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1.
Comparison of serological, culture, and bait methods for detection of Pythium and Phytophthora zoospores in water 总被引：1，自引：1，他引：1
Two immunodiagnostic detection assay procedures were compared with two conventional assays for their sensitivity in detecting propagules of Pythium ultimum var. sporangiiferum , Pythium Group F, Phytophthora cactorum and P. cryptogea in dilution series in sterile distilled water. The most sensitive assay for all four species was the zoospore trapping immunoassay (ZTI). Conventional membrane filtration-dilution plating gave similar results to ZTI with the two Phytophthora spp., but was less sensitive in Pythium detection. Immunodiagnostic dipstick assays and conventional bait tests showed similar sensitivities in the dilution series, and were generally about two orders of magnitude less sensitive than ZTI. The four techniques were also compared for their detection efficacy with water samples collected from horticultural nurseries and in in situ tests of infected root zones of Chamaecyparis , tomato and Chrysanthemum . In these comparisons, ZTI was again the most sensitive test for water samples, although membrane filtration-dilution plating proved to be a more consistent test. Dipstick and baiting assays were the best techniques for in situ testing, and dipsticks provided epidemiologically valuable, quantitative data on pathogen propagule numbers. 相似文献
2.
Elucidation of the microbial complex having a causal role in the development of apple replant disease in washington 总被引：2，自引：0，他引：2
Mazzola M 《Phytopathology》1998,88(9):930-938
ABSTRACT Systematic studies were conducted to elucidate the role of different soil microbial groups in the development of apple replant disease. Populations of targeted microorganisms were reduced by the application of semiselective biocides and soil pasteurization. Bacteria were not implicated in the disease, because application of the antibiotic chloramphenicol reduced soil populations of bacteria but failed to improve growth of apple transplants, while enhanced growth was achieved at pasteurization temperatures that did not alter attributes of the bacterial community recovered from apple roots. Populations of Pratylenchus penetrans were below the damage threshold level in eight of nine orchards surveyed, and nematicide applications failed to enhance apple growth in four of five replant soils tested, indicating that plant parasitic nematodes have a minor role or no role in disease development. Application of the fungicide difenconazole or metalaxyl enhanced growth of apple in all five replant soils, as did fludioxinil in the two soils tested. Soil pasteurization enhanced growth of apple and resulted in specific changes in the composition of the fungal community isolated from the roots of apple seedlings grown in these treated soils. Cylindrocarpon destructans, Phytophthora cactorum, Pythium spp., and Rhizoctonia solani were consistently isolated from symptomatic trees in the field and were pathogenic to apple. However, the composition of the Pythium and Rhizoctonia component and the relative contribution of any one component of this fungal complex to disease development varied among the study orchards. These findings clearly demonstrate that fungi are the dominant causal agents of apple replant disease in Washington state. 相似文献
3.
Rajeshwar S. Utkhede 《Pest management science》1984,15(3):241-246
The mycelial growth of Phytophthora cactorum on corn meal agar was completely inhibited by metalaxyl or mancozeb at 50 or 100 μg ml?1, but not by fosetyl-aluminium at the same concentrations. Soil drenches with metalaxyl, followed by metalaxyl+mancozeb, around the base of naturally infected trees under orchard conditions, prevented the growth of P. cactorum in the infected bark of the tree. Soil drenchs with metalaxyl, followed by metalaxyl+mancozeb, also prevented the infection of apple trees with P. cactorum under orchard conditions. The trunk circumference and spur length were not significantly affected by any of the fungicide treatments in all the trials over 2 years, except in one of the four orchards used in the study. Soil drench with metalaxyl alone, or followed by metalaxyl+mancozeb, may provide an effective control method for crown rot of apple trees. 相似文献
4.
