Geographic location and year determine virulence,and year determines genetic change,in populations of Neopseudocercosporella capsellae

White leaf spot (Neopseudocercosporella capsellae) can be severe and problematic worldwide across both horticultural and oilseed Brassicaceae, including susceptible rapeseed. In this study, 82 isolates from 2015 and 106 isolates from across Australia in 2016 were first assessed for their virulence against three different rapeseed (Brassica napus) cultivars. For both years there were significant (P < 0.001) differences. Also, there were significant (all P < 0.001) differences between isolates in each year, and between cultivars. For 2016 isolates, there were also significant differences (P < 0.001) between isolates across three different Australian states, and a significant interaction (P < 0.001) between isolates with cultivars. Of the three Australian states, isolates from Victoria were most virulent. Among tested cultivars, cv. Scoop was most susceptible. Subsequently, phylogenetic analysis of 114 of these same 2015 and 2016 isolates showed current isolates clustered separately from the majority of 2005 N. capsellae isolates collected from Western Australia a decade earlier, confirming significant genetic change within N. capsellae populations over the past decade. However, isolate clusters showed no association with geographical location. The results suggest that phylogenetic association among 2005 and 2015–2016 N. capsellae isolates is complementary with pathogenicity variations explained by geographically different N. capsellae pathogen populations. Neopseudocercosporella capsellae populations are evolving rapidly, challenging management through host resistance at a time of increasing incidence and severity of white leaf spot disease over the past decade. The outcome is well illustrated by cv. Scoop, previously resistant to 2005 isolates but moderately susceptible to 2015 and highly susceptible to 2016 isolates.     

Survival of pathogens of Brussels sprouts (Brassica oleracea Gemmifera Group) in crop residues

Mycosphaerella brassicicola (ringspot), Alternaria brassicicola and A. brassicae (dark leaf spot) and Xanthomonas campestris pv. campestris (black spot) can infect leaves of Brussels sprouts resulting in yield losses. Infections of outer leaves of sprouts cause severe losses in quality. Crop residues can be a major primary inoculum source of the pathogens. Their population dynamics were followed in residues of leaves and stalks of crops of Brussels sprouts during 24 months using real‐time PCR assays. Leaf residues on the soil surface or buried in soil decomposed within 4 months. However, residues of stalks were present in the field after 24 months. In such residues, M. brassicicola populations increased during the first 2 months, but decreased thereafter and the pathogen was found only occasionally in the second year. Alternaria brassicicola multiplied on stalks exposed on the surface of field soil and was present on such residues after 24 months. Survival was less on residues buried in soil. Alternaria brassicae population increased in stalks exposed on the soil surface during the first months but decreased thereafter under the detection limit. Xanthomonas campestris cv. campestris populations fluctuated in time but 1 × 10⁴ cells mg^?1 stalk residue were still found after 24 months. Additionally, the four pathogens were present in residues of 11 commercial rapeseed crops that were analysed. The observed variation in population sizes of the pathogens between individual pieces of crop residues indicates a stochastic spread of pathogens. Unravelling the underlying processes will support the development of novel methods for sustainable disease prevention.     

Incidence,pathogenicity and diversity of Alternaria spp. associated with alternaria leaf spot of canola (Brassica napus) in Australia

