Description of Bursaphelenchus taphrorychi sp. n. (Nematoda: Parasitaphelenchidae), the second Bursaphelenchus species from larval galleries of the beech bark beetle, Taphrorychus bicolor (Herbst.) (Coleoptera: Curculionidae: Scolytinae), in European beech, Fagus sylvatica L.

in Nematology
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Bursaphelenchus taphrorychi sp. n. is described from the bark of European beech, Fagus sylvatica. All propagative stages of the nematode are numerous in larval galleries of the beech bark beetle, Taphrorychus bicolor, while dauer juveniles are transmitted to new breeding trees under the elytra of adult beetles. The new species is characterised by the body length of 782 (717-858) μm in female and 638 (475-789) μm in male, moderately slender body (a = 35.0 (31.7-36.5) and 35.5 (31.4-37.1) in female and male, respectively), spicules 12.0-16.0 μm long, lateral fields with four incisures (i.e., three bands), and the arrangement of the seven male caudal papillae (i.e., a single precloacal ventromedian papilla (P1), one pair of adcloacal ventrosublateral papillae (P2), one postcloacal pair (P3) located at ca 60% of the tail length, posterior to the cloacal aperture, and one pair (P4) of subventral papillae of a similar size as the previous pair, but with somewhat sunken tips, located near base of bursa). In the number and arrangement of caudal papillae, stout and curved spicules with prominent rostrum and condylus, small vulval flap, body narrowed posterior to vulva, four incisures in the lateral fields, and long post-uterine sac, B. taphrorychi sp. n. shares most of the key morphological characters with members of the sexdentati-group. However, the newly described species is unique amongst Bursaphelenchus species of this group by the combination of shape of female tail, shape of spicules, and some other morphometric characters. The close relation of B. taphrorychi sp. n. with members of the sexdentati-group has been confirmed by DNA sequencing and phylogenetic analysis of the 28S rDNA region. The taxonomic separation of the new species is also confirmed by the unique molecular profile of the ITS region (ITS-RFLP). In laboratory rearing, B. taphrorychi sp. n. can develop and reproduce on Botrytis cinerea cultures.

Description of Bursaphelenchus taphrorychi sp. n. (Nematoda: Parasitaphelenchidae), the second Bursaphelenchus species from larval galleries of the beech bark beetle, Taphrorychus bicolor (Herbst.) (Coleoptera: Curculionidae: Scolytinae), in European beech, Fagus sylvatica L.

in Nematology



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    Bursaphelenchus taphrorychi sp. n. A: Female; B: Male; C: Dauer juvenile; D: Anterior region (female); E: Female mid-body region with spermatheca, crustaformeria, vulva, and post-uterine sac; F-H: Female tail (range of forms). I, J: Male tail with spicules, bursa and positions of caudal papillae (lateral view); K: Male tail with bursa and positions of cloacal lips and caudal papillae (ventral view, somewhat flattened).

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    Light (DIC) micrographs of Bursaphelenchus taphrorychi sp. n. A-N: Adults. A: Female (entire); B: Male (entire); C-D: Anterior region: C: Head with stylet, median bulb, nerve ring, excretory pore (Ep) and dorsal pharyngeal gland; D: Head with stylet, median bulb, excretory pore (Ep) and nerve ring; E-H: Mid-body region: E: Spermatheca (Sp), crustaformeria (Cf), vulva, and post-uterine sac; F: Spermatheca, crustaformeria, and vulva; G: Spermatheca, uterus with egg, vulva, and post-uterine sac; H: Lateral field with four incisures; I-K: Male tail with spicules (lateral view); L: Male tail with bursa and caudal papillae in partially ventral view (somewhat flattened); M, N: Female tail (M: inserted micrograph of occasional female tail showing mucro-like terminus); O-Q: Dauer juveniles. O: Juvenile (entire view); P: Anterior region with faintly marked median bulb; Q: Tail.

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    SEM micrographs of Bursaphelenchus taphrorychi sp. n. A, B: Female vulva and lateral field with four incisures (i.e., three bands) (lateral view); C, D: Male tail: C: Cloacal lips, caudal papillae and bursa (partially ventral view); D: Cloacal lips and spicule tip (lateral view).

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    ITS-RFLP profile of Bursaphelenchus taphrorychi sp. n. Lanes 1 = DNA marker (100 bp ladder, Sigma-Aldrich); Lane 2-6 = Restriction fragments: Lane 2 = RsaI; Lane 3 = HaeIII; Lane 4 = MspI; Lane 5 = HinfI; Lane 6 = AluI; Lane 7 = rDNA amplification product.

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    Maximum likelihood (ML) analysis of Bursaphelenchus taphrorychi sp. n. and 76 other Bursaphelenchus species based on partial 28S rDNA. Aphelenchoides fujianensis served as outgroup species. Phylogenetic trees were generated with 1000 bootstrap replications.

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    Neighbour joining (NJ) analysis of Bursaphelenchus taphrorychi sp. n. and 76 other Bursaphelenchus species based on partial 28S rDNA. Aphelenchoides fujianensis served as outgroup species. Phylogenetic trees were generated with 1000 bootstrap replications.

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    Maximum parsimony (MP) analysis of Bursaphelenchus taphrorychi sp. n. and 76 other Bursaphelenchus species based on partial 28S rDNA. Aphelenchoides fujianensis served as outgroup species. Phylogenetic trees were generated with 1000 bootstrap replications.


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