Genome-Wide Association Study Reveals a QTL Region for Ashy Stem Blight Resistance in PRA154 Andean Common Bean.

Ashy stem blight (ASB) caused by Macrophomina phaseolina (Tassi) Goidanich affects the common bean (Phaseolus vulgaris L.) at all growing stages. Higher levels of resistance were observed in Andean common beans, but specific resistant quantitative trait loci (QTLs) conferring resistance to this pathogen have not been reported in this gene pool. The objectives of this research were to: (i) conduct a genome-wide association study (GWAS) and QTL mapping for resistance in the Andean breeding line PRA154; and (ii) identify single nucleotide polymorphism (SNP) markers and candidate genes for ASB resistance. Phenotyping was conducted under greenhouse conditions by inoculating the 107 F6:7 recombinant inbred lines (RILs) derived from the cross between the susceptible cultivar 'Verano' and the partial-resistant breeding line PRA154 twice with the M. phaseolina isolate PRI21. Genotyping was performed with 109,040 SNPs distributed across all 11 P. vulgaris chromosomes. A novel major QTL was located between 28,761,668 and 31,263,845 bp, extending 2.5 Mbp on chromosome Pv07, and the highest significant SNP markers were Chr07_28761668_A_G, Chr07_29131720_G_A, and Chr07_31263845_C_T with the highest LOD (more than 10 in most of the cases) and R-squared values, explaining 40% of the phenotypic variance of the PRI21 isolate. The gene Phvul.007G173900 (methylcrotonyl-CoA carboxylase alpha chain and mitochondrial 3-methylcrotonyl-CoA carboxylase 1 [MCCA]) with a size of 10,891 bp, located between 29,131,591 and 29,142,481 bp on Pv07, was identified as one candidate for ASB resistance in PRA154, and it contained Chr07_29131720_G_A. The QTL and genetic marker information could be used to assist common bean breeders to develop germplasm and cultivars with ASB resistance through molecular breeding.

Screening a newly developed common bean germplasm with improved resistance to ashy stem blight in multiple environments.

Ashy stem blight (ASB) caused by the necrotrophic fungus Macrophomina phaseolina (Tassi) Goidanich is an important disease in common bean (Phaseolus vulgaris L.) in the Americas and worldwide. Low to intermediate levels of ASB resistance exist in cultivated and landrace genotypes of the common bean and the tertiary gene pool. However, cultivars with higher levels of resistance are not yet available. Our objectives were to 1) pyramid higher levels of resistance from multiple parent populations within the primary gene pool and 2) compare the response of the newly developed breeding lines (BL) with known sources of resistance. The BL UPR-Mp-22, UPR-Mp-34, UPR-Mp-42, and UPR-Mp-48, known sources of resistance, and susceptible checks were inoculated twice per plant with the PRI21 M. phaseolina isolate in the greenhouse and field trials conducted in Isabela and Lajas, Puerto Rico. None of the genotypes tested were resistant (mean scores 1-3). However, the new black UPR-Mp-42 and white UPR-Mp-48 BL had an intermediate response (mean scores 4-6) compared to white common bean genotypes 'Bella,' NY6020-4, and 'Verano' and black bean TARS-MST1 that were susceptible (scores ≥7) in all environments. Andean genotypes A 195, PRA154, PRA155, and UPR-Mp-22 were intermediate in the greenhouse. In contrast, UPR-Mp-34 had significantly lower scores than BAT 477 that had a susceptible reaction in the greenhouse in Isabela and in the field in Lajas and SEA 5 that was susceptible in all environments. These new BL possess an enhanced ASB resistance and may be used to improve common bean cultivars or germplasms of different market classes.

Timely Application of Four Insecticides to Control Corn Earworm and Fall Armyworm Larvae in Sweet Corn.

