Polymyxa betae zoospores as vectors of beet necrotic yellow vein furovirus1

When beet seedlings exposed as bait plants in infested soil were placed in a nutrient solution, they released Polymyxa betae zoospores, infected with beet necrotic yellow vein furovirus. The roots produced the first zoospores 5 days after the start of soil baiting. When seedlings were inoculated with zoospore suspensions, infection occurred within 5 min and reached a maximum in 30 min. The suspensions remained infectious for at least 2 h after removal of the bait plants from which the zoospores were released. So many spores were released into the suspension that disease transmission could be obtained within half an hour from an infected plant to a healthy plant, placed together into fresh medium. Suspensions could be diluted 1/16 with nutrient solution without any loss of infectivity, whereas 1/4 dilution with tap water resulted in a complete loss of infectivity.     

Effects of inoculum density,plant age and temperature on disease severity caused by pythiaceous fungi on several plants

Alfalfa, maize, sorghum and sugarbeet plants were inoculated with zoospores ofPhytophthora andPythium species in order to assess the effects of inoculum density, plant age and temperature on disease severity. Seedlings were grown axenically in test tubes and inoculated with zoospore suspensions. Disease severity was assessed by measuring the root growth and discoloration of treated and control seedlings. The incremental root length of all plants decreased and root discoloration increased as inoculum concentration of the pathogen increased. Changes were more intensive among low levels of zoospore concentrations and no significant differences in disease severity were found for inoculum densities higher than 10⁴ zoospores ml^-1. Disease severity was negatively related to plant age. Disease development on sugarbeet seedlings infected withPythium andPhytophthora species was affected by temperature, but the pattern of response was determined by the pathogen’s temperature preferences. The incremental root length decreased as temperature increased up to 25°C. The effect ofPythium dissimile andPhytophthora cactorum on root length was significantly lower at 35°C than at 25°C, whereasPythium aphanidermatum andPhytophthora nicotianae caused significant damage to roots even at 35°C. http://www.phytoparasitica.org posting Dec. 3, 2001.     

Prevention of Phytophthora Root Rot in Gerbera by increasing copper ion concentration in the nutrient solution

Increased copper concentration in the nutrient solution was used as a means for reducing the severity of root rot caused by Phytophthora cryptogea in Gerbera jamesonii in three experiments. Plants were grown in pots on ebb-and-flow benches with separate systems for recirculating nutrient solutions. Eight nutrient solutions with two concentrations of copper ions (0.07 and 0.28 ppm), two electrical conductivity values (1.5 and 2.2 mScm^-1), and two iron sources (FeHEEDTA or FeSO₄) were combined in a factorial design. Plants were inoculated with zoospores of P. cryptogea via the recirculating nutrient solution. Disease incidence was significantly reduced in inoculated plants grown on nutrient solution with 0.28 ppm copper compared with 0.07 ppm copper, when FeSO₄was introduced as the iron source. No effects of increased copper concentration was observed when iron was added as FeHEEDTA. The change in electrical conductivity from 1.5 to 2.2 mScm^-1 without changing the Cu²⁺ concentration did not influence the disease severity in these experiments. The results suggest that increased copper ion concentration in the nutrient solution could be a component of disease management strategy for ebb-and-flow systems. Possible management of the cupric ion concentration in the nutrient solutions is discussed.     

Zu den Ausbreitungswegen von Phytophthora ramorum an Rhododendron und Viburnum auf Container-Stellflächen

In Europe, Phytophthora ramorum basically infects Rhododendron and Viburnum. To prevent the spread of the new quarantine organism in nurseries more knowledge about the transmission biology of this pathogen is necessary. For this reason the pathways of spread for P. ramorum on the two main host plants have been studied for the first time. Under practical field conditions inoculated plants were placed as sources of infections in a larger stock. Over 3?months the development of infestation was recorded. The pathogen showed a poor potential of spread. At the end of the trial only 1.0% of Rhododendron and 0.3% of Viburnum were infested with P. ramorum. Typical symptoms could be observed. On Rhododendron the pathogen caused a branch dieback. Stems showed a brown discoloration, which starts usually at the tip of the twig and moved towards the base. Infected Viburnum showed a stem base rot with wilting symptoms. Additionally rhododendrons were natural infested with Phytophthora citricola. This pathogen caused the same symptoms like P. ramorum and spread much faster. Investigations of leaf litter showed that both Phytophthora species had colonized the ground. This observation and the pattern of spread indicate that inoculum on the ground has been transmitted to arial plant parts via rainsplash. There is little evidence that P. ramorum has been transmitted directly from plant to plant via splashwater or air.     

