Suppression of clubroot by Clonostachys rosea via antibiosis and induced host resistance

The mechanism of the biofungicide Prestop^® (Clonostachys rosea) was investigated for control of clubroot (Plasmodiophora brassicae) on canola. The key product components were partitioned and assessed for their effect on pathogen resting spores, root hair infection (RHI) and disease development using light microscopy, quantitative PCR and different application treatments during infection. The whole product of Prestop was consistently more effective than the C. rosea conidial suspension or product filtrate alone in reducing RHI and clubroot development. This biofungicide showed little effect on germination or viability of resting spores. Two‐application treatments at seeding and 7–14 days after seeding achieved greater clubroot control than a single application of the biofungicide at either seeding or post‐seeding stage. This may indicate the need to maintain a high biofungicide dose in the soil during primary and secondary infection. This biocontrol fungus colonized the rhizosphere and interior of canola roots extensively, and possibly induced plant resistance based on up‐regulation of the genes that are involved in jasmonic acid (BnOPR2), ethylene (BnACO) and phenylpropanoid (BnOPCL, BnCCR) biosynthetic pathways. It is concluded that the biofungicide Prestop suppressed clubroot on canola at least via root colonization and induced systemic resistance (ISR), and the latter may be through the modulation of phenylpropanoid and jasmonic acid/ethylene metabolic pathways elicited by the fungus.     

Potential biological control of clubroot on canola and crucifer vegetable crops

Clubroot caused by Plasmodiophora brassicae is an emerging threat to canola (Brassica napus) production in western Canada, and a serious disease on crucifer vegetable crops in eastern Canada. In this study, seven biological control agents and two fungicides were evaluated as soil drenches or seed treatments for control of clubroot. Under growth cabinet conditions, a soil‐drench application of formulated biocontrol agents Bacillus subtilis and Gliocladium catenulatum reduced clubroot severity by more than 80% relative to pathogen‐inoculated controls on a highly susceptible canola cultivar. This efficacy was similar to that of the fungicides fluazinam and cyazofamid. Under high disease pressure in greenhouse conditions, the biocontrol agents were less effective than the fungicides. Additionally, all of the treatments delivered as a seed coating were less effective than the soil drench. In field trials conducted in 2009, different treatments consisting of a commercial formulation of B. subtilis, G. catenulatum, fluazinam or cyazofamid were applied as an in‐furrow drench at 500 L ha^?1 water volume to one susceptible and one resistant cultivar at two sites seeded to canola in Alberta and one site of Chinese cabbage in Ontario. There was no substantial impact on the susceptible canola cultivar, but all of the treatments reduced clubroot on the susceptible cultivar of Chinese cabbage, lowering disease severity by 54–84%. There was a period of 4 weeks without rain after the canola was seeded, which likely contributed to the low treatment efficacy on canola. Under growth cabinet conditions, fluazinam and B. subtilis products became substantially less effective after 2 weeks in a dry soil, but cyazofamid retained its efficacy for at least 4 weeks.     

Efficacy of Vapam fumigant against clubroot (Plasmodiophora brassicae) of canola

Clubroot, caused by Plasmodiophora brassicae, has become a serious threat to canola (Brassica napus) production in western Canada. Experiments were conducted under greenhouse and field conditions to assess the effect of Vapam fumigant (dithiocarbamate; sodium N‐methyldithiocarbamate) on primary and secondary infection by P. brassicae, clubroot severity, and growth parameters in canola. Preliminary trials showed a 12–16‐fold reduction in primary and secondary infection and clubroot severity at all of the Vapam application rates (0·4–1·6 mL L^?1 soil) assessed. Vapam was also found to be effective in reducing clubroot severity and improving seed yield of canola under field conditions. Application of Vapam at soil moisture levels in the range of 10–30% (v:v) had a large effect on both disease severity and infection rates and plant growth parameters. The results suggest that Vapam can effectively reduce clubroot severity and may be useful for the treatment of transplant propagation beds in brassica vegetable production, and for the containment of small, localized clubroot infestations in commercial canola crops.     

