Plant Biology

Irsogladine maleate (2,4-diamino-6-[2,5-dichlorophenyl]-s-triazine maleate; IM), an anti-peptic ulcer drug, may have a protective effect on the gastrointestinal mucosa. This study investigated the effects of IM on spontaneous colitis in interleukin-10 gene-deficient (IL-10(-/-)) mice. Five-week-old IL-10(-/-) mice were fed a control diet or one containing 100 ppm of IM for 10 weeks. Colonic tissues were evaluated morphologically and histologically. J774A.1 murine monocyte/macrophage cells were incubated with IM after lipopolysaccharide stimulation. mRNA expression was assessed by quantitative polymerase chain reaction (PCR) and protein concentration by enzyme-linked immunosorbent assay (ELISA). Colonic length, weight, and histological scores clearly demonstrated that spontaneous colitis was prevented in IL-10(-/-) mice fed a diet containing IM compared with those fed control diet. Levels of tumor necrosis factor-alpha (TNF-α) (-2.5-fold), IL-1β (-5.4), interferon-gamma (IFN-γ) (-4.5...

Preserving remaining North American grasslands requires a multiability approach. In this book, we investigate three aspects of an admittedly larger problem: (1) how we as humans perceive grasslands; (2) the ecology of grasslands, in order to define the framework within which conservation and preservation efforts must operate; and (3) conservation issues. Additional sociological, economic, philosophical, and cultural considerations will provide important additional insights to preserving and managing grasslands, but are not included here. By restricting our focus to only three issues, we feel that we can provide a basic, but appropriate, understanding of grassland ecosystems for the prairie enthusiast. This provides an essential framework required for what we perceive to be necessary quick action. PART I: PERCEPTIONS As humans, we are trapped by our own perceptions and experience of the world, and we act within these constraints. Our first concern in this book is to highlight this is...

North American grasslands have figured prominently in our North American heritage. Prairies first provided significant barriers to westward expansion, and then offered both economic and sociological opportunity, as well as heartache, for settlers. Many artists have gained significant inspiration from the beauty as well as the harshness of this region and its biota. And because of ideal climate and soil conditions, these grasslands have provided the agricultural foundation of which much of the economic growth and stability of the United States has historically depended. Yet many see North American prairies as beautiful only when manipulated or exploited: Green croplands or manicured park lawns are attractive; native grasslands are “those ugly weeds.” In the past, plowing virgin prairie could be easily defended on both economic and sociological grounds. And, historically, North American prairies must have seemed threatening in both their wildness and their endlessness. The preservatio...

Riassunto Vengono proposte tre nuove combinazioni per il genere Lactarius Pers. nom. cons. et typ. cons.: L. australiensis (Berk. & Broome) PierotÝ, L. claridgei (Trappe) PierotÝ e L. corkii (Trappe & Claridge) PierotÝ. Queste specie devono essere inquadrate nel sottogenere Russularia (Fr.) Kauffman.

We have used atomic force microscopy and scanning tunnelling microscopy to extract new information about the substructure of the Abiils, Berzilu, Fugus. Ljcopodium and Rhododendron pollen grain exine. Our scans of exines using atomic force microscopy and scanning tunnelling microscopy reveal somewhat similar substructures for Lycopodiuiii spores and pollen of Aliiiu, Betiila, Fugits and Rliododeridron. They show various levels of alignment and clustering of substructure components. Except for AINUS, which showed polygonal clustering of spheroids and weak alignment, there is pronounced alignment of helical units. In Betula, Fagus, Lycopodiuiiz and Rliododendroii the subunits appear to be helical or perhaps consisting of elongated spheroids, these spheroids are however arranged in a way that suggest that they are part of a helical structure. The diameter of these helical subunits range from 10-15 nm in Fugus, 20-25nm in Lycopodiwn, 35-9Onm in Rliododendroii up to 70-120nm in Betula. Our preparations graded from intact or fractured fresh pollen to pollen that was acetolyzed, chemically fixed and epoxy resin embedded. While our knowledge of the exact radialfiatera1 orientation of most of our scans is less than perfect there were in all cases substructures or cross connections of exine units. We found results from scanning and transmission electron microscopy to be helpful in understanding images from Atomic Force-and Scanning Tunnelling Microscopy.

Hepcidin is a cysteine-rich cationic antimicrobial peptide central to iron metabolism. We report a comparative analysis of the sequences, gene organization and expression of two hepcidin genes from olive flounder Paralichthys olivaceus. Both consist of two introns and three exons that encode a prepropeptide (81 amino acids for hepcidin I and 89 amino acids for hepcidin II). A TATA box and several consensus-binding motifs for transcription factors were found upstream of the transcriptional starting site. Hepcidin II was predominantly expressed in the liver and highly inducible under the effect of lipopolysaccharide (LPS), while a large amount of hepcidin I transcripts was detected in various tissues but did not appear to have a significant effect during LPS-stimulation.

A timeline of nitrogen (N) dynamics of the carrot crop was developed based on studies on the effects of N application on carrot growth, N uptake and partitioning, and interactions with Alternaria [ALB; A. dauci (Kühn) Groves and Skolko] and Cercospora [CLB; C. carotae (Pass.) Solheim] leaf blight. Preplant-applied fertilizer N affects yield only when residual N in the top 60 cm of the soil profile is insufficient. The majority of the N required by the crop is taken up beyond 50 d after seeding (DAS). Carrots have a root system that extends to 150-cm depth and >50% of the fibrous root system occurs below 30-cm depth. It is likely that N below 30 cm depth is required beyond 50 DAS, since the majority of the fibrous roots are below 30 cm depth after that stage. There is insufficient time for sidedressed N to leach into the active root system beyond 50 DAS, unless its application is timed before irrigation or rain events. Regardless of the amount of residual N, preplant-applied N is essential for minimizing leaf blight severity. It is possible that leaf blight defences are conferred upon leaf emergence and may benefit from abundant N at this time. A preplant soil nitrate-N test from both the top 30 cm and the 30-to 60-cm depth could be useful for determining residual N levels and for adjusting N fertilizer application rates accordingly.

