Characterisation of populations of Longidorus orientalis Loof, 1982 (Nematoda: Dorylaimida) from date palm (Phoenix dactylifera L.) in the USA and other countries and incongruence of phylogenies inferred from ITS1 rRNA and coxI genes

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Needle nematode populations of Longidorus orientalis associated 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 of L. orientalis contained a small number of males that were not reported in the original description and are herein described. Longidorus orientalis was 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 of L. orientalis with L. africanus was observed in all of the surveyed sites, indicating that date palm is a host of both nematodes. Phylogenetic relationships of L. orientalis with closely related Longidorus species, in addition to relationships between populations of L. orientalis from the USA, Greece, Iran and Spain, were inferred from the analyses of D2-D3 of 28S rRNA, ITS1 rRNA and partial coxI gene sequences. The PCR-D2-D3 expansion segments of 28S rDNA-RFLP diagnostic profile is provided. Longidorus orientalis populations display a high level of intraspecific variation (up to 15.5%) in coxI mtDNA sequences. Analysis of phylogenetic relationships of nematode populations revealed incongruence of the ITS1 rRNA and coxI mtDNA 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.

Characterisation of populations of Longidorus orientalis Loof, 1982 (Nematoda: Dorylaimida) from date palm (Phoenix dactylifera L.) in the USA and other countries and incongruence of phylogenies inferred from ITS1 rRNA and coxI genes

in Nematology



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    Light micrographs of Longidorus orientalis female from California. A-C: Anterior region; D, E: Lateral and ventral view of vulval region; F, G: Tail shape variation. (Scale bar: A, D, E = 11 μm; B, C = 5.5 μm; F, G = 12 μm.)

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    Light micrographs of Longidorus orientalis female and male from Arizona. A: Portion of female gonad showing ovary (ov), a pear-like spermatheca (spe), sphincter (sph) and distal portion of uterus (U); B: Posterior body region of male showing spicules (sp); C: Middle body portion of male showing junction of both testes with vas deferens (Jt); D: Posterior body portion of males showing spicules (sp) and adcloacal (as) and four ventro-median pairs of supplements (s) arranged in a staggered row. (Scale bar = 20 μm.)

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    PCR-D2-D3-28S-RFLP diagnostic profile for Longidorus orientalis. M: 100 bp DNA marker (Promega, USA); 1: AluI; 2: HinfI; 3: Bsp143I; 4: Tru1I; 5: RsaI.

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    Phylogenetic relationships of Longidorus orientalis with other related species: Bayesian majority rule consensus tree reconstructed under the GTR + I + G model and inferred from the D2-D3 of 28S rRNA gene sequence.

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    Putative secondary structures of the D3 expansion segment of 28S rRNA gene for longidorid nematodes. Stem-loop structure codes are given according to He et al. (2005b).

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    Putative secondary structures of the ITS1 for Longidorus orientalis. A: Putative secondary structure of the ITS1 with indication of a longest stem-loop structure; B: Alignment of sequences of a longest stem-loop structure with insertions/deletions; C: Putative secondary structure of a longest ITS1 stem-loop reconstructed for nematodes from the USA with indication of the position of insertions by arrows; D: Putative secondary structure types (II-IV) of insertions for sequences obtained from nematodes collected from other geographical regions.

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    Phylogenetic relationships between Longidorus orientalis populations: Bayesian majority rule consensus tree reconstructed under the GTR + I + G model and inferred from the ITS1 rRNA gene sequences (A) and partial the coxI gene sequences (B). This figure is published in colour in the online edition of this journal, which can be accessed via

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    Statistical parsimony network showing the phylogenetic relationships between coxI haplotypes of Longidorus orientalis. Small black cycles represent missing haplotypes. Pie chart sizes are proportional to the number of samples with a particular haplotype.

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