Fusarium head blight (FHB) is one of the most significant causes of economic loss in cereal crops, resulting in a loss of $50–300 million for Canadian agriculture. The infected grain (containing Fusarium-damaged kernels (FDKs)) is often both lower in quality and kernel weight, and it may be unsuitable for human and animal consumption due to mycotoxin presence. However, it still contains a considerable amount of nutrients. A method to recover the nutrients without the mycotoxins should be beneficial for the agricultural economy. In this study, our objective was to examine recovery methods of the nutrients in relation to mycotoxin accumulation in the insect. The FDKs were fermented with Aspergillus oryzae and/or Lactobacillus plantarum (solid-state fermentation (SSF)). The SSF kernels were then provided to 50 young, black soldier fly larvae (BSFL) for 12 days. Weight gain, chemical composition, and mycotoxin bioaccumulation of BSFL and spent feed were evaluated. After 12 days of insect culture, the BSFL grew 5–6 times their initial weight. While the overall weights did not significantly vary, the proteins and lipids accumulated more in SSF FDK-fed insects. During the active growth period, the larval biomass contained deoxynivalenol (DON), a mycotoxin, at detectable levels; however, by day 12, when the larvae were in the pre-pupal stage, the amount of DON in the insect biomass was nearly negligible, i.e., BSFL did not accumulate DON. Thus, we conclude that the combination of BSFL and SSF can be employed to recover DON-free nutrients from FHB-infected grain to recover value from unmarketable grain.

The reemergence and spread of Xanthomonas translucens, the causal agent of bacterial leaf streak in cereal crops and wilt in turfgrass and forage species, is a concern to growers in the United States and Canada. The pathogen is seedborne and listed as an A2 quarantine organism by EPPO, making it a major constraint to international trade and exchange of germplasm. The pathovar concept of the X. translucens group is confusing due to overlapping of plant host ranges and specificity. Here, comparative genomics, phylogenomics, and 81 up-to-date bacterial core gene set (ubcg2) were used to assign the pathovars of X. translucens into three genetically and taxonomically distinct clusters. The study also showed that whole genome-based digital DNA–DNA hybridization unambiguously can differentiate the pvs. translucens and undulosa. Orthologous gene and proteome matrix analyses suggest that the cluster consisting of graminis, poae, arrhenatheri, phlei, and phleipratensis is very divergent. Whole-genome data were exploited to develop the first pathovar-specific TaqMan real-time PCR tool for detection of pv. translucens on barley. Specificity of the TaqMan assay was validated using 62 Xanthomonas and non- Xanthomonas strains as well as growth chamber-inoculated and naturally infected barley leaves. Sensitivity levels of 0.1 pg (purified DNA) and 23 CFUs per reaction (direct culture) compared favorably with other previously reported real-time PCR assays. The phylogenomics data reported here suggest that the clusters could constitute novel taxonomic units or new species. Finally, the pathovar-specific diagnostic tool will have significant benefits to growers and facilitate international exchange of barley germplasm and trade.

Two probe-based quantitative PCR (qPCR) systems, namely P-Xtt and P-Xtu, were developed to diagnose cereal bacterial leaf streak pathogens Xanthomonas translucens pv. translucens and pv. undulosa, respectively. P-Xtt is specific to pv. translucens, and P-Xtu is specific to pv. undulosa, pv. cerealis, pv. secalis, and pv. pistaciae. P-Xtt and P-Xtu worked on all accessible strains of pv. translucens and pv. undulosa, respectively. Both systems could detect 100 copies of the target gBlock DNA. The two systems could be used in both singleplex qPCR and duplex qPCR with similar efficiencies. On genomic DNA from strains of various X. translucens pathovars, both singleplex and duplex qPCR could specifically detect and differentiate pv. translucens and pv. undulosa. The duplex qPCR could detect pv. translucens and pv. undulosa from genomic DNA of 1,000 bacterial cells. On infected barley and wheat grain samples and on one infected wheat leaf sample, the duplex qPCR showed similar efficiency compared to a previously published qPCR system but with the additional capability of pathovar differentiation. The duplex qPCR system developed in this study will be useful in studies on bacterial leaf streak and detection/differentiation of the pathogens.

Fusarium head blight (FHB), caused by Fusarium spp., is a destructive disease of cereal grains. Apart from grain yield loss, a major quality concern is contamination with Fusarium-produced mycotoxi...

A triplex quantitative PCR (qPCR) system was developed for the simultaneous detection of the three most prevalent wheat leaf spot diseases: Septoria nodorum blotch caused by Parastagonospora nodorum, Septoria tritici blotch caused by Zymoseptoria tritici, and tan spot caused by Pyrenophora tritici-repentis. In this system, the primer set for P. tritici-repentis targets a species-specific multicopy genomic region, whereas the primer sets for the other two pathogens target the ribosomal DNA (rDNA) region. The specificity of the system was validated through sequence analysis using the currently available database and by testing against 24 DNA samples from nontarget species. Sensitivity testing on serial DNA dilutions from the three target species demonstrated that the system can detect as little as 2 fg of DNA of each species in a 20-μl reaction. For P. nodorum, the system was capable of detecting DNA extracted from a conidia suspension containing as few as 100 conidia. The system was further evaluated on 145 wheat leaf samples (45 symptomatic and 100 asymptomatic) collected from various fields in Alberta, Canada. At least one of the three pathogens was detected in 112 out of the 145 samples, with P. nodorum and/or P. tritici-repentis identified in 74 of the 100 asymptomatic samples. This triplex qPCR system offers a powerful tool for the diagnosis of wheat leaf spot diseases, surveillance, breeding for disease resistance, and research in epidemiology, population genetics, and host-pathogen interactions.