22 results on '"Castillo, Pablo"'
Search Results
2. Recombinase Polymerase Amplification assay for detection of the British root-knot nematode, Meloidogyne artiellia
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Subbotin Sergei A., Palomares-Rius Juan E., and Castillo Pablo
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chickpea ,diagnostics ,its rrna ,rpa ,species-specific primer ,Biology (General) ,QH301-705.5 - Abstract
Recombinase polymerase amplification (RPA) is an isothermal in vitro nucleic acid amplification technique that has been adopted for simple, robust, rapid, reliable diagnostics of nematodes. In this study, the real-time RPA assay and RPA assay combined with lateral flow dipsticks (LF-RPA) have been developed targeting the ITS rRNA gene of the British root-knot nematode, Meloidogyne artiellia. The assay provided specific and rapid detection of this root-knot nematode species from crude nematode extracts without a DNA extraction step with a sensitivity of 0.125 second-stage juvenile (J2) specimen per a reaction tube for real-time RPA during 11 min and a sensitivity of 0.5 J2 specimens per a reaction tube for LF-RPA during 25 min. The RPA assays were validated with a wide range of non-target root-knot nematodes. The LF-RPA assay has great potential for nematode diagnostics in the laboratory having minimal available equipment.
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- 2024
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3. Erratum to: Morphological and Molecular Characterization of Filenchus multistriatus n. sp. (Tylenchomorpha: Tylenchidae) and Data on a Known Species of the Genus from Bushehr Province, Southern Iran
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Monemi Somayeh, Atighi Mohammad Reza, Abolafia Joaquín, Castillo Pablo, and Pedram Majid
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Biology (General) ,QH301-705.5 - Published
- 2023
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4. Morphological and Molecular Characterization of Filenchus multistriatus n. sp. (Tylenchomorpha: Tylenchidae) and Data on a Known Species of the Genus from Bushehr Province, Southern Iran
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Monemi Somayeh, Atighi Mohammad Reza, Abolafia Joaquín, Castillo Pablo, and Pedram Majid
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filenchus sandneri ,lsu rdna d2-d3 ,new species ,ssu rdna ,taxonomy ,tylenchinae ,Biology (General) ,QH301-705.5 - Abstract
During a nematological survey in southern Iran, a population belonging to the family Tylenchidae was recovered from a tomato field in Bushehr province. The recovered population belongs to the genus Filenchus, was described and illustrated herein as F. multistriatus n. sp. It is mainly characterized by having a wide and low annulated lip region continuous with adjacent body; amphidial openings confined to the labial plate; four lines in lateral fields forming three bands, with the two outer bands broken by transverse, and the inner one broken by both transverse and longitudinal lines; and median bulb oval with visible valve and elongate-conoid tail uniformly and gradually narrowing toward the distal region, ending in a widely rounded tip. Its morphological and morphometric differences with three closely similar species were discussed. The phylogenetic relationships of the new species with other relevant genera and species were reconstructed using partial sequences of small, and large subunit ribosomal DNA (SSU and LSU rDNA) sequences. Morphometric and morphological data were also provided for an Iranian population of F. sandneri recovered from Bushehr province. Both populations were characterised using SEM data.
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- 2023
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5. Morphological and Molecular Characterization of Ottolenchus isfahanicus n. sp. (Tylenchomorpha: Tylenchoidea) from Rhizosphere of Gramineous Plants in Isfahan Province, Iran
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Mahboubi Zeinab, Keramat Mojtaba, Pourjam Ebrahim, Reza Atighi Mohammad, Clavero-Camacho Ilenia, Abolafia Joaquín, Castillo Pablo, and Pedram Majid
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new species ,ottolenchus facultativus ,phylogeny ,ssu rdna ,taxonomy ,Biology (General) ,QH301-705.5 - Abstract
A new species from the family Tylenchidae was recovered in the grasslands of Isfahan province, Iran, and is herein described based on morphological and molecular data. Ottolenchus isfahanicus n. sp. is mainly characterized by having a faintly annulated cuticle; elongated, slightly sigmoid amphidial apertures developed metacorpus with distinct valve under LM; vulva located at 69.472.3% of body length; large spermatheca about 2.75 times of corresponding body width; and elongated conoid tail with broadly rounded tip. SEM observations showed that the lip region is smooth; the amphidial apertures are elongated, slightly sigmoid slits; and the lateral field is a simple band. It is further characterized by 477-to-515-μm-long females with delicate 5.7-to-6.9-μm-long stylets with small, slightly posteriorly sloping knobs, as well as functional males, in the population. The new species closely resembles O. facultativus, but is separated from it based on morphological and molecular data. It was further morphologically compared with O. discrepans, O. fungivorus, and O. sinipersici. The phylogenetic relationships of the new species with other relevant genera and species were reconstructed using near-full-length sequences of small subunit and D2-D3 expansion segments of large subunit (SSU and LSU D2-D3). In the inferred SSU phylogeny, the newly generated sequence of Ottolenchus isfahanicus n. sp. formed a clade with two sequences of O. sinipersici and sequences assigned to O. facultativus and O. fungivorus. In the inferred LSU phylogeny, the three newly generated sequences of the new species and LSU sequences of O. sinipersici and O. discrepans formed a clade.