Studies were undertaken to determine Alternaria spp. associated with leaf spot symptoms on canola (Brassica napus) in two cropping seasons (2015, 2016) across southern Australia. Major allergen Alt a1 and plasma membrane ATPase genes were used to identify Alternaria spp. In 2015, 112 isolates of seven Alternaria spp. were obtained, with A. metachromatica predominating. In 2016, 251 isolates of 12 Alternaria spp. were obtained, with A. infectoria predominating. Alternaria spp. isolates were morphologically and phylogenetically identified and studies to determine their pathogenicity on both B. napus (cv. Thunder TT) and B. juncea (cv. Dune) confirmed 10 species (A. alternata, A. arborescens, A. brassicae, A. ethzedia, A. hordeicola, A. infectoria, A. japonica, A. malvae, A. metachromatica and A. tenuissima) as pathogenic on both Brassica species. Alternaria ethzedia, A. hordeicola and A. malvae were recorded for the first time in Australia on any host and the record of A. arborescens was the first for New South Wales (NSW) and South Australia (SA). Other first records included A. infectoria on B. napus in NSW; A. japonica on B. napus in NSW and Western Australia (WA); A. metachromatica on any host in NSW, Victoria (VIC), WA and SA; and A. tenuissima on B. napus in NSW, SA and WA. It is evident that alternaria leaf spot on canola across southern Australia is not solely caused by A. brassicae, but that a range of other Alternaria spp. are also involved to varying degrees, depending upon the year and the geographic locality.     

Temperature and relative humidity shape white leaf spot (Neopseudocercosporella capsellae) epidemic development in rapeseed (Brassica napus)

Studies were undertaken to determine the combined environmental effects of two temperature regimes (14/11 ℃, 17/14 ℃ day/night) and duration of postinoculation high humidity on progression of white leaf spot, caused by Neopseudocercosporella capsellae, in rapeseed (Brassica napus) at each of two different plant growth stages. Overall, percentage disease indices were significantly affected by temperature, high humidity duration, plant growth stage, and host cultivar (all p < 0.001). There were significant two-way interactions of temperature regime with plant age (p < 0.001) and with host cultivar (p < 0.01), and significant three-way interactions of high humidity duration with growth stage and host cultivar (p = 0.01) and with temperature and host cultivar (p = 0.03). At cotyledon stage, mean percentage cotyledon disease index was 38.2 at 17/14 ℃ day/night, and 33.1 at 14/11 ℃. At fourth leaf stage, mean percentage leaf disease index was 32.0 at 14/11 ℃ but 17.5 at 17/14 ℃. Disease severity increased with increasing duration of high humidity. Results explain why this disease is more prevalent and severe in Australia following longer durations of high humidity; why disease at cotyledon stage is largely independent of temperature if seasonal temperature ranges between 11 and 17 ℃; and why more severe leaf infection (e.g., at fourth leaf stage) can be attributed to cooling winter conditions between autumn and winter. Studies suggest current/predicted climate changes across southern Australia of increasingly warmer autumn-winter temperatures and decreasing growing-season (May–August) precipitation will lessen severity of future epidemics.     

Alternaria leaf spot of basil caused by <Emphasis Type="Italic">Alternaria alternata</Emphasis> in Japan

An outbreak of black mottle and dieback on basil (Ocimum basilicum L.; Lamiaceae) was recorded in a greenhouse in Okinawa Prefecture, Japan during 2004. The causal agent was identified as Alternaria alternata (Fr.) Keissler based on morphological characters and growth temperature. This report is the first of Alternaria leaf spot of basil caused by A. alternata.     

Induced systemic resistance of Chinese cabbage to bacterial leaf spot and <Emphasis Type="Italic">Alternaria </Emphasis>leaf spot by the root endophytic fungus, <Emphasis Type="Italic">Heteroconium chaetospira</Emphasis>

 The root endophytic fungus Heteroconium chaetospira isolate OGR-3 was tested for its ability to induce systemic resistance in Chinese cabbage against bacterial leaf spot caused by Pseudomonas syringae pv. maculicola and Alternaria leaf spot caused by Alternaria brassicae of the foliar diseases. Chinese cabbage seedlings planted in soil infested with an isolate of H. chaetospira were incubated in a growth chamber for 32 days. The first to fourth true leaves of the seedlings were challenge-inoculated with P. syringae pv. maculicola or A. brassicae. Chinese cabbage planted in soil infested with H. chaetospira showed significant decreases in the number of lesions of bacterial leaf spot or Alternaria leaf spot when compared to the control plants not treated with H. chaetospira. The results indicated that colonization of roots by H. chaetospira could induce systemic resistance in Chinese cabbage and reduce the incidence of bacterial leaf spot and Alternaria leaf spot. Received: April 24, 2002 / Accepted: August 9, 2002     

