Description and molecular characterisation of Paralongidorus litoralis sp. n. and P. paramaximus Heyns, 1965 (Nematoda: Longidoridae) from Spain

Nematology ◽  
2008 ◽  
Vol 10 (1) ◽  
pp. 87-101 ◽  
Author(s):  
Juan E. Palomares-Rius ◽  
Sergei A. Subbotin ◽  
Blanca B. Landa ◽  
Nicola Vovlas ◽  
Pablo Castillo

Abstract Paralongidorus litoralis sp. n., a new bisexual species of the genus, is described and illustrated by light microscopy, scanning electron microscopy and molecular studies from specimens collected in a coastal sand dune soil around roots of lentisc (Pistacia lentiscus L.) from Zahara de los Atunes (Cadiz), southern Spain. Paralongidorus litoralis sp. n. is characterised by the large body size (7.5-10.0 mm), a rounded lip region, clearly offset from the body by a collar-like constriction, and bearing a very large stirrup-shaped, amphidial fovea, with conspicuous slit-like aperture, a very long and flexible odontostyle ca 190 μm long, guiding ring located at 35 μm from anterior end, and males with spicules ca 70 μm long. In addition, identification data of a Spanish population of P. paramaximus Heyns, 1965 recovered from sandy soil of a commercial citrus orchard at Alcala de Guadaira (Seville), southern Spain, agree very well with the original description of the species from South Africa. The 18S rRNA and D2 and D3 expansion regions of 28S rRNA gene sequences were obtained for P. litoralis sp. n. and P. paramaximus. Phylogenetic analyses of P. litoralis sp. n. and P. paramaximus rRNA gene sequences and of Longidoridae sequences published in GenBank were done using maximum likelihood and Bayesian inference. In trees generated from the 18S data set Paralongidorus clustered as an external clade from Longidorus, and in trees generated from D2-D3 of 28S dataset Paralongidorus was monophyletic and nested within Longidorus. Maximum likelihood test supported the hypothesis of validity of the Paralongidorus genus.

Nematology ◽  
2012 ◽  
Vol 14 (4) ◽  
pp. 427-443 ◽  
Author(s):  
Majid Pedram ◽  
Ebrahim Pourjam ◽  
Somayeh Namjou ◽  
Mohammad Reza Atighi ◽  
Carolina Cantalapiedra-Navarrete ◽  
...  

Paralongidorus iranicus n. sp., a new bisexual species of the genus, is described and illustrated by light microscopy, scanning electron microscopy and molecular studies from specimens collected in the rhizosphere of Scots pine (Pinus sylvestris) from the Caspian (Khazar) seashore, Nour, northern Iran. Paralongidorus iranicus n. sp. is characterised by the large body size (7.8-11.4 mm), a rounded lip region, clearly set off by a collar-like constriction at level of, or slightly posterior to, the amphidial aperture, and bearing a very large, stirrup-shaped, amphidial fovea, with conspicuous slit-like aperture, a very long and flexible odontostyle ca 170 μm long, guiding ring located at 34 μm from anterior end and males with spicules ca 80 μm long. In addition, data from an Iranian population of P. bikanerensis recovered from the rhizosphere of palm (Phoenix dactylifera L.) in Bam, Kerman province, south-eastern Iran, agree very well and are very close to the original description of the species from India. The D2 and D3 expansion regions of 28S rRNA gene, ITS1, and 18S rRNA sequences were obtained for P. iranicus n. sp. and P. bikanerensis. Phylogenetic analyses of P. iranicus n. sp. and P. bikanerensis rRNA gene sequences and of Longidorus spp. sequences published in GenBank were done using Maximum Likelihood and Bayesian inference. Paralongidorus species (including P. iranicus n. sp.) clustered together; however, P. bikanerensis clustered within Longidorus spp. and was clearly separated from all other Paralongidorus spp. in trees generated from the D2-D3 expansion segments of 28S and partial 18S data set, respectively.