Symptom severity on differential host lines is currently used to characterize and identify pathotypes of Plasmodiophora brassicae, which is an obligate, soil-borne chromist pathogen that causes clubroot disease on canola (Brassica napus) and other brassica crops. This process is slow, variable and resource intensive; development of molecular markers could make identification of important pathotypes faster and more consistent for deployment of cultivars with pathotype-specific resistance. In the current study, a variant of gene 9171 was identified in the whole-genome sequences of only the highly virulent pathotypes of P. brassicae from around the world, including the new cohort of virulent pathotypes in Canada; its presence was confirmed using three KASP marker pairs. The gene was not present in the initial cohort of pathotypes identified in Canada. The putative structure, domains, and gene ontogeny of the protein product of gene 9171 were assessed using on-line software resources. Structural analysis of the putative protein produced by gene 9171 indicated that it was localized in the cytosol, and likely involved in cellular processes and catalytic activity. Identification of gene 9171 represents a potentially useful step toward molecular identification of the pathotypes of P. brassicae.

Aerial surveillance could be a useful tool for early detection and quantification of plant diseases, however, there are often confounding effects of other types of plant stress. Stemphylium leaf blight (SLB), caused by the fungus Stemphylium vesicarium, is a damaging foliar disease of onion. Studies were conducted to determine if near-infrared photographic images could be used to accurately assess SLB severity in onion research trials in the Holland Marsh in Ontario, Canada. The site was selected for its uniform soil and level topography. Aerial photographs were taken in 2015 and 2016 using an Xnite-Canon SX230NDVI with a near-infrared filter, mounted on a modified Cine Star-8 MK Heavy Lift RTF octocopter UAV. Images were taken at 15-20 m above the ground, providing an average of 0.5 cm/pixel and a field of view of 15 × 20 m. Photography and ground assessments of disease were carried out on the same day. NDVI (normalized difference vegetation index), green NDVI, chlorophyll index and plant senescence reflective index (PSRI) were calculated from the images. There were differences in SLB incidence and severity in the field plots and differences in the vegetative indices among the treatments, but there were no correlations between disease assessments and any of the indices.

Shanghai pak choy [Brassica rapa L. subsp. chinensis (Rupr.) var. communis Tsen and Lee] and Chinese flowering cabbage [Yow choy; B. rapa L. subsp. chinensis (Rupr.) var. utilis Tsen and Lee] were seeded into organic (muck) soil naturally infested with the clubroot pathogen (Plasmodiophora brassicae Woronin) at the

Field trials were conducted from 2008 to 2010 to assess the disease reaction to clubroot, caused by Plasmodiophora brassicae Woronin, in selected lines of Brassica spp., including short-season vegetable crops [Shanghai pak choy (B. rapa subsp. Chinensis var. communis)], Chinese flowering cabbage (B. rapa subsp. Chinensis var. utilis), and napa cabbage (B. rapa subsp. Pekinensis), the Rapid Cycling Brassica Collection (RCBC), also known as Wisconsin Fast Plants, and spring canola (B. napus). The trials were conducted on naturally infested soil with P. brassicae at the Muck Crops Research Station in Ontario, Canada, where pathotype 6 is predominant. Clubroot incidence and severity were higher in 2008 and 2010 compared with 2009. The lines of Shanghai pak choy and Chinese flowering cabbage were highly susceptible to clubroot, but each of the clubrootresistant cultivars of napa cabbage, 'Deneko', 'Bilko', and 'Yuki', was highly resistant to pathotype 6. Among the RCBC lines, B. carinata and B. juncea were highly susceptible and could be used as susceptible models for further studies. Two RCBC lines, B. napus and R. sativus, were resistant to pathotype 6. Two of the canola cultivars, 46A76 and 46A65, were susceptible, but two others, '45H21' and 'Invigor 5020LL', were highly resistant to pathotype 6. This difference in response can be exploited in future studies of clubroot reaction in canola.

A study was conducted to assess the effect of temperature on infection and development of Plasmodiophora brassicae in the root cortex of Shanghai pak choy (Brassica rapa subsp. chinensis) and on subsequent clubroot severity. Ten-day-old seedlings were grown individually, inoculated with resting spores, and maintained in growth cabinets at 10, 15, 20, 25, and 30°C. Seedlings were harvested at 2-day intervals, starting 8 days after inoculation (DAI) and continuing until 42 DAI. Roots were assessed at 4-day intervals for the incidence of cortical infection and stage of infection (young plasmodia, mature plasmodia, and resting spores

Untreated 15DSV (4 apps) 30DSV (2 apps) *Treatments are significantly different (Tukey's HSD, P≤0.05, n=16). **Fungicide treatments are significantly different from the untreated control (Tukey's HSD, P≤0.05, n=16). Data were transformed, if required, to satisfy normality assumptions. Means and standard errors are non-transformed. Brady soil (sandy) site Spray Timing Trial Untreated 15DSV (9 apps) 30DSV (5 apps) *Treatments are significantly different (Tukey's HSD, P≤0.05, n=16). **Fungicide treatments are significantly different from the untreated control (Tukey's HSD, P≤0.05, n=16). Data were transformed, if required, to satisfy normality assumptions. Means and standard errors are non-transformed Wauseon/Berrien soil (sandy loam) site Disease severity (%) 0 kg N/ha 120 kg N/ha 180 kg N/ha 120 kg N/ha + 20 L/ha SRN *Treatments are significantly different (Tukey's HSD, P≤0.05, n=4). **Fungicide treatments are significantly different from the untreated control (Tukey's HSD, P≤0.05, n=4). Data were transformed, if required, to satisfy normality assumptions. Means and standard errors are non-transformed.

