Jianfeng Gu

Redescription of Bursaphelenchus parapinasteri (Tylenchina: Aphelenchoididae) isolated from Pinus thunbergii in China with a key to the hofmanni-group

Bursaphelenchus parapinasteri is redescribed morphologically and with new molecular characterisation. It was isolated from a dead Pinus thunbergii on Changgang Mountain, Zhoushan Islands, Zhejiang Province, China. Detailed morphology of the spicule, female gonad, hemizonid position, arrangement of male caudal papillae, and female tail terminus shape are documented. The ITS-RFLP patterns and the ITS1/2, partial 28S D2-D3 and partial 18S gene sequences were characterised. The phylogenetic analyses revealed that B. parapinasteri belongs to the hofmanni-group sensu Braasch and is close to B. anamurius, B. hofmanni, B. mazandaranense, B. paracorneolus, B. pinasteri and B. ulmophilus. A diagnostic key to species of the hofmanni-group is presented.

Bursaphelenchus spp. in Wood Packaging Intercepted in China

Solid wood packaging material (including dunnage) made of unprocessed wood is known to be an important pathway for the introduction and spread of pests, but we may not know how serious this problem is. Another question is: Can a quarantine treatment certificate or the “HT” (“MB”) mark on the wood really assure that the wooden packaging is free of pests? In February of 2006, many living pinewood nematodes (PWN) were detected in wooden packaging exported from Portugal to China. The black mark “PT 048 HT” could clearly be seen on the wood (PT representing Portugal, 048 representing the number of the heat treatment site). Was the wood cut from the PWN affected zone in Portugal and not properly treated? At Ningbo Entry-exit Inspection and Quarantine Bureau, China, almost all wooden packaging imported through this harbour have been sampled and inspected since 1997. In recent years, the quarantine pest B. xylophilus was detected many times in large numbers (sometimes thousands of specimens) in wood samples from different countries, and a considerable number of other Bursaphelenchus species, among them several undescribed species, were also found. The morphological and molecular diagnostic work was stimulated and supported by a training course for identification of Bursaphelenchus species given by Helen Braasch and Wolfgang Burgermeister in Shanghai, China, in October 2002. From January, 2003 to June, 2006, wooden packaging material of 8720 batches was nematologically inspected in the Ningbo Entry-exit Inspection and Quarantine Bureau. Living nematodes were detected in 1772 batches, accounting for 20.3%. Most of them were identified as Rhabditida, Tylenchida and Aphelenchida (previously Aphelenchoides spp. and Bursaphelenchus spp., but also Aphelenchus spp., Paraphelenchus sp., Cryptaphelenchus spp. and Ruehmaphelenchus sp.). Bursaphelenchus spp. was detected in 343 batches from 26 different countries (Table 1). The following species were identified on the basis of their morphology and their ITSRFLP patterns: B. xylophilus, B. fungivorus, B. rainulfi, B. hylobianum, B. thailandae, B mucronatus, B. aberrans, B. lini, B. singaporensis, B. doui, B. conicaudatus,

Variation in ITS and 28S rDNA of Bursaphelenchus Species (Nematoda: Parasitaphelenchidae)

Internal transcribed spacers and 28S D2/D3 domain of rDNA were used to infer the phylogenetic relationships among Bursaphelenchus species, with special emphasis on members of the xylophilus and fungivorus groups. Sequence alignments and phylogenetic analysis using neighbour-joining and maximum parsimony algorithms resulted in trees with similar topologies. The 17 Bursaphelenchus species examined can be separated into two main branches: the first includes members of the xylophilus group and the second includes the species of the fungivorus group, separated from the remaining species B. eremus, B. hofmanni, B. rainulfi and B. yongensis. As far as known, the species of the xylophilus group are phoretically associated with longhorn beetles. The phylogenetic analysis revealed the digger bee associated B. abruptus as the basal taxon of the species investigated or at least of the xylophilus group. The remainder of the species are, as far as known, associated with bark beetles and a soil-dwelling bee in case of B. seani. The significantly supported groups are largely

