European Journal of Plant Pathology

, Volume 153, Issue 2, pp 441–464 | Cite as

An integrative approach to the study of Xiphinema brevicolle Lordello and Da Costa 1961, supports its limited distribution worldwide (Nematoda: Longidoridae)

  • Stela Lazarova
  • Claudio Marcelo G. Oliveira
  • Thomas Prior
  • Vlada PenevaEmail author
  • Shesh Kumari


Xiphinema americanum group has a cosmopolitan distribution, with several species having particular importance as virus-vectors of four economically important nepoviruses naturally occurring in the USA with IAI quarantine status for Europe. The morphological diagnosis of the species is problematic due to the limited diagnostic characters, overlapping morphometrics and incomplete taxonomic descriptions. An ongoing issue within the X. americanum group is the determination of species belonging to the X. brevicolle subgroup. By using a phylogenetic framework, molecular and morphometric data of the topotype population we demonstrate that certain of the published DNA sequences have been misidentified and belong to closely related species such as X. taylori, X. diffusum or unknown/unidentified species. Studying again the morphology and morphometrics of populations identified as X. brevicolle from the Czech Republic the identity, phylogenetic position and genetic variability of these populations have been clarified. These belong to a different species that is described herein as X. parataylori n. sp. Multi-gene phylogenies were reconstructed using nuclear ribosomal (18S rRNA gene, ITS1 and ITS2 regions, D2-D3 expansion segments of 28S rRNA gene) and mitochondrial DNA (partial cox1 and nad4) and the position of X. brevicolle (topotype population) to the most closely related species is shown. Morphology and species specific sequences were linked and the worldwide distribution discussed. The validity of X. brevicolle and the long-standing controversies about the taxonomic status of X. diffusum and X. taylori have been clarified. Xiphinema taylori and X. diffusum may represent complexes of cryptic species, however more comprehensive studies involving additional populations and materials from the type localities are needed to support this hypothesis.


Diagnostics Mislabeled sequences Phylogeny Taxonomy Type population Xiphinema americanum group 



The authors are grateful to Dr. Marta Lišková (Slovak Academy of Sciences) for providing nematodes and Markéta Vítámvásová, Dr. Milka Elshishka and Ms. Lyudmila Lozanova (IBER-BAS) for their technical help. The work was supported by the Ministry of Agriculture of the Czech Republic, Project number RO0418 and ANIDIV-3 project supported by the BAS.

Compliance with ethical standards

Disclosure of potential conflicts of interest

All the molecular experiments were carried out at Crop Research Institute, Czech Republic and supported by the Ministry of Agriculture of the Czech Republic, Project number RO0418. Taxonomic studies, phylogenetic analyses, as well as the preparation of the first draft of the manuscript were carried out at the Institute of Biodiversity and Ecosystem Research, Bulgaria and supported by the project ANIDIV-3 project, Bulgarian Academy of Sciences. Soil sampling in Brazil was carried out by Oliveira C. M. G. D. All authors were invoved in text reviewing. Type material and voucher specimens studied were formally loaned from international collections, and the holotype specimen and paratype series deposited with accession numbers in curated nematode collections.

Research involving human participants and/or animals

This article does not contain any studies with human participants or animals performed by any of the authors.

Informed consent

Informed consent was obtained from all individual participants included in the study.