A phylogenetically distinct lineage of Pyrenopeziza brassicae associated with chlorotic leaf spot of Brassicaceae in North America

Light leaf spot, caused by the ascomycete Pyrenopeziza brassicae, is an established disease of Brassicaceae in the United Kingdom (UK), continental Europe, and Oceania (OC, including New Zealand and Australia). The disease was reported in North America (NA) for the first time in 2014 on Brassica spp. in the Willamette Valley of western Oregon, followed by detection in Brassica juncea cover crops and on Brassica rapa weeds in northwestern Washington in 2016. Preliminary DNA sequence data and field observations suggest that isolates of the pathogen present in NA might be distinct from those in the UK, continental Europe, and OC. Comparisons of isolates from these regions using genetic (multilocus sequence analysis, MAT gene sequences, and rep-PCR DNA fingerprinting), pathogenic (B. rapa inoculation studies), biological (sexual compatibility), and morphological (colony and conidial morphology) analyses demonstrated two genetically distinct evolutionary lineages. Lineage 1 comprised isolates from the UK, continental Europe, and OC, and included the P. brassicae type specimen. Lineage 2 contained the NA isolates associated with recent disease outbreaks in the Pacific Northwest region of the USA. Symptoms caused by isolates of the two lineages on B. rapa and B. juncea differed, and therefore “chlorotic leaf spot” is proposed for the disease caused by Lineage 2 isolates of P. brassicae. Isolates of the two lineages differed in genetic diversity as well as sensitivity to the fungicides carbendazim and prothioconazole.     

Effects of R gene‐mediated resistance in Brassica napus (oilseed rape) on asexual and sexual sporulation of Pyrenopeziza brassicae (light leaf spot)

The phenotype of the R gene‐mediated resistance derived from oilseed rape (Brassica napus) cv. Imola against the light leaf spot plant pathogen, Pyrenopeziza brassicae, was characterized. Using a doubled haploid B. napus mapping population that segregated for resistance against P. brassicae, development of visual symptoms was characterized and symptomless growth was followed using quantitative PCR and scanning electron microscopy on leaves of resistant/susceptible lines inoculated with suspensions of P. brassicae conidia. Initially, in controlled‐environment experiments, growth of P. brassicae was unaffected; then from 8 days post‐inoculation (dpi) some epidermal cells collapsed (‘black flecking’) in green living tissue of cv. Imola and from 13 to 36 dpi there was no increase in the amount of P. brassicae DNA and no asexual sporulation (acervuli/pustules). By contrast, during this period there was a 300‐fold increase in P. brassicae DNA and extensive asexual sporulation in leaves of the susceptible cv. Apex. However, when leaf tissue senesced, the amount of P. brassicae DNA increased rapidly in the resistant but not in the susceptible cultivar and sexual sporulation (apothecia) was abundant on senescent tissues of both. These results were consistent with observations from both controlled condition and field experiments with lines from the mapping population that segregated for this resistance. Analysis of results of both controlled‐environment and field experiments suggested that the resistance was mediated by a single R gene located on chromosome A1.     

Destruxin B produced by Alternaria brassicae does not induce accessibility of host plants to fungal invasion

It has been reported that Alternaria brassicae, the causal agent of gray leaf spot in Brassica plants, produces a host-specific or host-selective toxin (HSTs) identified as destruxin B. In this study, the role of destruxin B in infection of the pathogen was investigated. Destruxin B purified from culture filtrates (CFs) of A. brassicae induced chlorosis on host leaves at 50–100 μg ml⁻¹, and chlorosis or necrosis on non-host leaves at 250–500 μg ml⁻¹. Destruxin B was detected in spore germination fluids (SGFs) on host and non-host leaves, but not in a sufficient amount to exert toxicity to host plants. When spores of non-pathogenic A. alternata were combined with destruxin B at 100 μg ml⁻¹ and inoculated on the leaves, destruxin B did not affect the infection behavior of the spores. Interestingly, SGF on host leaves allowed non-pathogenic spores to colonize host leaves. Moreover, a high molecular weight fraction (>5 kDa) without destruxin B obtained by ultrafiltration of SGF had host-specific toxin activity and infection-inducing activity. From these results, we conclude that destruxin B is not a HST and does not induce the accessibility of the host plant which is essential for colonization of the pathogen. In addition, the results with SGF imply that a high molecular weight HST(s) is involved in the host–pathogen interaction.     