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- 2023
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6. First report of Longidorus leptocephalus Hooper, 1961 (Nematoda: Longidoridae) from Greece
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Clavero-Camacho Ilenia, Cantalapiedra-Navarrete Carolina, Kormpi Maria, Palomares-Rius Juan E., Tzortzakakis Emmanuel A., Castillo Pablo, and Archidona-Yuste Antonio
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cytochrome oxidase c subunit 1 ,d2-d3 of 28s rdna ,description ,its1 rdna ,needle nematodes ,taxonomy ,Biology (General) ,QH301-705.5 - Abstract
Longidorid nematodes comprise more than 500 species, and Longidorus and Xiphinema are the most diversified, prevalent, and cosmopolitan genera within plant-parasitic nematodes. The genus Longidorus comprise a group of species, some of which are vectors of plant viruses. New sampling for needle nematodes was carried out in a grapevine area in Thessaloniki, northern Greece, and one nematode species of Longidorus (L. leptocephalus) was recovered. Nematodes were extracted from soil samples by modified sieving and a decanting method. Extracted specimens were processed using glycerol, mounted on permanent slides, and subsequently identified morphologically. Nematode DNA was extracted from individual, live specimens, and PCR (Polymerase Chain Reaction) assays were performed for D2-D3 expansion segments of 28S rRNA, ITS1 rRNA, and partial mitochondrial COI regions. Morphology and morphometric data obtained from this population were consistent with the original description and reported populations of L. leptocephalus. To our knowledge, this is the first report of L. leptocephalus in Greece and the second in the Mediterranean Basin after the record of the species from Slovenia, extending the geographical distribution of this species in Europe.
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- 2022
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7. Morpho-Molecular and Ultrastructural Characterization of Discocriconemella parasinensis n. sp. from Zhejiang Province, China
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Li Junxia, Munawar Maria, Castillo Pablo, and Zheng Jingwu
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camellia sinensis ,dna sequencing ,morphology ,morphometrics ,nematode ,new record ,phylogeny ,species ,Biology (General) ,QH301-705.5 - Abstract
During a recent inventory survey of the nematofauna of tea plantation at Zhejiang Province, China, Discocriconemella parasinensis n. sp. was detected in the rhizosphere of Camellia sinensis. The new species can be characterized by having the uninterrupted rounded labial disc, en face view showing rectangular-rounded labial plate without submedian lobes, R = 82.6 (80–86), Rex = 22 (21–23), stylet length of 68.3 (59–76) μm, excretory pore located 1–2 annuli posterior to the esophageal bulb, vulva open, postvulval body elongated conoid, and tail conoid with bilobed terminus. Morphologically, the species shares the same lip-type with D. discolabia, D. mauritiensis, D. mineira, D. perseae, and D. sinensis. Phylogenetic relationships of the new species based on D2–D3 expansion segments of 28S, ITS, and 18S rRNA genes revealed that D. parasinensis n. sp. formed a separated clade from other criconematid species, thereby supporting its status as a new species of the genus. The new species showed close phylogenetic relationships with Criconemoides geraerti.
