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Palmer amaranth is one of the most economically important and widespread weeds of arable land in the United States. Although no populations are currently known to exist in Canada, its distribution has expanded northward such that it is present in many of the states bordering Canada and multiple pathways exist for its introduction. In this short communication, we report on the transport of viable Palmer amaranth seed on imported sweet potato slips. A reproductive pair of Palmer amaranth seedlings were identified from soil accompanying imported sweet potato slips in 2018. Identification was confirmed using species-specific single nucleotide polymorphisms.

Photosystem II (PS II)-inhibitor herbicide resistance in Ontario waterhemp [Amaranthus tuberculatus (Moq.) Sauer] populations is conferred via target-site resistance (TSR) and non–target site resistance (NTSR) mechanisms. Metribuzin-resistant (MR) A. tuberculatus is due to TSR. Conversely, in other populations of PS II–inhibitor resistant A. tuberculatus, plants are resistant to atrazine but metribuzin sensitive (MS). The objective of this study was to determine the biologically effective dose of metribuzin applied preemergence and postemergence for the control of MS and MR A. tuberculatus. Ten field experiments were conducted in 2019 and 2020 to determine the effective doses of metribuzin for 50%, 80%, and 95% control of MS and MR A. tuberculatus. Metribuzin applied preemergence at the calculated doses of 133, 350, and 1,070 g ai ha−1 controlled MS A. tuberculatus 50%, 80%, and 95%, respectively, whereas the calculated doses of 7,868 and 17,533 g ai ha−1 controlled MR A. tuberculatus 50% and 80%, respectively, at 12 wk after application (WAA). Metribuzin applied postemergence at the calculated doses of 245 and 1,480 g ai ha−1 controlled MS A. tuberculatus 50% and 80%, respectively; the calculated dose for 50% MR A. tuberculatus control was greater than the highest dose (17,920 g ai ha−1) included in this study. Metribuzin at 560 and 1,120 g ha−1 and pyroxasulfone/flumioxazin (240 g ai ha−1) applied preemergence controlled MS A. tuberculatus 88%, 95%, and 98%, respectively, at 12 WAA, whereas the same treatments only controlled MR A. tuberculatus 0%, 4%, and 93%, respectively, at 12 WAA. Metribuzin at 560 and 1,120 g ha−1 and fomesafen (240 g ai ha−1) applied postemergence controlled MS A. tuberculatus 65%, 70%, and 78%, and MR A. tuberculatus 0%, 1%, and 49%, respectively, at 12 WAA. Based on these results, PS II–inhibitor resistant A. tuberculatus with NTSR (enhanced metabolism) is controlled with metribuzin applied preemergence and postemergence; in contrast, PS II–inhibitor resistant A. tuberculatus with TSR (glycine-264-serine altered target site) is not controlled with metribuzin.