Redescription of Bursaphelenchus eucarpus Rühm, 1956 (Nematoda: Aphelenchoididae) associated with the apple bark beetle, Scolytus mali Bechstein, and the shothole borer, S. rugulosus Müller

New morphological, molecular and bionomic data are added to update the description of Bursaphelenchus eucarpus. The nematode was isolated from larval galleries and adults of the apple bark beetle, Scolytus mali, and the shothole borer, S. rugulosus, present under the bark of the bird cherry, Prunus padus, and the common European plum, P. domestica, in Poland. Detailed male, female and dispersal juvenile morphology and morphometrics are documented and compared with the original description. From partial 18S, ITS1/2 and partial 28S D2-D3 sequences and morphological features (three lateral lines, position of caudal papillae, shape of the delicate spicules with dorsally bent condylus, lack of vulval flap, a long post-uterine branch and a hyaline region at the female tail tip) it can be concluded that B. eucarpus belongs to the eremus group of Bursaphelenchus. The differentiation from related species of the eremus group is discussed.

Bursaphelenchus moensi n. sp. (Tylenchina: Aphelenchoididae) found in packaging wood from the USA

Bursaphelenchus moensi n. sp., isolated in Tianjin Port, China, from Pinus packaging wood imported from the USA, is described. It is characterised by a lateral field with three lines (two bands), stylet 13.0-14.9 μ m long, excretory pore situated at same level as, or slightly posterior to, the nerve ring, spicules 13.0-15.5 μ m long, mitten-shaped, condylus high and broad with squared, rounded or pointed end, rostrum triangular or conical with bluntly rounded tip, and cucullus absent, bursa small, starting posterior to P4, narrow oblong to irregular in shape and with an oval central projection, female vulval flap absent, and tail conical with a finely rounded or mucronate terminus. The new species belongs to the hofmanni -group and is most similar to B. paraparvispicularis , B. parvispicularis and B. wuae . It is distinguished from closely related species by morphology, ITS-RFLP patterns and partial 18S, ITS and 28S D2-D3 rDNA gene sequencing results.

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.

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) μ m in female and 638 (475-789) μ m 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 μ m 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 rockyi n. sp. (Nematoda: Aphelenchoididae) in peat moss from Russia

Bursaphelenchus rockyi n. sp., isolated in Peking, China, from peat moss imported from Russia, is described. It is characterised by a lateral field with four lines, excretory pore located at the base of the nerve ring or slightly posterior, spicule 15.9 (15.1-17.3) μm long along the chord and with high condylus and a rounded tip, rostrum triangular or conical with bluntly pointed tip, lamina curvature becoming more pronounced at 60% of total length, six caudal papillae with P3 and P4 adjacent to each other, bursa small, starting posterior to P4, vulval lips hemispherical and protruding, and tail conical with finely rounded or mucronated terminus. The new species belongs to the fungivorus-group and is most similar to B. arthuri, B. arthuroides, B. fungivorus and B. seani, but can be distinguished from related species by morphological and sequencing results.

Radopholoides japonicus n. sp. (Nematoda: Pratylenchidae) found in rhizosphere soil associated with Podocarpus macrophyllus from Japan

Radopholoides japonicus n. sp. isolated from rhizospheric soil samples associated with Podocarpus macrophyllus imported from Japan into Ningbo Port of China is described and illustrated. It is characterised by four lateral lines, lip region low, convex with three annuli. Stylet 16.5-18.5 μ m long with anteriorly sloping rounded knobs. Excretory pore at the level of pharyngo-intestinal junction. Vulva posteriorly located, vulval lips not protruding, spermatheca squarish. Post-vulval uterine sac 35-44 μ m long. Tail conoid with broadly rounded terminus, phasmid 6-8 annuli posterior to anus. Among five species of genus Radopholoides , the new species is close to R. antoni but can be differentiated from it by lip and tail morphology. This is the first Radopholoides species that provided detailed morphological and molecular characterisation. Maximum Likelihood analysis using the Shimodaira-Hasegawa test for the validity of Radopholoides was performed and showed the validity of the genus using the partial 18S and D2-D3 expansion segments of 28S.