Fungicide and Cultivar Management of Leaf Spot Diseases of Winter Wheat in Western Canada.

A complex of leaf-spotting diseases-tan spot, the Septoria complex, spot blotch, and powdery mildew-are frequently observed on winter wheat in western Canada; however, there are few studies indicating varietal differences in reaction to these diseases or the benefit of fungicide application. To determine the benefit of varietal improvement and multiple fungicide treatments and application timings, field experiments were conducted at six site-years in western Canada. Two cultivars varying in reaction to leaf spot diseases were used in combination with fungicide treatments. Disease severity ranged from trace to 64% of the combined flag and penultimate leaf area diseased, differed between cultivars, and was reduced from the check by some fungicide treatments. Yield improvement by fungicide treatment varied from 3.3 to 13.2% greater than the nontreated check. At two site-years, the split application of two half rates of fungicide resulted in the greatest yield; however, in both cases, the yield benefit did not differ from a single application at the flag leaf growth stage. Cultivar selection and fungicide use under environments conducive to disease are beneficial components of an integrated leaf spot disease management program for winter wheat in western Canada.

A Quantitative PCR System for Measuring Sclerotinia sclerotiorum in Canola (Brassica napus).

Sclerotinia stem rot, caused by Sclerotinia sclerotiorum, is an economically important disease of canola (Brassica napus) commonly managed by routine application of fungicides. Petal infestation has been demonstrated to be an important stage of the disease cycle in canola and has been the focus of previously developed Sclerotinia stem rot risk assessment methods. Quantitative polymerase chain reaction (qPCR) analysis can provide a more rapid and accurate assessment of petal infestation levels. Primers and a hydrolysis probe were designed to amplify a 70-bp region of an S. sclerotiorum-specific gene, SS1G_00263. A hydrolysis probe-based qPCR assay was developed that had a detection limit of 8.0 × 10-4 ng of S. sclerotiorum DNA and only amplified S. sclerotiorum DNA. Evaluation of petals collected at five sampling points in each of 10 commercial canola fields on each of two sampling dates (corresponding to 20 to 30% bloom and 40 to 50% bloom) revealed S. sclerotiorum DNA infestation levels of 0 to 3.3 × 10-1 ng/petal. This qPCR assay can be used to reliably quantify petal infestation and, with further research, has the potential to serve as the basis for a Sclerotinia stem rot risk assessment tool or as a means to study Sclerotinia stem rot epidemiology.

Adaptation to Brassica Host Genotypes by a Single-Spore Isolate and Population of Plasmodiophora brassicae (Clubroot).

Plasmodiophora brassicae, the cause of clubroot of crucifers, is an increasingly important pathogen of canola (Brassica napus) in Alberta, Canada. In response, clubroot-resistant canola genotypes are being deployed to help reduce yield losses. Two experiments were conducted to examine the effect on P. brassicae virulence of repeated exposure of a population and single-spore isolate of the pathogen to the same host. The first experiment examined changes in the index of disease over five cycles of infection on seven Brassica hosts (European Clubroot Differential [ECD] 02, ECD 04, ECD 05, ECD 15, '45H26', '45H29', and 08N823R). The second experiment tested the virulence of five cycled populations ('45H29', 08N823R, ECD 05, and ECD 15) and three cycled single-spore isolates ('45H29', 08N823R, and ECD05) on four resistant canola genotypes ('73-77', '73-67', VT-SD-09, and '9558C'). The results from these experiments clearly demonstrate the ability of both single-spore isolates and populations of P. brassicae to rapidly erode the resistance present in the two canola genotypes, '45H29' and 08N823R. Although the index of disease increased on these two genotypes, the four resistant canola genotypes remained resistant to all the cycled populations and single-spore isolates in the second experiment. These results underscore the importance of crop rotation in the management of clubroot in Alberta.

Distribution of Pathotypes of Rhynchosporium secalis and Cultivar Reaction on Barley in Alberta.

