Abstract
Potato cyst nematode (PCN) is a damaging soilborne pest of potatoes which can cause major crop losses. In 2010, a new European Union directive (2007/33/EC) on the control of PCN came into force. Under the new directive, seed potatoes can only be planted on land which has been found to be free from PCN infestation following an official soil test. A major consequence of the new directive was the introduction of a new harmonized soil sampling rate resulting in a threefold increase in the number of samples requiring testing. To manage this increase with the same staffing resources, we have replaced the traditional diagnostic methods. A system has been developed for the processing of soil samples, extraction of DNA from float material, and detection of PCN by high-throughput real-time PCR. Approximately 17,000 samples are analyzed each year using this method. This chapter describes the high-throughput processes for the production of float material from soil samples, DNA extraction from the entire float, and subsequent detection and identification of PCN within these samples.
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References
Evans K, Trudgill DL (2008) A history of potato cyst nematode research. In: Webster JM, Eriksson KB, McNamara DG (eds) An anecdotal history of nematology. Pensoft Publishers, Sophia, Bulgaria, pp 107–128
Minnis ST, Haydock PPJ, Ibrahim SK et al (2000) The occurrence and distribution of potato cyst nematodes in England and Wales. Aspect Appl Biol 59:1–9
Haydock PPJ, Evans K (1998) Management of potato cyst nematodes in the UK: an integrated approach? Outlook Agr 27:253–260
Evans K, Haydock PPJ (2000) Potato cyst nematode management: present and future. Aspect Appl Biol 59:91–97
COUNCIL DIRECTIVE 2007/33/EC of 11 June 2007 on the control of potato cyst nematodes and repealing Directive 69/465/EEC. http://eur-lex.europa.eu/legal-content/en/ALL/?uri = CELEX:32007L0033
Pickup J (2008) New revised EU PCN Directive: implications for potato growers in Northern Britain. In: Proceedings of the Crop Protection in Northern Britain Conference 2008, p 199–204
Fenwick DW (1940) Methods for the recovery and counting of cysts of Heterodera schachtii from soil. J Helminthol 21:37–41
Golden AM (1986) Morphology and identification of cyst nematodes. In: Lamberti F, Taylor CE (eds) Cyst nematodes, p 23–45.
Reid A, Kenyon DM, Evans FF et al (2010) Development of a high-throughput method for the detection and species determination of PCN. Aspect Appl Biol 103:13–16
Bulman SR, Marshall JW (1997) Differentiation of Australasian potato cyst nematode (PCN) populations using the polymerase chain reaction (PCR). New Zeal J Crop Hort Sci 25:123–129
Fullaondo A, Barrena E, Viribay M et al (1999) Identification of potato cyst nematode species Globodera rostochiensis and G. pallida by PCR using specific primer combinations. Nematology 1:157–163
Mulholland V, Carde L, O’Donnell KJ et al (1996) Use of the polymerase chain reaction to discriminate potato cyst nematode at the species level. Diagnostics in Crop Production, BCPC Symposium Proceedings, 65: 247–252
Pylypenko LA, Uehara T, Phillips MS et al (2005) Identification of Globodera rostochiensis and G. pallida in the Ukraine by PCR. Eur J Plant Pathol 111:39–46
Vejl P, Skupinova S, Sedlak P et al (2002) Identification of PCN species (Globodera rostochiensis, G. pallida) by using of ITS-1 region’s polymorphism. Rostlinná Výroba 48:486–489
Bates JA, Taylor EJA, Gans PT et al (2002) Determination of relative proportions of Globodera species in mixed populations of potato cyst nematodes using PCR product melting peak analysis. Mol Plant Pathol 3:153–161
Madani M, Ward LJ, De Boer SH (2008) Multiplex real-time polymerase chain reaction for identifying potato cyst nematodes, Globodera pallida and Globodera rostochiensis, and the tobacco cyst nematode, Globodera tabacum. Can J Plant Pathol 30:554–564
Quader M, Nambiar L, Cunnington J (2008) Conventional and real-time PCR-based species identification and diversity of potato cyst nematodes (Globodera spp.) from Victoria, Australia. Nematology 10:471–478
Pickup J, Cole Y, Reid A et al (2012) Potato cyst nematodes: initial impressions of the impact of the new EU directive in Scotland. In: Proceedings of the Crop Protection in Northern Britain Conference 2012, p 227–232
Ladeveze LL, Anthoine G (2010) Outcomes of a seven-year proficiency test for the detection and identification of potato cyst nematodes. Aspect Appl Biol 103:1–9
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Reid, A., Evans, F., Mulholland, V., Cole, Y., Pickup, J. (2015). High-Throughput Diagnosis of Potato Cyst Nematodes in Soil Samples. In: Lacomme, C. (eds) Plant Pathology. Methods in Molecular Biology, vol 1302. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2620-6_11
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DOI: https://doi.org/10.1007/978-1-4939-2620-6_11
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2619-0
Online ISBN: 978-1-4939-2620-6
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