Conventional PCR and real time quantitative PCR detection of <Emphasis Type="Italic">Phytophthora cryptogea</Emphasis> on <Emphasis Type="Italic">Gerbera jamesonii</Emphasis>

A conventional PCR and a SYBR Green real-time PCR assays for the detection and quantification of Phytophthora cryptogea, an economically important pathogen, have been developed and tested. A conventional primer set (Cryp1 and Cryp2) was designed from the Ypt1 gene of P. cryptogea. A 369 bp product was amplified on DNA from 17 isolates of P. cryptogea. No product was amplified on DNA from 34 other Phytophthora spp., water moulds, true fungi and bacteria. In addition, Cryp1/Cryp2 primers were successfully adapted to real-time PCR. The conventional PCR and real-time PCR assays were compared. The PCR was able to detect the pathogen on naturally infected gerbera plants and on symptomatic artificially infected plants collected 21 days after pathogen inoculation. The detection limit was 5 × 10³ P. cryptogea zoospores and 16 fg of DNA. Real-time PCR showed a detection limit 100 times lower (50 zoospores, 160 ag of DNA) and the possibility of detecting the pathogen in symptomless artificially infected plants and in the re-circulating nutrient solution of closed soilless cultivation systems.     

Effects of soil moisture and sowing depth on the development of bean plants grown in sterile soil infested by <Emphasis Type="Italic">Rhizoctonia solani</Emphasis> and <Emphasis Type="Italic">Trichoderma harzianum</Emphasis>

The effects of soil moisture (varying from 15% to 42% (v/v)) and sowing depth (1.5–6.0 cm) on the development of bean plants grown in sterile soil infested by the pathogen Rhizoctonia solani and its antagonist Trichoderma harzianum were studied under greenhouse conditions. The four possible combinations of soil infestation with both fungi were tested. Disease severity, percentage of plants emerged, plant height and dry weight were evaluated 3 weeks after sowing. Emergence rate and growth of plants inoculated only with R. solani were not affected by soil moisture, but in the presence of both fungi, plant emergence, plant height and dry weight significantly decreased when soil moisture diminished. Deep sowing significantly reduced the emergence rate and growth of those plants that were inoculated with R. solani only. However, when the soil was infested with both fungi, the effect of sowing depth was not significant. At a sowing depth of 6.0 cm, the percentage of plants emerged was 50% in the presence of T. harzianum, but only 6.7% when the pathogen was inoculated alone. The antagonist protected bean seedlings from pre-emergence damping-off, reduced disease severity and increased plant growth in the presence of R. solani, especially in moist soil.     

韭菜和辣椒间作对辣椒疫病的防治效果及其化感机理

为明确韭菜和辣椒间作对辣椒疫病的防治效果及其化感机理,比较了韭菜和辣椒不同行比间作对辣椒疫病的防治效果,测定了韭菜茎、叶挥发物与浸提液对辣椒疫霉菌的抑制作用,并分析了韭菜根系对辣椒疫霉菌游动孢子侵染和传播行为的影响。结果表明,韭菜与辣椒行比为3∶1间作能显著控制辣椒疫病的扩展与传播;2.0 g/皿韭菜茎挥发物及0.15 m L/m L茎浸提液对辣椒疫霉菌菌丝生长的抑制活性可达33.33%和88.75%;辣椒疫霉菌游动孢子对韭菜根系具有明显的趋化活性,且于根围迅速休止并萌发,丧失在土壤中继续寻找寄主的能力,其传播侵染行为受到干扰。研究表明,韭菜和辣椒间作可以有效控制辣椒疫病的扩展蔓延,实现对辣椒疫病的生态防控。     

Bait method to detect Pythium species that grow at high temperatures in hydroponic solutions