Plasmodiophora brassicae, which causes clubroot of Brassica crops, persists in soil as long-lived resting spores. Quantitative polymerase chain reaction (qPCR) analysis is often used to quantify resting spores but does not distinguish between DNA of viable and nonviable spores. The impact of pretreating spores with propidium monoazide (PMA), which inhibits amplification of DNA from nonviable microorganisms, was assessed in several experiments. Spore suspensions from immature and mature clubs were heat treated; then, PMA-PCR analyses and bioassays were performed to assess spore viability. Prior to heat treatment, assessments comparing PMA-PCR to qPCR for mature spores were similar, indicating that most of these spores were viable. However, only a small proportion (<26%) of immature spores were amplified in PMA-PCR. Bioassays demonstrated that clubroot severity was much higher in plants inoculated with mature spores than with immature spores. Heat treatment produced little or no change in estimates of mature spores from qPCR but spore estimates from PMA-PCR and clubroot severity in bioassays were both substantially reduced. Estimates of spore concentration with PMA-PCR were less consistent for immature spores. To facilitate use of PMA-PCR on infested soil, a protocol for extracting spores from soil was developed that provided higher extraction efficiency than the standard methods.

Infested wooden boxes, previously used for carrot storage, were sampled in four commercial carrot production farms in Bradford Marsh, Ontario, and screened for fungal occurrence. At least 128 and 465 fungal isolates were recovered from these boxes in 2001 and 2002, respectively, and were classified into 10 taxonomic groups, including Alternaria spp., Aspergillus spp., Botrytis cinerea , Fusarium spp., Mucor spp., Penicillium spp., Rhizoctonia carotae , Rhizopus spp., Sclerotinia sclerotiorum and Trichoderma spp. A subsample of 27 putative pathogenic isolates was further tested for the ability to cause disease on carrots and to colonize wood surfaces under growth room and cold storage conditions. Approximately 60% of the taxa growing on wood caused lesions upon contact with intact carrots in cold storage. Isolates of S. sclerotiorum , B. cinerea and R. carotae caused the most severe diseases, developed most extensively on wooden surfaces in cold storage, and represented 12% of the recovered fungi. Isolates of Alternaria spp., Aspergillus spp., Fusarium spp., Mucor spp., Penicillium spp., Rhizopus spp. and Trichoderma spp. caused negligible or no disease on carrots and represented 88% of recovered fungi. Several of these fungi, however, showed potential to colonize wooden surfaces and cause disease on sliced carrots. This study suggests that pathogenic inocula occurring on used wooden boxes can initiate disease upon contact with healthy carrots and reusing infested boxes can affect carrots in storage.

Four canopy management treatments were evaluated in carrot (Daucus carota) production in Bradford Marsh, Ontario, in 2001 and 2002: (i) unclipped control; (ii) unclipped canopy with manual removal of collapsed senescing leaves at 2-week intervals following the first appearance on the soil; (iii) lateral clipping of the canopy at the initial emergence of apothecia, leaving the debris in the furrow, and (iv) lateral clipping of the canopy with manual removal of the debris from the furrow. Clipping reduced the canopy width by ca. 20% on both sides of the carrot bed by cutting off overlapping leaves above the furrow and senescing foliage on the soil surface. Maximum air and soil temperatures were up to 9.2 and 3.1°C lower, respectively, and relative humidity was up to 30% higher in unclipped canopies than in clipped canopies. The total number of apothecia in clipped plots was reduced by 74 and 76% compared with unclipped plots in 2001 and 2002, respectively. Canopy clipping reduced the quantity of apothecia in the crop by creating an unfavorable microclimate for the development of S. sclerotiorum without affecting the fresh foliar and root weights of carrot at harvest. The presence of clipped foliar debris in the furrow affected the number of apothecia in 2001; however, apothecia under the debris are unlikely to contribute to the overall inoculum in the crop. Lateral clipping also appeared to control Sclerotinia rot of carrot where it occurred (in 2001). This is the first report that documents and quantifies the effects of canopy architecture on the microclimate, development of apothecia of S. sclerotiorum, and Sclerotinia rot in carrot crops.

dence that the biofungicide Serenade (Bacillus subtilis) suppresses clubroot on canola via antibiosis and induced host resistance. Phytopathology 103:245-254. This study investigated how the timing of application of the biofungicide Serenade (Bacillus subtilis QST713) or it components (product filtrate and bacterial cell suspension) influenced infection of canola by Plasmodiophora brassicae under controlled conditions. The biofungicide and its components were applied as a soil drench at 5% concentration (vol/vol or equivalent CFU) to a planting mix infested with P. brassicae at seeding or at transplanting 7 or 14 days after seeding (DAS) to target primary and secondary zoospores of P. brassicae. Quantitative polymerase chain reaction (qPCR) was used to assess root colonization by B. subtilis as well as P. brassicae. The biofungicide was consistently more effective than the individual components in reducing infection by P. brassicae. Two applications were more effective than one, with the biofungicide suppressing infection completely and the individual components reducing clubroot severity by 62 to 83%. The biofungicide also reduced genomic DNA of P. brassicae in canola roots by 26 to 99% at 7 and 14 DAS, and the qPCR results were strongly correlated with root hair infection (%) assessed at the same time (r = 0.84 to 0.95). qPCR was also used to quantify the transcript activity of nine host-defense-related genes in inoculated plants treated with Serenade at 14 DAS for potential induced resistance. Genes encoding the jasmonic acid (BnOPR2), ethylene (BnACO), and phenylpropanoid (BnOPCL and BnCCR) pathways were upregulated by 2.2-to 23-fold in plants treated with the biofungicide relative to control plants. This induced defense response was translocated to the foliage (determined based on the inhibition of infection by Leptosphaeria maculans). It is possible that antibiosis and induced resistance are involved in clubroot suppression by Serenade. Activity against the infection from both primary and secondary zoospores of P. brassicae may be required for maximum efficacy against clubroot.

