B. D. Gossen

Genetic Diversity of Ascochyta rabiei in Canada

Assessment of variability of Ascochyta rabiei (teleomorph: Didymella rabiei) was based on virulence tests of 40 isolates and on random amplified polymorphic DNA (RAPD) analysis of 39 isolates from Canada. In addition, isolates of A. rabiei from other countries were assessed in the virulence (18 isolates) and RAPD (20 isolates) analyses. Seven isolates of A. lentis (teleo-morph: Didymella lentis) and two of A. pinodes (teleomorph: Mycosphaerella pinodes) also were included in the RAPD analysis. Significant line-isolate interactions in the virulence tests indicated that certain isolates were virulent only on certain lines. Canadian isolates were grouped into 14 pathotypes using eight chickpea differentials. These groupings also encompassed 17 of the 18 isolates from other countries. RAPD analysis of all 68 isolates using 8 primers produced 112 fragments, of which 96% were polymorphic. Similarities among A. rabiei isolates from Canada ranged from 20 to 100%. In the RAPD dendrogram, all five A. rabiei isolates from Australia, three of six from Syria, three of five from the United States, and one of two from India clustered within the major groups of Canadian isolates. There was a weak association between RAPD and pathotype groups. A. rabiei was 45% similar to A. lentis and only 14% similar to A. pinodes. The levels of DNA variability and virulence among isolates show that the population of A. rabiei in Canada is highly diverse.

Plasmodiophora brassicae: a review of an emerging pathogen of the Canadian canola (Brassica napus) crop

UNLABELLED Plasmodiophora brassicae causes clubroot disease in cruciferous plants, and is an emerging threat to Canadian canola (Brassica napus) production. This review focuses on recent studies into the pathogenic diversity of P. brassicae populations, mechanisms of pathogenesis and resistance, and the development of diagnostic tests for pathogen detection and quantification. TAXONOMY Plasmodiophora brassicae is a soil-borne, obligate parasite within the class Phytomyxea (plasmodiophorids) of the protist supergroup Rhizaria. DISEASE SYMPTOMS Clubroot development is characterized by the formation of club-shaped galls on the roots of affected plants. Above-ground symptoms include wilting, stunting, yellowing and premature senescence. DISEASE CYCLE: Plasmodiophora brassicae first infects the root hairs, producing motile zoospores that invade the cortical tissue. Secondary plasmodia form within the root cortex and, by triggering the expression of genes involved in the production of auxins, cytokinins and other plant growth regulators, divert a substantial proportion of plant resources into hypertrophic growth of the root tissues, resulting in the formation of galls. The secondary plasmodia are cleaved into millions of resting spores and the root galls quickly disintegrate, releasing long-lived resting spores into the soil. A serine protease, PRO1, has been shown to trigger resting spore germination. PHYSIOLOGICAL SPECIALIZATION: Physiological specialization occurs in populations of P. brassicae, and various host differential sets, consisting of different collections of Brassica genotypes, are used to distinguish among pathotypes of the parasite. DETECTION AND QUANTIFICATION: As P. brassicae cannot be cultured, bioassays with bait plants were traditionally used to detect the pathogen in the soil. More recent innovations for the detection and quantification of P. brassicae include the use of antibodies, quantitative polymerase chain reaction (qPCR) and qPCR in conjunction with signature fatty acid analysis, all of which are more sensitive than bioassays. RESISTANCE IN CANOLA: Clubroot-resistant canola hybrids, recently introduced into the Canadian market, represent an important new tool for clubroot management in this crop. Genetic resistance must be carefully managed, however, as it has been quickly overcome in other regions. At least three resistance genes and one or two quantitative trait loci are involved in conferring resistance to P. brassicae. Root hair infection still occurs in resistant cultivars, but secondary plasmodia often remain immature and unable to produce resting spores. Fewer cell wall breakages occur in resistant hosts, and spread of the plasmodium through cortical tissue is restricted. More information on the genetics of clubroot resistance in canola is needed to ensure more effective resistance stewardship. USEFUL WEBSITES http://www.canolacouncil.org/clubroot/resources.aspx, http://tu-dresden.de/die_tu_dresden/fakultaeten/fakultaet_mathematik_und_naturwissenschaften/fachrichtung_biologie/botanik/pflanzenphysiologie/clubroot, http://www.ohio.edu/people/braselto/plasmos/