  1. Andrássy, I. (1998). The genus Boreolaimus gen. n. and its six species (Dorylaimida: Qudsianematidae), nematodes from the European Arctic. Fundamental and Applied Nematology, 21, 553–567.Google Scholar
  2. Archidona-Yuste, A., Navas-Cortés, J. A., Cantalapiedra-Navarrete, C., Palomares-Rius, J. E., & Castillo, P. (2016). Cryptic diversity and species delimitation in the Xiphinema americanum-group complex (Nematoda: Longidoridae) as inferred from morphometrics and molecular markers. Zoological Journal of the Linnean Society, 176(2), 231–265. Scholar
  3. Barsalote, E. M., Tian, Z., & Zheng, J. (2017). Intra-species variability of Xiphinema brevicolle Lordello & Costa, 1961 (Nematoda: Longidoridae) from China. Russian Journal of Nematology, 25(1), 1–16.Google Scholar
  4. Barsi, L. (1989). The Longidoridae (Nematoda: Dorylaimida) in Yugoslavia. I. Nematologia Mediterranea, 17(2), 97–108.Google Scholar
  5. Barsi, L., & Lamberti, F. (2004). Xiphinema parasimile sp. n. from Serbia and X. simile, first record from Bosnia and Herzegovina (Nematoda, Dorylaimida). Nematologia Mediterranea, 32(1), 101–109.Google Scholar
  6. Chen, D. Y., Ni, H. F., Yen, J. H., Cheng, Y. H., & Tsay, T. T. (2005). Differentiation of the Xiphinema americanum-group nematodes X. brevicollum, X. incognitum, X. diffusum and X. oxycaudatum in Taiwan by morphometrics and nuclear ribosomal DNA sequences. Nematology, 7(5), 713–725. Scholar
  7. Coomans, A., Huys, R., Heyns, J., & Luc, M. (2001). Character analysis, phylogeny, and biogeography of the genus Xiphinema Cobb, 1973 (Nematoda, Longidoridae). Annales du Musée Royal de l’Afrique Centrale (Zoologie), Tervuren, Belgium, 287, 1–239.Google Scholar
  8. Erbenová, M. (1975). Ectoparasitic eelworms of the genus Xiphinema Cobb in the orchards of the CSR. Zahradnictvi, 2(2), 79–86.Google Scholar
  9. Gozel, U., Lamberti, F., Duncan, L., Agostinelli, A., Rosso, L., Nguyen, K., & Adams, B. J. (2006). Molecular and morphological consilience in the characterisation and delimitation of five nematode species from Florida belonging to the Xiphinema americanum - group. Nematology, 8(4), 521–532. Scholar
  10. Gutiérrez-Gutiérrez, C., Cantalapiedra-Navarrete, C., Decraemer, W., Vovlas, N., Prior, T., Rius, J. E. P., & Castillo, P. (2012). Phylogeny, diversity, and species delimitation in some species of the Xiphinema americanum-group complex (Nematoda: Longidoridae), as inferred from nuclear and mitochondrial DNA sequences and morphology. European Journal of Plant Pathology, 134(3), 561–597. Scholar
  11. He, Y., Subbotin, S. A., Rubtsova, T. V., Lamberti, F., Brown, D. J., & Moens, M. (2005). A molecular phylogenetic approach to Longidoridae (Nematoda: Dorylaimida). Nematology, 7(1), 111–124. Scholar
  12. Holterman, M., van der Wurff, A., van den Elsen, S., van Megen, H., Bongers, T., Holovachov, O., Bakker, J., & Helder, J. (2006). Phylum-wide analysis of SSU rDNA reveals deep phylogenetic relationships among nematodes and accelerated evolution toward crown clades. Molecular Biology and Evolution, 23(9), 1792–1800. Scholar
  13. Janssen, T., Karssen, G., Couvreur, M., Waeyenberge, L., & Bert, W. (2017a). The pitfalls of molecular species identification: A case study within the genus Pratylenchus (Nematoda: Pratylenchidae). Nematology, 19, 1179–1199. Scholar
  14. Janssen, T., Karssen, G., Orlando, V., Subbotin, S. A., & Bert, W. (2017b). Molecular characterization and species delimiting of plant-parasitic nematodes of the genus Pratylenchus from the penetrans group (Nematoda: Pratylenchidae). Molecular Phylogenetics and Evolution, 117, 30–48. Scholar