Pythium helicoides, P. aphanidermatum and P. myriotylum are important pathogens that cause root rot of several crops in hydroponic culture and in ebb-and-flow irrigation systems. These species belong to a group of Pythium species that can grow at temperatures higher than 40°C. We developed a method for baiting these high-temperature Pythium species and evaluated its practicality to monitor their presence in nutrient solutions. Seeds of cucumber, tomato, radish, hemp, perilla and millet and leaves of bent grass and rose were tested as baits in hydroponic systems. Hemp, perilla and radish seeds and bent grass and rose leaves were more effective than the other baits for Pythium zoospores, and bent grass leaves were the most effective. In a sensitivity test, bent grass leaf traps (BLTs) detected three Pythium species after only a 1 day exposure to suspensions of 40 zoospores per liter of water, and the frequency of detection increased with zoospore density and with baiting period. A temperature of 38°C was optimum for the selective reisolation of the high-temperature Pythium species from the BLTs. The BLT was also tested with inoculated and noninoculated miniature roses that shared a recirculating nutrient solution. The pathogen was detected in the nutrient solution 23 days before the disease spread to the noninoculated roses. In addition, P. helicoides was detected 30 days before the disease was evident in a commercial greenhouse. The baiting method described here will be useful for monitoring high-temperature Pythium species in recirculating hydroponic culture systems.     

Direct and host‐mediated interactions between Fusarium pathogens and herbivorous arthropods in cereals

Fusarium head blight and fusarium ear rot diseases of cereal crops are significant global problems, causing yield and grain quality losses and accumulation of harmful mycotoxins. Safety limits have been set by the European Commission for several Fusarium‐produced mycotoxins; mitigating the risk of breaching these limits is of great importance to crop producers as part of an integrated approach to disease management. This review examines current knowledge regarding the role of arthropods in disease epidemiology. In the field, diseased host plants are likely to interact with arthropods that may substantially impact the disease by influencing spread or condition of the shared host. For example, disease progress by Fusarium graminearum can be doubled if wheat plants are aphid‐infested. Arthropods have been implicated in disease epidemiology in several cases and the evidence ranges from observed correlations between arthropod infestation and increased disease severity and mycotoxin accumulation, to experimental evidence for arthropod infestation causing heightened pathogen prevalence in hosts. Fusarium pathogens differ in spore production and impact on host volatile chemistry, which influences their suitability for arthropod dispersal. Herbivores may allow secondary fungal infection after wounding a plant or they may alter host susceptibility by inducing changes in plant defence pathways. Post‐harvest, during storage, arthropods may also interact with Fusarium pathogens, with instances of fungivory and altered behaviour by arthropods towards volatile chemicals from infected grain. Host‐mediated indirect pathogen–arthropod interactions are discussed alongside a comprehensive review of evidence for direct interactions where arthropods act as vectors for inoculum.     

Effects of hydrostatic pressure,agitation and CO2 stress on Phytophthora nicotianae zoospore survival

BACKGROUND: Phytophthora nicotianae Breda de Haan is a common pathogen of ornamental plants in recycled irrigation systems. In a previous study, annual vinca (Catharanthus roseus Don) inoculated with zoospore suspensions using a CO₂‐pressurized sprayer had less foliage blight than plants inoculated using a hand sprayer. Here, the impact of hydrostatic pressure, agitation and aeration with CO₂ on the survival of P. nicotianae zoospores was examined. RESULTS: Exposure of zoospores to 840 kPa hydrostatic pressure for 8 min or agitation at a mixing intensity (G) of 6483 s^?1 for 4 min at 22–23 °C did not kill zoospores, but resulted in viable cysts. Motile and forcefully encysted zoospores of P. nicotianae were equally infectious on vinca or lupine (Lupinus polyphylus Lindl.). Bubbling CO₂ into zoospore‐infested water at 110.4 mL (0.2 g) min^?1 for 5 min caused 81% reduction in the number of germinated zoospores. Pressure at 630 kPa (16.3 g CO₂) or 70 kPa (3.85 g CO₂) facilitated CO₂ injection and shortened the zoospore inactivation time to 30 s. When air was bubbled through the suspension, germination was similar to the control. CONCLUSIONS: Exposure to CO₂ killed P. nicotianae zoospores in water. Neither pressure nor agitation had an effect on zoospore viability or infectivity. Based on results of this study, the authors designed a recycling CO₂ water treatment system that is currently under evaluation. Copyright © 2010 Society of Chemical Industry     

Root rot inCampanula carpatica caused byPhytophthora cryptogea

This is the first record of root rot inCampanula carpatica as well as in the genus ofCampanula caused byPhytophthora cryptogea. An attack was observed in potted plants grown in a greenhouse on ebb- and flow benches and with recirculation of the nutrient solution. The fungus caused wilting of the leaves together with discoloration and rotting of the roots. Pathogenicity tests showed that the isolate ofP. cryptogea fromC. carpatica attactedGerbera jamesonii and vice versa. Treatment with the fungicide furalaxyl was able to reduce disease development.     