In the last 5 years, asparagus acreage in Canada has increased by over 25%. Stemphylium leaf spot, caused by Stemphylium vesicarium, has emerged as the predominant foliar pathogen of asparagus. Typically, contact fungicides are applied every 14 days; however, regardless of the number of applications, growers are not achieving adequate control of the disease. The TOM-CAST forecasting model is used widely in Michigan asparagus fields, but it has never been assessed for suitability in Ontario or in the popular cultivar, Guelph Millennium. Six field trials were conducted in 2012 and 2013 to evaluate the TOM-CAST forecasting model in two asparagus cultivars. The fungicides chlorothalonil or azoxystrobin/difenoconazole were applied according to the forecasting model or on a 14-day interval. The effectiveness of the forecasting model differed between sites and cultivars. Even though TOM-CAST is used in all cultivars in Michigan, TOM-CAST was not effective on Guelph Millennium. In the cultivar Jersey Giant, however, TOM-CAST with a 20 disease severity value spray interval improved control of Stemphylium leaf spot without increasing the number of sprays, compared with a 14day treatment. The results in Guelph Millennium differed between sites. At one site, TOM-CAST maintained similar levels of Stemphylium leaf spot, but increased the number of applications, compared with a 14-day application interval. Of more concern, none of the fungicide treatments differed greatly from the untreated control at the other site. Our results show that forecasting models need to be validated locally in asparagus cultivars relevant to production today.

Field trials to determine the effect of carrot pigmentation and weather parameters on cavity spot (CS) of carrot were conducted in the Holland/Bradford Marsh region of Ontario between 2002 and 2009. In all, 23 colored carrot cultivars from the United States Department of Agriculture (USDA) Agricultural Research Service breeding program at the University of Wisconsin (n = 5) and commercial seed companies (n = 18) were seeded in organic soil (pH 6 to 7, 45 to 75% organic matter) in late May to early June and harvested in late October or early November. Carrot roots were assessed for CS severity midseason and postharvest. Evaluations postharvest indicated that the purple pigmented carrot from breeding line 'USDA 106-3' and cultivars 'Purple Rain' and 'Purple Haze' consistently had low CS severity. The orange-pigmented 'USDA 101-23', 'Cellobunch', 'YaYa', and 'Envy' had moderate CS; and the red-pigmented carrot breeding line 'USDA 104-3' and cultivars 'Atomic Red', 'Proline Red', 'Dragon', and an unnamed line from India had high CS. Differences in CS severity in carrot cultivars between evaluations at midseason and postharvest suggest that some carrot cultivars are more susceptible to Pythium spp. inoculum in soil (alloinfection) and others to secondary infection (autoinfection) that can be attributed to the Pythium sp. involved in CS. CS severity was positively correlated with total rainfall 2 and 3 months after seeding, and was negatively correlated with number of days with air temperature ≥30°C 3 and 4 months after seeding. Soil temperature and total rainfall were the best predictors of CS incidence and severity. These results could allow a forecast of disease incidence and severity at harvest.

Botrytis leaf blight (BLB) of onion (Allium cepa L.) is caused by Botrytis squamosa J.C. Walker. The disease has been reported on onion crops in several of the onion production areas of the world including North and South America, Europe, Asia, and Australia , although it is not a problem in arid production regions such as the western United States. In eastern Canada, the disease is generally present every year and is especially severe on cultivars of yellow globe onion (54). The pathogen biology and disease epidemiology have been intensively researched (1-

The influence of nitrogen and calcium fertilizers on septoria late blight and yield of celery. Can. J. Plant Sci. 94: 1391Á1399. Late blight (Septoria apiicola) reduces both yield and quality of fresh market and processing celery. Field trials were conducted over 4 site-years to evaluate the effect of nitrogen (N) rate and foliar calcium (Ca) fertilization on the development of late blight and yield on celery grown on muck soils, which have a high organic matter (OM) content. A treatment with the protectant fungicide chlorothalonil was included to determine if fertilizer could replace fungicide application in celery production. There was a cultivar)site-year interaction for each of the disease and yield variables. Repeated application of chlorothalonil fungicide reduced late blight and generally increased plant height, weight and marketable yield. The rate of N applied had a small effect on late blight, which varied among site-years. Area under the disease progress curve decreased with increasing rate of N at both sites in 2008. Similarly, an increasing rate of N resulted in an increase in yield in one of four site-years. Application of Ca did not affect late blight severity and increased yield in only 1 site-year. There was a negative correlation between late blight severity and yield. Tissue N concentrations varied among the treatments at sites with a relatively low OM content, but not at the high OM sites. The treatments had no effect on Ca tissue concentrations. Application of N and Ca alone did not adequately control late blight, but there was a slight reduction in late blight severity with increasing rates of N. Application of both N and Ca can improve yield and also contribute indirectly to improved late blight management in combination with fungicides and other integrated crop management practices.

Stemphylium leaf blight (SLB), caused by Stemphylium vesicarium, is a foliar disease of onion worldwide, and has recently become an important disease in the northeastern United States and Ontario, Canada. The symptoms begin as small, tan to brown lesions on the leaves that can progress to defoliate plants. Crop loss occurs through reduced photosynthetic area, resulting in smaller, lower-quality bulbs. Leaf necrosis caused by SLB also can compromise bulb storage, as green leaves are required for the uptake of sprout inhibitors applied prior to harvest. The pathogen can overwinter on infested onion residue and infected volunteer plants. Asymptomatic weedy hosts near onion fields may also be a source of inoculum. Production of ascospores of the teleomorph (Pleospora allii) peaks in early spring in northeastern North America, often before the crop is planted, and declines rapidly as daily mean air temperatures rise. Conidia are usually present throughout the growing season. Application of fungicides is a standard practice for management of the complex of fungi that can cause foliar diseases of onion in this region. Recent assessments have shown that populations of S. vesicarium in New York and Ontario are resistant to at least three single-site mode-of-action fungicides. Three disease prediction systems have been developed and evaluated that may enable growers to reduce the frequency and/or number of fungicide applications, but the loss of efficacious fungicides due to resistance development within S. vesicarium populations threatens sustainability. The lack of commercially acceptable onion cultivars with sufficient resistance to reduce the number of fungicides for SLB also limits the ability to manage SLB effectively. Integrated disease management strategies for SLB are essential to maintain profitable, sustainable onion production across eastern North America.