Pathotypes and phylogenetic variation determine downy mildew epidemics in Brassica spp. in Australia

Isolates of Hyaloperonospora brassicae inoculated onto cotyledons of 28 diverse Brassicaceae genotypes, 13 from Brassica napus, two from B. juncea, five from B. oleracea, two from Eruca vesicaria, and one each from B. nigra, B. carinata, B. rapa, Crambe abyssinica, Raphanus sativus and R. raphanistrum, showed significant effects (P ≤ 0.001) of isolate, host and their interaction. Host responses ranged from no visible symptom or a hypersensitive response, to systemic spread and abundant pathogen sporulation. Isolates were generally most virulent on their host of origin. Using an octal classification, six host genotypes were identified as suitable host differentials to characterize pathotypes of H. brassicae and distinguished eight distinct pathotypes. There were fewer, but more virulent, pathotypes in 2015–2016 isolates than 2006–2008 pathogen populations, probably explaining the increase in severity of canola downy mildew over the past decade. Phylogenetic relationships determined across 20 H. brassicae isolates collected in 2006–2008 and 88 isolates collected in 2015–2016 showed seven distinct clades, with 70% of 2006–2008 isolates distributed within clade I (bootstrap value (BVs) of 100%) and the remaining 30% in clade V (BVs 83.3%). This is the first study to define phylogenetic relationships of H. brassicae isolates in Australia, setting a benchmark for understanding current and future genetic shifts within pathogen populations; it is also the first to use octal classification to characterize pathotypes of H. brassicae, providing a novel basis for standardizing phenotypic characterization and monitoring of pathotypes on B. napus and some crucifer species in Australia.     

Multiple Alternaria species groups are associated with leaf blotch and fruit spot diseases of apple in Australia

Leaf blotch and fruit spot of apple caused by Alternaria species occur in apple orchards in Australia. However, there is no information on the identity of the pathogens and whether one or more Alternaria species cause both diseases in Australia. Using DNA sequencing and morphological and cultural characteristics of 51 isolates obtained from apple leaves and fruit with symptoms in Australia, Alternaria species groups associated with leaf blotch and fruit spot of apples were identified. Sequences of Alternaria allergen a1 and endopolygalacturonase gene regions revealed that multiple Alternaria species groups are associated with both diseases. Phylogenetic analysis of concatenated sequences of the two genes resulted in four clades representing A. arborescens and A. arborescens‐like isolates in clade 1, A. tenuissima/A. mali isolates in clade 2, A. alternata/A. tenuissima intermediate isolates in clade 3 and A. longipes and A. longipes‐like isolates in clade 4. The clades formed using sequence information were supported by colony characteristics and sporulation patterns. The source of the isolates in each clade included both the leaf blotch variant and the fruit spot variant of the disease. Alternaria arborescens‐like isolates were the most prevalent (47%) and occurred in all six states of Australia, while A. alternata/A. tenuissima intermediate isolates (14%) and A. tenuissima/A. mali isolates (6%) occurred mostly in Queensland and New South Wales, respectively. Implications of multiple Alternaria species groups on apples in Australia are discussed.     

