Detection of Polymyxa betae in sugar beet roots using microscopical, serological and molecular methods

Document Type : Research Paper

Authors

Abstract

In order to design a precise diagnostic and quantitative system to detect of P. betae in sugar beet roots, three procedures including microscopical, serological and molecular methods were compared on a sugar beet cultivar susceptible to causal agent and vector of rhizomania disease. At first, susceptible cultivar Regina was cultivated in infested soil in greenhouse condition. After 5 weeks, roots of plants were stained with fuchsin- acid- lactophenol solution. Quantitative ELISA test was optimized with specific antiserum against recombinant protein glutathione s- transferase (GST) of P. betae. Running ELISA test, extracts of health and infected plants were prepared and used in DAS- ELISA test. Presence of P. betae in roots of plants was detected with amplification of rDNA of Plasmodiophoromycetes and P. betae species, simultaneously. Existence of cystosori of P. betae was observed in all of the plants planting in infested soil. Results of ELISA test were distinguished healthy plants from infected plants. Average of optical density at 405 nanometer for healthy plants, infected plants, protein PBS (negative control) and GST (positive control) was 0.126, 0.75, 0.11 and 2.45 respectively. Duplex PCR method was amplified two fragments of 454bp and 170bp in infected samples relating to rDNA region and specific region of the P. betae, respectively. Based on the results, it seems that, rapid detection and identification of P. betae in sugar beet and other hosts of the vector could be done on the basis of PCR method. Evaluating of resistance germplasm of sugar beet to P. betae could be recognized using quantitative ELISA tests.