Onions and garlic are the most popular cultivated forms of the family Alliaceae and consumed by almost every culture on earth. According to the 2001 statistics of the FAO, onions are second only to tomatoes in value of vegetable crops cultivated worldwide. There are a number of pathogens attack onions and garlic throughout their developmental stages since the developmental process is important in the progression of diseases. Onions are normally propagated from seed, planted directly into the field, although onion sets and transplants are also used. Garlic has been vegetatively propagated for millennia, by planting cloves. Vegetative propagation results in additional disease management problems. This chapter covers disease management of many of the major bacterial, fungal and viral diseases of onions and garlic, with an emphasis on those diseases that have been the topic of disease management research or for which effective disease management systems have been put into practice. Short descriptions of the pathogens and symptoms are included to assist in identification of the specific diseases.

Brassica oleracea var. capitata, Brassica rapa ssp. pekinensis, Plasmodiophora brassicae, clubroot management SUMMARY. Field trials were conducted to evaluate resistance to clubroot (Plasmodiophora brassicae, pathotype 6) in green cabbage (Brassica oleracea var. capitata) and napa cabbage (Brassica rapa ssp. pekinensis) at sites in southern Ontario in 2009 and 2010. The reaction of green cabbage cultivars Kilaton, Tekila, Kilaxy, and Kilaherb and the commercial standard cultivars, Bronco or Atlantis, were evaluated on organic (two site-years) and mineral soils (two site-years) that were naturally infested with the clubroot pathogen. In addition, fluazinam fungicide was drench applied to one treatment of the commercial standard cultivar immediately after transplanting. The napa cabbage cultivars Yuki, Deneko, Bilko, and Mirako (in 2009) and Emiko, Mirako, Yuki, and China Gold (in 2010) were evaluated only on organic soils (two site-years). At harvest, the roots of each plant were assessed for clubroot incidence and severity. Also, plant and head characteristics of the resistant green cabbage cultivars were evaluated at one site in 2010. The green cabbage cultivars Kilaton, Tekila, Kilaxy, and Kilaherb were resistant to pathotype 6 (0% to 3.8% incidence), but 'Bronco' was susceptible (64% to 100% incidence). Application of fluazinam reduced clubroot severity on 'Bronco' by 6% at one of three sites. Resistance was more effective in reducing clubroot than application of fluazinam. Plant and head characteristics of the resistant cultivars were similar to those of 'Bronco' treated with fluazinam. Napa cabbage cultivars Yuki, Deneko, Bilko, Emiko, and China Gold were resistant to clubroot (0% to 13% incidence), and 'Mirako' was highly susceptible (87% to 92% incidence). We conclude that the clubroot resistance available in several cultivars of green and napa cabbage was effective against P. brassicae pathotype 6.

The clubroot reaction of five Rapid Cycling Brassica Collection (RCBC) lines (Brassica carinata, B. juncea, B. napus, B. oleracea, and B. rapa) and 84 lines of Arabidopsis thaliana to pathotypes 2, 3, 5, and 6 of Plasmodiophora brassicae (as classified on William's system) was assessed. Also, the reaction of the Arabidopsis lines to a single-spore isolate of each of pathotypes 3 and 6 was compared with that of a field isolate. Seedlings were inoculated with resting spores of P. brassicae, maintained at 25 and 20°C (day and night, respectively), and assessed for clubroot incidence and severity at 6 weeks after inoculation. Several lines of A. thaliana and RCBC exhibited a differential response to pathotype but none of the lines were immune. Among the RCBC lines, B. napus was resistant to all of the pathotypes; B. oleracea was re-sistant to pathotypes 2, 3, and 5; B carinata and B. rapa were resistant to pathotypes 2 and 5; and B. juncea was susceptible to pathotypes 5 and 6 and had an intermediate response to pathotypes 2 and 3. Line Ct-1 of A. thaliana was highly resistant to pathotype 2, Pu2-23 was highly resistant to pathotype 5, and Ws-2 and Sorbo were highly resistant to pathotype 6. These results indicate that the lines of RCBC and A. thaliana have potential for use as model crops for a wide range of studies on clubroot, and could be used to differentiate these four pathotypes of P. brassicae. The reaction of the RCBC lines to pathotype 6 was highly correlated with response under field conditions but the reaction to the single-spore isolates of pathotypes 3 and 6 was not strongly correlated with reaction to the field collections in the Arabidopsis lines.

Clubroot, caused by Plasmodiophora brassicae, is an important disease on Brassica species worldwide. A clubroot resistance gene, Rcr1, with efficacy against pathotype 3 of P. brassicae, was previously mapped to chromosome A03 of B. rapa in pak choy cultivar "Flower Nabana". In the current study, resistance to pathotypes 2, 5 and 6 was shown to be

The fungal genus Stemphylium (phylum Ascomycota, teleomorph Pleospora) includes plant pathogenic, endophytic, and saprophytic species with worldwide distributions. Stemphylium spp. produce prodigious numbers of airborne spores, so are a human health concern as allergens. Some species also produce secondary metabolites, such as glucosides, ferric chelates, aromatic polyketides, and others, that function as toxins that damage plants and other fungal species. Some of these compounds also exhibit a low level of mammalian toxicity. The high production of airborne spores by this genus can result in a high incidence of human exposure. Concern about toxin production appears to be the reason that Stemphylium vesicarium, which is a pathogen of several vegetable crops, was classified in Canada as a potential risk of harm to humans for many years. A detailed assessment of the risk of exposure was provided to the relevant regulatory body, the Public Health Agency of Canada, which then determined that Stemphylium spp. in nature or under laboratory conditions posed little to no risk to humans or animals, and the species was re-assigned as a basic (level 1) risk agent.