ABSTRACT Recovery of Phytophthora ramorum from soils throughout sudden oak death-affected regions of California illustrates that soil may serve as an inoculum reservoir, but the role of soil inoculum in the disease cycle is unknown. This study addresses the efficacy of soil baiting, seasonal pathogen distribution under several epidemiologically important host species, summer survival and chlamydospore production in soil, and the impact of soil drying on pathogen survival. The efficacy of rhododendron leaves and pears as baits for detection of soilborne propagules were compared. Natural inoculum associated with bay laurel (Umbellularia californica), tanoak (Lithocarpus densiflorus), and redwood (Sequoia sempervirens) were determined by monthly baiting. Summer survival and chlamydospore production were assessed in infected rhododendron leaf disks incubated under bay laurel, tanoak, and redwood at either the surface, the litter/soil interface, or in soil. Rhododendron leaf baits were superior to pear baits for sporangia detection, but neither bait detected chlamydospores. Most inoculum was associated with bay laurel and recovery was higher in soil than litter. Soil-incubated inoculum exhibited over 60% survival at the end of summer and also supported elevated chlamydospore production. P. ramorum survives and produces chlamydospores in forest soils over summer, providing a possible inoculum reservoir at the onset of the fall disease cycle. 相似文献
5.
G. Tuitert 《European journal of plant pathology / European Foundation for Plant Pathology》1990,96(6):331-341
Application of a bioassay on serial dilutions of rhizomania-infested soil provided adequate information on the level of infestation withPolymyxa betae and beet necrotic yellow vein virus (BNYVV). Different combinations of dilution ratios ratios and numbers of replicates (N) that had the same average precision were compared. A most probable number (MPN) computer programme was written to enable the comparison, because MPN tables available in literature are limited to certain dilution ratios and values of N. Most probable numbers of infective units per ml soil assessed for infested soil from the Noordoostpolder and from Tholen (the Netherlands) were 48 forP. betae with 7.1 for BNYVV and 16 forP. betae with 1.6 for BNYVV, respectively. So in these soils 10–15% of the infective population ofP. betae was viruliferous. The inoculum potential of stored soil samples was not affected by conditions during storage for 28 months (dry and warm or wet and cool). 相似文献
6.
The effects of several factors on the estimation of Verticillium dahliae in soil by the wet-sieving method were studied. The following factors were important for maximising recovery: the removal of soil particles of < 20 μm size from suspensions before plating; the medium used for plating; the amount of sieved soil inoculated to plates; the length of time of incubating plates; and the method of incubating soil on plates. There was no short-term effect of air-drying soil before analysis. Using sodium hexametaphos-phate to aid soil dispersion, treating soil suspensions with sodium hypochlorite, or making suspensions from different amounts of soil had no consistent effect on recovery. Excessive sample-to-sample variability in the results of some experiments was attributed to non-random distribution of fungus propagules in soil, or to the difficulty of standardizing the sieving part of the analysis, or both. Wet-sieving was generally more effective in detecting V. dahliae and gave higher counts than a sucrose-flotation method. Bait methods with eggplants or antirrhinums proved ineffective for estimating V. dahliae in naturally infested soil. 相似文献
7.
Treena I. Burgess Adrián López-Villamor Trudy Paap Briony Williams Rajah Belhaj Michael Crone William Dunstan Kay Howard Giles E. St. J. Hardy 《Plant pathology》2021,70(3):604-614
The genus Phytophthora contains species that are major pathogens worldwide, affecting a multitude of plant species across agriculture, horticulture, forestry, and natural ecosystems. Here, we concentrate on those species that are dispersed through soil and water, attacking the roots of the plants, causing them to rot and die. The intention of this study was to compare the soil baiting protocol developed by the Centre for Phytophthora Science and Management (CPSM) with two other baiting methods used in Australia. The aim was to demonstrate the effectiveness of each protocol for soil baiting Phytophthora species in different substrates. Three experiments were conducted: the first to test the sensitivity of each method to detect Phytophthora cinnamomi, the second to test the effect of substrate type (sand or loam), and the third to test the detection of species (P. cinnamomi, P. multivora, or P. pseudocryptogea). The specificity of different plant species baits was compared within and between the methods. Substrate type influenced isolation in all methods; however, the CPSM method was superior regardless of substrate, albeit slower than one of the other methods for one substrate. Comparing bait species between the three methods, Quercus ilex was the most attractive bait for P. cinnamomi, particularly in the CPSM method. The choice of protocol affected the isolation associated with each bait type. Overall, the multiple bait system used by CPSM was shown to provide the most sensitive and reliable detection of Phytophthora species from soil samples. 相似文献
8.