Nematology ◽  
2017 ◽  
Vol 19 (3) ◽  
pp. 333-350 ◽  
Author(s):  
Shigeyuki Sekimoto ◽  
Taketo Uehara ◽  
Takayuki Mizukubo

The Korean cyst nematode, Heterodera koreana, was recorded for the first time from Japan and characterised morphologically, morphometrically and molecularly. In total, 41 populations were detected from soil samples collected from the rhizosphere of four bamboo species in Japan: 31 populations from moso bamboo (Phyllostachys edulis), seven from madake (P. bambusoides), two from henon bamboo (P. nigra var. henonis) and one from fish pole bamboo (P. aurea). The morphology and morphometrics of the Japanese population were in agreement with those of the original description of H. koreana from South Korea and other subsequent descriptions from China and Iran, with the exception of some minor differences. The results of the phylogenetic analyses of the D2-D3 expansion segments of 28S rRNA gene and ITS rRNA gene sequences confirmed the species identification and phylogenetic relationship of H. koreana with other Heterodera species. The COI mtDNA gene sequences were obtained for the first time for H. koreana. Three COI haplotypes found in Japanese H. koreana populations showed a characteristic geographical distribution in Japan.


Nematology ◽  
2013 ◽  
Vol 15 (3) ◽  
pp. 363-378 ◽  
Author(s):  
Juan E. Palomares-Rius ◽  
Carolina Cantalapiedra-Navarrete ◽  
Carlos Gutiérrez-Gutiérrez ◽  
Gracia Liébanas ◽  
Pablo Castillo

Paralongidorus plesioepimikis n. sp. is described and illustrated by light microscopy, scanning electron microscopy and molecular studies from specimens collected in a sandy soil in the rhizosphere of stone pine (Pinus pinea) in Bonares (Huelva Province), southern Spain. The new species is characterised by a very long body (9.71-14.11 mm), an expanded and rounded lip region, with a clear constriction followed by a depression posterior to the amphidial aperture, a very long and flexible odontostyle (202-227 μm), guiding ring located at 36.5-44.0 μm from anterior end, dorsal pharyngeal gland nucleus in anterior part of bulb, one subventral pair of nuclei near middle of bulb, vulva at 33-38%, and a dorsally convex tail with rounded terminus (32-39 μm long). SEM observations showed a very large amphidial fovea with conspicuous aperture ca three-fourths as wide as lip region and cephalic papillae appearing as small apertures, each located just anterior to a distinct cephalic lobe. The sequences of the D2-D3 expansion segments of 28S rRNA, partial 18S rRNA and ITS rRNA gene for P. plesioepimikis n. sp. were obtained. Phylogenetic analyses of P. plesioepimikis n. sp. rRNA gene sequences and of Paralongidorus spp. and Longidorus spp. sequences published in GenBank were done using Maximum Likelihood and Bayesian inference. Paralongidorus species (including P. plesioepimikis n. sp.) clustered together, except for P. bikanerensis which clustered within Longidorus spp. and was clearly separated from all other Paralongidorus spp. in trees generated from the D2-D3 expansion segments of 28S and partial 18S data set. ML analysis using SH-test for the validity of Paralongidorus was performed and showed the validity of the genus using the D2-D3 expansion segment of 28S and partial 18S.


Plant Disease ◽  
2012 ◽  
Vol 96 (11) ◽  
pp. 1707-1707 ◽  
Author(s):  
D. S. Mollov ◽  
S. A. Subbotin ◽  
Carl Rosen