Interactions in the Brassica napus–Pyrenopeziza brassicae pathosystem and sources of resistance to P. brassicae (light leaf spot)

Pyrenopeziza brassicae, cause of light leaf spot (LLS), is an important pathogen of oilseed rape and vegetable brassicas and has a wide geographic distribution. Exploitation of host resistance remains the most sustainable and economically viable solution for disease management. This study evaluated 18 oilseed rape cultivars or breeding lines for host resistance against P. brassicae in glasshouse experiments. Selected cultivars/lines were inoculated with eight single-spore isolates of the pathogen obtained from three different regions in England. Analysis of P. brassicae infection-related changes on host plants identified leaf deformation as a characteristic feature associated with P. brassicae infection, this showed poor correlation to LLS severity measured as the amount of pathogen sporulation on infected plants. Resistant host phenotypes were identified by limitation of P. brassicae sporulation, with or without the presence of a necrotic response (black flecking phenotype). Investigation of this pathosystem revealed significant differences between cultivars/lines, between isolates, and significant cultivar/line-by-isolate interactions. In total, 37 resistant and 16 moderately resistant interactions were identified from 144 cultivar/line-by-isolate interactions using statistical methods. Most of the resistant/moderately resistant interactions identified in this study appeared to be nonspecific towards the isolates tested. Our results suggested the presence of isolate-specific resistant interactions for some cultivars. Several sources of resistance have been identified that are valuable for oilseed rape breeding programmes.     

Evaluation of virulence and pathogenicity of Alternaria patula on French marigold (Tagetes patula)

Alternaria patula, the cause of French marigold leaf black spot and flower blight, was first isolated from seeds of French marigold cv. Queen Sophia. It is described as a new species of Alternaria and has a considerable morphological variation with a preferential pathogenicity to Asteraceae, Solanaceae and Cucurbitaceae plants. Alternaria patula produces an array of pectin depolymerases that can break 1,4-α-glycosidic bonds either by hydrolysis of polygalacturonases (PG, E.C. 3.2.1.15) or via trans-elimination of pectate lyases (PL, E.C. 4.2.2.2) and pectin lyases (PNL, E.C. 4.2.2.10). This study is the first to emphasize the variability and significance to pathogenesis of the pectinolytic enzymes of A. patula that target various pectic polymers structures during host tissue invasion. Alternaria patula also produced zinniol derivatives as non-host-specific toxins (nHSTs), albeit without phytotoxic symptoms in French marigold. The management of Alternaria blight caused by A. patula via the application of pyrifenox and antagonistic Bacillus amyloliquefaciens effectively reduced disease severity, without adverse effects on French marigold in both in vitro and in vivo bioassays.     

Activity of carboxylic acid amide (CAA) fungicides against Bremia lactucae

Four carboxylic acid amide (CAA) fungicides, mandipropamid (MPD), dimethomorph (DMM), benthiavalicarb (BENT) and iprovalicarb (IPRO) were examined for their effects on various developmental stages of Bremia lactucae, the causal agent of downy mildew in lettuce, in vitro and in planta. Spore germination in vitro or on leaf surfaces was inhibited by all CAA fungicides (technical or formulated). MPD was more effective in suppressing germination than DMM or BENT, whereas IPRO was least effective. CAA induced no disruption of F-actin microfilament organisation in germinating spores of B. lactucae. CAA applied to germinating spores in vitro prevented further extension of the germ tubes. When applied to germinated spores on the leaf surface they prevented penetration. Preventive application of CAA to intact plants inhibited infection. MPD was more effective in suppressing infection than DMM or BENT, whereas IPRO was least effective. Curative application was effective at ≤3 h post-inoculation (hpi) but not at ≥18 hpi. CAA (except IPRO) applied to upper leaf surfaces inhibited spore germination on the lower surface and hence reduced infection. CAA suppressed sporulation of B. lactucae on floating leaf discs and when sprayed onto infected plants two days before onset of sporulation. BENT and DMM were more effective in suppressing sporulation than MPD or IPRO. Epidemics of downy mildew in shade-house grown lettuce were suppressed by CAA. A single spray applied to five-leaf plants before transplanting controlled the disease for 50 days. The results suggest that CAA are effective inhibitors of spore germination and therefore should be used as preventive agents against downy mildew of lettuce caused by B. lactucae.     