Effect of plant age on resistance to Ascochyta rabiei in chickpea

The impact of plant age on resistance to ascochyta blight of chickpea (Cicer arietinum) was assessed on kabuli cvs. UC27 (susceptible) and Sanford and B90 (partially resistant) and desi cv. Myles (partially resistant) using two isolates of Ascochyta rabiei. Tests were conducted under controlled conditions and in field trials in 1999 and 2000. Seed was sown at regular intervals to produce plants at different growth stages (seedling, vegetative, flowering, podding), which were all inoculated at the same time in randomized complete block experiments with four replications. Cultivar UC27 was highly susceptible at all stages of plant development. However, plant age had a large impact on the reaction of resistant cultivars to A. rabiei. In growth chamber studies, blight severity was low in seedlings of the resistant cultivars (range, 4-20%) and highest in podding plants (41-82%) 2 weeks after inoculation. In the field trials, severity at 2 weeks after inoculation showed a similar pattern, with mean severity among resistant cultivars ranging from 1 to 2% in seedlings and 3 to 28% in podding plants. Severity at 8 weeks after inoculation on plants of resistant cultivars inoculated as seedlings was low (6-14% in 1999, 10-23% in 2000) relative to those inoculated at podding (65-91% in 1999; 54-83% in 2000) in the field trials. These results indicate that resistance decreases with plant age in the partially resistant chickpea cultivars adapted to western Canada and that resistance alone may not provide adequate disease control.

Reaction of seedling roots of 14 crop species to Fusarium graminearum from wheat heads

To increase our understanding of the epidemiology of fusarium head blight of wheat and barley, a study was conducted under controlled conditions to determine whether Fusarium graminearum Schwabe from wheat can cause seedling blight or root rot in various crop species. Inoculum of F. graminearum, consisting of a wheat floret infected with the pathogen, was placed adjacent to surface-sterilized seed of each crop in a sterile potting mix. Wheat, barley, oat, rye, triticale, canaryseed, flax, canola (Brassica napus L. and Brassica rapa L.), mustard, bean, field pea, lentil, and chickpea were included in the study. Seedling emergence and root rot severity were scored at 3–4 weeks after seeding. The effect of temperature on seedling blight severity was also tested in barley cv. Brier. Inoculation reduced emergence in all crops, except canola, mustard, and field pea, and increased root rot severity in most crops. Emergence of seedlings was not affected at the lowest temperature (10:5°C day:night) and no root infection occurred. However, as the temperature increased from 10 to 30°C, seedling emergence and establishment were reduced and root rot severity increased. Infection of roots, crowns, and seedlings of the crops grown in rotation with wheat indicates that these crops may act as alternative hosts to F. graminearum.

Effect of Temperature on Cortical Infection by Plasmodiophora brassicae and Clubroot Severity

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), at 2-day intervals for symptom development and clubroot severity, and at 8-day intervals for the number of spores per gram of gall. Temperature affected every stage of clubroot development. Cortical infection was highest and symptoms were observed earliest at 25?C, intermediate at 20 and 30?C, and lowest and latest at 15?C. No cortical infection or symptoms were observed at 42 DAI in plants grown at 10?C. A substantial delay in the development of the pathogen was observed at 15?C. Resting spores were first observed at 38 DAI in plants at 15?C, 26 DAI at 20 and 30?C, and 22 DAI at 25?C. The yield of resting spores from galls was higher in galls that developed at 20 to 30?C than those that developed at 15?C over 42 days of assessment. These results support the observation in companion studies that cool temperatures result in slower development of clubroot symptoms in brassica crops, and demonstrate that the temperature has a consistent pattern of effect throughout the life cycle of the pathogen.

Identification of Lentil Germ Plasm Resistant to Colletotrichum truncatum and Characterization of Two Pathogen Races

ABSTRACT A total of 1,771 lentil accessions from the U.S. lentil collection (U.S. Department of Agriculture-Agricultural Research Service, Pullman, WA) and the Institut für Pflanzengenetik und Kulturpflanzenforschung (Gatersleben, Germany) were screened for resistance to Colletotrichum truncatum, the cause of anthracnose. About 95% of the accessions were susceptible when inoculated with a single isolate in the field. Retesting, under controlled conditions, of accessions rated as resistant or moderately resistant in the field resulted in identification of anthracnose resistance in four accessions from the U.S. collection (PI 320937, PI 320952 [cv. Indianhead], PI 345629, and 468901), and 12 accessions from the German collection (Lens 3, 102, 104, 106, 107, 119, 122, 134, 135, 177, 195, and 209). Seven of the accessions were used as host differentials to characterize pathogenic variability of 50 single-spore isolates collected in Manitoba and Saskatchewan, Canada. The presence of two distinct races was demonstrated. Isolates of C. truncatum avirulent on cv. Indianhead, PI 320937, PI 345629, PI 468901, Lens 102, Lens 104, and Lens 195 were designated race Ct1. Isolates that were virulent on these seven entries were designated race Ct0, indicating their lack of avirulence genes. Race Ct0 was isolated more frequently from commercial seed samples than race Ct1, but the two races were isolated with similar frequency from plants in commercial fields planted to susceptible cultivars. Race Ct0, to which no resistance has yet been identified, presents a high risk to lentil production in Canada and potentially worldwide.

