A novel method for detection of viable zoospores of <i>Pythium</i> in irrigation water

Wakeham, Alison; Pettitt, Timothy; White, J. G.

doi:10.1111/j.1744-7348.1997.tb05170.x

Cited by 21 publications

(20 citation statements)

References 18 publications

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“…in soil from citrus orchards, pepper and cucurbit crops, and in irrigation water using ELISA-kits (ALI-SHTAYEH et al 1991;TIMMER et al 1993;MILLER et al 1994;WAKEHAM et al 1997). These can be collected by centrifugation, ®ltration or via the infection of bait tissues.…”

Section: Discussionmentioning

confidence: 99%

A combination of baiting and PCR techniques for the detection of Phytophthora quercina and P. citricola in soil samples from oak stands

et al. 2001

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A description is given of the use of a combination of polymerase chain reaction (PCR) and baiting techniques for the speci®c detection of Phytophthora quercina and Phytophthora citricola from soil around declining oak trees. The soil was¯ooded with water and subjected to a speci®c baiting procedure using Quercus robur lea¯ets as baits. Single round or nested PCR, respectively, with species-speci®c primers allowed the detection of P. quercina and P. citricola in infected oak lea¯ets used as baits and in the water from the same bait tests. PCR detection of both fungi was also possible after soil samples had been thoroughly mixed with water and the¯oating organic debris had been collected. Phytophthora quercina and P. citricola could be readily detected in almost every case in the water from these tests by PCR but less frequently in the organic debris. The identities of P. quercina and P. citricola were con®rmed by restriction digests of the corresponding PCR amplicons. The presence of both fungi was also con®rmed in parallel in soil samples tested by baiting with oak lea¯ets. Nested PCR with the primers used allowed the detection of as few as ®ve zoospores of P. citricola and 300 zoospores of P. quercina in a volume of 100 ll. The methods presented here allow detection and identi®cation of species of Phytophthora in soil without the need for direct extraction of soil samples, and without speci®c knowledge of the morphological characteristics of the genus.

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Section: Discussionmentioning

confidence: 99%

A combination of baiting and PCR techniques for the detection of Phytophthora quercina and P. citricola in soil samples from oak stands

et al. 2001

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“…For Phytophthora spp., the modified BNPRA mixture [pimaricin 10 mg L −1 , rifamycin 10 mg L −1 , ampicillin (Sigma) 500 mg L −1 , nystatin (Sigma) 25 mg L −1 , pentachloronitrobenzene 25 mg L −1 , benomyl 10 mg L −1 ] was used at the same concentration as used for agar plates (Pettitt & Pegg, 1991). Following overnight incubation, membranes were air‐dried and processed by immunoassay, as described by Wakeham et al . (1997), using a polyclonal antiserum coded 95/10/2.…”

Section: Methodsmentioning

confidence: 99%

“…This antiserum had previously been raised to a soluble mycelial fraction collected from five Pythium spp. and characterized for reactivity with a range of other fungal spore types (Wakeham et al ., 1997).…”

Section: Methodsmentioning

confidence: 99%

“…These techniques take several days, require taxonomic expertise, and are often too slow to assist growers in disease management decisions. Technological advances in antibody‐based assays have enabled rapid detection of Pythium and Phytophthora species in plant tissues (Rittenburg et al ., 1988; Harrison et al ., 1990; Lyons & White, 1992; Beckman et al ., 1994); in soil (Miller et al ., 1992; White et al ., 1996; Miller et al ., 1997); and in water samples (Ali‐Shtayeh et al ., 1991; Cahill & Hardham, 1994; Themann & Werres, 1996; Wakeham et al ., 1997). Test formats vary, and employ either enzyme‐linked immunosorbent assay (ELISA), membrane trapping assay, or a dipstick format.…”

Section: Introductionmentioning

confidence: 99%

“…However, most of these tests are limited by their inability to differentiate between live and dead propagules of target pathogens. Tests developed by Cahill & Hardham (1994) and Wakeham et al . (1997) do detect viable propagules and have commercial potential for on‐site use.…”

Section: Introductionmentioning

confidence: 99%

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Comparison of serological, culture, and bait methods for detection of Pythium and Phytophthora zoospores in water

Pettitt¹,

Wakeham²,

Wainwright³

et al. 2002

Plant Pathology

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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.

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Detection of Fungal Pathogens in the Environment

Narayanasamy¹

2010

Microbial Plant Pathogens-Detection and Disease Diagnosis:

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The presence of fungal pathogens in the environment has to be detected and also quantified rapidly and precisely, because the inoculum for infection of crop plants comes from the pathogen propagules present in the soil, water, air and alternative host plant species. Soilborne pathogens may have different degrees of saprophytic ability, utilizing the organic matter present in the soil for their survival in the absence of crop plants. They produce different structures such as chlamydospores and sexual spores that are long-lived and are capable of surviving in the soil for several years. Fungal structures may be carried by irrigation water or rain water from one part of the field to other parts or to different fields. It has been possible to detect and identify the fungal pathogens in the irrigation water, recycled water used for growing hydroponic plants and also in wash water in the storage facilities for fruits and vegetables. The pathogens infecting aerial plant parts/organs are generally disseminated by wind to different locations. Traditionally spore traps have been used for assessing the spore load of air. Various biological, immunological and nucleic acid-based techniques have been employed for the detection, identification and quantification of pathogen propagules in the environment. Significant improvements have been made in the sensitivity and specificity of detection of fungal pathogens by applying immunoassys and nucleic acid-based methods that are capable of providing reproducible results rapidly and reliably. The relative usefulness and limitations of the detection techniques applied for the detection of fungal pathogens in the environment are discussed.Studies on the ecology of plant hosts provide information on the influence of environment on the growth and development of plants. Likewise, the influence of the environment on the microbial plant pathogens is studied to understand the extent of population build up resulting in the incidence of diseases in different agroecological conditions. Crop husbandry techniques aim to increase the crop yield to the maximum levels. But some of these techniques like monoculture and excessive application of nutrients may provide favorable conditions for pathogen development. Epidemiology deals with effects of biotic and abiotic environments on disease development in plant populations. Microbial pathogens multiply at different rates in a given set of environmental conditions and exhibit wide variations in their pathogenic potential (aggressiveness). The crop plants also have varying growth

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A novel method for detection of viable zoospores of Pythium in irrigation water

Cited by 21 publications

References 18 publications

A combination of baiting and PCR techniques for the detection of Phytophthora quercina and P. citricola in soil samples from oak stands

A combination of baiting and PCR techniques for the detection of Phytophthora quercina and P. citricola in soil samples from oak stands

Comparison of serological, culture, and bait methods for detection of Pythium and Phytophthora zoospores in water

Detection of Fungal Pathogens in the Environment

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