Clubroot [Plasmodiophora brassica] severity in canola (Brassica napus) can be suppressed by addition of boron (B), but excess B produces phytotoxicity. However, some lines of a closely related species, B. rapa, are relatively insensitive to high levels of B. Assessment of 150 accessions of B. napus and B. rapa treated with 0, 8 and 16 kg B ha-1 as disodium octaborate tetrahydrate (trade name Solubor) identi ed several lines that were relatively insensitive to high levels of B. In 2015, 88 lines were planted in a clubroot-infested plot in Ontario, Canada in a replicated split-plot design, where the subplot treatments were 8 kg ha-1 of B versus a non-treated control. Boron-insensitive lines generally had slightly lower clubroot severity than sensitive lines, even in the control with no added B. Application of B reduced clubroot slightly in the most sensitive lines, but produced a large and signi cant reduction in severity in the most insensitive lines. Assessment of ve insensitive lines under controlled conditions supported the observation that application of B substantially reduced clubroot severity in insensitive lines. Comparison of two sensitive and two insensitive lines each of B. napus and B. rapa using Boron K-edge X-ray absorption near-edge structure (XANES) spectra from a synchrotron-based soft-X-ray beamline showed that some insensitive lines were able to extract B e ciently from a soil that contained very low levels of B. The spectra of sensitive and insensitive lines differed when additional B was applied, which demonstrated that the insensitive lines stored B (and other elements such as S and P) differently than the sensitive lines. Differences in the amount and type of storage likely affect the susceptibility to clubroot, and merits further study.

A Basidiomycota endomycorrhizal fungus, Piriformospora indica Verma, colonizes and promotes the growth of canola (Brassica napus L.) and other Brassica crops and can reduce diseases of other crops. Clubroot is an important disease of Brassica crops caused by the obligate, soil-borne pathogen Plasmodiophora brassicae Woronin. The effect of P. indica on clubroot severity in canola was assessed in replicated growth room studies. Seed was treated with P. indica using a proprietary process. Microscopic observation confirmed that canola roots grown from treated seed were colonized by P. indica; however, P. indica did not consistently reduce clubroot severity and did not promote the growth of canola.

Background: Clubroot is an important disease of brassica crops world-wide. The causal agent, Plasmodiophora brassicae, has been present in Canada for over a century but was first identified on canola (Brassica napus) in Alberta, Canada in 2003. Genetic resistance to clubroot in an adapted canola cultivar has been available since 2009, but resistance breakdown was detected in 2013 and new pathotypes are increasing rapidly. Information on genetic similarity among pathogen populations across Canada could be useful in estimating the genetic variation in pathogen populations, predicting the effect of subsequent selection pressure on changes in the pathogen population over time, and even in identifying the origin of the initial pathogen introduction to canola in Alberta. The genomic sequences of 43 strains (34 field collections, 9 single-spore isolates) of P. brassicae from Canada, the United States, and China clustered into five clades based on SNP similarity. The strains from Canada separated into four clades, with two containing mostly strains from the Prairies (provinces of Alberta, Saskatchewan, and Manitoba) and two that were mostly from the rest of Canada or the USA. Several strains from China formed a separate clade. More than one pathotype and host were present in all four Canadian clades. The initial pathotypes from canola on the Prairies clustered separately from the pathotypes on canola that could overcome resistance to the initial pathotypes. Similarly, at one site in central Canada where resistance had broken down, about half of the genes differed (based on SNPs) between strains before and after the breakdown. Clustering based on genome-wide DNA sequencing demonstrated that the initial pathotypes on canola on the Prairies clustered separately from the new virulent pathotypes on the Prairies. Analysis indicated that these 'new' pathotypes were likely present in the pathogen population at very low frequency, maintained through balancing selection, and increased rapidly in response to selection from repeated exposure to host resistance.

Each year, downy mildew caused by Bremia lactucae occurs in lettuce-producing regions of Quebec and Ontario. Trials were conducted to determine if the disease could be managed with fewer fungicide applications, using disease forecasting. Disease-forecasting programs designed to time the application of fungicide sprays were evaluated for control of downy mildew of lettuce in 1997 and 1998 at 9 field sites. Disease forecasting was based on lesion development following a recorded sporulation-infection period (SIP), with leaf wetness between 3:00 AM and 10:00 AM and temperature between 5 and 20 °C. Presymptom sprays with the protectant fungicide mancozeb were applied before lesions were expected; they were based on 110 accumulated degree-days following a SIP, but not within 7 days of a previous spray. Post-latent-period sprays with the systemic fungicide metalaxyl-M in combination with mancozeb were applied after forecasted lesion development; they were based on 135 accumulated degree-days, but not within 14 days of a previous spray. The effects of forecasted sprays were compared with those of weekly applications of mancozeb and with those of metalaxyl-M or metalaxyl applied in furrow at the time of seeding or transplanting. Downy mildew developed at 7 of the 9 sites. The number of SIPs recorded at all sites ranged from 7 to 17. In 8 of the 9 sites, there were fewer mancozeb applications with the presymptom program than with the weekly sprays, that is a reduction of one, two, and three sprays at six, one, and one sites, respectively. This resulted in a 16%-60% reduction in applied fungicide. When metalaxyl-M plus mancozeb was applied according to the post-latent-period forecast, one or two sprays were required, that is a reduction of two, three, and four applications at four, three, and two sites, respectively, compared with the weekly application of mancozeb alone. Metalaxyl-M or metalaxyl applied in furrow provided control for 30-40 days after seeding or transplanting. The number of fungicide applications can be reduced with acceptable disease control if a forecasting program is followed.

The efficacy of two sclerotia germination stimulants, diallyl disulfide (DADS, 85.5%; related compounds, 4.5%) and di-N-propyl disulfide (DPDS, 88%; related compounds, 2%) were evaluated in naturally infested field plots and in the greenhouse for reduction in populations of sclerotia and control of onion white rot in organic soil. Field trials evaluated one or two applications of DADS or DPDS injected into soil at 10 L/ha in 500 L/ha of water. To determine the treatment effects on the survival of sclerotia, sacs of 100 sclerotia were exposed to treated and untreated soil in the field and greenhouse. In controlled greenhouse studies, survival of sclerotia was significantly decreased when sclerotia were exposed for 1 to 3 months to soil treated with DADS or DPDS compared with untreated soil. DADS was considerably more effective than DPDS. In field trials, survival of sclerotia decreased after 3 months exposure to DADS-treated soil. Single DADS applications reduced disease incidence on onions at harvest compared with the untreated checks at three of four sites. Following two DADS applications, disease was reduced to below 1% at all four sites. DADS applied to organic soil provided effective control of white rot under conditions of moderate disease pressure.