Flooding affects the development of Plasmodiophora brassicae in Arabidopsis roots during the secondary phase of infection

Clubroot, a disease of Brassicaceae species, is caused by the soilborne pathogen Plasmodiophora brassicae. High soil water content was previously described to favour the motility of zoospores and their penetration into root cells. In this study, the effect of irrigation regimes on clubroot development during the post‐invasive secondary phase of infection was investigated. Three irrigation regimes (low, standard, high) were tested on two Arabidopsis accessions, Col‐0 (susceptible) and Bur‐0, a partially resistant line. In Col‐0, clubroot symptoms and resting spore content were higher under the ‘low irrigation’ regime than the other two regimes, thus enhancing the phenotypic contrast between the two Arabidopsis accessions. Clubroot severity under high and low irrigation regimes was evaluated in near‐isogenic lines derived from a Col‐0 ×  Bur‐0 cross, to assess the effect of soil moisture on the expression of each of four quantitative trait loci (QTL) controlling partial resistance. The presence of the Bur‐0 allele at the QTL PbAt5.2 resulted in reduced severity only under low irrigation, whereas the Bur‐0 allele at QTL PbAt5.1 was associated with partial resistance only under high irrigation. QTL PbAt4 reduced the number of resting spores in infected roots, but was not associated with reduced clubroot symptoms. The results indicated that soil moisture could have consequences for the secondary phase of clubroot development, depending on plant genotype. Future genetic studies may benefit from using combinations of watering conditions during the secondary stage of infection, thus opening up the possibility of identifying genetic factors expressed under specific environmental conditions.     

Variation in pathotypes and virulence of Plasmodiophora brassicae populations in Germany

Between 2012 and 2015, 49 new clubroot‐infested fields were identified in 12 German federal states. Clubroot disease incidence varied within these fields from 22% to 92%. Field information revealed that in 85% of fields, oilseed rape was grown in rotation once every 2 or 3 years. Frequency of OSR in the rotation was significantly correlated with the incidence and prevalence of clubroot disease. The disease was detected in fields with soil pH ranging from 5.1 to 8.3, and a significant negative correlation was found between soil pH and the disease incidence of infested fields. Furthermore, more cases of disease and severe incidences were observed in sandy loam and loamy sand as compared with other soil types. Pathotype classification of the 49 Plasmodiophora brassicae populations was conducted on two differential sets, the European Clubroot Differential set and the set of Somé (1996). Additionally, the degree of virulence of the collected isolates was analysed on the clubroot‐resistant oilseed rape cv. Mendel. The results showed variation in pathotype distribution in different regions in Germany. The majority of isolates according to Somé were pathotypes 1 and 3, respectively, with pathotypes 2 and 5 in the minority. Detailed classification according to Buczacki showed the dominance of 16/31/31, 16/14/30 and 16/14/31 populations among 20 distinct virulence patterns of collected isolates. From all populations tested for virulence on cv. Mendel, 15 isolates were found to be moderately or highly virulent. These virulent populations were not restricted to a small geographical area in the country.     

Reducing the build‐up of Plasmodiophora brassicae inoculum by early management of oilseed rape volunteers

Clubroot of oilseed rape (OSR), caused by Plasmodiophora brassicae, is a disease of increasing economic importance worldwide. Previous studies indicated that OSR volunteers, Brassica crops and weeds play a critical role in the predisposition of the disease. To determine the effect of timing of foliar application of the herbicide glyphosate or mechanical destruction of OSR volunteers in reduction of clubroot severity and resting spore production, a series of studies was conducted under controlled conditions with a susceptible OSR cultivar and an isolate of P. brassicae. Plants were inoculated by injecting a spore suspension beside the root hairs at growth stage 11–12 (BBCH scale) and were terminated at 7 (early) or 21 (late) days post‐inoculation (dpi). Under controlled conditions, the first symptoms on roots were observed as early as 7 dpi. The early application of glyphosate as well as early mechanical destruction resulted in significant (P ≤ 0.05) reduction in the development of clubroot symptoms, root fresh weight and the number of resting spores?g root. Furthermore, the effect of volunteer management on clubroot severity in the succeeding OSR was studied by inoculating plants with the resting spores obtained from treated clubbed roots. Inoculated OSR exhibited root clubs similar to the initial symptoms after 35 dpi. Plants that were inoculated with spore suspension from early treated roots resulted in significant reductions in clubroot incidence and severity. Conversely, plants inoculated with the spore suspension from the late treated roots displayed levels of clubroot similar to the plants inoculated with the spore solutions of positive controls.     