  15. Katoh, K., & Standley, D. M. (2013). MAFFT multiple sequence alignment software version 7: Improvements in performance and usability. Molecular Biology and Evolution, 30(4), 772–780. Scholar
  16. Khan, S. H., & Ahmad, S. (1975a). Longidoridae (Thorne 1935) n. rank. (Nematoda: Dorylaimina) with the description of Xiphinema neoamericanum n. sp. from India and proposal of a new name for X. americanum sensu Carvalho (1956) non Cobb, 1913. Nematologia Mediterranea 3, 23–28.Google Scholar
  17. Khan, S. H., & Ahmad, S. (1975b). Xiphinema inaequale nom. nov. (syn.: X. neoamericanum Khan et Ahmad, 1975). Nematologia Mediterranea, 5, 93.Google Scholar
  18. Kumari, S., & Di Cesare, A. (2013). Nicotinamide dehydrogenase subunit 4 analysis of Xiphinema diversicaudatum and Xiphinema simile (Nematoda: Longidoridae). European Journal of Plant Pathology, 136(4), 803–810. Scholar
  19. Kumari, S., & Subbotin, S. A. (2012). Characterization of Longidorus helveticus (Nematoda: Longidoridae) from the Czech Republic. European Journal of Plant Pathology, 133(4), 923–933. Scholar
  20. Kumari, S., Polák, J., & Choutka, R. (2005). Plant-parasitic nematodes of the genus Xiphinema (Nematoda: Longidoridae) in the vineyards of the Czech Republic. Nematology, 7(1), 81–93. Scholar
  21. Kumari, S., Decraemer, W., Traversa, D., & Lišková, M. (2009). Molecular and morphological delineation of Longidorus poessneckensis Altherr, 1974 (Nematoda: Dorylaimida). European Journal of Plant Pathology, 123(2), 125–137. Scholar
  22. Kumari, S., Decraemer, W., & De Luca, F. (2010). Molecular characterization of Xiphinema brevicollum (Nematoda: Longidoridae) from the Czech Republic. European Journal of Plant Pathology, 128(2), 243–250. Scholar
  23. Lamberti, F., & Bleve-Zacheo, T. (1979). Studies on Xiphinema americanum sensu lato with descriptions of fifteen new species (Nematoda, Longidoridae). Nematologia Mediterranea, 7(1), 51–106.Google Scholar
  24. Lamberti, F. D., Ciancio, A., Agostinelli, A., & Coiro, M. I. (1991). Relationship between Xiphinema brevicolle and X diffusum, with a redescription of Xiphinema brevicolle and description of three new species of Xiphinema (Nematoda, Dorylaimida). Nematologia Mediterranea, 19(2), 311–326.Google Scholar
  25. Lamberti, F., Molinari, S., Moens, M., & Brown, D. J. (2000). The Xiphinema americanum group. I. Putative species, their geographical occurrence and distribution, and regional polytomous identification keys for the group. Russian Journal of Nematology, 8(1), 65–84.Google Scholar
  26. Lamberti, F., De Luca, F., Molinari, S., Duncan, L. W., Agostinelli, A., Coiro, M. I., et al. (2002a). Studies on some species of the Xiphinema americanum group (Nematoda, Dorylaimida) occurring in Florida. Nematologia Mediterranea, 30(1), 31–44.Google Scholar
  27. Lamberti, F., Molinari, S., Moens, M., & Brown, D. J. (2002b). The Xiphinema americanum-group. 11. Morphometric relationships. Russian Journal of Nematology, 10, 99–112.Google Scholar
  28. Lamberti, F., Hockland, S., Agosttinelli, A., Moens, M., & Brown, D. J. F. (2004). The Xiphinema americanum group. III. Keys to species identification. Nematologia Mediterannea, 32(1), 53–56.Google Scholar
  29. Lazarova, S. S., Malloch, G., Oliveira, C. M., Hübschen, J., & Neilson, R. (2006). Ribosomal and mitochondrial DNA analyses of Xiphinema americanum-group populations. Journal of Nematology, 38(4), 404–410.Google Scholar