In the summer of 2011, two independent garlic samples from Morrison and Dakota counties and in 2012 one garlic sample from Carver county in Minnesota were submitted by commercial growers to the University of Minnesota Plant Disease Clinic for disease analyses. Symptoms of the above-ground plant parts were stunting and chlorosis. Symptoms of bulbs were necrosis, underdevelopment, and distortion. Upon microscopic examination, phytonematodes exuded into the surrounding water droplet. Nematodes were present in the protective leaves, abscission zone, and cloves in all submitted bulbs (n = 18) for analyses. Morphometric examination of females, males, and juveniles determined that they were Ditylenchus dipsaci. Nematodes extracted from garlic cloves were fixed in TAF (97 ml formalin [40%], 2 ml triethanolamine, and 91 ml dH2O). Morphological observations and measurements were made under an Olympus BX51 microscope equipped with a Nomarski differential interference contrast. Female (n = 6) measurements were: L = 1.411 to 1.636 mm, a = 38 to 44, b = 5.8 to 8.0, c = 14 to 17, stylet = 11.5 to 12.3 μm, V = 79 to 81%, and tail = 95 to 105 μm. The body was almost straight, when heat relaxed, lip region flattened, median bulb oval, and isthmus elongate and slender. The basal pharyngeal bulb overlapped the intestine. The post-vulval uterine branch was about half of vulva-anus distance. The tail was conoid with a pointed terminus. Male (n = 9) measurements were: L = 1.372 to 1.558 mm, a = 40 to 50, b = 6.5 to 7.0, c = 14 to 16, stylet = 11.5 to 12.3 μm, spicules = 22 to 27 μm, and gubernaculum = 9 to 10 μm. The bursa was leptoderan and spicules were curved with simple gubernaculum. Morphology and morphometrics of females and males of D. dipsaci from Minnesota generally fit the descriptions provided for the type and other populations by Hopper (1) and other authors. Several specimens were also taken for molecular identification. DNA extraction, PCR, and sequencing protocols were as described by Subbotin et al. (2). The TW81 and AB28 primers were used for amplification of ITS-rRNA region and the D2A and D3B primers were used for amplification of the D2-D3 expansion segments of 28S rRNA gene. Comparison of the ITS and D2-D3 of 28 rRNA gene sequences showed 100 and 99% identity with corresponding gene sequences of D. dipsaci published in the GenBank (2). The sequences were submitted in the GenBank under accession numbers JX123258 and X123259. This nematode problem has not been known to occur in either of these locations previously. The most likely source of introduction of D. dipsaci are imported garlic seed bulbs. To our knowledge, this is the first report of D. dipsaci affecting garlic or any other crops in Minnesota. The garlic produced in these locations was considered unmarketable and complete loss to the farmers. The presence of D. dipsaci could have a significant economic impact in the emerging multi-million dollar garlic industry in Minnesota. References: (1) D. J. Hooper. Ditylenchus dipsaci. CIH Descriptions of Plant-Parasitic Nematodes Set 1, No. 14, 1972. (2) S. A. Subbotin et al. Phytopathology 95:1308, 2005.


Zootaxa ◽  
2020 ◽  
Vol 4819 (2) ◽  
pp. 295-315
Author(s):  
HIROSHI KAJIHARA

The heteronemertean Cerebratulus orochi sp. nov. is described based on material collected intertidally at a muddy beach in Akkeshi, northern Japan. For the last 80 years, the species has been confused with Cerebratulus marginatus Renier, 1804; the latter was originally described from the Adriatic and once believed to occur in many places in the northern hemisphere including Japan. Cerebratulus orochi sp. nov. is morphologically different from all the congeners including C. marginatus by the following combination of characters: several layers of diagonal-muscle meshwork coated with connective tissue, proximo-distally distributed in cross section from the distal portion of the body-wall outer longitudinal muscle layer to the cutis-gland zone throughout the anterior portion of the body from the precerebral to the foregut regions; the cephalic vascular system consisting of lateral and mid-dorsal vessels; and the sub-rhynchocoelic vessel possessing a pair of antero-lateral diverticula before the former forks posteriorly into a pair of lower lateral vessels in the post-cerebral, pre-oral region. Previous records of C. marginatus from Japanese waters are no longer considered to be substantiated. Multi-locus phylogenetic analyses based on the mitochondrial 16S rRNA and cytochrome c oxidase subunit I (COI), as well as the nuclear 18S rRNA, 28S rRNA, and histone H3 genes among heteronemerteans comprising the “Cerebratulus clade” indicated that C. orochi sp. nov. was closely related to C. cf. marginatus from the US Pacific coast. A MegaBLAST search at the NCBI website with the 16S rRNA gene sequence from C. orochi sp. nov. followed by a couple of species delimitation analyses suggests that larvae of the species are also distributed in Vostok Bay, Far East Russia.


Nematology ◽  
2017 ◽  
Vol 19 (6) ◽  
pp. 681-695 ◽  
Author(s):  
László Barsi ◽  
Francesca De Luca

Paralongidorus francolambertii sp. n., a bisexual species found in the rhizosphere of silver lime (Tilia tomentosa) and common juniper (Juniperus communis), is described. The species is characterised by its medium to large size (L = 5.86-8.29 mm) and slender body (a = 143-197), a lip region flattened with rounded profile, clearly offset from the body by a deep constriction, a narrow neck between the head and body, a shoulder-like body posterior to the neck, a stirrup-shaped amphidial fovea, with conspicuous slit-like aperture, lying on the lateral cuticular collar, a moderately long odontostyle ca 140 μm long, a guide ring located at ca 28 μm from anterior end, a tail terminus with thickened outer cuticular layer in both sexes, and males with spicules ca 52 μm long. The D2-D3 expansion domains of the 28S rRNA gene and the ITS-containing region of P. francolambertii sp. n. were amplified and sequenced. Phylogenetic analyses by using the Maximum Likelihood method showed that P. francolambertii sp. n. had a sister relationship with P. rex and that all Paralongidorus species formed a well-supported group.