Pathogenic variation of Mycosphaerella species infecting banana and plantain in Nigeria

Mycosphaerella species that cause the ‘Sigatoka disease complex’ account for significant yield losses in banana and plantain worldwide. Disease surveys were conducted in the humid forest (HF) and derived savanna (DS) agroecological zones from 2004 to 2006 to determine the distribution of the disease and variation among Mycosphaerella species in Nigeria. Disease prevalence and severity were higher in the HF than in the DS zone, but significant (P < 0·001) differences between agroecological zones were only observed for disease severity. A total of 85 isolates of M. fijiensis and 11 isolates of M. eumusae were collected during the survey and used to characterize the pathogenic structure of Mycosphaerella spp. using a putative host differential cultivar set consisting of Calcutta‐4 (resistant), Valery (intermediate) and Agbagba (highly susceptible). Area under disease progress curve (AUDPC) was higher on all cultivars when inoculated with M. eumusae than with M. fijiensis, but significant (P < 0·05) differences between the two species were only observed on Valery. Based on the rank‐sum method, 8·3% of the isolates were classified as highly aggressive and 46·9% were classified as aggressive. About 11·5% of all the isolates were classified as least aggressive, and all of these were M. fijiensis. The majority of M. eumusae isolates (seven out of 11; 64%) were classified as aggressive. A total of nine pathotype clusters were identified using cluster analysis of AUDPC. At least one M. fijiensis isolate was present in all the nine pathotype clusters, while isolates of M. eumusae were present in six of the nine clusters. Isolates in pathotype clusters III and V were the most aggressive, while those in cluster VIII were the least aggressive. Shannon’s index (H) revealed a more diverse Mycosphaerella collection in the DS zone (H = 1·81) than in the HF (H = 1·50) zone, with M. fijiensis being more diverse than M. eumusae. These results describe the current pathotype structure of Mycosphaerella in Nigeria and provide a useful resource that will facilitate screening of newly developed Musa genotypes for resistance against two important leaf spot diseases of banana and plantain.     

Effect of host and non‐host crops on Plasmodiophora brassicae resting spore concentrations and clubroot of canola

Plasmodiophora brassicae, causal agent of clubroot of crucifers, poses a serious threat to Canadian canola production. The effects of fallow (F) periods and bait crops (clubroot‐susceptible canola (B) and perennial ryegrass (R)) on clubroot severity and P. brassicae resting spore populations were evaluated in five sequences: R–B, B–R, R–F, B–F and F–F. Both host and non‐host bait crops reduced clubroot severity in a subsequent crop of a susceptible canola cultivar compared with fallow. Resting spore and P. brassicae DNA concentrations decreased in all treatments, but were lowest for the R–B and B–R bait crop sequences. In addition, two studies were conducted in mini‐plots under field conditions to assess the effect of rotation of susceptible or resistant canola cultivars on clubroot severity and P. brassicae resting spore populations. One study included three crops of susceptible canola compared with a 2‐year break of oat–pea, barley–pea, wheat–wheat or fallow–fallow. The other study assessed three crops of resistant canola, two crops of resistant canola with a 1‐year break, one crop of resistant canola and a 2‐year break, and a 3‐year break with barley followed by a susceptible canola. The rotations that included non‐host crops of barley, pea or oat reduced clubroot severity and resting spore concentrations, and increased yield, compared with continuous cropping of either resistant or susceptible canola. Growing of a susceptible canola cultivar contributed 23–250‐fold greater gall mass compared with resistant cultivars.     