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- 2022
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8. First report of Xiphinema ifacolum Luc, 1961 (Dorylaimida: Longidoridae) from Nigeria
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Bello Tesleem T., Fabiyi Oluwatoyin A., Clavero-Camacho Ilenia, Cantalapiedra-Navarrete Carolina, Palomares-Rius Juan E., Castillo Pablo, and Archidona-Yuste Antonio
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africa ,cytochrome oxidase c subunit 1 ,d2-d3 expansion segments of 28s rrna gene ,dagger nematodes ,taxonomy ,Biology (General) ,QH301-705.5 - Abstract
A population of a species of dagger nematode (Xiphinema) belonging to the non-americanum group was recovered from the fields of kola nut (Cola sp.) in southern Nigeria. The morphological and morphometric data obtained from this population were consistent with the characteristics of the species Xiphinema ifacolum. In addition, molecular identification based on D2-D3 expansion segments of 28S rRNA and partial mitochondrial COI gene regions confirmed its identity. According to our knowledge, this is the first report of the species from Nigeria, and the second report from Africa, after the original description from Foulaya, Guinea.
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- 2022
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9. Dorylaimida Mundi (Nematoda)
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Castillo Pablo
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Biology (General) ,QH301-705.5 - Published
- 2022
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10. Integrative taxonomy of Xiphinema histriae and Xiphinema lapidosum from Spain
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Cai Ruihang, Archidona-Yuste Antonio, Cantalapiedra-Navarrete Carolina, Palomares-Rius Juan E., Zheng Jingwu, and Castillo Pablo
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first record ,juvenile stages ,molecular ,morphology ,morphometrics ,phylogeny ,xiphinema ,x. histriae ,x. lapidosum ,Biology (General) ,QH301-705.5 - Abstract
Three populations of Xiphinema non-americanum group species were detected in agricultural and natural ecosystems, during routine surveys for plant-parasitic nematodes in Spain. Based on morphological and molecular analyses, the species were identified as Xiphinema histriae and Xiphinema lapidosum, being this the first record and molecular characterization of both species in Spain. The morphometrics and morphology of the Spanish populations agree with those of the original description and paratype specimens and the present study provided a first description of the second to fourth juvenile stages of both species. A detailed study on the morphology in the Spanish populations of X. histriae, as well as in paratypes, showed a pseudo-Z-organ with weakly muscularized wall and containing numerous small dense granular bodies, which was different to the original description by Lamberti et al. (1993). This new finding suggests that X. histriae must be considered a member of the morphospecies Group 5 of X. non-americanum. Phylogenetic analysis based on D2 to D3 expansion segments of 28S gene, ITS1 and partial CoxI gene indicated that X. histriae and X. lapidosum are phylogenetically related with other Xiphinema non-americanum group spp. reported from Spain. Considering the pathological and economic importance of this group of nematodes, the combination of morphological characters, measurements, and molecular analysis is crucial for accurate identification of these species.
- Published
- 2019
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11. First report of cultivated Cretan mountain tea (Sideritis syriaca) as a host of Meloidogyne hapla and M. javanica in Crete, with some additional records on the occurrence of Meloidogyne species in Greece
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Tzortzakakis Emmanuel A., Cantalapiedra-Navarrete Carolina, Archidona-Yuste Antonio, Kormpi Maria, Palomares-Rius Juan E., and Castillo Pablo
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meloidogyne hispanica ,meloidogyne incognita ,corn ,soybean ,aloe ,Biology (General) ,QH301-705.5 - Abstract
Cultivated Cretan mountain tea or Malotira (Sideritis syriaca L.) was found to be infected by Meloidogyne hapla and Meloidogyne javanica in the island of Crete. The authors provide the first molecular characterization of M. hapla in Greece and the first report of Cretan mountain tea or Malotira as a host of Meloidogyne species worldwide. In addition, Meloidogyne hispanica was found infecting aloe (Andros island) and corn (Drama, North Greece) consisting the first reports of natural infection of these plants by M. hispanica in Europe. Furthermore, infection of corn by M. incognita and soybean by M. javanica (Drama, North Greece) are reported for the first time in Greece. Integrative taxonomical approach based on perineal pattern and EP/st ratio, as well as the region of the mitochondrial genome between the cytochrome oxidase subunit II (coxII) and 16S rRNA mitochondrial DNA (mtDNA) genes was used to differentiate Meloidogyne species.
