Bursaphelenchus tiliae sp. n. (Nematoda: Parasitaphelenchidae), a nematode associate of the bark beetle Ernoporus tiliae (Panz.) (Coleoptera: Curculionidae, Scolytinae), in small-leaved lime, Tilia cordata Mill.

in Nematology
Restricted Access
Get Access to Full Text
Rent on DeepDyve

Have an Access Token?

Enter your access token to activate and access content online.

Please login and go to your personal user account to enter your access token.


Have Institutional Access?

Access content through your institution. Any other coaching guidance?


Bursaphelenchus tiliae sp. n. is described from the bark of dead branches of the small-leaved lime, Tilia cordata, infested with the lime bark beetle, Ernoporus tiliae. The nematode adults and propagative juveniles were present in larval galleries, while dispersal dauer juveniles colonised the Malpighian tubules of adult beetles. Bursaphelenchus tiliae sp. n. is characterised by the body length of 664 (568-817) μm in female and 622 (515-754) μm in male, moderately slender body (a = 41.0 (34.7-47.9) and 43.1 (37.8-51.9) in female and male, respectively), and spicules 20.0 (18.1-21.8) μm long. The extended anterior vulval lip in female, lateral fields with four incisures, long post-uterine sac, and number (7) and arrangement of male caudal papillae, may indicate that B. tiliae sp. n. is closely related to the xylophilus group. It differs from this group by smaller, arcuate spicules with relatively short rostrum, narrow capitulum (5.2-6.9 μm), and indistinct condylus in an almost continuous line with dorsal lamina. Distal third of spicule dorsal contour is usually almost straight. The spicules have a distinct, disk-like cucullus at the terminus. The status of the new species is confirmed by the unique molecular profile of the ITS region (ITS-RFLP) and its close relation to the xylophilus group by DNA sequencing and phylogenetic analysis of the 28S rDNA region. The presence of B. tiliae sp. n. dauer juveniles in Malpighian tubules of their vector bark beetle, E. tiliae, caused limited pathogenicity expressed as a local expansion of the tubule basement membrane and partial degradation of the cellular epithelium.

Bursaphelenchus tiliae sp. n. (Nematoda: Parasitaphelenchidae), a nematode associate of the bark beetle Ernoporus tiliae (Panz.) (Coleoptera: Curculionidae, Scolytinae), in small-leaved lime, Tilia cordata Mill.

in Nematology



BraaschH. (2008). The enlargement of the xylophilus group in the genus Bursaphelenchus. In: MotaM.VieiraP. (Eds). Pine wilt disease: a worldwide threat to forest ecosystems. Berlin, GermanySpringer pp.  139-150.

BraaschH.GuJ.BurgermeisterW.BrandstetterM.MetgeK. (2007). Bursaphelenchus africanus sp. n. (Nematoda: Parasitaphelenchidae) – found in packaging wood exported from South Africa to Ningbo/China. Journal of Nematode Morphology and Systematics 971-81.

BraaschH.BurgemeisterW.GuJ. (2009). Revised intra-generic grouping of Bursaphelenchus Fuchs, 1937 (Nematoda: Aphelenchoididae). Journal of Nematode Morphology and Systematics 1265-88.

BurgermeisterW.MetgeK.BraaschH.BuchbachE. (2005). ITS-RFLP patterns for differentiation of 26 Bursaphelenchus species (Nematoda: Parasitaphelenchidae) and observations on their distribution. Russian Journal of Nematology 1329-42.

BurgermeisterW.BraaschH.MetgeK.GuJ.SchröderT.WoldtE. (2009). ITS-RFLP analysis, an efficient tool for differentiation of Bursaphelenchus species. Nematology 11649-668.

De LeyP.FélixM.A.FrisseL.M.NadlerS.A.SternbergP.W.ThomasW.K. (1999). Molecular and morphological characterisation of two reproductively isolated species with mirror-image anatomy (Nematoda: Cephalobidae). Nematology 1591-612.

EndaN.MamiyaY. (1972). [ The effect of maturation feeding of Monochamus alternatus contaminated with Bursaphelenchus lignicolus on pine trees.] Transactions of the Meeting of the Japanese Forestry Society 83320-322.

FerrisV.R.FerrisJ.M.FaghihiJ. (1993). Variation in spacer ribosomal DNA in some cyst-forming species of plant parasitic nematodes. Fundamental and Applied Nematology 16177-184.