Severity of phytophthora root rot and pre‐emergence damping‐off in subterranean clover influenced by moisture,temperature, nutrition,soil type,cultivar and their interactions

Studies were carried out in controlled environment rooms reflecting field situations. In the presence of the devastating soilborne pathogen Phytophthora clandestina, subterranean clover (Trifolium subterraneum) seedling emergence was significantly affected by moisture, soil type, temperature and cultivar. The level of rotting of tap and lateral roots was significantly affected by nutrition, soil type, temperature and cultivar. There were significant interactions involving temperature, moisture, soil type and cultivar; cultivar resistance, high moisture, high or medium temperature, high nutrition and sand soil all contributed towards less pre‐emergence damping‐off and tap and lateral root disease and to greater clover productivity. Host resistance of subterranean clover cultivars was critical for reducing disease severity and increasing productivity, even when favourable environmental conditions for severe disease occurred. In the presence of P. clandestina, the most resistant cultivar, Seaton Park, performed best under a high temperature, high nutrition and high moisture combination, but showed lower productivity under conditions of low nutrition or lower temperature, even when moisture level was high. In contrast, less resistant cultivars Riverina and Meteora had less disease and greater productivity under low moisture conditions. Findings reflect field observations that pre‐emergence damping‐off and root disease from P. clandestina in subterranean clover is particularly severe under colder conditions and in nutritionally impoverished sandy soils, and demonstrate how variations in soil type, nutrition, moisture, temperature and cultivar have profound effects on the expression and severity of phytophthora pre‐emergence damping‐off and root disease and the productivity of subterranean clover forages.     

Mechanisms of resistance in Brassica carinata,B. napus and B. juncea to Pseudocercosporella capsellae

Studies were undertaken to compare susceptible and resistant host responses to Pseudocercosporella capsellae in cotyledons of Brassica carinata, B. juncea and B. napus in order to define the mechanisms of resistance in these three species. On both resistant and susceptible hosts, hyphal penetration was always through stomatal openings and without infection pegs or appressoria. On resistant B. carinata ATC94129P, up to 72% of spores disintegrated and, generally, germination (<22%) and germ tube lengths (<25 μm) were comparatively low. Resistant B. napus Hyola 42 had the lowest germination (8%) and susceptible B. carinata UWA#012 had the highest (51%). On resistant B. carinata ATC94129P, germ tube extension was impeded across 24–60 h post‐inoculation (hpi) and percentage stomatal penetration lower (4%) at 60 hpi compared with susceptible B. carinata UWA#012 (26%). Stomatal densities (stomata/14 757 μm²) on resistant B. juncea Dune (2·12) and B. napus Hyola 42 (1·62) were lower than for susceptible B. juncea Vardan (2·40) and B. napus Trilogy (2·03). Resistant B. carinata ATC94129P had greater stomatal density (1·89) than susceptible B. carinata UWA#012 (1·58). Overall, B. juncea had greater stomatal density (2·26) compared with B. napus (1·83) and B. carinata (1·74). In resistant B. carinata ATC94129P, P. capsellae induced 28% stomata to close, while in susceptible B. carinata UWA#012 no such closure was induced. Epicuticular wax crystalloids were present only on resistant B. carinata ATC94129P and probably also contribute towards resistance.     

Impacts of soil moisture level and organic matter content on growth of two Juncus species and Poa pratensis grown under acid soil conditions

The abundance of Juncus effusus (soft rush) and Juncus conglomeratus (compact rush) has increased in coastal grasslands in Norway over recent decades, and their spread has coincided with increased precipitation in the region. Especially in water‐saturated, peaty soils, it appears from field observations that productive grasses cannot compete effectively with such rapidly growing rush plants. In autumn–winters of 2012–2013 and 2013–2014, a four‐factor, randomised block greenhouse experiment was performed to investigate the effect of different soil moisture regimes and organic matter contents on competition between these rush species and smooth meadow‐grass (Poa pratensis). The rush species were grown in monoculture and in competition with the meadow‐grass, using the equivalent of full and half the recommended seed rate for the latter. After about three months, above‐ and below‐ground dry matter was measured. J. effusus had more vigorous growth, producing on average 23–40% greater biomass in both fractions than J. conglomeratus. The competitive ability of both rush species declined with decreasing soil moisture; at the lowest levels of soil moisture, growth reductions were up to 93% in J. conglomeratus and 74% in J. effusus. Increasing water level in peat–sand mixture decreased competivitiveness of meadow‐grass, while pure peat, when moist, completely impeded its below‐ground development. These results show that control of rush plants through management may only be achieved if basic soil limitations have been resolved.     