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- 2019
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12. First Report of Xiphinema Ifacolum Luc, 1961 (Dorylaimida: Longidoridae) from Nigeria
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Bello, Tesleem T., Fabiyi, Oluwatoyin A., Clavero-Camacho, Ilenia, Cantalapiedra-Navarrete, C., Palomares-Rius, Juan E., Castillo, Pablo, and Archidona-Yuste, Antonio
- Subjects
Cytochrome oxidase c subunit 1 ,D2-D3 expansion segments of 28S rRNA gene ,Dagger nematodes ,Africa ,Taxonomy - Abstract
A population of a species of dagger nematode (Xiphinema) belonging to the non-americanum group was recovered from the fields of kola nut (Cola sp.) in southern Nigeria. The morphological and morphometric data obtained from this population were consistent with the characteristics of the species Xiphinema ifacolum. In addition, molecular identification based on D2-D3 expansion segments of 28S rRNA and partial mitochondrial COI gene regions confirmed its identity. According to our knowledge, this is the first report of the species from Nigeria, and the second report from Africa, after the original description from Foulaya, Guinea.
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- 2022
13. A new stunt nematode, Geocenamus chengi n. sp. (Nematoda: Merliniinae) in the rhizosphere of tea (Camellia sinensis) from Zhejiang Province, China.
- Author
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Maria, Munawar, Wentao Miao, Castillo, Pablo, and Jingwu Zheng
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TEA ,RHIZOSPHERE ,TEA plantations ,NEMATODES ,HOST plants ,TEA trade - Abstract
The tea plant is native to China; the country has the greatest tea production areas in the world. In an attempt to investigate the nematode biodiversity associated with the tea plantations of Hangzhou, Zhejiang Province, a population of stunt nematode was detected. This group of nematodes is comprised of migratory ecto-parasites of roots and can subsist on a variety of host plants. Therefore, the detected population was studied carefully using the integrative taxonomy approach and identified as a new species of genus Geocenamus. Geocenamus chengi n. sp. can be characterized by females having six incisures in the lateral field; labial region is dome shaped and slightly offset from the rest of the body having four to five annuli; head framework is weakly developed; deirids are absent; excretory pore is located at the anterior region of basal pharyngeal bulb. Under SEM, the vulva is a transverse slit, vulval lips are elongated and ellipsoidal with epiptygma. The tail is annulated, elongated, and conical having bluntly pointed tip and a terminal hyaline region that forms 21 to 33% of the tail length. Spicule is 22 to 25 μm long, gubernaculum is saucer shaped; bursa is crenated covering the tail until the hyaline tail region. Morphologically, the species is close to G. circellus, G. joctus, G. loofi, G. ordinarius, G. processus, G. tetyllus, and G. tortilis. Phylogenetic relationships of the new species based on D2-D3 expansion domains of 28 S, ITS, and 18 S rRNA genes indicated that G. chengi n. sp. clustered in a separate clade with G. vietnamensis. [ABSTRACT FROM AUTHOR]
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- 2020
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14. A new rare nematode Nothocriconemoides hangzhouensis n. sp. (Nematoda: Criconematidae) from Hangzhou, China.
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Maria, Munawar, Wentao Miao, Ruihang Cai, Castillo, Pablo, and Jingwu Zheng
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PLANT parasites ,CHLOROPLAST DNA ,VULVA - Abstract
The Family Criconematidae is commonly referred as ring nematodes that include some members with economic importance as plant parasites. During a recent nematode inventory survey at Zhejiang Province, China, a new species of genus Nothocriconemoides was detected in the rhizosphere of elm tree. Nothocriconemoides hangzhouensis n. sp. can be characterized by the female body having annuli with fine longitudinal striations and 2 to 3 anastomoses at the posterior half of the body. The first cephalic annulus is rounded and expanded enclosing the lip region, and the second annulus is narrow, offset, collar like. En face view shows a central elevated labial disk bearing four distinct equal-sized submedian lobes and “I†shaped oral aperture. Excretory pore is located 3–4 annuli posterior to esophageal bulb. Vagina is straight and vulva closed. The ventral side of postvulval annuli is inverted, in majority of individuals. Anus is indistinct and located on the next annuli posterior to vulva. Tail is short, conoid, with forked or branched terminus. Juveniles are devoid of collar-shaped annuli in the lip region. The cephalic region has two rounded annuli where the first annulus shows slight depression in the middle. Body annuli are finely crenated. Anus is indistinct and located 3 to 4 annuli from tail terminus. Tail is short ending in a single lobed terminus. Phylogenetic studies based on analysis of the D2–D3 expansion segments of the 28 S rRNA, ITS rRNA, partial 18 S rRNA, and coxI gene revealed that the new species formed a separate clade from other criconematid species, thereby supporting its status as a new species of the genus. The new species showed close relationships with Discocriconemella sinensis. Additionally, this is the first record of genus Nothocriconemoides from China. [ABSTRACT FROM AUTHOR]
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- 2020
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15. Integrative taxonomy of Xiphinema histriae and Xiphinema lapidosum from Spain.