BACKGROUND Poa annua is a pervasive grassy, self‐pollinating, weed that has evolved resistance to 10 different herbicide modes‐of‐action, third most of all weed species. We investigated constitutive overexpression of genes associated with non‐target site resistance (NTSR) in POAAN‐R3 and the response of those genes when treated with trifloxysulfuron despite the biotype having a known target site mutation in acetolactate synthase (ALS). RESULTS Despite having an ALS target site mutation, POAAN‐R3 still had a transcriptomic response to herbicide application that differed from a susceptible biotype. We observed differential expression of genes associated with transmembrane transport and oxidation–reduction activities, with differences being most pronounced prior to herbicide treatment. CONCLUSIONS In the P. annua biotype we studied with confirmed target site resistance to ALS inhibitors, we also observed constitutive expression of genes regulating transmembrane transport, as well as differential expression of genes associated with oxidative stress after treatment with trifloxysulfuron. This accumulation of mechanisms, in addition to the manifestation of target site resistance, could potentially increase the chance of survival when plants are challenged by different modes of action., In Poa annua with target‐site resistance to ALS inhibitors, we observed constitutive expression of genes regulating transmembrane‐transport, and differential expression of genes associated with oxidative stress after treatment with trifloxysulfuron. © 2021 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Bacterial leaf spot of lettuce, caused by Xanthomonas hortorum pv. vitians, is an economically important disease worldwide. For instance, it caused around 4 million CAD in losses in only a few months during the winter of 1992 in Florida. Because only one pesticide is registered to control this disease in Canada, the development of lettuce cultivars tolerant to bacterial leaf spot remains the most promising approach to reduce the incidence and severity of the disease in lettuce fields. The lack of information about the genetic diversity of the pathogen, however, impairs breeding programs, especially when disease resistance is tested on newly developed lettuce germplasm lines. To evaluate the diversity of X. hortorum pv. vitians, a multilocus sequence analysis was performed on 694 isolates collected in Eastern Canada through the summers of 2014 to 2017 and two isolates in 1996 and 2007. All isolates tested were clustered into five phylogroups. Six pathotypes were identified following pathogenicity tests conducted in greenhouses, but when phylogroups were compared with pathotypes, no correlation could be drawn. However, in vitro production of xanthan and xanthomonadins was investigated, and isolates with higher production of xanthomonadins were generally causing less severe symptoms on the tolerant cultivar Little Gem. Whole-genome sequencing was undertaken for 95 isolates belonging to the pathotypes identified, and de novo assembly made with reads unmapped to the reference strain’s genome sequence resulted in 694 contigs ranging from 128 to 120,795 bp. Variant calling was performed prior to genome-wide association studies computed with single-nucleotide polymorphisms (SNPs), copy-number variants and gaps. Polymorphisms with significant p-values were only found on the cultivar Little Gem. Our results allowed molecular identification of isolates likely to cause bacterial leaf spot of lettuce, using two SNPs identified through genome-wide association study.

Herbicide resistance is selected for within a weed population through the recurrent use of an herbicide. Once the use of the herbicide is reduced or discontinued, the trait may persist in the population unless resistance endows a fitness penalty. Few studies have examined the long-term persistence of an herbicide resistance trait in a weed population once the selection pressure that led to its prevalence has been removed. The objective of this research was to re-survey the locations described by A.J.J. Smisek in her 1995 study of a paraquat resistant biotype of C. canadensis on the grounds of the Harrow Research and Development Centre. Results indicate that, ∼20 yr after paraquat use was significantly reduced, a glyphosate resistant biotype had replaced the paraquat resistant biotype at all but one of the original locations surveyed by Smisek. The sole location where the paraquat resistant biotype was observed was an old, un-treed orchard row currently managed by mowing. This location not only served as a refuge for the paraquat resistant biotype, ensuring its persistence in the face of strong selection for glyphosate resistance at all other locations, but it facilitated the co-occurrence of glyphosate and paraquat resistant biotypes. The observation of a multiple resistant biotype at this location alone, with resistance to both paraquat and glyphosate, suggests a role for refugia in the stacking of herbicide resistance traits. Given that this stacking occurred in the absence of either herbicide, we hypothesize that the multiple resistant biotype arose through pollen mediated gene flow among biotypes.

Erwinia amylovora and Pseudomonas syringae are bacterial phytopathogens responsible for considerable yield losses in commercial pome fruit production. The pathogens, if left untreated, can compromise tree health and economically impact entire commercial fruit productions. Historically, the choice of effective control methods has been limited. The use of antibiotics was proposed as an effective control method. The identification of these pathogens and screening for the presence of antibiotic resistance is paramount in the adoption and implementation of disease control methods. Molecular tests have been developed and accepted for identification and characterization of these disease-causing organisms. We improved existing molecular tests by developing methods that are equal or superior in robustness for identifying E. amylovora or P. syringae while being faster to execute. In addition, the real-time PCR-based detection method for E. amylovora provided complementary information on the susceptibility or resistance to streptomycin of individual isolates. Finally, we describe a methodology and results that compare the aggressiveness of the different bacterial isolates on four apple cultivars. We show that bacterial isolates exhibit different behaviors when brought into contact with various apple varieties and that the hierarchical clustering of symptom severity indicates a population structure, suggesting a genetic basis for host cultivar specificity.