FutaiK. (2013). Pine wood nematode, Bursaphelenchus xylophilus. Annual Review of Phytopathology 5161-83.

GuJ.ZhengW.BraaschH.BurgermeisterW. (2008). Description of Bursaphelenchus macromucronatus sp. n. (Nematoda: Parasitaphelenchidae) in packaging wood from Taiwan and India – a new species of the xylophilus group. Journal of Nematode Morphology and Systematics 1131-40.

KanzakiN.FutaiK. (2003). Description and phylogeny of Bursaphelenchus luxuriosae n. sp. (Nematoda: Aphelenchoididae) isolated from Acalolepta luxuriosa (Coleoptera: Cerambycidae). Nematology 5565-572.

KanzakiN.TsudaK.FutaiK. (2000). Description of Bursaphelenchus conicaudatus sp. n. (Nematoda: Aphelenchoididae), isolated from the yellow-spotted longicorn beetle, Psacothea hilaris (Coleoptera: Cerambycidae) and fig trees, Ficus carica. Nematology 2165-168.

KanzakiN.AikawaT.MaeharaN.MatsumotoK. (2008). Bursaphelenchus doui Braasch, Gu, Burgermeister & Zhang, 2005 (Aphelenchida: Parasitaphelenchidae), an associate of Monochamus subfasciatus Bates (Coleoptera: Cerambycidae) and Pinus densiflora Sieb. & Zucc. Nematology 1069-78.

KanzakiN.AikawaT.Giblin-DavisR.M. (2009). Bursaphelenchus tokyoensis n. sp. (Nematoda: Parasitaphelenchinae) isolated from dead wood of the Japanese red pine, Pinus densiflora Sieb. & Zucc., in Japan. Nematology 11171-180.

KanzakiN.MaeharaN.AikawaT.MatsumotoK. (2012). Bursaphelenchus firmae n. sp. (Nematoda: Aphelenchoididae), isolated from Monochamus grandis Waterhouse that emerged from dead firs, Abies firma Sieb. et Zucc. Nematology 14395-404.

MamiyaY.EndaN. (1979). Bursaphelenchus mucronatus n. sp. (Nematoda: Aphelenchoididae) from pine wood and its biology and pathogenicity to pine trees. Nematologica 25353-361.

MotaM.BraaschH.BravoM.A.PenasA.C.BurgermeisterW.MetgeK.CousaE. (1999). First report of Bursaphelenchus xylophilus in Portugal and in Europe. Nematology 1727-734.

RyssA.VieiraP.MotaM.KulinichO. (2005). A synopsis of the genus Bursaphelenchus Fuchs, 1937 (Aphelenchida: Parasitaphelenchidae) with keys to species. Nematology 7393-458.

SeinhorstJ.W. (1959). A rapid method for the transfer of nematodes from fixative to anhydrous glycerin. Nematologica 467-69.

SulstonJ.HodgkinJ. (1988). Methods. In: WoodB. (Ed.). The nematode Caenorhabditis elegans. Plainview, NY, USACold Spring Harbor Laboratory Press pp.  587-606.

TamuraK.PetersonD.PetersonN.StecherG.NeiM.KumarS. (2011). MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Molecular Biology and Evolution 282731-2739.

TanakaS.E.TanakaR.AkibaM.AikawaT.MeaharaN.TakeuchiY.KanzakiN. (2014). Bursaphelenchus niphades sp. n. (Tylenchida: Aphelenchoididae) amensally associated with Niphades variegatus (Roelofs) (Coleoptera: Curculionidae). Nematology 16259-281.

TomalakM.FilipiakA. (2010). Description of Bursaphelenchus populi sp. n. (Nematoda: Parasitaphelenchidae), a new member of the xylophilus group from aspen, Populus tremula L., in Europe. Nematology 12399-416.

TomalakM.FilipiakA. (2011). Bursaphelenchus trypophloei sp. n. (Nematoda: Parasitaphelenchinae) – an associate of the bark beetle, Trypophloeus asperatus (Gyll.) (Coleoptera: Curculionidae, Scolytinae), in aspen, Populus tremula L. Nematology 13619-636.

