the relationship between resistance to BLS and resistance to three foliar diseases of maize, which had been mapped previously. The only significant correlation we found for BLS was with northern corn leaf blight [caused by Exserohilum turcicum (Pass.) K. J. Leonard & Suggs] in one of the populations, although two of the five BLS regions were involved in resistance to other diseases. These data will be useful for developing maize varieties resistant to BLS to mitigate the impact of bacterial leaf streak on maize production.This is a manuscript of an article published as Qiu, Yuting, Christopher Kaiser, Clarice Schmidt, Kirk Broders, Alison E. Robertson, and Tiffany M. Jamann. "Identification of quantitative trait loci associated with maize resistance to bacterial leaf streak." Crop Science 60, no. 1 (2020): 226-237. doi:10.1002/csc2.20099. Posted with permission.</p