Potential loss of clubroot resistance genes from donor parent Brassica rapa subsp. rapifera (ECD 04) during doubled haploid production

Clubroot resistance derived from the oilseed rape/canola Brassica napus ‘Mendel’ has been overcome in some fields in Alberta, Canada, by the emergence of ‘new’ strains of the protist Plasmodiophora brassicae. Resistance to the pathogen was assessed in 112 doubled haploid (DH) lines, derived from B. rapa subsp. rapifera (European clubroot differential (ECD) 04). The lines were evaluated against five single‐spore isolates representing the ‘old’ pathotypes 2, 3, 5, 6 and 8, and 15 field populations representing new strains of P. brassicae. The disease severity index (ID%) data revealed that none of the DH lines were resistant or moderately resistant to the new pathotype 5X (field populations L‐G1, L‐G2, L‐G3) and D‐G3, while 3–42% were resistant or moderately resistant to the other 11 new strains. Using the mean ID induced by the old pathotype 3 (approx. 13.5%) as the baseline, clubroot severity increased by 300–600% when inoculated with the new pathotypes. A significant finding of this study was the fact that ECD 04 showed absolute resistance to all of the old and new P. brassicae strains while the B. napus ‘Mendel’, although resistant to all of the old pathotypes, was resistant to only about 50% of the new strains. Similarly, all of the selected clubroot‐resistant commercial canola cultivars evaluated in this study were susceptible to 87% of the new P. brassicae strains. The molecular data revealed that the breakdown of clubroot resistance in Mendel and the canola cultivars was in part due to the non‐inheritance of the Crr1 gene on the A08 chromosome from ECD 04.     

Colonization of lettuce cultivars and rotation crops by Fusarium oxysporum f. sp. lactucae,the cause of fusarium wilt of lettuce

The severity of fusarium wilt is affected by inoculum density in soil, which is expected to decline during intervals when a non‐susceptible crop is grown. However, the anticipated benefits of crop rotation may not be realized if the pathogen can colonize and produce inoculum on a resistant cultivar or rotation crop. The present study documented colonization of roots of broccoli, cauliflower and spinach by Fusarium oxysporum f. sp. lactucae, the cause of fusarium wilt of lettuce. The frequency of infection was significantly lower on all three rotation crops than on a susceptible lettuce cultivar, and the pathogen was restricted to the cortex of roots of broccoli. However, F. oxysporum f. sp. lactucae was isolated from the root vascular stele of 7·4% of cauliflower plants and 50% of spinach plants that were sampled, indicating a greater potential for colonization and production of inoculum on these crops. The pathogen was also recovered from the root vascular stele of five fusarium wilt‐resistant lettuce cultivars. Thus, disease‐resistant plants may support growth of the pathogen and thereby contribute to an increase in soil inoculum density. Cultivars that were indistinguishable based on above‐ground symptoms, differed significantly in the extent to which they were colonized by F. oxysporum f. sp. lactucae. Less extensively colonized cultivars may prove to be superior sources of resistance to fusarium wilt for use in breeding programmes.     

Measuring lesion attributes and analysing their spatial patterns at the leaf scale using digital image analysis