TomalakM.FilipiakA. (2014). Bursaphelenchus fagi sp. n. (Nematoda: Parasitaphelenchidae), an insect pathogenic nematode in Malpighian tubules of the bark beetle, Taphrorychus bicolor (Herbst.) (Coleoptera: Curculionidae, Scolytinae), in European beech, Fagus silvatica L. Nematology 16591-606.

TomalakM.WelchH.E.GallowayT.D. (1989). Pathology of Aphelenchoides pityokteini (Nematoda: Aphelenchoididae) in the Malpighian tubules of Pityokteines sparsus (Coleoptera: Scolytidae). Journal of Invertebrate Pathology 53140-141.

TomalakM.WorrallJ.FilipiakA. (2013). Bursaphelenchus masseyi sp. n. (Nematoda: Parasitaphelenchinae) – a nematode associate of the bark beetle Trypophloeus populi Hopkins (Coleoptera: Curculionidae: Scolytinae), in aspen, Populus tremuloides Michx., affected by sudden aspen decline in Colorado. Nematology 15907-921.

VrainT.C. (1993). Restriction fragment length polymorphism separates species of the Xiphinema americanum group. Journal of Nematology 25361-364.


  • View in gallery

    Bursaphelenchus tiliae sp. n. A: Female; B: Male; C: Anterior region (female); D: Vulva region (female); E, F: Male tail with spicules (range of forms), bursa and positions of caudal papillae (lateral view); G, H: Female tail (range of forms); I-K: Male tail showing shape of bursa (range of forms) and position of caudal papillae (partially ventral view, somewhat flattened); L: Dauer juvenile (dissected from a Malpighian tubule of vector bark beetle, Ernoporus tiliae).

  • View in gallery

    Light (DIC) and SEM micrographs of Bursaphelenchus tiliae sp. n. A-L: Adults. A: Head with stylet, median bulb and nerve ring; B: Head and median bulb showing position of excretory pore; C: Vulva with extended anterior lip (flap) and spermatheca, crustaformeria (anterior) and post-uterine sac (posterior); D: Vulva with extended anterior lip; E: Lateral field with four incisures; F, G: Range of spicule forms; H: Positions of male caudal papillae (SEM); I, J: Modifications of otherwise ovoid bursa (partially ventral view – somewhat flattened); K, L: Female tail (range of forms); M: Dauer juvenile (entire); N: Dauer anterior region with median bulb; O: Dauer tail.

  • View in gallery

    ITS-RFLP profile of Bursaphelenchus tiliae sp. n. Lanes 1 and 8 = DNA marker (100 bp, Invitrogen, with lowest band at 200 bp and prominent bands at 600, 1500, 2072 bp). Lane 2 = uncleaved PCR product; Lanes 3-7 = restriction fragments obtained with: Lane 3 = RsaI; Lane 4 = HaeIII; Lane 5 = MspI; Lane 6 = HinfI; Lane 7 = AluI.

  • View in gallery

    Maximum likelihood (ML) analysis of Bursaphelenchus tiliae sp. n. and 39 other Bursaphelenchus species based on partial 28S rDNA. Aphelenchoides fragariae served as outgroup species.

  • View in gallery

    Neighbour-joining (NJ) analysis of Bursaphelenchus tiliae sp. n. and 39 other Bursaphelenchus species based on partial 28S rDNA. Aphelenchoides fragariae served as outgroup species.

  • View in gallery

    Maximum parsimony (MP) analysis of Bursaphelenchus tiliae sp. n. and 39 other Bursaphelenchus species based on partial 28S rDNA. Aphelenchoides fragariae served as outgroup species.

  • View in gallery

    Light micrographs of Malpighian tubules of adult Ernoporus tiliae (DIC). A: Entire tubule with dauer juveniles of Bursaphelenchus tiliae sp. n. in distal part; B: Uninfested tubule with cellular epithelium and distinct nuclei; C-E: Tubules colonised by dauer juveniles of B. tiliae sp. n.; C, D: Overall view of tubule filled with nematodes showing local expansion of tubule width; E: Detailed view of tubule filled with nematodes showing epithelial tissue only partially damaged with cells and distinct nuclei still present.


Content Metrics

Content Metrics

All Time Past Year Past 30 Days
Abstract Views 33 33 12
Full Text Views 90 90 44
PDF Downloads 6 6 4
EPUB Downloads 0 0 0