Evidence that the Biofungicide Serenade (Bacillus subtilis) Suppresses Clubroot on Canola via Antibiosis and Induced Host Resistance

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.

Crop rotation, cultivar resistance, and fungicides/biofungicides for managing clubroot (Plasmodiophora brassicae) on canola

Abstract Select biofungicides and fungicides, used alone or with cultivar resistance or crop rotation, were assessed for their potential in integrated management of clubroot disease. The synthetic fungicides pentachloronitrobenzene, fluazinam and cyazofamid showed activities against Plasmodiophora brassicae. The biofungicides Serenade® and Prestop® also suppressed the disease on canola via antibiosis and induced host resistance under controlled-environment conditions. Granular and seed-treatment formulations were developed to facilitate the delivery of biofungicide in field trials. Where P. brassicae resting spore populations were large in the soil, neither biofungicides nor synthetic fungicides were sufficiently effective when applied in the seed furrow. They occasionally reduced clubroot severity on Chinese cabbage. More than 5000 soil microbial isolates indigenous to the Canadian prairies were screened for potential clubroot control, but none showed consistent efficacy. Resistant cultivars reduced clubroot severity and canola yield losses significantly. A 2-year break from canola reduced P. brassicae resting spore concentrations by 90% relative to growing continuous canola or only a 1-year break in heavily infested field plots. This 2-year break alleviated disease impact on plant growth and development in a susceptible canola cultivar. Despite the substantial inoculum reduction after 2 years, the levels were still too high to obtain commercially acceptable yields in a susceptible cultivar. In a resistant cultivar, >2-year breaks increased yields by up to 25% relative to continuous growing of canola. A 2-year interval with non-hosts between canola crops, together with use of resistant cultivars, is recommended to reduce the inoculum load of P. brassicae in soil and achieve maximum yields of canola.

Foliar fungicides to manage ascochyta blight [Ascochyta rabiei] of chickpea in Canada

Ascochyta blight, caused by Ascochyta rabiei, is a major constraint to chickpea production in Saskatchewan. Foliar fungicides were evaluated at various rates and timings for managing blight epidemics over 18 station years from 1998 to 2000. Dry weather in 1998 resulted in low disease pressure, and fungicide application had no effect on blight severity and yield. In 1999 and 2000, wet conditions favored the development of epidemics. Under high disease pressure, a single fungicide application often reduced disease severity, but had no effect on yield. Two applications (early + mid flowering) of chlorothalonil (Bravo® 500) at 1 kg active ingredients (a.i.)/ha, two applications of azoxystrobin (Quadris®) at 125 g a.i./ha, or chlorothalonil + azoxystrobin reduced ascochyta blight and increased yield. An alternative formulation of chlorothalonil, Bravo Ultrex®, was less effective than Bravo 500, but more effective than mancozeb (Dithane®). Fungicide application had a substantial impact on seed yield; at one site, yield in the untreated control was less than 5% of the best fungicide treatment. Regression analysis showed a strong relationship between disease severity and seed yield. With high disease pressure in 2000, the incidence of seed-borne A. rabiei was 30–;48%. Applying azoxystrobin at early + mid flowering reduced it to 7–9%. Fungicide application had no impact on other seed-borne pathogens such as Botrytis cinerea, Sclerotinia sclerotiorum, and Fusarium spp. These results indicate that fungicide application can complement partial resistance to reduce blight severity and increase seed yield and quality in chickpea.

Effect of temperature, seeding date, fungicide seed treatment and inoculation with Fusarium avenaceum on seedling survival, root rot severity and yield of lentil.

Early seeding of lentil is necessary on the northern prairies to ensure that the crop has time to mature. However, planting into cold soils in spring results in slow germination, which may predispose seedlings to infection by soil-borne pathogens. In a controlled-environment study of the impact of temperature on infection of lentil seedlings (cv. Eston) byFusarium avenaceum, root rot symptoms were most severe at warm temperatures (20° to 27.5 °C) and declined in warmer or cooler soils. Field plots were seeded on three dates, which were spaced about 2 wk apart between early May and early June in four station years. Seedling emergence was most consistent and seed yield was highest in three of four sites at the second seeding date. Emergence and yield were substantially lower for the latest seeding date. Inoculation with F. avenaceum reduced establishment and seed yield. Seed treatment with Crown (thiabendazole and carbathiin) improved seedling survival, reduced root rot severity and increased seed yield rel...