Digital image analysis was used to quantify size, shape and relative positions of individual plant disease lesions to determine their spatial distribution pattern at the leaf scale. Rice brown spot was used as a necrotrophic pathogen causing numerous discrete lesions. A 50‐leaf subsample was selected from an existing data set of 350 images of leaves taken from the field, and analysed for disease severity using image analysis. Further measurements included size, shape and the relative positions of lesions for all leaves with severity > 8% (n = 25) and an additional 25‐leaf sample with severity <8%. A total of 3964 necrotic and/or halo areas were selected using a manually defined threshold in the computer program Assess . There were significant and positive associations (Pearson's r > 0.81; P < 0.001) between the size‐related measurements (lesion area, longest and shortest axis). Coalesced areas, formed by interconnection of lesions and associated haloes, and a high number of small lesions were found with an increase in severity, suggesting a secondary cycle and autoinfection process. Results from quadrat‐based (Poisson distribution and Spatial Analysis by Distance IndicEs) and distance‐based (point‐process Poisson) spatial methods were in good agreement and, together with a Taylor power law model, suggested a shift from random to predominantly aggregated patterns of lesions at severities approaching 10%. This framework, which is applicable to other foliar diseases, proved useful in providing quantitative knowledge of epidemic processes at the leaf scale. Finally, these results may be useful in improving simulation models and disease assessment methods.     

Quantification of Mirafiori lettuce big‐vein virus and its vector,Olpidium virulentus,from soil using real‐time PCR

Big vein disease of lettuce (Lactuca sativa) is an economically important disease transmitted through soil by Olpidium virulentus, and has occurred in most production areas worldwide. The disease is assumed to be caused by Mirafiori lettuce big‐vein virus (MiLBVV). To understand the dynamics of the virus and its vector, MiLBVV and O. virulentus were directly detected in soil. DNA and RNA were extracted from 5 g soil using a bead beating method, followed by purification using adsorption to a column. Detection and quantification were performed using real‐time PCR and a TaqMan probe that was prepared based on the CP region of MiLBVV and the rDNA‐ITS region of O. virulentus, respectively. Furthermore, using a visual assessment of the incidence rate of big vein disease on lettuce in agricultural fields, the C_t values of MiLBVV and O. virulentus from soil were also determined using real‐time PCR. The results showed that MiLBVV concentrations in the soil were high in the field, as also determined by a visual assessment of the incidence rate of big vein disease on lettuce. However, the amount of O. virulentus in soil was not directly correlated with the incidence of MiLBVV. From these results, it is suggested that the risk of lettuce crops developing big vein disease can be estimated using an index of the amount of MiLBVV in the soil.     

Soil management in a mountain agroecosystem and clubroot disease

Clubroot, caused by Plasmodiophora brassicae, is a disease limiting the production of Brassica species. Its severity varies according to the density of the pathogen's resting spores and environmental conditions. Although the soil environment (including its physical, chemical and biological characteristics) is determinant in the development of the disease, little is known regarding these characteristics in tropical regions, such as Brazil. The objective of this study was to investigate the relationships between soil attributes, bacterial population composition and clubroot intensity in a tropical mountain agroecosystem. Soil and plant samples were collected from 17 cauliflower-producing localities in Nova Friburgo, Brazil. The physical and chemical attributes, the bacterial population composition, the number of pathogen resting spores in the soil samples, and the clubroot severity were evaluated. Based on bacterial population composition, two distinct groups of soils were identified, which varied in attributes such as organic matter, potential acidity, pH and disease severity. The organic matter level was associated with differences in soil acidity, bacterial population composition and disease intensity, but there was not a cause and effect relationship between bacterial population composition and clubroot severity. Under the conditions of this agroecosystem, soils with increased organic matter presented higher acidity potential (H + Al), which was strongly correlated to clubroot severity. Thus, high acidity potential could be a key factor leading to clubroot development in mountain regions of Brazil.     

Reduction of clubroot (Plasmodiophora brassicae) formation in Arabidopsis thaliana after treatment with prohexadione‐calcium,an inhibitor of oxoglutaric acid‐dependent dioxygenases

