Molecular phylogeny, diagnostics, and diversity of plant-parasitic nematodes of the genus Hemicycliophora (Nematoda: Hemicycliophoridae)

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.Fil: Subbotin, Sergei A.. California Department of Food and Agriculture; Estados Unidos. Institute of Ecology and Evolution of the Russian Academy of Sciences; RusiaFil: Chitambar, John J.. California Department of Food and Agriculture; Estados UnidosFil: Chizhov, Vladimir N.. Institute of Ecology and Evolution of the Russian Academy of Sciences; RusiaFil: Stanley, Jason D.. Florida Department of Agriculture and Consumer Services; Estados UnidosFil: Inserra, Renato N.. Florida Department of Agriculture and Consumer Services; Estados UnidosFil: Doucet, Marcelo Edmundo. Universidad Nacional de Cordoba. Facultad de Cs.exactas Fisicas y Naturales. Centro de Zoologia Aplicada; Argentina. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnologico Cordoba. Instituto de Diversidad y Ecologia Animal; ArgentinaFil: Mcclure, Michael. University Of Arizona; Estados UnidosFil: Ye, Weimin. North Carolina Department of Agriculture & Consumer Services; Estados UnidosFil: Yeates, George.Fil: Mollov, Dimitre S.. University Of Minnesota; Estados UnidosFil: Cantalapiedra Navarrete, Carolina. Consejo Superior de Investigaciones Científicas. Instituto de Agricultura Sostenible; EspañaFil: Vovlas, Nicola. Istituto per la Protezione delle Piante; ItaliaFil: Van Den Berg, Esther. ARC-Plant Protection Research Institute; SudáfricaFil: Castillo, Pablo. Consejo Superior de Investigaciones Científicas. Instituto de Agricultura Sostenible; Españ

Molecular and morphological characterisation of Xiphinema americanum-group species (Nematoda: Dorylaimida) from California, USA, and other regions, and co-evolution of bacteria from the genus Candidatus Xiphinematobacter with nematodes

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

Morphological and molecular characterisation of two new Hemicycliophora species (Tylenchida: Hemicycliophoridae) with a revision of the taxonomic status of some known species and a phylogeny of the genus

Hemicycliophora presently contains 132 valid species of sheath nematodes. Within several samples obtained from surveys in Canada, South Africa, Spain and the USA, we distinguished six valid and six putative unidentified species by integrating the results of morphological and molecular analyses. Valid species included: H. californica, H. gracilis, H. parvana, H. poranga, H. raskii, and H. signata. The putative unidentified species were indicated as Hemicycliophora sp. 10, sp. 12, sp. 15, sp. 16, sp. 17, and sp. 18. Two new species of sheath nematodes from Spain and the USA were described and named as H. onubensis sp. n. and H. robbinsi sp. n., respectively. Hemicycliophora wyei is proposed as a junior synonym of H. parvana and H. ripa is proposed as a junior synonym of H. poranga. Eighteen valid and 13 unidentified species of sheath nematodes were characterised using the partial COI mtDNA gene. A total of 94 new sequences of which 77 were for the COI mtDNA gene were obtained in this study. Phylogenetic relationships within Hemicycliophora, using the D2-D3 expansion segments of 28S rDNA, ITS rRNA and COI gene sequences, are presented as inferred from Bayesian analysis.The sixth author thanks financial support from grant 201740E042, ‘Análisis de diversidad molecular, barcoding, y relaciones filogenéticas de nematodos fitoparásitos en cultivos mediterráneos’ from the Spanish National Research Council (CSIC).Peer reviewe

Caracterización morfológica y molecular de nematodes picadores de la raíz (Longidorus spp.) sobre palmera datilera en Arizona, California y Florida

Trabajo presentado en la 47th Annual Meeting of the Organization of Nematologists of Tropical America, celebrada en Varadero (Cuba) del 17 al 22 de mayo de 2015.For decades, ornamental growers in Florida have imported date palms (Phoenix dactylifera) from Arizona and California. Needle nematodes (Longidorus spp.) with conoid and hemispherical tails have been detected consistently in these imported palms in Florida. The populations with conoid tails have been identified routinely as Longidorus africanus and those with hemispherical tails as L. belloi, L. belondiroides, and L. orientalis. Recent molecular and morphological analyses have confirmed the identification of L. africanus and also that of L. orientalis, reported by R. Esser, in Florida, in 1995 on date palms from California. These findings disprove the reports of detection of L. belloi and L. belondiroides, not found again, and validate that of L. orientalisas a continental record in the United States and the Americas. Longidorus africanus is a known parasite of vegetables in Southern California, whereas L. orientalis is a parasite of date palm in the Middle East. The populations of L. orientaliscontained a few males, not reported in the original description. Longidorus orientalis was commonly found in association with L. africanus and was able to survive for at least four years, at low densities in warm and humid environments of Florida, on transplanted date palms imported from California and Arizona. Phylogenetic relationships of L. orientalis with closely related Longidorus species were reconstructed using D2-D3 of 28S rRNA, ITS1 rRNA, and partial coxI gene sequences. The PCR-D2-D3 of 28S rDNA-RFLP diagnostic profile was obtained and proposed for the identification of this species. Longidorus orientalis shows high intraspecific variation (up to 15.5%) in coxI mtDNA sequences. Incongruence between ITS1 rRNA and coxImtDNA gene trees was noticed from the analysis of phylogenetic relationships between L. orientalispopulations, indicating selective introgression of mtDNA through gene flow as a consequence of hybridization of populations from different origin.N

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

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.Peer reviewe