Cover cropping is a soil conservation practice that may reduce the impacts of the economically important pathogen Pseudomonas syringae on crops including squash (Cucurbita pepo). To date, no studies have directly quantified the effect of rye cover crops on P. syringae populations, nor on the bacterial community of squash leaves. In this work, we tested the hypothesis that the protective effects of cover cropping on squash may be mediated by cover cropping effects on the plant’s microbiota that in turn protects against P. syringae. Using combined 16S sequencing and culture-based approaches, we showed that rye cover cropping protects squash against P. syringae, by decreasing pathogen population size on squash leaves and increasing fruit health and marketability at harvest. We also found evidence of a strong effect of rye cover crops on bacterial communities of the squash phyllosphere. Those findings were more striking early in the growing season. Finally, we identified numerous phyllosphere bacteria belonging to the genera Sphingomonas, Methylobacterium and Pseudomonas that were promoted by rye cover crops. Overall, our findings suggest cover cropping is effective for the sustainable management of P. syringae on squash and may provide a reservoir of potential microbial biocontrol agents colonizing the phyllosphere.

Wild buckwheat (Polygonum convolvulus L.) is a problem weed and ALS-inhibitors (e.g., chlorsulfuron) are commonly used for its management. Recently, a population of wild buckwheat (KSW-R) uncontrolled with ALS-inhibitors was found in a wheat field in Kansas, USA. The objectives of this research were to determine the level and mechanism of resistance to chlorsulfuron and cross resistance to other ALS-inhibitors in the KSW-R population. In response to chlorsulfuron rates ranging from 0 to 16x (x = 18 g ai/ha), the KSW-R wild buckwheat was found &gt, 100-fold more resistant compared to a known ALS-inhibitor susceptible (KSW-S) wild buckwheat. Also, &gt, 90% of KSW-R plants survived field recommended rates of sulfonylurea but not imidazolinone family of ALS-inhibitors. A portion of the ALS gene covering all previously reported mutations known to bestow resistance to ALS-inhibitors was sequenced from both KSW-R and KSW-S plants. The Pro-197-Ser substitution that confers resistance to the sulfonylurea herbicides was found in KSW-R plants. Our results support the evolution of high level of chlorsulfuron resistance as a result of a mutation in the ALS-gene in KSW-R buckwheat. This is the first case of resistance to any herbicides in wild buckwheat in the US.

In this short communication, we report the observation of a tricotyledenous giant ragweed (Ambrosia trifida L.) individual. In contrast to the normal dicotyledenous pattern of phyllotaxy, this individual was observed to express three axillary meristems at each node that intersected the main stem at angles of 120°.

Glyphosate-resistant populations ofConyza canadensishave been spreading at a rapid rate in Ontario, Canada, since first being documented in 2010. Determining the genetic relationship among existing Ontario populations is necessary to understand the spread and selection of the resistant biotypes. The objectives of this study were to: (1) characterize the genetic variation ofC. canadensisaccessions from the province of Ontario using simple sequence repeat (SSR) markers and (2) investigate the molecular mechanism (s) conferring resistance in these accessions. Ninety-eightC. canadensisaccessions were genotyped using 8 SSR markers. Germinable accessions were challenged with glyphosate to determine their dose response, and the sequences of 5-enolpyruvylshikimate-3-phosphate synthase genes 1 and 2 were obtained. Results indicate that a majority of glyphosate-resistant accessions from Ontario possessed a proline to serine substitution at position 106, which has previously been reported to confer glyphosate resistance in other crop and weed species. Accessions possessing this substitution demonstrated notably higher levels of resistance than non–target site resistant (NTSR) accessions from within or outside the growing region and were observed to form a subpopulation genetically distinct from geographically proximate glyphosate-susceptible and NTSR accessions. Although it is unclear whether other non–target site resistance mechanisms are contributing to the levels of resistance observed in target-site resistant accessions, these results indicate that, at a minimum, selection for Pro-106-Ser has occurred in addition to selection for non–target site resistance and has significantly enhanced the levels of resistance to glyphosate inC. canadensisaccessions from Ontario.