ABSTRACT Traditional methods of quantifying Pythium spp. rely on the use of selective media and dilution plating. However, high variability is inherent in this type of enumeration and counts may not be representative of the pathogenic population of Pythium spp. Variable regions of the internal transcribed spacer of the rDNA were used to design species-specific primers for detection and quantification of nine Pythium spp. from soils in eastern Washington. Primer pairs were designed for Pythium abappressorium, P. attrantheridium, P. heterothallicum, P. irregulare group I, P. irregulare group IV, P. paroecandrum, P. rostratifingens, P. sylvaticum, and P. ultimum and used with real-time polymerase chain reaction. Standard curves were generated for each of the species using SYBR Green I fluorescent dye for detection of amplification. Seventy-seven isolates of Pythium were screened to confirm specificity of each primer set. DNA was extracted from soil and standard curves were generated for P. irregulare group I, P. irregulare group IV, and P. ultimum to correlate populations of each species in the soil with quantities of DNA amplified from the same soil. Examination of raw field soils revealed results similar to those observed in previous studies. This new technique for the quantification of Pythium spp. is rapid and accurate, and will be a useful tool in the future study of these pathogenic Pythium spp. 相似文献
9.
恶疫霉有性杂交后代的生物学研究 总被引：1，自引：1，他引：1
将2个带有不同抗药性标记的恶疫霉菌株配对,培养36 d后诱导其卵孢子萌发,从3760个单卵孢株中,获得50株带有双亲标记的杂交个体。对其中32株杂交个体的生长速率、有性和无性繁殖能力、卵孢子和游动孢子萌发率、致病力及对高温的耐受力等生物学性状进行测定。结果显示,测试的32株杂交个体在LBA培养基上均能较好地生长,有24株的杂交个体的生长速率介于2亲本之间;在LBA和SL培养基上,大多数杂交个体均能产生较大数量的卵孢子,有37%的杂交个体在SL培养基上产卵孢子能力显著大于双亲,仅有1个杂交个体不产生卵孢子;在人工培养条件下,大多数供试杂交个体可以产生较大数量的孢子囊,其中15株杂交个体产孢子囊能力处于双亲之间;被测的杂交个体产生的卵孢子或游动孢子均可以萌发形成有效的单孢株,有22株的杂交个体的卵孢子萌发率大于50%;被测的杂交个体接种在苹果上都有较强的致病力,有16株杂交个体的致病力显著大于双亲。表明在群体水平上恶疫霉有性杂交后代具有较强的生活能力,提示同宗配合恶疫霉不同菌株间的有性重组,对该种的群体遗传多样性可能具重要作用。 相似文献
10.
A rapid method for direct detection of Polymyxa DNA in soil 总被引：1，自引：0，他引：1
Polymyxa spp. are vectors for a number of economically important soilborne plant viruses. The development of a technique to detect virus and vectors directly in soil would be useful for epidemiological studies and assessment of disease risk prior to planting. A rapid method was developed to extract and quantify Polymyxa spp. DNA from soils. DNA was extracted from three soils infested with Polymyxa betae and three infested with P. graminis using an EDTA lysis buffer in combination with a MagneSil™ DNA extraction kit and Kingfisher™ magnetic particle processor. Primers and probes designed to correspond to sequences within the internal transcribed spacer region 2 (ITS2) of ribosomal DNA enabled recovery and amplification of P. betae and P. graminis DNA using real-time PCR and TaqMan chemistry. For the P. graminis- infested soils, the purity of DNA obtained was sufficient to allow Polymyxa DNA to be amplified without dilution to remove inhibitors, but with P. betae- infested soils, amplification was only achieved if the DNA was diluted 1:10. Using TaqMan PCR, a standard curve was constructed from uninfested soil spiked with known numbers of P. betae cystosori; the quantity of P. betae inoculum from naturally infested soil was then extrapolated from the curve. This technique offers a sensitive method of extracting, detecting and quantifying Polymyxa spp. DNA in soil. 相似文献
11.
