Development of PCR for the detection of Polymyxa betae in sugar beet roots and its application in field studies

Mutasa, E.S.; Chwarszczynska, D. M.; Adams, Michael J.; Ward, Elaine; Asher, M. J. C.

doi:10.1006/pmpp.1995.1060

Cited by 34 publications

(15 citation statements)

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“…This method is laborious and involves use of such hazardous chemicals as phenol. More recently, to facilitate rapid detection of Polymyxa spp., molecular techniques have been developed (35,36,47). These techniques, even though very specific, are expensive and require special skills and equipment not readily available in developing countries.…”

Section: Discussionmentioning

confidence: 99%

Serological Methods for Detection of Polymyxa graminis, an Obligate Root Parasite and Vector of Plant Viruses

Delfosse¹,

Reddy²,

Legrève³

et al. 2000

Phytopathology®

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Delfosse, P., Reddy, A. S., Legrève, A., Thirumala Devi, K., Abdurahman, M. D., Maraite, H., and Reddy, D. V. R. 2000. Serological methods for detection of Polymyxa graminis, an obligate root parasite and vector of plant viruses. Phytopathology 90:537-545.A purification procedure was developed to separate Polymyxa graminis resting spores from sorghum root materials. The spores were used as immunogen to produce a polyclonal antiserum. In a direct antigen coating enzyme-linked immunosorbent assay (DAC ELISA), the antiserum could detect one sporosorus per well of the ELISA plate. In spiked root samples, the procedure detected one sporosorus per mg of dried sorghum roots. The majority of isolates of P. graminis from Europe, North America, and India reacted strongly with the antiserum. Interestingly, P. graminis isolates from the state of Rajasthan (northern India), from Pakistan, and an isolate from Senegal (West Africa) reacted weakly with the antiserum. The cross-reactivity of the serum with P. betae isolates from Belgium and Turkey was about 40% of that observed for the homologous isolate. There was no reaction with common fungi infecting roots or with the obligate parasite Olpidium brassicae. However, two isolates of Spongospora subterranea gave an absorbance similar to that observed with the homologous antigen. The DAC ELISA procedure was successfully used to detect various stages in the life cycle of P. graminis and to detect infection that occurred under natural and controlled environments. A simple procedure to conjugate antibodies to fluorescein 5-isothiocyanate (FITC) is described. Resting spores could be detected in root sections by using FITC-labeled antibodies. The potential for application of the two serological techniques for studying the epidemiology of peanut clump disease and for the characterization of Polymyxa isolates from various geographical origins is discussed.Polymyxa spp. are soilborne, obligate intracellular parasites of roots, classified under the order Plasmodiophorales. Currently, two species, P. graminis Ledingham and P. betae Keskin, are recognized. They transmit at least 12 different plant viruses in the genera Benyvirus, Bymovirus, Furovirus, and Pecluvirus (1,30,32). Peanut clump virus (PCV) and Indian peanut clump virus (IPCV) are members of the genus Pecluvirus (40) that affect the production of peanut and monocotyledonous crops in West Africa and in the Indian subcontinent (3,9,40,43). In the absence of cost-effective methods for the management of peanut clump disease (40), it was essential to study its epidemiology with emphasis on identification of the hosts for the virus and P. graminis and their influence on the perpetuation and spread of clump disease. Epidemiological studies on an obligate parasite such as Polymyxa spp. are comparatively difficult and time consuming, because its detection has traditionally relied on light microscopic inspection of roots. However, results from these studies are expected to aid in the formulation of costeffective cultural practices ...