Nematology ◽  
2021 ◽  
pp. 1-15
Author(s):  
Ignacio Cid del Prado Vera ◽  
Sergei A. Subbotin ◽  
Wilfrida Decraemer

Summary A new species of stubby root nematode, Trichodorus lownsberyi sp. n., collected from soil around Buxus sempervirens at Montecillo Campus, Colegio de Postgraduados, Mexico, is described and illustrated. Trichodorus lownsberyi sp. n. is characterised by the abundant males with a characteristic spicule shape, the narrower part about mid-blade having visible bristles in most specimens, and the presence and position of the three ventromedian cervical papillae, which are all anterior to the secretory-excretory pore. Females possess a rhomboid-shaped vagina and oval to rounded triangular vaginal sclerotised pieces with a pore-like vulva. The phylogenetic relationships of T. lownsberyi sp. n. with related species were constructed using the ITS2 rRNA and the D2-D3 expansion segments of 28S rRNA gene sequences. Trichodorus lownsberyi sp. n. was a sister species to T. viruliferus in the phylogenetic trees. A Mexican population of Nanidorus minor from a peach orchard is also described and illustrated. The new needle nematode, Longidorus quercus sp. n., was recovered from soil around roots of oak, Quercus crassipes, from Cerro Jusda ‘El Diablo’ in Mexico State. Longidorus quercus sp. n. females are characterised by the C-shaped posterior end of the body after fixation, L = 4.9 (3.9-5.6) mm, a = 71 (60.8-93.2), lip region rounded, sometimes slightly flattened, marked by depression, odontostylet 170 (144-206) μm long, hemispherical to bluntly conoid tail and no males. The new species was characterised using the D2-D3 of 28S rRNA and COI gene sequences. The phylogenetic relationships of L. quercus sp. n. with other Longidorus species were reconstructed using the D2-D3 of 28S rRNA gene sequences.


Nematology ◽  
2011 ◽  
Vol 13 (7) ◽  
pp. 773-785 ◽  
Author(s):  
Sergei A. Subbotin ◽  
Abbas Mohammad Deimi ◽  
Jingwu Zheng ◽  
Vladimir N. Chizhov

Abstract Seventy-eight ITS rRNA gene sequences obtained from the potato rot nematode, Ditylenchus destructor, collected across the world from different hosts were compared and analysed. The ITS rRNA gene sequences showed significant length variation between populations. The differences in this rRNA fragment length were due to the presence of repetitive elements in the ITS1, which were characterised by relatively higher rates of substitution changes. Reconstruction of secondary structure for the ITS1 revealed that these minisatellites formed a stem structure. Phylogenetic analyses of ITS rRNA and D2-D3 expansion segments of 28S rRNA gene sequences showed that all studied populations clustered in two major clades: a group of populations having the ITS sequences with the repetitive elements and a group of populations without the repetitive elements in the ITS. We propose to distinguish seven ITS rRNA haplotypes within potato rot nematode populations. PCR-ITS-RFLP diagnostic profiles are presented for these ITS haplotypes and usefulness of recently developed PCR methods with species-specific primers for D. destructor are analysed and discussed.


Nematology ◽  
2020 ◽  
Vol 22 (7) ◽  
pp. 815-824
Author(s):  
Sedighe Azimi ◽  
Majid Pedram

Summary A population of Hemicriconemoides phoenicis was recovered from Khuzestan province, south-western Iran, in association with date palm. The recovered population was characterised by 518-645 μm long females having a 76-82 μm long stylet, rounded to oblong spermatheca filled with sperm, a 28.0-39.8 μm long tail, juveniles common, with 14 longitudinal rows of rounded scales, and males absent. Compared to the original data, no morphological and morphometric differences were observed. In molecular phylogenetic analyses using the D2-D3 expansion segments of the 28S rRNA gene and a near-full-length fragment of the 18S rRNA gene sequences using Bayesian inference (BI) and maximal number of species of the genus, the two newly generated 28S sequences of the Iranian population formed a maximally supported clade with two original sequences of the species; and the 18S sequence formed a maximally supported clade with an unidentified isolate of the genus in the corresponding phylogeny. This is the second report of the species since its original description, Iran representing a new geographical record and supporting the suggestion that date palm could be its preferred host.


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