Insecticide sprays are a common practice to control corn earworm, Helicoverpa zea (Boddie), and fall armyworm, Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae), in corn (Zea mays L.) at reproductive stages. Our objectives were to determine (1) the most appropriate time for insecticide applications and (2) the effect of four insecticides on the survival of larvae as well as their weight. ß-cyfluthrin (0.4 mL/L), chlorantraniliprole (0.6 mL/L), emamectin benzoate (0.2 g/L), and spinetoram (1.5 mL/L) were sprayed on silks of sweet corn planted in Isabela and Lajas, Puerto Rico 3 h before and 24 and 48 h after pollination. The number of kernels produced and the damage of larvae on kernels were quantified at harvest. In addition, percentages of mortality and changes on larval weight were noted at 96 h after insecticide applications. Insecticide sprays at 3 h before pollination reduced the number of kernels or were similar to the control in all treatments. However, emamectin benzoate sprayed in Lajas and chlorantraniliprole applied in Isabela at 48 h after pollination increased the number of kernels (281−294) and reduced the damage of larvae on kernels (<0.5%) compared to the control (201−229; >7%). Furthermore, applications of emamectin benzoate caused higher percentages of fall armyworm larval mortality (>70%). Conversely, ß-cyfluthrin and chlorantraniliprole caused lower percentages of mortality (<30%) and only chlorantraniliprole and spinetoram reduced the weight of corn earworm and fall armyworm larvae collected in both locations. This information may help pest management programs and corn breeders to schedule insecticide sprays and pollination in the field.

Genetic Screening to Identify Candidate Resistance Alleles to Cry1F Corn in Fall Armyworm Using Targeted Sequencing.

Evolution of practical resistance is the main threat to the sustainability of transgenic crops producing insecticidal proteins from Bacillus thuringiensis (Bt crops). Monitoring of resistance to Cry and Vip3A proteins produced by Bt crops is critical to mitigate the development of resistance. Currently, Cry/Vip3A resistance allele monitoring is based on bioassays with larvae from inbreeding field-collected moths. As an alternative, DNA-based monitoring tools should increase sensitivity and reduce overall costs compared to bioassay-based screening methods. Here, we evaluated targeted sequencing as a method allowing detection of known and novel candidate resistance alleles to Cry proteins. As a model, we sequenced a Cry1F receptor gene (SfABCC2) in fall armyworm (Spodoptera frugiperda) moths from Puerto Rico, a location reporting continued practical field resistance to Cry1F-producing corn. Targeted sequencing detected a previously reported Cry1F resistance allele (SfABCC2mut), in addition to a resistance allele originally described in S. frugiperda populations from Brazil. Moreover, targeted sequencing detected mutations in SfABCC2 as novel candidate resistance alleles. These results support further development of targeted sequencing for monitoring resistance to Bt crops and provide unexpected evidence for common resistance alleles in S. frugiperda from Brazil and Puerto Rico.

Whole genome comparisons reveal panmixia among fall armyworm (Spodoptera frugiperda) from diverse locations.

BACKGROUND: The fall armyworm (Spodoptera frugiperda (J.E. Smith)) is a highly polyphagous agricultural pest with long-distance migratory behavior threatening food security worldwide. This pest has a host range of > 80 plant species, but two host strains are recognized based on their association with corn (C-strain) or rice and smaller grasses (R-strain). The population genomics of the United States (USA) fall armyworm remains poorly characterized to date despite its agricultural threat. RESULTS: In this study, the population structure and genetic diversity in 55 S. frugiperda samples from Argentina, Brazil, Kenya, Puerto Rico and USA were surveyed to further our understanding of whole genome nuclear diversity. Comparisons at the genomic level suggest a panmictic S. frugiperda population, with only a minor reduction in gene flow between the two overwintering populations in the continental USA, also corresponding to distinct host strains at the mitochondrial level. Two maternal lines were detected from analysis of mitochondrial genomes. We found members from the Eastern Hemisphere interspersed within both continental USA overwintering subpopulations, suggesting multiple individuals were likely introduced to Africa. CONCLUSIONS: Our research is the largest diverse collection of United States S. frugiperda whole genome sequences characterized to date, covering eight continental states and a USA territory (Puerto Rico). The genomic resources presented provide foundational information to understand gene flow at the whole genome level among S. frugiperda populations. Based on the genomic similarities found between host strains and laboratory vs. field samples, our findings validate the experimental use of laboratory strains and the host strain differentiation based on mitochondria and sex-linked genetic markers extends to minor genome wide differences with some exceptions showing mixture between host strains is likely occurring in field populations.