Prodiamine is a dinitroaniline herbicide labeled for PRE control of goosegrass in warm- and cool-season turfgrass. In 2013, several golf course roughs in Maryville, TN reported poor goosegrass control (< 20%) following prodiamine treatment at 1,120 g ai ha-1. We harvested suspected prodiamine-resistant (PR) and prodiamine-susceptible (S) goosegrass phenotypes from the field and exposed them to a range of increasing prodiamine concentrations in hydroponic culture. Exposure to prodiamine at 0.001 mM reduced root growth of the S phenotype to 11% of the non-treated check. By comparison, exposure to 0.001 mM prodiamine had minimal effect on the PR phenotype, as root growth was 94% of the non-treated check. Molecular analyses revealed that PR plants contained a threonine (Thr) to isoleucine (Ile) substitution at position 239 on the α-tubulin 1 (TUA1) protein. The substitution, found in all PR plants, is the mechanism of prodiamine resistance in this phenotype. In field studies, topramezone controlled PR goosegrass 72% to 89% by 50 d after treatment (DAT) compared to only 22% to 23% for foramsulfuron. Topramezone treatment injured bermudagrass 34% to 60% from 7 to 14 DAT; however, injury was≤6% 28 DAT and 0% by the end of the study. Our results indicate that POST applications of topramezone can control dinitroaniline-resistant goosegrass. In addition, we established an easy-to-use genotyping assay to quickly screen goosegrass phenotypes for a target-site mutation (Thr-239-Ile) on TUA1 associated with resistance to dinitroaniline herbicides such as prodiamine. Future research should work to expand this assay for use with other weed species and herbicidal modes of action.

BACKGROUND The occurrence of herbicide-resistant weed biotypes is increasing and this report of an acetyl-CoA carboxylase (ACCase) inhibitor-resistant Digitaria sanguinalis L. Scop. from southwestern Ontario is another example. The identified weed escaped control in an onion and carrot rotation in which graminicides were used for several consecutive years. Our goal was to characterize the level and mechanism of resistance of the biotype. RESULTS The biotype was resistant to all five ACCase inhibitor herbicides tested. Gene-expression profiling was performed because none of the mutations known to confer resistance in the ACCase gene were detected. RNASeq and quantitative reverse-transcriptase PCR (qRT-PCR) results indicated that transcription of ACCase was 3.4–9.3 times higher in the resistant biotype than the susceptible biotype. ACCase gene copy number was determined by qPCR to be five to seven times higher in the resistant compared with the susceptible biotype. ACCase gene overexpression was directly related to the increase of the ACCase gene copy number. CONCLUSION Our results are consistent with the hypothesis that overexpression of the herbicide target gene ACCase confers resistance to the herbicide. This is the first reported case of target gene duplication conferring resistance to a herbicide other than glyphosate. © 2017 Society of Chemical Industry See related Article

Background Due to the accessibility of underlying technologies the 'Omics', in particular genomics, are becoming commonplace in several fields of research, including the study of agricultural pests. The weed community is starting to embrace these approaches; genome sequences have been made available in the past years, with several other sequencing projects underway, as promoted by the International Weed Genome Consortium. Chromosome-scale sequences are essential to fully exploit the power of genetics and genomics. Results We report such an assembly for Conyza canadensis, an important agricultural weed. Third-generation sequencing technology was used to create a genome assembly of 426 megabases, of which nine chromosome-scale scaffolds cover more than 98% of the entire assembled sequence. As this weed was the first to be identified with glyphosate resistance, and since we do not have a firm handle on the genetic mechanisms responsible for several herbicide resistances in the species, the genome sequence was annotated with genes known to be associated with herbicide resistance. A high number of ABC-type transporters, cytochrome P450 and glycosyltransferases (159, 352 and 181, respectively) were identified among the list of ab initio predicted genes. Conclusion As C. canadensis has a small genome that is syntenic with other Asteraceaes, has a short life cycle and is relatively easy to cross, it has the potential to become a model weed species and, with the chromosome-scale genome sequence, contribute to a paradigm shift in the way non-target site resistance is studied. © 2020 Her Majesty the Queen in Right of CanadaPest Management Science © 2020 Society of Chemical Industry.