Forty-four barley accessions and commercial cultivars with different levels of resistance to scald caused by Rhynchosporium secalis were evaluated for scald reaction from 1997 to 1999 at various sites in Alberta. The accessions Hudson, Atlas, Atlas 46, Atlas 68, Abyssinian, and Kitchin that have the major resistance genes were resistant to pathotypes of R. secalis at all sites. Although scald levels were low for these accessions, they were significantly different among years. Pathotypes of R. secalis and environmental conditions affected diseases levels on 32 commercial cultivars, resulting in significantly different scald reactions among sites and seasons. Resistance in commercial cultivars, AC Stacy, Kasota, and Seebe, held up at most sites with the majority of cultivars being intermediate to moderately susceptible. Cultivars that were previously considered resistant were intermediate in reaction and became increasingly susceptible at some sites from 1997 to 1999. Pathogen virulence was more diverse at the sites where the cultivars became increasingly susceptible compared with sites where the same cultivars were resistant. Scald reactions of the commercial cultivars depended on location, which reflected the presence of different pathotypes, as well as variation in environmental conditions. Consequently, scald management via cultivar choice will be dependent on location.

A PCR-Based Assay to Detect Rhynchosporium secalis in Barley Seed.

A polymerase chain reaction (PCR)-based diagnostic assay was developed to detect Rhynchosporium secalis, the barley scald fungus, in barley seed. Species-specific primers were designed based on sequence data of a region consisting of the 5.8S RNA gene and internal transcribed spacers 1 and 2 of R. secalis. The sequenced regions showed 100% homology between the two R. secalis isolates and 93% homology between R. secalis and R. orthosporum. Five sets of synthesized oligonucleotide primers were tested for their specificity using 29 isolates of R. secalis of diverse geographic origins and from different barley cultivars. In addition, DNA extracts from 22 species of microbes either taxonomically related to or from the same niche as R. secalis were tested as negative controls. Among five sets of primers, a primer set, RS8 and RS9, was selected for use in detecting R. secalis because it amplified a 264-bp fragment from the DNA of all R. secalis isolates but not the DNA from other species used for validation of the specificity of this primer set. This primer set was also used to detect R. secalis in barley seed and successfully amplified the predicted size of the DNA fragment in the infected material. PCR detection of as little as 1 to 10 pg of R. secalis DNA was possible. The method described here requires 1 day for completion, compared to 10 days required for the cultural method.

Histopathological Study of Barley Cultivars Resistant and Susceptible to Rhynchosporium secalis.

ABSTRACT Differences in the penetration process by Rhynchosporium secalis were compared in resistant and susceptible barley cultivars at the seedling stage. Percent penetration and percent host cell wall alteration (HCWA) differed significantly among cultivars and isolates as revealed by light microscopy. Based on these two variables, the cultivars were statistically separated into two groups that corresponded to their disease reactions. The resistant cultivars, Johnston and CDC Guardian, showed 81.2 to 99.4% HCWA and 0.1 to 20.1% penetration at encounter sites, whereas the susceptible cultivars, Harrington, Argyle, and Manley, had 30.1 to 78.3% HCWA and 31.8 to 81.8% penetration. In the current study, cv. Leduc, which is susceptible at the seedling stage and resistant at the adult stage, showed the same percent HCWA and penetration as did susceptible cultivars. A significant negative correlation (P < 0.01) was found between percent penetration and percent HCWA for cultivars inoculated with two isolates of the pathogen. Isolate 1 was less virulent than isolate 2 with respect to percent penetration and induced significantly fewer HCWA. Scanning electron microscopy showed various shapes of fungal appressoria but no apparent difference in host reaction between resistant and susceptible cultivars. Transmission electron microscopy revealed interactions between the host and pathogen at various stages of penetration. The resistant cv. Johnston responded by producing appositions, as evidenced by a layer of compact osmiophilic material deposited on the inner side of the cell wall. Infection pegs produced by conidia were unable to penetrate the cuticle where an apposition had formed inside. When penetration occurred in the susceptible cv. Argyle, cytoplasmic aggregates and separation of the plasmalemma were visible from the host cell wall, but the layer of compact osmiophilic material was not always present. Data based on light microscopic observations suggested that HCWA may be one of the mechanisms responsible for resistance that is characterized as penetration prevention rather than as a slow rate of mycelial growth after successful penetration. HCWA occurred in response to attempted cuticle penetration, suggesting that HCWA may produce chemical barriers that help to prevent penetration.