Use of zoospores of Polymyxa betae in screening beet seedlings for resistance to beet necrotic yellow vein virus

A system to culture viruliferousPolymyxa betae and to produce zoospores is described. The zoo spores were used for inoculation of beet seedlings, grown in nutrient solution, in tests for resistance to beet necrotic yellow vein virus (BNYVV). On most occasions in a time course experiment, and with various zoospore cultures, the partially resistant cultivar Rima and the accession Holly-1–4 had virus concentrations similar to the susceptible cultivar Regina, but the virus concentration inBeta vulgaris ssp.maritima accession WB42 was significantly lower (P<0.05). ‘Regina’ could be distinguished from various resistant accessions by a significantly higher virus concentration (P<0.05) shortly after inoculation, or after transplanting the seedlings from the nutrient solution into sand. Results of screening for resistance to BNYVV, using zoospores for inoculation, did not correspond with results of a test in which infested soil was used.Tests in which seedlings are grown in nutrient solution and inoculated with zoospores are suitable for the detection of accessions with a high level of resistance to BNYVV. To obtain virus infection in all plants, the optimal density of the zoospore suspension should first be determined and plants should be assayed shortly after inoculation.     

Infection by <Emphasis Type="Italic">Pythium aphanidermatum</Emphasis> increases production of phenolics in hydroponically grown peppers and predisposes healthy plants to root rot

The type and concentration of phenolics found in the cytosol (free), cell wall (bound), and nutrient solution of hydroponic pepper (Capsicum annuum L.) inoculated with Pythium aphanidermatum (Edson) Fitzp. were compared with non-inoculated controls. The quantities and types of free phenolics found in the roots and nutrient solution were greater in inoculated than in control plants. With regard to bound phenolics, while there was no difference between the types of phenolics in inoculated and control roots, quantities were greater in inoculated roots. High Pressure Liquid Chromatography analyses revealed two compounds in root extracts, and one in nutrient solution extracts that were contributors to root discolouration caused by Pythium aphanidermatum colonization. The compounds were subsequently isolated and identified as 4-hydroxybenzoic acid and 3-methoxy-4-hydroxybenzoic acid (vanillic acid) by their High Pressure Liquid Chromatography retention times, UV signature and Nuclear Magnetic Resonance spectroscopic comparisons with pure standards. Addition of various concentrations of 4-hydroxybenzoic acid or vanillic acid to the nutrient solution predisposed healthy plants to infection by zoospores of Pythium aphanidermatum when compared to plants treated with only Pythium aphanidermatum. 4-hydroxybenzoic acid and vanillic acid had little or no effect on colony growth of Pythium aphanidermatum in vitro, but respectively increased and decreased numbers of sporangia produced. These results are the first demonstration of host predisposition to infection by Pythium aphanidermatum caused by phenolic compounds that accumulate in inoculated roots and leach into the nutrient system of hydroponic systems.     

Monitoring by real-time PCR of three water-borne zoosporic Pythium species in potted flower and tomato greenhouses under hydroponic culture systems

The high-temperature-tolerant Pythium species P. aphanidermatum, P. helicoides, and P. myriotylum cause serious diseases in many crops under hydroponic culture systems in Japan. Control of the diseases is difficult because these zoosporic pathogens spread quickly. In this study, a real-time PCR method was developed for monitoring the spread of zoospores of the three pathogens. Specific primers and TaqMan probes were established using the internal transcribed spacer regions of the rDNA. Specificity was confirmed using known isolates of each species and closely related non-target species. The sensitivity of DNA detection was 10 f. for each pathogen. 10 f. DNA corresponded to 4 P. aphanidermatum, 3 P. myriotylum, and 4 P. helicoides zoospores, respectively. Therefore, this real-time PCR method was used to evaluate and monitor zoospores in the nutrient solutions of ebb-and-flow irrigation systems for potted flower production and closed hydroponic culture systems for tomato production. The results indicated that the pathogens were present in the hydroponic culture systems throughout the year, and spread before disease occurrence.     