Keywords

Full Text

 Rhizomania, Glutathione- S- transferase, ELISA, PCR, ITS

References

Asher MJC, Grimmert MK, Mutasa- Goettgens ES (2008). Selection and characterization of resistance to Polymyxa betae, vector of Beet Necrotic Yellow Vein Virus, derived from wild sea beet. Plant pathology 58: 250-260.
Barr KJ, Asher MJC, Lewis BG (1995). Resistance to Polymyxa betae in wild Beta species. Plant Pathology 44: 301-307.
Braselton, J P (1995). Current status of the plasmodiophorids. Current Reviews in Microbiology 21: 263-275.
Darabi S, Masoomi M, Izadpanah K (2003). Rhizomania of sugar beet. Publication of Plant Virology Research Center of Shiraz Agriculture University (In Farsi).
Delfosse P, Reddy AS, Legrève A, Thirumala Devi K, Abdurahman M D, Maraite H, Reddy DVR (2000). Serological Methods for Detection of Polymyxa graminis, an Obligate Root Parasite and Vector of Plant Viruses. The American Phytopathological Society 90: 537- 545.
Dick MW (2001). Straminipilous Fungi: Systemics of the Pyrenomycetes including accounts of the marine Straminipilous protists, the plasmodiophorids  and similar organisms. Dordrecht: Kluwer Academic Publishers.
Draycott, AP (2006). Sugar Beet (World Agriculture Series). Wiley-Blackwell. London. 496pp.
Henry C (1996). Rhizomania- its effect on sugar beet yield in the UK. Breeding sugar beet review 64: 24-26.
Izadpanah K, Hashemi P, Kamran R, Pakniat M, Sahandpour A, Masoomi M (1996). Wide existence of rhizomania sugar beet in Fars. Plant Pathology 32: 200-206 (In Farsi).
Jalali S, Afzali H, Ravanloo A, Karimipourfard H, Baykzadeh N, Almasi H (2009). Distribution of fungus Polymyxa betae vector of root madness virus of sugar beet in Esfahan,Azarbayjan Gharbi and Razavi Khorasan provinces, Journal of Research in Agriculture Science 5(1): 69- 79 (In Farsi).
Kingsnorth CS, Kingsnorth AJ, Lyons PA, Chwarszczynska DM, Asher MJC (2003a). Real-time analysis of Polymyxa betae GST expression in infected sugar beet. Molecular Plant Pathology 4:171-176.
Kingsnorth CS, Asher MJC, Keane GJP, Chwarszczynska DM, Luterbacher MC, Mutasa-Göttgens ES (2003b). Development of a recombinant antibody ELISA test for the detection of Polymyxa betae and its use in resistance screening. Plant Pathology 52:673–680.
Lamey HA (1992) .The rhizomania disease of sugar beet, Sugar Beet Research and Extension Reports USDA. 23: 149-152.
Legreve A, Delfosse P, Van HV, Bragard C, Maraite H (2003). Broad- spectrum detection of Polymyxa species and form species by polymerase chain reaction. In Proceedings of the 5th symposium of the internal working group on plant viruses with fungal vectors ; Eds. C. M. Rush & U. Merz; Zurich, Switzerland.
Lennefors BL (2006) Molecular breeding for resistance to rhizomonia in sugar beets. Ph. D. thesis. Swedish University of Agricultural Sciences. Uppsala. 44pp.
Liu HY, Lewellen RT (2007). Distribution and molecular characterization of resistance-breaking isolates of Beet necrotic yellow vein virus in the United States. Plant Disease 91: 847–851
McGrann GRD, Grimmer MK, Mutasa- Gottgens EF, Steven M (2009). Progress towards the understanding and control of sugar beet rhizomania disease. Molecular Plant Pathology 10: 129- 141.
Mesbah M, Scolten OE, DeBock TSM, Lange W (1997). Chromosome localization of genes for resistance to Heterodera schachtii, Cercospora beticola, and Polymyxa betae using sets of Beta procumbens and B. pattelaris derived monosomic additions in B. vulgaris. Euphytica, 97: 117- 127.
Meunier A, Schmit J F, Stas A, Kutluk N, Bragard C (2003). Multiple reverse transcription –PCR for simultaneous detection of beet necrotic yellow vein virus, Beet soil born virus , and Beet virus Q and their vector Polymyxa betae on sugar beet. Applied and Environmental Microbiology 69: 2356- 2360.
Mouhanna AM, Langen G, Schlosser E (2008). Weeds as alternative hosts for BSBV, BNYVV, and the vector Polymyxa betae (German Isolate). Journal of Plant Disease and Protection 115: 193- 198
Mutasa-Göttgens EF, Chwarszczynska D, Adams M, Ward E, Asher MJC (1995). Development of PCR for the detection of Polymyxa betae in sugar beet roots and its application in field studies. Physiological and Molecular Plant Pathology 47: 303-313
Mutasa-Göttgens EF, Chwarszczynska D, Halsey K, Asher MJC (2000). Specific polyclonal antibodies for the obligate plant parasite Polymyxa – a targeted recombinant DNA approach. Plant Pathology 49: 276-287.
Paul H, Henken B, De Bock TSM, Lange W (1992). Resistance to Polymyxa betae in Beta species of the section procumbentes, in hybrids with Beta vulgaris and in monosomic chromosome additions of Beta procumbens in Beta vulgaris Plant Breeding 109: 265–273.
Paul H, Henken B, Scolten OE, De Bock TSM, Lange W (1993). Variation in the level of infection with Polymyxa betae and its effect on infection with Beet Necrotic Yellow Vein Virus in beet accession of the sections Beta and Corollinae. Proceedings of the 2nd Symposium of the International Working Group on Plant Viruses with Fungal Vectors. Montreal. Canada, pp. 133-136. 
Pavli OL, Stevanato P, Biancardi E, Skaracis GN (2011). Achievements and prospects in breeding for rhizomania resistance in sugar beet. Field Crops Research 122: 165- 172.
Rush CM (2003). Ecology and epidemiology of Benyvirus and plasmodiophorid vectors. Annual Review of Phytopathology 41: 567–592
Rush CM, Lewellen RT, Acosta- leal R (2006). The continuing saga of rhizomania of sugar beet in the United States. Plant Disease 90: 4- 15
Safarpour H, Safarnejad MR, Tabatabaie M, Mohsenifar A, Rad F, Shahryari F, Hasanzadeh F (2012). Development of a quantum dots FRET-Based biosensor for efficient detection of Polymyxa betae. Canadian Journal of Plant Pathology. In Press.
Verchot- Lubicz J, Rush CM, Payton M, Colberg T (2007). Beet necrotic yellow vein virus accumulates inside resting spores and zoosporangia of its vector Polymyxa betae BNYVV infects P. betae. Virology Journal. 4:37.
Ward LI, Fenn MGE, Henry CM (2004). A rapid method for detection of Polymyxa DNA in soil. Plant Pathology 53: 485-490.
 
Agricultural Biotechnology Journal
Volume 4, Issue 2 - Serial Number 2
Vol 4 No 2 autumn and winter 1391
December 2012
Pages 117-128

Files

Share

How to cite

Statistics