The potato cyst nematode (PCN), Globodera pallida, originates from South America and is considered one of the most severe agricultural pests of potato crops and other Solanaceae plants globally. Based on their virulence and ability to reproduce on various potato cultivars, the populations of G. pallida are divided into three pathotypes, Pa1– Pa3. In this study, comparative sequence analyses of the fragment of mitochondrial cytochrome c oxidase subunit II (mtCOII) gene for eight populations of G. pallida, representing three pathotypes, Pa1, Pa2 and Pa3, indicated genetic diversity between them. However, we did not identify significant mutations distinguishing Pa2 from Pa3. Interestingly, two single nucleotide substitutions, T441C and A468G, were characteristic only for populations assigned to Pa1. On this basis, we developed high resolution melting (HRM) PCR protocol. As a result, the melting curves obtained for samples of Pa1 populations varied from those obtained for populations designed as Pa2 and Pa3, allowing their differentiation. Thus, the HRM protocol developed here enables a rapid, very sensitive and low-cost screening assay for SNPs identification in mtCOII of G. pallida pathotypes. In effect, it might also be a helpful molecular tool in pathotype differentiation. However, further verification of the correlation of the occurrence of single nucleotide mutations in mtCOII in particular pathotypes should be carried out on a much larger number of samples of G. pallida, to determine if these mutations are characteristic only for this pathotype.
I wish to thank Dr. Renata Dobosz from the Department of Entomology and Animal Pests IPP–NRI (Poznan, Poland) for kindly providing cysts of G. pallida populations of three pathotypes. I also wish to thank Dr A. Obrępalska-Stęplowska from the Department of Molecular Biology and Biotechnology IPP-NRI (Poznań, Poland) for her valuable suggestions during preparation of the manuscript.
Natasza Borodynko-Filas
The authors have declared that no conflict of interests exist.
Bakker J., Bouwman-Smits L., Gommers F.J. 1992. Genetic relationships between Globodera pallida pathotypes in Europe assessed by using two dimensional gel electrophoresis of proteins. Fundamental and Applied Nematology 15: 481–490.
Bates J.A., Taylor E.J.A., Gans P.T., Thomas J.E. 2002. Determination of relative proportions of Globodera species in mixed populations of potato cyst nematodes using PCR product melting peak analysis. Molecular Plant Pathology 3 (3): 153–161. DOI: https://doi.org/10.1046/j.1364....
Bulman S.R., Marshall J.W. 1997. Differentiation of Australasian potato cyst nematode (PCN) populations using the polymerase chain reaction (PCR). New Zealand Journal of Crop and Horticultural Science 25 (2): 123–129.
Burrows P.R., Boffey S.A. 1986. A technique for the extraction and restriction endonuclease digestion of total DNA from Globodera rostochiensis and Globodera pallida second stage juveniles. Revue de Nématologie 9 (2): 199–200.
EPPO. 2017. 7/40 (4) Globodera rostochiensis and Globodera pallida. EPPO Bulletin 47: 174–197.
Folkertsma R.T., der Voort J., van Gent-Pelzer M.P.E., De Groot K.E., van Den Bos W.J., Schots A., Bakker J., Gommers F.J. 1994. Inter-and intraspecific variation between populations of Globodera rostochiensis and G. pallida revealed by random amplified polymorphic DNA. Phytopathology 84 (8): 807–811.
Fox P.C., Atkinson H.J. 1984. Isoelectric focusing of general protein and specific enzymes from pathotypes of Globodera rostochiensis and G. pallida. Parasitology 88 (1): 131–139. DOI: https://doi.org/10.1017/S00311....
Hinch J.M., Alberdi F., Smith S.C., Woodward J.R., Evans K. et al. 1998. Discrimination of European and Australian Globodera rostochiensis and G. pallida pathotypes by high performance capillary electrophoresis. Fundamental and Applied Nematology 21 (2): 123–128.
Hoolahan A.H., Blok V.C., Gibson T., Dowton M. 2012. A comparison of three molecular markers for the identification of populations of Globodera pallida. Journal of Nematology 44 (1): 7.
Kort J., Ross H., Rumpenhorst H.J., Stone A.R. 1977. An international scheme for identifying and classifying pathotypes of potato cyst-nematodes Globodera rostochiensis and G. pallida. Nematologica 23 (3): 333–339.
Madani M., Subbotin S.A., Moens M. 2005. Quantitative detection of the potato cyst nematode, Globodera pallida, and the beet cyst nematode, Heterodera schachtii, using real-time PCR with SYBR green I dye. Molecular and Cellular Probes 19 (2): 81–86.
Nakhla M.K., Owens K.J., Li W., Wei G., Skantar A.M., Levy L. 2010. Multiplex real-time PCR assays for the identification of the potato cyst and tobacco cyst nematodes. Plant Disease 94 (8): 959–965.
Nowaczyk K., Dobosz R., Budziszewska M., Kamasa J., Obrępalska-Stęplowska A. 2011. Analysis of diversity of golden potato cyst nematode (Globodera rostochiensis) populations from Poland using molecular approaches. Journal of Phytopathology 159 (11–12): 759–766.
Nowaczyk K., Dobosz R., Kornobis S., Obrepalska-Steplowska A. 2008. TaqMan REAL-Time PCR-based approach for differentiation between Globodera rostochiensis (golden nematode) and Globodera artemisiae species. Parasitology Research 103 (3): 577–581.
Phillips M.S., Trudgill D.L. 1983. Variations in the ability of Globodera pallida to produce females on potato clones bred from Solanum vernei or S. tuberosum ssp. andigena CPC 2802. Nematologica 29 (2): 217–226.
Phillips M.S., Trudgill D.L. 1998. Variation of virulence, in terms of quantitative reproduction of Globodera pallida populations, from Europe and South America, in relation to resistance from Solanum vernei and S. tuberosum ssp. andigena CPC 2802. Nematologica 44 (4): 409–423.
Saenz M.C., De Scurrah M.M. 1977. Races of the potato cyst nematode in the Andean region and a new system of classification. Nematologica 23 (3): 340–349.
Schnick D., Rumpenhorst H.J., Burgermeister W. 1990. Differentiation of closely related Globodera pallida (Stone) populations by means of DNA restriction fragment length polymorphisms (RFLPs). Journal of Phytopathology 130 (2): 127–136.
Sedlák P., Melounová M., Skupinová S., Vejl P., Domkarova J. 2004. Study of European and Czech populations of potato cyst nematodes (Globodera rostochiensis and G. pallida) by RAPD method. Plant Soil and Environment 50 (2): 70–74.
Subbotin S.A., Franco J., Knoetze R., Roubtsova T.V., Bostock R.M., Del Prado Vera I.C. 2020. DNA barcoding, phylogeny and phylogeography of the cyst nematode species from the genus Globodera (Tylenchida: Heteroderidae). Nematology 22 (3): 269–297.
Thiéry M., Fouville D., Mugniéry D. 1997. Intra-and interspecific variability in Globodera, parasites of Solanaceous plants, revealed by Random Amplified Polymorphic DNA (RAPD) and correlation with biological features. Fundamental and Applied Nematology 20 (5): 495–504.
Vejl P., Skupinová S., Sedlák P., Domkarova J. 2002. Identification of PCN species (Globodera rostochiensis, G. pallida) by using of ITS-1 region’s polymorphism. Rostlinna Vyroba 48 (11): 486–489.
Zouhar M., Ryšánek P., Kočová M. 2000. Detection and differentiation of the potato cyst nematodes Globodera rostochiensis and Globodera pallida by PCR. Plant Protection Science 36 (3): 81–84.
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