The fungal genus Stemphylium (phylum Ascomycota, teleomorph Pleospora) includes plant pathogenic, endophytic, and saprophytic species with worldwide distributions. Stemphylium spp. produce prodigious numbers of airborne spores, so are a human health concern as allergens. Some species also produce secondary metabolites, such as glucosides, ferric chelates, aromatic polyketides, and others, that function as toxins that damage plants and other fungal species. Some of these compounds also exhibit a low level of mammalian toxicity. The high production of airborne spores by this genus can result in a high incidence of human exposure. Concern about toxin production appears to be the reason that Stemphylium vesicarium, which is a pathogen of several vegetable crops, was classified in Canada as a potential risk of harm to humans for many years. A detailed assessment of the risk of exposure was provided to the relevant regulatory body, the Public Health Agency of Canada, which then determined that Stemphylium spp. in nature or under laboratory conditions posed little to no risk to humans or animals, and the species was re-assigned as a basic (level 1) risk agent.

research departments of the Crop Production Chemical Industry, numerous Seed Companies and Growers. This report consists of two sections, the first contains highlights of research projects which were conducted in 2002 under the supervision of Research Scientist Dr. Mary Ruth McDonald. The second section contains highlights of various Muck Crops cultivar evaluations in 2002 on both field and storage trials, under the supervision of Research Station Manager, Shawn Janse. The results published in this report should be treated as a progress report. Some of the chemicals used in the trials are not registered for use on the crops they were applied to. Additional trials may be necessary before firm conclusions and recommendations can be made. The Muck Crops Research Station is an active participant in the training of new researchers on Muck Vegetables through the Graduate Student Program of the University of Guelph. Presently the Muck Crops Research Station has one Master's Graduate Students and three Ph. D. student working on various research projects involving Muck Vegetables. The Muck Crops Research Station recieved certifiation in 200 1 in the Good Laboratory Practices (GLP) program which will allow us to continue in the participation of future registration of chemicals for muck vegetables. Past participation has helped in the registration of such products as Dithane DG in furrow treatment for the control of onion smut; Select for annual bluegrass control in onions, Admire for Aphid control on lettuce. Apron XL for pythium control in onion seedlings. We would like to take this opportunity to express our sincere appreciation to the staff for their efforts in conducting these research projects; cultivar evaluation trials and producing this report. Many thanks also to all the co-operating researchers, technicians, industry personal, and growers for their continued support and interest in muck crops.

In the last 5 years, asparagus acreage in Canada has increased by over 25%. Stemphylium leaf spot, caused by Stemphylium vesicarium, has emerged as the predominant foliar pathogen of asparagus. Typically, contact fungicides are applied every 14 days; however, regardless of the number of applications, growers are not achieving adequate control of the disease. The TOM-CAST forecasting model is used widely in Michigan asparagus fields, but it has never been assessed for suitability in Ontario or in the popular cultivar, Guelph Millennium. Six field trials were conducted in 2012 and 2013 to evaluate the TOM-CAST forecasting model in two asparagus cultivars. The fungicides chlorothalonil or azoxystrobin/difenoconazole were applied according to the forecasting model or on a 14-day interval. The effectiveness of the forecasting model differed between sites and cultivars. Even though TOM-CAST is used in all cultivars in Michigan, TOM-CAST was not effective on Guelph Millennium. In the cultivar Jersey Giant, however, TOM-CAST with a 20 disease severity value spray interval improved control of Stemphylium leaf spot without increasing the number of sprays, compared with a 14day treatment. The results in Guelph Millennium differed between sites. At one site, TOM-CAST maintained similar levels of Stemphylium leaf spot, but increased the number of applications, compared with a 14-day application interval. Of more concern, none of the fungicide treatments differed greatly from the untreated control at the other site. Our results show that forecasting models need to be validated locally in asparagus cultivars relevant to production today.

The clubroot reaction of five Rapid Cycling Brassica Collection (RCBC) lines (Brassica carinata, B. juncea, B. napus, B. oleracea, and B. rapa) and 84 lines of Arabidopsis thaliana to pathotypes 2, 3, 5, and 6 of Plasmodiophora brassicae (as classified on William's system) was assessed. Also, the reaction of the Arabidopsis lines to a single-spore isolate of each of pathotypes 3 and 6 was compared with that of a field isolate. Seedlings were inoculated with resting spores of P. brassicae, maintained at 25 and 20°C (day and night, respectively), and assessed for clubroot incidence and severity at 6 weeks after inoculation. Several lines of A. thaliana and RCBC exhibited a differential response to pathotype but none of the lines were immune. Among the RCBC lines, B. napus was resistant to all of the pathotypes; B. oleracea was re-sistant to pathotypes 2, 3, and 5; B carinata and B. rapa were resistant to pathotypes 2 and 5; and B. juncea was susceptible to pathotypes 5 and 6 and had an intermediate response to pathotypes 2 and 3. Line Ct-1 of A. thaliana was highly resistant to pathotype 2, Pu2-23 was highly resistant to pathotype 5, and Ws-2 and Sorbo were highly resistant to pathotype 6. These results indicate that the lines of RCBC and A. thaliana have potential for use as model crops for a wide range of studies on clubroot, and could be used to differentiate these four pathotypes of P. brassicae. The reaction of the RCBC lines to pathotype 6 was highly correlated with response under field conditions but the reaction to the single-spore isolates of pathotypes 3 and 6 was not strongly correlated with reaction to the field collections in the Arabidopsis lines.