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Ruihang Cai, Archidona-Yuste, Antonio, Cantalapiedra-Navarrete, Carolina, Palomares-Rius, Juan E., Jingwu Zheng, and Castillo, Pablo
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TAXONOMY ,MORPHOLOGY ,NEMATODES ,SPECIES ,ECOSYSTEMS ,MORPHOMETRICS ,SOUTHERN root-knot nematode - Abstract
Three populations of Xiphinema non- americanum group species were detected in agricultural and natural ecosystems, during routine surveys for plant-parasitic nematodes in Spain. Based on morphological and molecular analyses, the species were identified as Xiphinema histriae and Xiphinema lapidosum, being this the first record and molecular characterization of both species in Spain. The morphometrics and morphology of the Spanish populations agree with those of the original description and paratype specimens and the present study provided a first description of the second to fourth juvenile stages of both species. A detailed study on the morphology in the Spanish populations of X. histriae, as well as in paratypes, showed a pseudo- Z-organ with weakly muscularized wall and containing numerous small dense granular bodies, which was different to the original description by Lamberti et al. (1993). This new finding suggests that X. histriae must be considered a member of the morphospecies Group 5 of X. non-americanum. Phylogenetic analysis based on D2 to D3 expansion segments of 28S gene, ITS1 and partial CoxI gene indicated that X. histriae and X. lapidosum are phylogenetically related with other Xiphinema non-americanum group spp. reported from Spain. Considering the pathological and economic importance of this group of nematodes, the combination of morphological characters, measurements, and molecular analysis is crucial for accurate identification of these species. [ABSTRACT FROM AUTHOR]
- Published
- 2019
- Full Text
- View/download PDF
16. First Report of Bitylenchus hispaniensis, Pratylenchoides alkani, and Helicotylenchus vulgaris in Association with Cultivated and Wild Olives in Crete, Greece and Molecular Identification of Helicotylenchus microlobus and Merlinius brevidens.
- Author
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Tzortzakakis, Emmanuel A., Cantalapiedra-Navarrete, Carolina, Kormpi, Maria, Lazanaki, Maria S., Castillo, Pablo, and Archidona-Yuste, Antonio
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PLANT nematodes ,OLIVE diseases & pests ,MOLECULAR biology ,NEMATODE morphology - Abstract
Nematode samplings in cultivated and wild olive in Crete, Greece, yielded the presence of Bitylenchus hispaniensis, Helicotylenchus microlobus, Helicotylenchus vulgaris, Merlinius brevidens, and Pratylenchoides alkani. With the exception of H. microlobus and M. brevidens, reports of these plant-parasitic nematode species constitute new records for Greece. Bitylenchus hispaniensis is also reported for first time in a country outside of Spain, where it was originally described. Pratylenchoides alkani is herein reported for the second time in the Mediterranean area and for the first time in association with olive. Two further populations of H. microlobus and H. vulgaris, from walnut and goji berry from Greece, were identified. Molecular data for all of these nematode species are provided, resulting in the first integrative identification of these Greek populations. [ABSTRACT FROM AUTHOR]
- Published
- 2018
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17. Morphological and Molecular Characterization of Paralongidorus sali Siddiqi, Hooper, and Khan, 1963 with a Description of the First-Stage Juvenile and Male of Longidorus jonesi Siddiqi, 1962 from China.