Phenotypic and phylogenetic studies associated with the crucifer white leaf spot pathogen,Pseudocercosporella capsellae,in Western Australia

Pseudocercosporella capsellae (white leaf spot disease) is an important disease on crucifers. Fifty‐four single‐conidial isolates collected from Brassica juncea (Indian mustard), B. napus (oilseed rape), B. rapa (turnip), and Raphanus raphanistrum (wild radish) across Western Australia were investigated for differences in pathogenicity and virulence using cotyledon screening tests, genetic differences using internal transcribed spacer (ITS) sequencing and phylogenetic analysis, and growth rates on potato dextrose, V8 juice and malt extract agars. All isolates from the four crucifer hosts were pathogenic on the three test species: B. juncea, B. napus and R. raphanistrum, but showed differences in levels of virulence. Overall, isolates from B. juncea, B. napus and B. rapa showed greatest virulence on B. juncea, least on R. raphanistrum and intermediate virulence on B. napus. Isolates from R. raphanistrum showed greatest virulence on B. juncea, least on B. napus and intermediate virulence on R. raphanistrum. Growth and production of a purple‐pink pigment indicative of cercosporin was greatest on malt extract agar and cercosporin production on V8 juice agar was positively correlated with virulence of isolates on B. juncea and B. napus. ITS sequencing and phylogenetic analysis showed that isolates collected from B. napus, B. juncea and B. rapa, in general and with few exceptions, had a high degree of genetic similarity. In contrast, isolates from R. raphanistrum were clearly differentiated from isolate groups collected from Brassica hosts. Pseudocercosporella capsellae reference isolates from other countries generally grouped into a single separate cluster, highlighting the genetic distinctiveness of Western Australian isolates.     

Characterization of resistance mechanisms activated by Trichoderma harzianum T39 and benzothiadiazole to downy mildew in different grapevine cultivars

Downy mildew, caused by Plasmopara viticola, is one of the most destructive diseases of grapevine and is controlled with intense application of chemical fungicides. Treatment with Trichoderma harzianum T39 (T39) or benzothiadiazole‐7‐carbothioic acid S‐methyl ester (BTH) has been previously shown to activate grapevine resistance to downy mildew and reduce disease symptoms in the Pinot noir cultivar. However, enhancement of plant resistance can be affected by several factors, including plant genotype. In order to further extend the use of resistance inducers against downy mildew, the physiological and molecular properties of T39‐ and BTH‐activated resistance in different cultivars of table and wine grapes were characterized under greenhouse conditions. T39 treatment reduced downy mildew symptoms, but the degree of efficacy differed significantly among grapevine cultivars. However, efficacy of BTH‐activated resistance was consistently high in the different cultivars. Expression profiles of defence‐related genes differed among cultivars in response to resistance inducers and to pathogen inoculation. T39 treatment enhanced the expression of defence‐related genes in the responsive cultivars, before and after P. viticola inoculation. A positive correlation between the efficacy of T39 and the expression level of defence‐related genes was found in Primitivo and Pinot noir plants, while different genes or more complex processes were probably activated in Sugraone and Negroamaro. The data reported here suggest that the use of a responsive cultivar is particularly important to maximize the efficacy of resistance inducers and new natural inducers should be explored for the less responsive cultivars.     

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.     

Plasmodiophora brassicae resting spore dynamics in clubroot resistant canola (Brassica napus) cropping systems