Age‐related susceptibility of Eucalyptus species to Phytophthora boodjera

Phytophthora boodjera is a newly described pathogen causing damping off and mortality of Eucalyptus seedlings in Western Australian nurseries. This study evaluated the age‐related susceptibility of several taxa of mallee Eucalyptus to P. boodjera in sterilized washed river sand‐infestation pot trials. Phytophthora cinnamomi and P. arenaria were included for comparison. Seedlings of Eucalyptus taxa were inoculated at 0, 2, 4, 12 and 88 weeks with individual Phytophthora isolates. Pre‐emergent mortality in the presence of Phytophthora was almost 100%. Post‐emergent mortality was 50–100%, depending on isolate, compared to 0% for the control. Mortality was also high for inoculated 1 month‐old seedlings (46–68%) and root length of surviving seedlings was severely reduced. Death from root infection was not observed for seedlings inoculated at 12 and 88 weeks, but they developed root necrosis and reduced root dry weight compared to non‐inoculated controls. Phytophthora boodjera is a pre‐ and post‐emergent pathogen of mallee eucalypts. These eucalypts are susceptible to P. boodjera at all life stages tested, but the mortality rates declined with plant age. Similar results were obtained for P. cinnamomi and P. arenaria. The events leading to its recent appearance in the nurseries remain unknown and further investigations are underway to determine if this is an introduced or endemic pathogen. The approach used here to understand the impact of a Phytophthora species on multiple hosts at different seedling ages is novel and sets a benchmark for future work.     

Effects of inoculum density,leaf wetness duration and nitrate concentration on the occurrence of lettuce leaf spot

In Ehime Prefecture, Japan, lettuce leaf spot (Septoria lactucae) caused huge losses in marketable lettuce yields. To explore potential measures to control disease outbreaks, the effects of inoculum density, leaf wetness duration and nitrate concentration on the development of leaf spot on lettuce (Lactuca sativa) were evaluated. Conidia were collected from diseased plants in an infested field by single-spore isolation and were used to inoculate potted lettuce plants with different conidial concentrations. Lesions developed on inoculated lettuce plants at inoculum concentrations from 10⁰ to 10⁶ conidia/ml. The disease was more severe when the inoculum exceeded 10² conidia/ml, and severity increased with increasing concentrations. Assessment of the relationship between disease development and the duration of postinoculation leaf wetness revealed that symptoms appeared when the inoculated plants remained wet for 12 h or longer. The number of lesions and total nitrogen content in the lettuce leaves both increased when nitrate was applied.     

Detection and Quantification of Spongospora subterranea in Soil, Water and Plant Tissue Samples Using Real-Time PCR

A sensitive real-time polymerase chain reaction (PCR) assay was developed for the quantification of Spongospora subterranea, the cause of powdery scab and root galling in potato, and the vector of Potato mop top virus. A specific primer pair and a fluorogenic TaqMan® probe were designed to perform a quantitative assay for the detection of S. subterranea in soil, water and plant tissue samples. The assay was tested using DNA from cystosori, zoospores, plasmodia and zoosporangia of the pathogen. DNA was extracted directly from cystosori suspended in water and from clay soil with varying levels of added cystosori. DNA obtained from zoospores released into nutrient solution by cystosori in the presence of tomato bait plants was also tested, as was DNA from plasmodia and zoosporangia in infected tomato roots. In many cases, detection was successful even at low inoculum levels. This specific quantitative assay could therefore be a useful tool for studying the biology of S. subterranea, and for the optimisation of disease avoidance and control measures.     