Validated protocols for DNA purification and PCR amplification are reported for detection of Phytophthora cactorum in diseased strawberry plants. To remove PCR inhibitors, necrotic strawberry tissues were soaked in 5% alconox solution for >12 h before DNA extraction, and the extracted genomic DNA was embedded in an agarose gel chamber and subjected to electrophoresis. The purified DNA was amplified reliably by PCR. Nested PCR was used to detect a portion of the rRNA gene of P. cactorum in samples. In the first round of PCR, primers ITS1 and ITS4 amplified fragments of varying sizes from total genomic DNA from diseased strawberry plants. In the second round of PCR, a 1:25 dilution of the first-round PCR products was used as template with two P. cactorum- specific primer pairs (BPhycacL87FRG and BPhycacR87RRG, which amplified a 340-bp fragment and a 480-bp fragment from the rRNA gene; and BPhycacL89FRG and BPhycacR176RRG, which amplified a 431-bp fragment). Validation tests using culture-based isolations as a standard for comparison indicated that the DNA purification and PCR primers and amplification protocols were reliable and specifically amplified a portion of the rRNA gene of P. cactorum from necrotic root, crown and petiole tissues of strawberry naturally infected by the pathogen. 相似文献
12.
T.C. Paulitz 《European journal of plant pathology / European Foundation for Plant Pathology》2000,106(5):401-413
Understanding the dynamics between a pathogen and a biocontrol agent (BCA) in soil or in an infection court such as the rhizosphere is crucial for predicting the success of biological control. This is especially true for biological control using the strategy of reduction of initial inoculum prior to infection. By studying the population density fluxes over time, one can observe how the BCA and pathogen influence each other's population and life cycles, and how the biological and physical environment influence this relationship. Methods for quantifying fungi and bacteria in soil, including classical (dilution plating, baiting, bioassays), immunological (ELISA, IFC, immunomagnetic) and molecular (DNA hybridization, PCR, marker and reporter genes) methods, are discussed and critiqued. Finally, the value of mathematical modeling of population dynamics as a means of providing important information about the tripartite relationship between the pathogen, BCA, and host plant is reviewed. 相似文献
13.
Crown rot of strawberry, caused by Phytophthora cactorum , was detected for the first time in Norway in 1992. This paper reports on surveys for P. cactorum in Norwegian certified strawberry plant production and on the distribution of the pathogen in regular strawberry production. In 1996 and 1997, samples of plant material from all certified strawberry plant growers in the country were investigated by isolation on artificial growth medium and using an enzyme-linked immunosorbent assay (ELISA). P. cactorum was not detected in any of the samples. A total of 171 isolations from plants with symptoms resembling crown rot were made from plants in a survey of the distribution of Phytophthora fragariae var. fragariae and from other samples. P. cactorum was detected at 35 different strawberry-producing farms in 11 of the 19 counties of Norway. The fungus was most frequently isolated from cv. Korona (at 18 locations), followed by cv. Inga (at 10 locations). 相似文献
14.
ABSTRACT Studies were conducted to assess the impact of short-term rotations of wheat on microbial community composition and growth of apple in soils from replant orchard sites. Soils from two orchards were cultivated with three successive 28-day growth cycles of 'Eltan', 'Penewawa', or 'Rely' wheat in the greenhouse and subsequently planted to 'Gala' apple seedlings. Cultivation of orchard replant soils with any of the three wheat cultivars enhanced growth of apple relative to that achieved in untreated soils. Improved growth was associated with a marked reduction in apple root infection by species of Rhizoctonia and Pythium. Populations of plant-parasitic nematodes were below damage threshold levels in these orchard soils; however, apple seedlings grown in wheat-cultivated soils had significantly lower root populations of Pratylenchus spp. than did seedlings grown in untreated soils. Growth of apple in 'Penewawa'-cultivated soils often was superior to that observed in soils planted with 'Eltan' or 'Rely'. In untreated orchard soils, fluorescent pseudomonad populations isolated from soil and the apple rhizosphere were dominated by Pseudomonas fluorescens biotype C and Pseudomonas syringae. Cultivation of replant soils with wheat induced a characteristic transformation of the fluorescent pseudomonad population, and Pseudomonas putida dominated the population of this bacterial group recovered from wheat-cultivated replant orchard soils. Results from this study suggest that use of short-term wheat cropping sequences during orchard renovation could be useful in management of replant disease and that this disease-control option may operate, in part, through modification of the fluorescent pseudomonad community. 相似文献
15.