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Section: Discussionmentioning

confidence: 99%

Serological Methods for Detection of Polymyxa graminis, an Obligate Root Parasite and Vector of Plant Viruses

Delfosse¹,

Reddy²,

Legrève³

et al. 2000

Phytopathology®

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“…Either bait plant bioassays using soil dilutions are applied to estimate the most probable numbers (MPN) of infective propagules (Tuitert 1990) or polymerase chain reaction (PCR) is used to estimate P. betae concentration in plant material and soil (Mutasa et al 1995;Mutasa-Gottgens et al 2000;Kingsnorth et al 2003). In a soil sample from The Netherlands, Tuitert (1990) estimated a proportion of 10-15% of the rootinfecting P. betae population to be viruliferous.…”

Section: Introductionmentioning

confidence: 99%

Breaking of Beet necrotic yellow vein virus resistance in sugar beet is independent of virus and vector inoculum densities

Pferdmenges¹,

Varrelmann²

2008

Eur J Plant Pathol

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Beet necrotic yellow vein virus (BNYVV) is transmitted by Polymyxa betae to sugar beet, causing rhizomania disease. Resistance-breaking strains of BNYVV, overcoming single (Rz1) or double (e.g. Rz1+Rz2) major resistance genes in sugar beet have been observed in France and recently in the USA and Spain. To demonstrate if resistance-breaking is dependent on inoculum density, the inoculum concentration of BNYVV and P. betae in soil samples where resistance-breaking had been observed was estimated using the most probable number (MPN) method. The MPN-values obtained displayed highly significant differences with respect to the virus concentration in various soils and did not correlate with the ability to overcome resistance. Virus quantification in susceptible plants demonstrated that soils containing resistance-breaking isolates of BNYVV did not produce higher virus concentrations. The MPN assay was repeated with Rz1+Rz2 partiallyresistant sugar beets to see if the resistance-breaking is concentration-dependent. There was no correlation between soil dilution and increased virus concentration in Rz1+Rz2 plants produced by BNYVV resistancebreaking strains. Determination of the absolute P. betae concentration by ELISA demonstrated that all resistance-breaking soil samples contained elevated concentrations. However, the calculation of the proportion of viruliferous P. betae did not show a positive correlation with the resistance-breaking ability. Finally resistance-breaking was studied with susceptible, Rz1 and Rz1+Rz2 genotypes and standardised rhizomania inoculum added to sterilised soil. Results from these experiments supported the conclusion that resistance-breaking did not correlate with virus concentration or level of viruliferous P. betae in the soil.

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“…Evaluation methods were developed specifically for the mass screening of diverse germplasm which differs in many characteristics, eg germination rate, plant architecture, pigmentation etc; these are reported elsewhere 20. 21 For all resistance tests, it was important that inoculation was uniform across, and within, all accessions being tested. For some diseases, this was achieved by artificial inoculation; eg for BMYV, viruliferous aphids were produced on Capsella bursa‐pastoris (L) Medic plants and then Beta plants were inoculated by transferring five or more aphids to the heart leaves on a segment of Capsella leaf.…”

Section: Identification and Exploitation Of Novel Resistance Sourcesmentioning

confidence: 99%

Identification and exploitation of novel disease resistance genes in sugar beet

Francis¹,

Luterbacher²

2003

Pest Management Science

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Sugar beet is a valuable arable crop providing a large proportion of world sugar supplies. It is susceptible to a range of fungal and viral diseases, to most of which there is inadequate genetic resistance. Control instead relies largely on the use of pesticides. To increase the sustainability of sugar-beet cultivation, strong and durable genetic resistance is urgently needed. This review describes an IACR-Broom's Barn programme devoted to evaluating Beta germplasm for novel resistance to major viral and fungal diseases, and its progress in elucidating the numbers and locations of the genes responsible.

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Development of PCR for the detection of Polymyxa betae in sugar beet roots and its application in field studies

Cited by 34 publications

References 0 publications

Serological Methods for Detection of Polymyxa graminis, an Obligate Root Parasite and Vector of Plant Viruses

Serological Methods for Detection of Polymyxa graminis, an Obligate Root Parasite and Vector of Plant Viruses

Breaking of Beet necrotic yellow vein virus resistance in sugar beet is independent of virus and vector inoculum densities

Identification and exploitation of novel disease resistance genes in sugar beet

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