Relationships of aphid and mite infestations to control ofBotrytis cinerea byClonostachys rosea in rose(Rosa hybrida) leaves

Infestations of aphids(Macrosiphum rosae L.) and of twospotted spider mites(Tetranychus urticae Koch) were examined in relation to growth and sporulation ofClonostachys rosea andBotrytis cinerea, and to suppression of the pathogen by the agent, in green rose leaves. Leaves were infested artificially with 10 aphids/leaflet for 3 h, or naturally with 15-30 aphids/leaflet for 7-12 days or with undetermined numbers of mites for 10-12 days. Leaves that had or had not been infested were inoculated withC. rosea, withB. cinerea, or withC. rosea plusB. cinerea. Germination incidence and germ tube growth ofC. rosea andB. cinerea on the phylloplane in most instances were much greater in leaves previously infested with aphids or mites compared with noninfested leaves. After combined inoculation,C. rosea suppressed germination ofB. cinerea from 47% to 19% in noninfested leaves, but in leaves that had been infested the agent was ineffective and germination incidence of the pathogen increased to 75-93%. Previous infestation with naturally introduced aphids or mites, but not brief infestations of artificially introduced aphids, markedly increased sporulation ofC. rosea after the leaves died during an initial 7-15 days of incubation on a paraquat agar medium, regardless of whether or notB. cinerea was present. Sporulation ofB. cinerea was similarly increased when inoculated alone. After 15-20 days, however, conidiophores of the agent or pathogen covered most of the leaf surface in these treatments. In leaves inoculated withC. rosea plusB. cinerea, the agent suppressed sporulation of the pathogen almost completely in both previously infested and noninfested leaves. Thus, aphid and mite infestations did not compromise the ability ofC. rosea to suppress inoculum production byB. cinerea in the leaves. Increased nutrient availability on the phylloplane through exudation or as honeydew or frass is proposed as a basis to explain effects of the pest infestations onC. rosea andB. cinerea.     

Phytophthora agathidicida: research progress,cultural perspectives and knowledge gaps in the control and management of kauri dieback in New Zealand

Kauri (Agathis australis), which is one of the world's largest and longest-living conifer species, is under threat from a root and collar dieback disease caused by the oomycete pathogen Phytophthora agathidicida. The noted incidence of kauri dieback has increased in the past decade, and even trees >1000 years old are not immune. This disease has profound effects on both forest ecosystems and human society, particularly indigenous Māori, for whom kauri is a taonga or treasure of immense significance. This review brings together existing scientific knowledge about the pathogen and the devastating disease it causes, as well as highlighting important knowledge gaps and potential approaches for disease management. The life cycle of P. agathidicida is similar to those of other soilborne Phytophthora pathogens, with roles for vegetative hyphae, zoospores and oospores in the disease. However, there is comparatively little known about many aspects of the biology of P. agathidicida, such as its host range and disease latency, or about the impact on the disease of abiotic and biotic factors such as soil health and co-occurring Phytophthora species. This review discusses current and emerging tools and strategies for surveillance, diagnostics and management, including a consideration of genomic resources, and the role these play in understanding the pathogen and how it causes this deadly disease. Key aspects of indigenous Māori knowledge, which include rich ecological and historical knowledge of kauri forests and a holistic approach to forest health, are highlighted.     

Population Dynamics of Fusarium Oxysporum f. Sp. Radicis-lycopersici in Relation to the Onset of Fusarium Crown and Root Rot of Tomato

Fusarium oxysporum f. sp. radicis-lycopersici the causal agent of crown and root rot in tomato comprises two overlapping separate phases: monocyclic and polycyclic. Oversummering inoculum is the source of primary infection (the monocyclic phase) and the spread from plant to plant via root-to-root contact is the source of the secondary infection (the polycyclic phase). In the present work, relationships between initial inoculum density, population dynamics of the pathogen in the root zone of diseased plants, and disease onset were studied. For the monocyclic phase, 55.1% of the variance of disease onset was attributed to the rate of pathogen proliferation in the root zone of plants, and only 12.8% of the variance was attributed to the amount of initial inoculum density. For the polycyclic phase, disease onset was not related to either initial inoculum density or the rate of pathogen proliferation in the root zone. At disease onset, the inoculum density of the pathogen in the root zone of plants infected from oversummering inoculum reached an average of 4.08 log cfu g soil^–1. The inoculum density of the pathogen in the root zone of plants infected by their diseased neighbors was 3.23 log cfu g soil^–1. A large variation in pathogen proliferation rate in the root zone was found among individual plants, suggesting that differences in the level of soil suppressiveness may occur not only between fields, but even in the same field over short distances.