The Xiphinema americanum-group is a large species complex containing more than 50 nematode species. They are economically important because they are vectors of nepoviruses. The species differentiation of X. americanum-group is problematic because the species share similar morphological characters. In the present study we collected nematode samples from different locations in the USA, Italy and Russia. Six valid species, X. americanum s. str., X. brevicolle, X. californicum, X. pachtaicum, X. rivesi and X. simile, and four unidentified putative Xiphinema species were characterised by morphology and sequencing of D2-D3 of 28S rRNA, ITS1 rRNA and mitochondrial COI genes. New nematode sequences generated totalled 147. Phylogenetic relationships of the X. americanum-group species reconstructed by Bayesian inference for D2-D3 of 28S rRNA gene sequences did not provide clear species delimitation of the samples studied, although the mtDNA presented interspecific variations useful for demarcation among species. Xiphinema americanum s. str., X. californicum, X. pachtaicum, X. rivesi, and two unidentified Xiphinema species were found in 72 soil samples from California. We also reconstructed the phylogenetic relationships using partial 16S rRNA gene sequences within endosymbiotic bacteria of the genus Candidatus Xiphinematobacter and provided solid evidence for distinguishing 17 species of this genus based on the analysis of new and previously published sequences. Fifty-five new bacterial sequences were obtained in the present study and comparison of the bacterial 16S rRNA and nematode COI phylogenies revealed a high level of co-speciation events between host and symbiont.
The genus Hemicycliophora (Nematoda: Hemicycliophoridae) contains 132 valid species of plant‐parasitic nematodes, collectively known as ‘sheath nematodes’. Hemicycliophora spp. are characterized morphologically by a long stylet with rounded basal knobs and a cuticular sheath, present in juvenile and adult stages. Populations of 20 valid and 14 putative species of Hemicycliophora and Loofia from several countries were characterized morphologically using light (LM) and scanning electron microscopy (SEM) and molecularly using the D2‐D3 segments of 28S rRNA and internal transcribed spacer (ITS) rRNA gene sequences. LM and SEM observations provided new details on the morphology of these species. PCR‐restriction fragment length polymorphisms (PCR‐RFLPs) of the D2‐D3 of 28S rDNA were proposed for identification of the species. Phylogenetic relationships within populations of 36 species of the genus Hemicycliophora using 102 D2‐D3 of 28S rDNA and 97 ITS rRNA gene sequences as inferred from Bayesian analysis are reconstructed and discussed. Ancestral state reconstructions of diagnostic characters (body and stylet length, number of body annuli, shape of vulval lip and tail), using maximum parsimony and Bayesian inference, revealed that none of the traits are individually reliable characters for classifying the studied sheath nematode. The Shimodaira–Hasegawa test rejected the validity of the genus Loofia. This is the most complete phylogenetic analysis of Hemicycliophora species conducted so far. © 2014 The Linnean Society of London
Needle nematode populations ofLongidorus orientalisassociated with date palm,Phoenix dactylifera, and detected during nematode surveys conducted in Arizona, California and Florida, USA, were characterised morphologically and molecularly. The nematode species most likely arrived in California a century ago with propagative date palms from the Middle East and eventually spread to Florida on ornamental date palms that were shipped from Arizona and California. This is the first validated continental record of this needle nematode species in the USA and the Americas. The USA populations ofL. orientaliscontained a small number of males that were not reported in the original description and are herein described.Longidorus orientaliswas able to survive for at least 4 years at very low numbers in the warm and humid environment of Florida on date palms imported from California and Arizona. Association ofL. orientaliswithL. africanuswas observed in all of the surveyed sites, indicating that date palm is a host of both nematodes. Phylogenetic relationships ofL. orientaliswith closely relatedLongidorusspecies, in addition to relationships between populations ofL. orientalisfrom the USA, Greece, Iran and Spain, were inferred from the analyses of D2-D3 of 28S rRNA, ITS1 rRNA and partialcoxIgene sequences. The PCR-D2-D3 expansion segments of 28S rDNA-RFLP diagnostic profile is provided.Longidorus orientalispopulations display a high level of intraspecific variation (up to 15.5%) incoxImtDNA sequences. Analysis of phylogenetic relationships of nematode populations revealed incongruence of the ITS1 rRNA andcoxImtDNA gene trees, which might be the result of selective introgression of mtDNA through gene flow between previously isolated populations introduced simultaneously into new geographical regions.
The cystoid sedentary nematodes of the Heteroderidae include more than 30 recognised species belonging to from nine to 11 genera, depending on the opinion of the authority. In the present study, we collected nematode populations of species belonging to some of these genera, as well as other cyst-forming species from different locations in the USA, Vietnam, Germany and Russia. The populations of cystoid nematodes represent seven valid species, Atalodera carolynae, Cryphodera sinensis, Meloidodera astonei, M. floridensis, M. mexicana, M. sikhotealiniensis, Rhizonemella sequoiae, two unidentified species of Atalodera, six unidentified species of Cryphodera, and three putative new species of Rhizonemella. We also obtained samples of cyst-forming nematodes that include an unidentified species of Betulodera from California, and Heterodera guangdongensis from Vietnam. A population of Rotylenchulus sp. from Arizona, intercepted in Florida, and a population of an unidentified species of Verutus from Germany were also added to this study. All of these populations were characterised using sequences of the D2-D3 of 28S rRNA, ITS rRNA or mitochondrial COI genes. A total of 89 new sequences were obtained from these analyses. Phylogenetic relationships within the family Heteroderidae were reconstructed based on the D2-D3 of 28S rRNA, ITS rRNA and mitochondrial COI gene sequences. Results revealed that the subfamily Ataloderinae was paraphyletic. Ekphymatodera thomasoni, a non-cyst-forming species, clustered with the cyst-forming nematodes. Representatives of the subfamily Verutinae formed clades within Heteroderidae. The genus Meloidodera was non-monophyletic and distributed within two clades: i) M. sikhotealiniensis and Cryphodera spp. from Asia and Europe; and ii) M. astonei, M. floridensis and M. mexicana from North America. Based on comparative molecular analysis Meloidodera alni syn. n. is proposed as a junior synonym of M. sikhotealiniensis. Problems of taxonomy and phylogeography of cystoid nematodes are also discussed.
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