Potato producers in the Atlantic Canadian provinces of New Brunswick (NB) and Prince Edward Island (PE) rely on the photosystem II-inhibiting herbicide metribuzin for weed management. Recently, potato producers in the region have reported unacceptable common lambsquarters control following an application of metribuzin. Tissue and seed samples were collected from escaped common lambsquarters populations from across the potato producing regions of NB and PE and screened for the Ser264Gly mutation in psbA. Overall, 46% of sampled populations possessed the Ser264Gly mutation across the region. Cross-resistance testing to atrazine, metribuzin and linuron confirmed populations with the Ser264Gly were resistant to triazines and triazinones but remained susceptible to linuron. Dose response analysis determined a moderate level of resistance to metribuzin in common lambsquarters which would not be controlled in producers fields. A field experiment was conducted in Fredericton, NB and Harrington, PE, to determine if currently registered and unregistered products and tank-mixes would control PSII-inhibitor-resistant common lambsquarters in potato. All evaluated products, with the exception of S-metolachlor, provided control equivalent to the weed-free check without compromising potato yield or quality. This study demonstrates that PSII-inhibitor-resistant common lambsquarters are found in Atlantic Canadian potato production systems, but can be controlled with currently registered herbicides and rates with alternative modes of action.

Common ragweed (Ambrosia artemisiifolia L.) is frequently observed in Quebec carrot fields. Carrot growers essentially rely on linuron, a photosystem II inhibitor, to control this broadleaf weed. A...

The two forms of cytoplasmic male sterility (CMS) native to the oilseed rape or canola species Brassica napus, nap and pol, have novel features that may provide insight into the molecular mechanisms through which CMS/nuclear restorer systems evolve. One such feature is the finding that the distinct nuclear restorer genes for the two systems represent different alleles or haplotypes of the same nuclear locus. Improved understanding of how these systems have evolved will require molecular cloning and characterization of this novel locus. We have employed an approach that exploits the regional co-linearity between the Arabidopsis and Brassica genomes to construct a high-resolution genetic map of the nuclear restorer for the pol system, Rfp. Specifically, Arabidopsis-derived sequences have been used as a set of ordered RFLP probes to localize Rfp to a region of the B. napus genome equivalent to a 115 kb interval on Arabidopsis chromosome 1. Based on the known relationship of physical distances between orthologous segments of Arabidopsis and Brassica chromosomes, it is anticipated that the B. napus restorer locus is now mapped to sufficient resolution to permit its isolation and characterization.

A single radish nuclear gene, Rfo, restores Ogura (ogu) cytoplasmic male sterility (CMS) in Brassica napus. A map-based cloning approach relying on synteny between radish and Arabidopsis was used to clone Rfo. A radish gene encoding a 687-amino-acid protein with a predicted mitochondrial targeting pre-sequence was found to confer male fertility upon transformation into ogu CMS B. napus. This gene, like the recently described Petunia Rf gene, codes for a pentatricopeptide repeat (PPR)-containing protein with multiple, in this case 16, PPR domains. Two similar genes that do not appear to function as Rfo flank this gene. Comparison of the Rfo region with the syntenic Arabidopsis region indicates that a PPR gene is not present at the Rfo-equivalent site in Arabidopsis, although a smaller and related PPR gene is found about 40 kb from this site. The implications of these findings for the evolution of restorer genes and other PPR encoding genes are discussed.

Profilin is a ubiquitous actin-monomer-binding protein. The protistPhysarum polycephalum contains two profilins, ProA and ProP, present in amoebae and plasmodia, respectively. We have used mutantSaccharomyces cerevisiae cells in an attempt to observe distinct functions for the two profilins. Profilin-deficient yeast cells (Δpfy1) have delocalized actin cortical patches, do not contain visible actin cables, have reduced mating efficiency and do not grow at 37 °C or in the presence of caffeine. Deletion of theSRV2 gene (Δsrv2), coding for the adenylyl cyclase-associated protein, also results in an altered actin distribution and an inability to survive on rich medium. We found that the Δpfy1 and Δsrv2 mutant phenotypes were corrected equally well by the overexpression of Physarum ProA or yeast Pfy1p profilins. The Δpfy1 cells overexpressing ProP have improved mating efficiency and a normal distribution of actin cortical patches. These cells, however, have barely detectable actin cables, do not grow at 37 °C, and are sensitive to caffeine. Also, the expression of ProP does not correct the growth defect of the Δsrv2 cells. These results suggest that the two Physarum proteins are not functionally equivalent in yeast cells. No difference was detected in the affinity of ProA and ProP for poly-L-proline, while ProA has a slightly greater affinity than ProP for phosphatidylinositol 4,5-biphosphate.