Characterizing the mechanism of biological control of postharvest diseases on fruits with a simple method to study competition for nutrients 总被引：1，自引：0，他引：1
ABSTRACT Biocontrol agents may compete with pathogens for nutrients and space to delay or prevent decay of fruits after harvest. These mechanisms of biological control have been difficult to study because no method has been available to determine the significance of each of the components of competition. We developed a nondestructive method using tissue culture plates with cylinder inserts containing defusing membrane at one end to study competition for nutrients without competition for space. Other biocontrol mechanisms in which direct contact between an antagonist and a pathogen is not required also can be studied. The method was used to determine the competition between the yeastlike biocontrol agent, Aureobasidium pullulans, and Penicillium expansum for limited nutrients in apple juice during 24 h incubation, simulating a fruit wound. The antagonist depleted amino acids and inhibited germination of P. expansum conidia. Exposing these conidia to fresh apple juice increased conidial germination to the level comparable to that exhibited by conidia which were not exposed to the antagonist. Because the culture plate method was nondestructive, follow-up experiments in an agar diffusion test were conducted. Juice in which the antagonist grew did not inhibit germination of P. expansum conidia that were seeded on the plates. This corroborates findings from the culture plate method that inhibition of the conidia germination resulted from competition for nutrients. The new method can be coupled with existing techniques to improve understanding of antagonist-pathogen interaction for biological control of postharvest diseases. 相似文献
16.
ABSTRACT Hybrid isolates of Phytophthora nicotianae x P. cactorum from five different hosts (Cyclamen, Lavandula, Lewisia, Primula, and Spathiphyllum spp.) were identified by their atypical morphology and their well-defined heterozygous isozyme patterns. The hybrid nature of these isolates was tested by restriction fragment length polymorphism analysis of the internal transcribed spacer (ITS) region of rDNA, generating fragments typical for both P. nicotianae and P. cactorum. In hybrid isolates, polymerase chain reactions (PCR) with primers derived from unique parts of the ITS region (ITS-PCR) of both species yielded a combination of unique amplicons typical of both parental species. Eleven hybrid isolates, three isolates of each parental species and two atypical isolates from Rhododendron and Idesia spp. close to P. cactorum, were analyzed for amplified fragment length polymorphisms (AFLP). Consistent differences in AFLP patterns existed among the hybrid isolates, strongly indicating that these hybrids have arisen from independent hybridization events between P. nicotianae and P. cactorum. The two atypical isolates morphologically resembling P. cactorum were identical to the latter species in ITS-restriction fragment length polymorphism and response to the specific PCR primers but were intermediate between P. nicotianae x P. cactorum and P. cactorum in isozyme profiles and AFLP patterns. Since the introduction of hydroponic systems in greenhouses in the Netherlands, outbreaks of Phytophthora diseases are occurring in previously unaffected host species. This may be due to interspecific hybridization events resulting in novel pathogenic behavior. 相似文献
17.
Journal of Plant Diseases and Protection - Phytophthora cinnamomi and P. cactorum were detected in soil samples from a Vaccinium corymbosum stand in southern Germany. Both species were virulent in... 相似文献
18.