  30. Lazarova, S. S., Brown, D. J., Oliveira, C. M. G., Fenton, B., MacKenzie, K., Wright, F., et al. (2016a). Diversity of endosymbiont bacteria associated with a non-filarial nematode group. Nematology, 18(5), 615–623. Scholar
  31. Lazarova, S., Peneva, V., & Kumari, S. (2016b). Morphological and molecular characterisation, and phylogenetic position of X. browni sp. n., X. penevi sp. n. and two known species of Xiphinema americanum-group (Nematoda, Longidoridae). ZooKeys, (574), 1–42.
  32. Lišková, M. (1995). Faunistic and ecological notes on Xiphinema (Nematoda: Dorylaimida) in Slovakia. Sect. Zoology, 50, 125–131.Google Scholar
  33. Loof, P. A. A., & Coomans, A. (1972). The oesophageal gland nuclei of Longidoridae (Dorylaimida). Nematologica, 18, 213–233. Scholar
  34. Lordello, L. G. E., & Da Costa, C. (1961). A new nematode parasite of coffee roots in Brazil. Revista Brasileira de Biologia, 21(4), 363–366.Google Scholar
  35. Luc, M., Coomans, A., Loof, P. A., & Baujard, P. (1998). The Xiphinema americanum-group (Nematoda: Longidoridae). 2. Observations on Xiphinema brevicollum Lordello & Da Costa, 1961 and comments on the group. Fundamental and Applied Nematology, 21(5), 475–490.Google Scholar
  36. Monteiro, A. R. (2010). Nomenclatorial note on Xiphinema brevicolle Lordello & Costa, 1961 (Nematoda: Longidoridae). Nematologia Brasileira, 34, 1–2.Google Scholar
  37. Nunn, G. B. (1992). Nematode molecular evolution: An investigation of evolutionary patterns among nematodes based upon DNA sequences (Doctoral dissertation, University of Nottingham).Google Scholar
  38. Oliveira, C. M., Hübschen, J., Brown, D. J., Ferraz, L. C., Wright, F., & Neilson, R. (2004). Phylogenetic relationships among Xiphinema and Xiphidorus nematode species from Brazil inferred from 18S rDNA sequences. Journal of Nematology, 36(2), 153–159.Google Scholar
  39. Oliveira, C. M., Fenton, B., Malloch, G., Brown, D. J., & Neilson, R. (2005). Development of species-specific primers for the ectoparasitic nematode species Xiphinema brevicolle, X. diffusum, X. elongatum, X. ifacolum and X. longicaudatum (Nematoda: Longidoridae) based on ribosomal DNA sequences. Annals of Applied Biology, 146(3), 281–288. Scholar
  40. Orlando, V., Chitambar, J. J., Dong, K., Chizhov, V. N., Mollov, D., Bert, W., & Subbotin, S. A. (2016). Molecular and morphological characterisation of Xiphinema americanum-group species (Nematoda: Dorylaimida) from California, USA, and other regions, and co-evolution of bacteria from the genus Candidatus Xiphinematobacter with nematodes. Nematology, 18(9), 1015–1043. Scholar
  41. Palomares-Rius, J. E., Cantalapiedra-Navarrete, C., Archidona-Yuste, A., Subbotin, S. A., & Castillo, P. (2017a). The utility of mtDNA and rDNA for barcoding and phylogeny of plant-parasitic nematodes from Longidoridae (Nematoda, Enoplea). Scientific Reports, 7(1), 10905. Scholar
  42. Palomares-Rius, J. E., Cantalapiedra-Navarrete, C., Archidona-Yuste, A., Blok, V. C., & Castillo, P. (2017b). Mitochondrial genome diversity in dagger and needle nematodes (Nematoda: Longidoridae). Scientific Reports, 7, 41813. Scholar
  43. Peneva, V., & Choleva, B. (1992). Nematodes of the family Longidoridae from forest nurseries in Bulgaria. II. The genus Xiphinema Cobb, 1913. Helminthology, 32, 46–58.Google Scholar
  44. Sakai, H., Takeda, A., & Mizukubo, T. (2011). First report of Xiphinema brevicolle Lordello et Costa, 1961 (Nematoda, Longidoridae) in Japan. ZooKeys, 135, 21–40.
