Diagnosis of potato root nematode Ditylenchus destructor in phytosanitary practice

Among over 90 nematode species of the genus Ditylen­ chus, Ditylenchus dipsaci (Kühn, 1857) Filipjev, 1936 and Ditylenchus destructor Thorne, 1945 are of great economic importance for agricultural crops. These nematodes are migratory endoparasites and polyphages. The species D. destructor is most common and harmful for potato culture in Russia, which has a quarantine status for some countries importing Russian products. Therefore, its rapid and accurate diagnosis is of great importance for phytosanitary practice. International and regional diagnostic protocols focus on D. dipsaci, and for the identification of D. destructor, quite sophisticated tests for laboratories are presented. In order to optimize the diagnosis of D. destructor, the classical polymerase chain reaction method was chosen, proposed by Polish scientists – Arnika Jeszke with co-authors (Jeszke et al., 2013), with universal  and  species-specific DITuniF/DITdesR primers designed for the region of internal transcribed spacer 1 (ITS1) and 5.8S rDNA. The test was verified with Russian commercial kits for DNA extraction and amplification, and the composition of the working mixture was also optimized to obtain the best result when working with these kits. In the verification process, Russian potato populations of D. de­ structor of various geographic origins were tested, and their belonging to the E haplotype was established. The applicability of the test was assessed successfully. It is recommended for research in Russian diagnostic laboratories for phytosanitary and plant quarantine.

1. Efremenko V.P., Burshtein Kh.S. Potato stem nematode and control measures in the Lithuanian SSR [Steblevaya nematoda kartofelya i mery borby s neyu v Litovskoy SSR] // In: Nematode diseases of agricultural crops and control measures. 1972; 97–98. (In Russ.)

2. Protection of potatoes from diseases, pests and weeds [Zashchita kartofelya ot bolezney, vrediteley i sornyakov ] / Anisimov B.V., Belov G.L., Varitsev Yu.A., Elansky S.N., Zhuromsky G.K., Zavriev S.K., Zeiruk V.N., Ivanyuk V.G., Kuznetsova M.A., Plyakhnevich M.P., Pshechenkov K.A., Simakov E.A., Sklyarova N.P., Stashevsky Z., Uskov A.I., Yashina THEM. // Potato grower. 2009; 272. (In Russ.)

3. ISPM 27 DP 8: Ditylenchus dipsaci and Ditylenchus destructor (2015).

4. Applied nematology / Butorina N.N., Zinovieva S.V., Kulinich O.A. and etc.; [res. ed. Zinovieva S.V., Chizhov V.N.]; Institute of Parasitology RAS. – M.; Nauka, 2006. 350 p.: – ISBN 5-02-034307-2 (in transl.). (In Russ.)

5. Phytoparasitic nematodes of Russia [Fitoparaziticheskiye nematody Rossii] / Zinovieva S.V., Chizhov V.N., Pridannikov M.V., Subbotin S.A., Ryss A.Yu., Khusainov R.V. // Association of scientific publications KMK. 2012; 242–248. (In Russ.)

6. A comparative and phylogenetic study of the Ditylenchus dipsaci, Ditylenchus destructor and Ditylenchus gigas populations occurring in Poland (Short Communication) / Jeszke A., Budziszewska M., Dobosz R., Stachowiak A., Protasewicz D., Wieczorek P. & Obrępalska-Stęplowska A. // Journal of Phytopathology. 2013. 162: 61–67.

7. Molecular diagnostics, taxonomy and phylogeny of the stem nematode Ditylenchus dipsaci species complex based on the sequences of the ITS-rDNA / Subbotin S.A., Madani M., Krall E., Sturhan D. & Moens M. // Phytopathology. 2005. 95: 1308–1315.

8. Molecular identification of Ditylenchus destructor nematode with PCR Species-Specific primers in the Moscow region / Mahmoudi N., Naserzadeh Y., Pakina E.N., Limantceva L.A., Nejad D.K. // RUDN Journal of Agronomy and Animal Industries. 2019. 14 (4): 430–436.

9. PM 7/87 (2) Ditylenchus destructor and Ditylenchus dipsaci (2017) // Bulletin OEPP/EPPO Bulletin. 2017. 47 (3), 401–419.

10. Wendt K.R., Vrain T.C. & Webster J.M. Separation of three species of Ditylenchus and some host races of D. dipsaci by restriction fragment length polymorphism // Journal of Nematology. 1993. 25: 555–563.