Shashika Shivanthi Hewavitharana David Ruddell Mark Mazzola 《European journal of plant pathology / European Foundation for Plant Pathology》2014,140(1):39-52
Anaerobic soil disinfestation (ASD) has been shown to be effective in the control of a wide range of soil–borne plant pathogens but has not been examined as a means for disease control in perennial fruit crops such as apple. Since ASD has demonstrated a broad spectrum of biological activity, it may be well suited as an alternative to current fumigation–based control of apple replant disease (ARD) which is caused by a diverse pathogen complex. The efficacy of ASD for control of ARD pathogens was evaluated in growth chamber experiments using soils from two orchard sites having a history of the disease. Suppression of Pratylenchus penetrans apple root densities was dependent upon carbon source utilized during the ASD process. Volatiles emitted during the anaerobic phase from soils treated with ethanol, grass residues, or Brassica juncea seed meal as the carbon input effectively retarded growth of Rhizoctonia solani AG–5, Pythium ultimum and Fusarium oxysporum. Each carbon amendment generated a unique volatile profile produced in the treated orchard soil during ASD. Allyl isothiocyanate (AITC) and dimethyl trisulphide (DMTS) were emitted from B. juncea SM treated soils whereas the latter and 2–ethyl–1–hexanol were detected in soils treated with grass residues. When assayed individually using pure standards, Decanal, DMTS, and AITC retarded in vitro growth of all three fungal/oomycete pathogens. Nonanal was inhibitory toward only P. ultimum and R. solani AG–5, whereas 2–ethyl–1–hexanol only suppressed growth of P. ultimum. AITC and DMTS caused significantly higher mortality of P. penetrans compared to other tested volatiles. These findings demonstrate that carbon source–dependent volatile chemistries contribute significantly but not exclusively to suppression of certain ARD pathogens during the ASD process. 相似文献
19.
Yared Tesfai Tewoldemedhin Mark Mazzola Wilhelm J. Botha Christoffel F. J. Spies Adéle McLeod 《European journal of plant pathology / European Foundation for Plant Pathology》2011,130(2):215-229
Several species of fungi and oomycetes including Fusarium, Rhizoctonia, Phytophthora and Pythium have been reported as root pathogens of apple where they contribute to a phenomenon known as apple replant disease. In South
Africa, little is known about specific species in these genera and their pathogenicity toward apple. Therefore, these aspects
were investigated along with the development and optimization of qPCR tests for detection and quantification of the most virulent
oomycete species. In eight investigated orchards, the oomycete Phythophthora cactorum was widely distributed, while nine Pythium species were differentially distributed among the orchards. Pythium irregulare was the most widely distributed and the most virulent species along with P. sylvaticum, P. vexans and Ph. cactorum. Seven binucleate Rhizoctonia anastomosis groups (AGs) were also differentially distributed among the orchards, with the majority appearing to be non-pathogenic
while certain AG-I and AG-F isolates exhibited low virulence on apple. In the genus Fusarium, F. oxysporum was widely distributed, but isolates were non-pathogenic. Fusarium solani and F. avenaceum were less frequently encountered, with only some isolates having low virulence. qPCR data obtained from seedling roots inoculated
with the most virulent Pythium species (P. irregulare, P. sylvaticum and P. vexans) and the genus Phytophthora were not always reproducible between trials, or isolates of the same species. In general, seedling growth inhibition was
associated with the presence of a low amount of pathogen DNA (±40 fg μl−1 to 2 pg μl−1) in roots. Pythium irregulare, although having the lowest DNA concentrations in roots, was the only species for which a significant negative correlation
was found between seedling weight and pathogen DNA concentration. 相似文献
20.
Rajeshwar S. Utkhede 《Pest management science》1987,19(4):289-295
Four fungicides were evaluated as combined soil drench and trunk treatments for the control of crown rot, caused by Phytophthora cactorum, in naturally infected apple trees. In a 3-year preliminary study, metalaxyl at 1 and fosetyl-aluminium at 8 g a. i. per tree, applied twice a year, prevented death of infected trees showing only initial, but not more severe, symptoms of crown rot. This indicates that these fungicides have the ability to arrest further symptom development in naturally infected trees if they are treated as soon as symptoms appear. Healthy trees remained free of infection if they were treated with these fungicides, suggesting a preventative action. Repeated bi-annual application of metalaxyl or fosetyl-aluminium should be sufficient to prevent the loss of trees to crown rot. 相似文献