Alternaria leaf blight (ALB) and Cercospora leaf spot (CLS) are economically important diseases of carrot in Ontario. Field experiments were conducted in the Holland Marsh, Ontario, to determine the effect of nitrogen (N) application rates on both diseases. Five rates of N were applied to organic and mineral soils in which two carrot cultivars, Idaho and Fontana, were grown in each of 2002, 2003, and 2004. Both diseases were rated every 2 weeks on a scale of 0 (healthy) to 10 (tops destroyed), and the number of live (green) leaves per plant was assessed at harvest. In addition, three N rates were applied to carrot plants grown in the greenhouse, and the plants were inoculated with Alternaria dauci (Ku ¨hn) Groves and Skolko. Disease severity, senescence, and sap nitrate-N concentration were assessed. In the field trials, the response of ALB and CLS to N application rate was relatively consistent across cultivar, soil type, and year. Area-under-the-diseaseprogress curves typically increased with decreasing N rate for both diseases. In lower N treatments, this resulted in fewer live leaves per plant at harvest. In the greenhouse, ALB severity increased with increasing amount of leaf senescence at final assessment. The results suggest that N application rate could be used to reduce the need for fungicide applications to control these diseases in the field.

A Basidiomycota endomycorrhizal fungus, Piriformospora indica Verma, colonizes and promotes the growth of canola (Brassica napus L.) and other Brassica crops and can reduce diseases of other crops. Clubroot is an important disease of Brassica crops caused by the obligate, soil-borne pathogen Plasmodiophora brassicae Woronin. The effect of P. indica on clubroot severity in canola was assessed in replicated growth room studies. Seed was treated with P. indica using a proprietary process. Microscopic observation confirmed that canola roots grown from treated seed were colonized by P. indica; however, P. indica did not consistently reduce clubroot severity and did not promote the growth of canola.

Plasmodiophora brassicae, which causes clubroot of Brassica crops, persists in soil as long-lived resting spores. Quantitative polymerase chain reaction (qPCR) analysis is often used to quantify resting spores but does not distinguish between DNA of viable and nonviable spores. The impact of pretreating spores with propidium monoazide (PMA), which inhibits amplification of DNA from nonviable microorganisms, was assessed in several experiments. Spore suspensions from immature and mature clubs were heat treated; then, PMA-PCR analyses and bioassays were performed to assess spore viability. Prior to heat treatment, assessments comparing PMA-PCR to qPCR for mature spores were similar, indicating that most of these spores were viable. However, only a small proportion (<26%) of immature spores were amplified in PMA-PCR. Bioassays demonstrated that clubroot severity was much higher in plants inoculated with mature spores than with immature spores. Heat treatment produced little or no change in estimates of mature spores from qPCR but spore estimates from PMA-PCR and clubroot severity in bioassays were both substantially reduced. Estimates of spore concentration with PMA-PCR were less consistent for immature spores. To facilitate use of PMA-PCR on infested soil, a protocol for extracting spores from soil was developed that provided higher extraction efficiency than the standard methods.

Untreated 15DSV (4 apps) 30DSV (2 apps) *Treatments are significantly different (Tukey's HSD, P≤0.05, n=16). **Fungicide treatments are significantly different from the untreated control (Tukey's HSD, P≤0.05, n=16). Data were transformed, if required, to satisfy normality assumptions. Means and standard errors are non-transformed. Brady soil (sandy) site Spray Timing Trial Untreated 15DSV (9 apps) 30DSV (5 apps) *Treatments are significantly different (Tukey's HSD, P≤0.05, n=16). **Fungicide treatments are significantly different from the untreated control (Tukey's HSD, P≤0.05, n=16). Data were transformed, if required, to satisfy normality assumptions. Means and standard errors are non-transformed Wauseon/Berrien soil (sandy loam) site Disease severity (%) 0 kg N/ha 120 kg N/ha 180 kg N/ha 120 kg N/ha + 20 L/ha SRN *Treatments are significantly different (Tukey's HSD, P≤0.05, n=4). **Fungicide treatments are significantly different from the untreated control (Tukey's HSD, P≤0.05, n=4). Data were transformed, if required, to satisfy normality assumptions. Means and standard errors are non-transformed.

Brassica oleracea var. capitata, Brassica rapa ssp. pekinensis, Plasmodiophora brassicae, clubroot management SUMMARY. Field trials were conducted to evaluate resistance to clubroot (Plasmodiophora brassicae, pathotype 6) in green cabbage (Brassica oleracea var. capitata) and napa cabbage (Brassica rapa ssp. pekinensis) at sites in southern Ontario in 2009 and 2010. The reaction of green cabbage cultivars Kilaton, Tekila, Kilaxy, and Kilaherb and the commercial standard cultivars, Bronco or Atlantis, were evaluated on organic (two site-years) and mineral soils (two site-years) that were naturally infested with the clubroot pathogen. In addition, fluazinam fungicide was drench applied to one treatment of the commercial standard cultivar immediately after transplanting. The napa cabbage cultivars Yuki, Deneko, Bilko, and Mirako (in 2009) and Emiko, Mirako, Yuki, and China Gold (in 2010) were evaluated only on organic soils (two site-years). At harvest, the roots of each plant were assessed for clubroot incidence and severity. Also, plant and head characteristics of the resistant green cabbage cultivars were evaluated at one site in 2010. The green cabbage cultivars Kilaton, Tekila, Kilaxy, and Kilaherb were resistant to pathotype 6 (0% to 3.8% incidence), but 'Bronco' was susceptible (64% to 100% incidence). Application of fluazinam reduced clubroot severity on 'Bronco' by 6% at one of three sites. Resistance was more effective in reducing clubroot than application of fluazinam. Plant and head characteristics of the resistant cultivars were similar to those of 'Bronco' treated with fluazinam. Napa cabbage cultivars Yuki, Deneko, Bilko, Emiko, and China Gold were resistant to clubroot (0% to 13% incidence), and 'Mirako' was highly susceptible (87% to 92% incidence). We conclude that the clubroot resistance available in several cultivars of green and napa cabbage was effective against P. brassicae pathotype 6.