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Ruihang Cai, Maria, Munawar, Nan Qu, Castillo, Pablo, and Jingwu Zheng
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NEMATODE morphology ,MOLECULAR biology ,LONGIDORIDAE ,PLANT nematodes ,NEMATODE phylogeny - Abstract
Longidorids are economically important plant-parasitic nematodes because several species are virus vectors. Populations of Paralongidorus sali and Longidorus jonesi, isolated from woody perennials of Hangzhou, Zhejiang, China, were characterized molecularly and morphologically. The morphometric data of the Chinese populations of both species were compared with other populations. The present study provided a first record of the occurrence of Paralongidorus in China coupled with description of the first-stage juvenile and male of L. jonesi. Phylogenetic analysis based on 18S and D2-D3 expansion segments of 28S gene indicated that L. jonesi clustered with L. jonesi reported from Japan and P. sali grouped with P. bikanerensis from Iran. Considering the pathological and economic importance of this group of nematodes, the study emphasized the need of updated descriptions from accurately identified specimens, isolation of sufficient material for examination, and molecular and phylogenetic analysis for a better understanding and diagnostics of Longidorid nematodes. [ABSTRACT FROM AUTHOR]
- Published
- 2018
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18. First Reports, Morphological, and Molecular Characterization of Longidorus caespiticola and Longidorus poessneckensis (Nematoda: Longidoridae) from Ukraine.
- Author
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SUSULOVSKA, SOLOMIA, CASTILLO, PABLO, and ARCHIDONA-YUSTE, ANTONIO
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LONGIDORIDAE , *DAGGER nematodes , *NEMATODES , *GLYCERIN , *DNA - Abstract
Seven needle nematode species of the genus Longidorus have been reported in Ukraine. Nematological surveys for needle nematodes were carried out in Ukraine between 2016 and 2017 and two nematode species of Longidorus (L. caespiticola and L. poessneckensis) were collected from natural and anthropogenically altered habitats on the territory of Opillia and Zakarpattia in Ukraine. Nematodes were extracted from 500 cm3 of soil by modified sieving and decanting method. Extracted specimens were processed to glycerol and mounted on permanent slides and subsequently identified morphologically and molecularly. Nematode DNA was extracted from single individuals and PCR assays were conducted as previously described for D2-D3 expansion segments of 28S rRNA. Sequence alignments for D2-D3 from L. caespiticola showed 97%-99% similarity to other sequences of L. caespiticola deposited in GenBank from Belgium, Bulgaria, Czech Republic, Russia, Slovenia, and Scotland. Similarly, D2-D3 sequence alignments from L. poessneckensis, showed 99% to other sequences of L. poessneckensis deposited in GenBank from Slovakia and Czech Republic. Morphology, morphometry, and molecular data obtained from these samples were consistent with L. caespiticola and L. poessneckensis identification. To our knowledge, these are the first reports of L. caespiticola and L. poessneckensis in Ukraine, extending the geographical distribution of these species. [ABSTRACT FROM AUTHOR]
- Published
- 2017
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19. Morphological and Molecular Identification of Longidorus euonymus and Helicotylenchus multicinctus from the Rhizosphere of Grapevine and Banana in Greece.
- Author
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TZORTZAKAKIS, EMMANUEL A., CANTALAPIEDRA-NAVARRETE, CAROLINA, CASTILLO, PABLO, PALOMARES-RIUS, JUAN E., and ARCHIDONA-YUSTE, ANTONIO
- Subjects
PLANT nematodes ,INSECT morphology ,RHIZOSPHERE microbiology ,GRAPE diseases & pests ,BANANA diseases & pests ,RECOMBINANT DNA - Abstract
Plant-parasitic nematodes such as Longidorus euonymus and Helicotylenchus multicintctus are species widely distributed in central Europe as well as in Mediterranean area. In Greece, both species have been previously reported but no morphometrics or molecular data were available for these species. Nematode surveys in the rhizosphere of grapevines in Athens carried out in 2016 and 2017, yielded a Longidorus species identified as Longidorus euonymus. Similarly, a population of Helicotylenchus multicinctus was detected infecting banana roots from an outdoor crop in Tertsa, Crete. For both species, morphometrics and molecular data of Greek populations were provided, resulting in the first integrative identification of both nematode species based on morphometric and molecular markers, confirming the occurrence of these two nematodes in Greece as had been stated in earlier reports. [ABSTRACT FROM AUTHOR]
- Published
- 2017
- Full Text
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20. First Report of the Spiral Nematode Rotylenchus incultus (Nematoda: Hoplolaimidae) from Cultivated Olive in Tunisia, with Additional Molecular Data on Rotylenchus eximius.