  45. Sakai, H., Takeda, A., & Mizukubo, T. (2012). Intra-specific variation of Xiphinema brevicolle Lordello et Costa, 1961 (Nematoda: Longidoridae) in Japan. Nematological Research, 42(1), 1–7.
  46. Sela, I., Ashkenazy, H., Katoh, K., & Pupko, T. (2015). GUIDANCE2: Accurate detection of unreliable alignment regions accounting for the uncertainty of multiple parameters. Nucleic Acids Research, 43(W1), W7–W14. Scholar
  47. Shah, F. A., Bell, N. L., & Bulman, S. R. (2009). Morphological and molecular confirmation of Xiphinema brevicollum, a nematode from the virus-vectoring X. americanum group, from New Zealand. Australasian Plant Pathology, 38(5), 500–504. Scholar
  48. Stanton, J. M., McNicol, C. D., & Steele, V. (1998). Non–manual lysis of second–stage Meloidogyne juveniles for identification of pure and mixed samples based on the polymerase chain reaction. Australasian Plant Pathology, 27(2), 112–115. Scholar
  49. Stothard, P. (2000). The sequence manipulation suite: JavaScript programs for analyzing and formatting protein and DNA sequences. Biotechniques, 28, 1102–1104.Google Scholar
  50. Tamura, K., Stecher, G., Peterson, D., Filipski, A., & Kumar, S. (2013). MEGA6: Molecular evolutionary genetics analysis version 6.0. Molecular Biology and Evolution, 30(12), 2725–2729. Scholar
  51. Troccoli, A., Subbotin, S. A., Chitambar, J. J., Janssen, T., Waeyenberge, L., Stanley, J. D., et al. (2016). Characterisation of amphimictic and parthenogenetic populations of Pratylenchus bolivianus Corbett, 1983 (Nematoda: Pratylenchidae) and their phylogenetic relationships with closely related species. Nematology, 18(6), 651–678. Scholar
  52. Vandekerckhove, T. T., Willems, A., Gillis, M., & Coomans, A. (2000). Occurrence of novel verrucomicrobial species, endosymbiotic and associated with parthenogenesis in Xiphinema americanum-group species (Nematoda, Longidoridae). International Journal of Systematic and Evolutionary Microbiology, 50(6), 2197–2205. Scholar
  53. Vrain, T. C., Wakarchuk, D. A., Levesque, A. C., & Hamilton, R. I. (1992). Intraspecific rDNA restriction fragment length polymorphism in the Xiphinema americanum group. Fundamental and Applied Nematology, 15(6), 563–573.Google Scholar
  54. Zasada, I. A., Peetz, A., Howe, D. K., Wilhelm, L. J., Cheam, D., Denver, D. R., & Smythe, A. B. (2014). Using mitogenomic and nuclear ribosomal sequence data to investigate the phylogeny of the Xiphinema americanum species complex. PLoS One, 9(2), e90035. Scholar
  55. Zeng, Y., Ye, W., Zhang, Z., Sun, H., Yong, L., Huang, Y., Zhao, K., Liang, H., & Kerns, J. (2016). Morphological and molecular characterization of Xiphinema species from Shenzhen, China. Helminthologia, 53(1), 62–75. Scholar
  56. Zhao, L., Ge, J., Wang, W., Gu, S., Zhang, Y., Hu, X., et al. (2012). Morphological and molecular characterization of Xiphinema brevicollum and X. himalayense. Journal of South China Agricultural University, 33(4), 488–492.Google Scholar

Copyright information

© Koninklijke Nederlandse Planteziektenkundige Vereniging 2018

Authors and Affiliations

  • Stela Lazarova
    • 1
  • Claudio Marcelo G. Oliveira
    • 2
  • Thomas Prior
    • 3
  • Vlada Peneva
    • 1
    Email author
  • Shesh Kumari
    • 4
  1. 1.Department of Animal BiodiversityInstitute of Biodiversity and Ecosystem ResearchSofiaBulgaria
  2. 2.Instituto BiológicoCampinasBrazil
  3. 3.Plant Pest & Disease ProgrammeYorkUK
  4. 4.Division of Plant HealthCrop Research InstitutePrague 6Czech Republic

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