Bursaphelenchus sakishimanus n. sp. (Tylenchomorpha: Aphelenchoididae) isolated from a stag beetle, Dorcus titanus sakishimanus Nomura (Coleoptera: Lucanidae), on Ishigaki Island, Japan

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An undescribed Bursaphelenchus species was isolated and cultured from Dorcus titanus sakishimanus collected during a field survey of the insect-associated nematodes in subtropical Japan. The stag beetles were collected from Ishigaki Island, Okinawa, Japan, and dissected to examine their nematode associates. Then the dissected bodies were individually transferred to 2.0% agar plates, and nematode propagation on the plates was periodically examined. Nematodes were first recovered from the agar plate, i.e., the number of nematodes carried by the beetle was low, and infection was not confirmed during dissection. The new species was morphologically and phylogenetically (on a molecular basis) close to B. gerberae, which was isolated from the palm weevil, Rhynchophorus palmarum (Curculionidae), from tropical America, and to other weevil-associated Bursaphelenchus species. However, the new species can be distinguished from its close relatives by its typological characters, e.g., long and slender female tail and male spicule morphology as well as phylogenetic status inferred from the near-full-length of the small subunit (SSU) of the ribosomal RNA gene. The new species is described and illustrated herein as B. sakishimanus n. sp. and its molecular profiles, near-full-length SSU, and D2-D3 expansion segments of the large subunit of ribosomal RNA are described. This is the second Bursaphelenchus species associated with stag beetles (Lucanidae).

Bursaphelenchus sakishimanus n. sp. (Tylenchomorpha: Aphelenchoididae) isolated from a stag beetle, Dorcus titanus sakishimanus Nomura (Coleoptera: Lucanidae), on Ishigaki Island, Japan

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Figures

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    Bursaphelenchus sakishimanus n. sp. A: Adult male in right lateral view; B: Adult female in right lateral view; C: Anterior part of adult female; D: Female gonad in right lateral view; E: Vulval opening in ventral view; F: Female tail in right lateral view; G: Female anal region in ventral view. H: Male tail in right lateral view; I-K: Variation in male spicule in lateral view; L: Male tail in ventral view.

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    Bursaphelenchus sakishimanus n. sp. A: Anterior part of adult female in left lateral view (ep: excretory pore; h: hemizonid); B: Body surface showing three-lined lateral field and surface coarse annulation; C: Female gonad in left lateral view (ov: ovary; od: oviduct; sp: spermatheca; cr: crustaformeria; ut: uterus; v: vulval opening; pus: post uterine sac); D: Ventral view of female vulval region in two different focal planes (v: vulval opening; sp: spermatheca; cr: crustaformeria; ut: uterus); E: Right lateral view of female tail (a: anal opening); F: Ventral view of female tail region in two different focal planes (a: anal opening; int: intestine; rec: rectum); G: Ventral view of male tail in two different focal planes (c: cloacal opening; P + number: genital papillae); H: Left lateral view of male tail in three different focal planes (c: cloacal opening; P + number: genital papillae).

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    Oviduct and spermatheca of Bursaphelenchus sakishimanus n. sp. A-C: Dissected and extracted female gonad in three different focal planes (ov: ovary; od: oviduct; sp: spermatheca), anterior end of spermatheca forming blind sac is shown with arrowheads in A; D: Schematic drawing showing relationships between ovary (ov), oviduct (od), spermatheca (sp), crustaformeria (cr) and uterus (ut). Oviduct is indicated in grey.

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    Phylogenetic relationships among Bursaphelenchus spp. Bayesian tree inferred from near-full-length of SSU under GTR + I + G model (lnL=10210.3799; freqA = 0.2613; freqC = 0.1756; freqG = 0.2574; freqT = 0.3057; R(a) = 1.3809; R(b) = 2.8639; R(c) = 1.2013; R(d) = 0.6333; R(e) = 5.3136; R(f) = 1; Pinvar = 0.3897; Shape = 0.4301). Posterior probability values exceeding 50% are given for appropriate clades. The new species and B. tadamiensis, a previously reported stag beetle associate, are indicated with arrows.

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