Recently, flavonoids were shown to modulate the outcome of clubroot development in Arabidopsis thaliana after infection with the obligate biotrophic pathogen Plasmodiophora brassicae. Therefore, the development of clubroot disease was investigated in Arabidopsis after treatment with prohexadione‐calcium (ProCa), an inhibitor of ascorbic acid/2‐oxoglutaric acid‐dependent dioxygenases such as flavanone‐3‐hydroxylase. The treatment resulted in a reduction of the flavonols quercetin and kaempferol in clubroots, whereas the precursor naringenin highly accumulated. The root system of ProCa‐treated plants was better developed although galls were still visible. Thus, ProCa treatment resulted in reduced gall size. Flavonoids are thought to inhibit polar auxin transport by modulating auxin efflux carriers. It was investigated whether the auxin response might change as a consequence of the accumulation of naringenin in ProCa‐treated plants. In the areas of gall development an auxin response was indicated by the auxin‐responsive promoter DR5 coupled to the reporter β‐glucuronidase (GUS), whereas very little staining was found in healthy root parts. No differences in GUS activity were found between P. brassicae‐infected and ProCa‐treated plants, and plants only infected with P. brassicae, indicating that the effect of ProCa treatment on clubroot reduction is not via changes in auxin responses. As ProCa is also an inhibitor of late steps in gibberellin biosynthesis, a specific gibberellin biosynthesis inhibitor, chlormequatchloride (CCC), was tested on club development. However, CCC did not reduce disease symptoms, indicating that the observed reduced gall development was not because of gibberellin biosynthesis inhibition by ProCa.     

The effect of post‐harvest storage conditions on the development of black dot (Colletotrichum coccodes) on potato in crops grown for different durations

The effects of post‐harvest curing and storage temperature on severity of black dot, caused by Colletotrichum coccodes, were investigated for potato crops grown for different crop durations (days from 50% emergence to harvest) in soils that posed a low, medium and high risk of disease. In field trials over four growing seasons (2005–8), black dot severity at harvest increased with increasing crop duration, within the range 103–146 days from 50% emergence to harvest (P < 0.05). In field trials over three growing seasons (2006–8), black dot severity on tubers at harvest increased significantly with increasing soil inoculum in each year, within the range 43–4787 pg C. coccodes DNA/g soil (P < 0.05). Storage trials were conducted to measure the influence of accumulated post‐harvest temperature on black dot. In 2005, no difference in black dot severity was observed on tubers stored for 20 weeks at 2.5 and 3.5 °C. In 2006 (but not 2007), increasing the duration of curing after harvest from 4 to 14 days increased black dot severity on tubers from 8.9 to 11.2% (P < 0.01) in long duration crops (>131 days after 50% emergence) grown under high (>1000 pg C. coccodes DNA/g soil) soil inoculum. The number of days of curing did not affect disease severity for shorter duration crops grown at high soil inoculum, or on crops grown at medium or low (100–1000 and <100 pg C. coccodes DNA/g soil, respectively) soil inoculum concentrations. Soil inoculum and crop duration together provided a reasonable prediction of black dot severity at harvest and after a 20‐week storage period.     

Sulphate and sulphurous acid alter the relative susceptibility of wheat to Phaeosphaeria nodorum and Mycosphaerella graminicola

The relative abundances of DNA of Mycosphaerella graminicola and Phaeosphaeria nodorum in archived wheat samples are closely correlated with UK anthropogenic emissions of oxidized sulphur over the last 160 years. To test whether this could be a causal relationship, possible modes of action of sulphur on the two fungi were examined. Mycelial growth of the two fungi in solutions of sulphurous acid was similar. Sulphurous acid at pH 4 reduced percentage germination of P. nodorum conidia more strongly than M. graminicola conidia. In spray inoculations of wheat cv. Squarehead's Master, Cappelle Desprez and Riband with water or sulphurous acid (pH 4), the ratio of leaves infected by P. nodorum to leaves infected by M. graminicola was increased by factors of 2·5, 2·1 and 0·6, respectively at pH 4. The same three cultivars of wheat were grown in sand and vermiculite and fertilized with nutrient solution containing 2·5 or 0·5 mm sulphate. Both pathogens infected less frequently at 2·5 mm sulphate, by a factor of about 2. The severity of infection by M. graminicola was reduced on all three cultivars by a factor of about 4·5 at 2·5 mm sulphate, but severity of P. nodorum was reduced only by a factor of about 2. Both elevated free sulphate concentrations in soil and sulphite in rainwater could therefore increase the prevalence of P. nodorum relative to M. graminicola, which is consistent with the historical changes in abundance.