Genome-wide association mapping of rust resistance in Aegilops longissima.

The rust diseases, including leaf rust caused by Puccinia triticina (Pt), stem rust caused by P. graminis f. sp. tritici (Pgt), and stripe rust caused by P. striiformis f. sp. tritici (Pst), are major limiting factors in wheat production worldwide. Identification of novel sources of rust resistance genes is key to developing cultivars resistant to rapidly evolving pathogen populations. Aegilops longissima is a diploid wild grass native to the Levant and closely related to the modern bread wheat D subgenome. To explore resistance genes in the species, we evaluated a large panel of Ae. longissima for resistance to several races of Pt, Pgt, and Pst, and conducted a genome-wide association study (GWAS) to map rust resistance loci in the species. A panel of 404 Ae. longissima accessions, mostly collected from Israel, were screened for seedling-stage resistance to four races of Pt, four races of Pgt, and three races of Pst. Out of the 404 accessions screened, two were found that were resistant to all 11 races of the three rust pathogens screened. The percentage of all accessions screened that were resistant to a given rust pathogen race ranged from 18.5% to 99.7%. Genotyping-by-sequencing (GBS) was performed on 381 accessions of the Ae. longissima panel, wherein 125,343 single nucleotide polymorphisms (SNPs) were obtained after alignment to the Ae. longissima reference genome assembly and quality control filtering. Genetic diversity analysis revealed the presence of two distinct subpopulations, which followed a geographic pattern of a northern and a southern subpopulation. Association mapping was performed in the genotyped portion of the collection (n = 381) and in each subpopulation (n = 204 and 174) independently via a single-locus mixed-linear model, and two multi-locus models, FarmCPU, and BLINK. A large number (195) of markers were significantly associated with resistance to at least one of 10 rust pathogen races evaluated, nine of which are key candidate markers for further investigation due to their detection via multiple models and/or their association with resistance to more than one pathogen race. The novel resistance loci identified will provide additional diversity available for use in wheat breeding.

Role of Yeasts in the Cranberry Fruit Rot Disease Complex.

Cranberry fruit rot (CFR) is an economically important disease caused by at least 10 species of filamentous fungi. Despite the application of fungicides, incidence of CFR is sometimes high, raising the possibility of a role for microbes other than fungi in the CFR complex. Isolation of microbes from rotten berries on culture media that favor either bacteria or yeasts resulted in mucoid colonies from <15% of dry-harvested rotten berries but up to 60% of wet-harvested berries. The mucoid colonies were identified as yeasts, primarily in the family Saccharomycetaceae. Inoculation of sound berries with three yeasts, Hanseniaspora uvarum, Pichia fermentans, and Pichia terricola, resulted in significantly higher incidence and severity of rot symptoms compared with mock-inoculated controls; these yeasts were recovered from inoculated berries, providing evidence of their pathogenicity. The minimum concentrations of azoxystrobin, chlorothalonil, and prothioconazole that resulted in 80% inhibition of growth compared with untreated controls (MIC80) were determined for a subset of yeasts. In general, MIC80s were higher for azoxystrobin and prothioconazole (usually >64 µg/ml) than for chlorothalonil (usually ≤1 µg/ml). To complement culture-dependent studies, DNA was isolated from wet- and dry-harvested rotten berries, and fungi were identified to the level of family by high-throughput sequencing of the fungal internal transcribed spacer region. There were no fungal families consistently detected among samples by one method (culturing or high-throughput sequencing) and missed by the other that have not previously been reported in cranberry; however, some fungal families were found to be more abundant by one method versus the other. Harvest method (wet or dry) had a significant effect on the composition of fungal communities of rotten berries (P < 0.001), and operational taxonomic units representing the Saccharomycetaceae were more abundant in wet- than dry-harvested berries. Taken together, the results suggest that some yeasts are pathogenic to cranberry and may be especially relevant in wet-harvested berries.

Hyperspectral imaging and improved feature variable selection for automated determination of deoxynivalenol in various genetic lines of barley kernels for resistance screening.

Fusarium head blight (FHB), a fungus disease of small grain cereal crops, results in reduced yields and diminished value of harvested grain due to the presence of deoxynivalenol (DON), a mycotoxin produced by the causal pathogen Fusarium graminearum. DON and other tricothecene mycotoxins pose serious health risks to both humans and livestock, especially swine. Due to these health concerns, barley used for malting, food or feed is routinely assayed for DON levels. Various methods are available for assaying DON levels in grain samples including enzyme-linked immunosorbent assay (ELISA) and gas chromatography-mass spectrometry (GC-MS). ELISA and GC-MS are very accurate; however, assaying grain samples by these techniques are laborious, expensive and destructive. In this study, we explored the feasibility of using hyperspectral imaging (382-1030 nm) to develop a rapid and non-destructive protocol for assaying DON in barley kernels. Samples of 888 and 116 from various genetic lines were selected for calibration and prediction. Full-wavelength locally weighted partial least squares regression (LWPLSR) achieved high accuracy with the coefficient of determination in prediction (R2P) of 0.728 and root mean square error of prediction (RMSEP) of 3.802. Competitive adaptive reweighted sampling (CARS) was used to choose potential feature wavelengths, and these selected variables were further optimized using the iterative selection of successive projections algorithm (ISSPA). The CARS-ISSPA-LWPLSR model developed using 7 feature variables yielded R2P of 0.680 and RMSEP of 4.213 in DON content prediction. Based on the 7 wavelengths selected by CARS-ISSPA, partial least square discriminant analysis (PLSDA) discriminated barley kernels having lower DON (less than1.25 mg/kg) levels from those with higher levels (including 1.25-3 mg/kg, 3-5 mg/kg, and 5-10 mg/kg), with Matthews correlation coefficient in cross-validation (M-RCV) of as high as 0.931. The results demonstrate that hyperspectral imaging have potential for accelerating non-destructive DON assays of barley samples.

Characterization of Blueberry shock virus, an Emerging Ilarvirus in Cranberry.

Blueberry shock virus (BlShV), an Ilarvirus sp. reported only on blueberry, was associated with scarring, disfigurement, and premature reddening of cranberry fruit. BlShV was detected by triple-antibody sandwich enzyme-linked immunosorbent assay and reverse-transcription polymerase chain reaction, and isometric virions of 25 to 28 nm were observed in cranberry sap. The virus was systemic, although unevenly distributed in plants. The coat protein of BlShV from cranberry shared 90% identity compared with BlShV accessions from blueberry on GenBank. Phylogenetic analysis of isolates of BlShV from cranberry collected from Wisconsin and Massachusetts did not indicate grouping by state. BlShV was detected in cranberry pollen, and seed transmission of up to 91% was observed. Artificial inoculation of cranberry flowers by pollination did not cause virus transmission. In some Nicotiana spp., rub inoculation of leaves with homogenized BlShV-positive cranberry flowers resulted in systemic infection. Cranberry plants recovered from symptoms the year after berry scarring occurred but continued to test positive for BlShV. The virus caused significant reduction in the average number of marketable fruit and average berry weight in symptomatic cranberry plants but recovered plants yielded comparably with healthy plants. Although recovery may limit the immediate economic consequences of BlShV, long-term implications of single- or mixed-virus infection in cranberry is unknown.