The soilborne pathogen Plasmodiophora brassicae, causal agent of clubroot of canola (Brassica napus), is difficult to manage due to the longevity of its resting spores, ability to produce large amounts of inoculum, and the lack of effective fungicides. The cropping of clubroot resistant (CR) canola cultivars is one of the few effective strategies for clubroot management. This study evaluated the impact of the cultivation of CR canola on P. brassicae resting spore concentrations in commercial cropping systems in Alberta, Canada. Soil was sampled pre-seeding and post-harvest at multiple georeferenced locations within 17 P. brassicae-infested fields over periods of up to 4 years in length. Resting spore concentrations were measured by quantitative PCR analysis, with a subset of samples also evaluated in greenhouse bioassays with a susceptible host. The cultivation of CR canola in soil with quantifiable levels of P. brassicae DNA resulted in increased inoculum loads. There was a notable lag in the release of inoculum after harvest, and quantifiable P. brassicae inoculum peaked in the year following cultivation of CR canola. Rotations that included a ≥2-year break from P. brassicae hosts resulted in significant declines in soil resting spore concentrations. A strong positive relationship was found between the bioassays and qPCR-based estimates of soil infestation. Results suggest that CR canola should not be used to reduce soil inoculum loads, and crop rotations in P. brassicae infested fields should include breaks of at least 2 years away from B. napus, otherwise the risk of selecting for virulent pathotypes may increase.     

Alternaria Leaf Spot on Three Species of Pelargonium Caused by Alternaria alternata in Japan

A new disease of pelargonium (Pelargonium domesticum Bailey), ivy geranium (P. peltatum (L.) L'Hér. ex Ait.) and scented geranium (P. graveolens L'Hér.), primarily causing brown spots on leaves, was found in Kawasaki-shi in Kanagawa Prefecture and Tachikawa-shi in Tokyo. An Alternaria sp. was consistently isolated from these diseased leaves, and the isolates were pathogenic to their host leaves. Based on morphological characteristics, the causal fungus in all three cases was identified as Alternaria alternata (Fr.) Keissler. Because Alternaria leaf spot of geranium by A. alternata has already been reported, the pathogenicity of isolates from four groups of genus Pelargonium was investigated. The isolates from scented geranium were pathogenic only to their original host, but the isolates from pelargonium, ivy geranium and geranium were pathogenic to all groups of pelargonium. This is the first report of this disease on pelargonium, ivy geranium and scented geranium caused by A. alternata in Japan. We propose the names for these diseases as Alternaria leaf spot of pelargonium (kappan-byo), Alternaria leaf spot of ivy geranium (kappan-byo) and Alternaria leaf spot of scented geranium (kappan-byo). Received 11 December 2000/ Accepted in revised form 19 July 2001     

Phoma black stem severity and phytoestrogen production in annual Medicago spp. is primarily determined by interaction of cultivar and pathogen isolate

Phoma black stem and leaf spot disease (caused by Phoma medicaginis) not only diminishes forage and seed yield but stimulates production of detrimental phytoestrogens in annual Medicago spp. This study aimed to evaluate relationships between disease development from five isolates of P. medicaginis on 16 cultivars with production of coumestrol and 4′-O-methylcoumestrol. In the presence of P. medicaginis, Sava had the highest coumestrol and 4′-O-methylcoumestrol (640 and 85 mg/kg, respectively) followed by Caliph (253 and 15 mg/kg, respectively). In the absence of P. medicaginis, Jemalong and Paragosa showed highest and lowest coumestrol (137 and 0 mg/kg, respectively). 4′-O-methylcoumestrol was not produced in disease-free plants, but coumestrol was. Disease incidence and severity on leaves and on petiole/stems, and consequent leaf yellowing severity ranged from 5%–98.7%, 0%–100%, 4.4%–98.7%, 1.7%–100%, and 0%–85%. Sava, Paraponto, Harbinger, and Serena were most susceptible, while Tornafield and Caliph were least susceptible. There was significant overall positive correlation of disease incidence/severity factors across cultivars (p < 0.01) with both coumestrol and 4′-O-methylcoumestrol. Jemalong was least responsive and Paragosa and Sava most responsive to coumestrol production following P. medicaginis inoculation. Coumestrol in inoculated Paragosa increased to 373 mg/kg in comparison with 0 mg/kg in controls. These findings are of critical importance towards developing less disease-susceptible annual Medicago spp. producing less detrimental phytoestrogens. Least susceptible cultivars like Tornafield and Caliph can be used to manage yield loss, whilst least responsive cultivars to phytoestrogen production like Caliph also can help to reduce phytoestrogen production.