- Author
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GUESMI-MZOUGHI, ILHEM, ARCHIDONA-YUSTE, ANTONIO, CANTALAPIEDRA-NAVARRETE, CAROLINA, REGAIEG, HAJER, HORRIGUE-RAOUANI, NAJET, PALOMARES-RIUS, JUAN E., and CASTILLO, PABLO
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NEMATODES ,ROTYLENCHUS ,OLIVE ,PLANT nematodes ,RHIZOSPHERE - Abstract
Spiral nematode species of the genus Rotylenchus have been reported on olive (Olea europaea L.) in several Mediterranean countries (Castillo et al., 2010; Ali et al., 2014). Nematological surveys for plant-parasitic nematodes on olive trees were carried out in Tunisia between 2013 and 2014, and two nematode species of Rotylenchus were collected from the rhizosphere of olive cv. Chemlali in several localities of Tunisia (Tables 1,2). Twenty-two soil samples of 3 to 4 kg were collected with a shovel from the upper 50 cm of soil from arbitrarily chosen olive trees. Nematodes were extracted from 500 cm³ of soil by centrifugal flotation method (Coolen, 1979). Specimens were heat killed by adding hot 4% formaldehyde solution and processed to pure glycerin using the De Grisse's (1969) method. Measurements were done using a drawing tube attached to a Zeiss III compound-microscope. Nematode DNA was extracted from single individuals and PCR assays were conducted as described by Castillo et al. (2003). Moderate-to-low soil populations of these spiral nematodes were detected (5.5-11.5, 1.5-5.0 individuals/500 cm³ of soil, respectively). This prompted us to undertake a detailed morphological and molecular comparative study with previous reported data. Morphological and molecular analyses of females identified these species as Rotylenchus eximius Siddiqi, 1964, and Rotylenchus incultus Sher, 1965. The morphology of R. eximius females (five specimens studied) was characterized by having a hemispherical lip region clearly off set, with four to five annuli, body without longitudinal striations, lateral fields areolated in the pharyngeal region only, stylet 32 to 36 mm long, and broadly rounded tail. The morphology of R. incultus females (51 females and 16 males; Table 2) was characterized by a hemispherical lip region with the basal annulus subdivided by irregular longitudinal striations, with three, rarely four annuli; stylet 21.5 to 27.5 mm long, female tail hemispherical with terminus regularly annulated; phasmids anterior to anus level (3-6 annuli above). The morphology of the isolated nematodes agreed with previous descriptions of R. eximius (Siddiqi, 1964; Castillo and Vovlas, 2005) and R. incultus (Sher, 1965; Castillo and Vovlas, 2005; Vovlas et al., 2008), respectively. A single individual was used for DNA extraction. Primers and PCR conditions used in this research were specified in Cantalapiedra-Navarrete et al. (2013), and a single amplicon of 800, 1,100, and 450 bp was obtained and sequenced for D2 to D3, ITS1, and cytochrome c oxidase subunit 1 (coxI), respectively. Sequence alignments for D2 to D3 (KX669231-KX669233), ITS1 (KX669238-KX669240), and cox I (KX669244-KX669245) from R. eximius, showed 99% to 97%, 98% to 94%, 93% similarity to other sequences of R. eximius deposited in GenBank (EU280794-DQ328741, EU373663-EU373664, JX015401-JX015402, respectively). Similarly, D2 to D3 (KX669234-KX669237), ITS1 (KX669241-KX669243), and coxI (KX669246-KX669249) sequence alignments from R. incultus, showed 99%, 99% to 95%, 99% to 90% similarity, respectively, to other sequences of R. incultus deposited in GenBank (EU280797, EU373672-EU373673, JX015403, respectively). The best fitted model of DNA evolution was obtained using µ Model Test v. 2.1.7 (Darriba et al. 2012) with the Akaike information criterion. BI analyses were performed under the general time reversible (GTR) with invariable sites and a gamma-shaped distribution of substitution rates (GTR + I + G) model for ITS1 and coxI. Phylogenetic analyses of ITS1 and coxI using Bayesian inference (BI) placed R. eximius and R. incultus from Tunisia in subclades that included all R. eximius and R. incultus sequences deposited in GenBank (Fig. 1), which agrees with previous results (Cantalapiedra-Navarrete et al., 2013). Morphology, morphometry, and molecular and phylogenetic data obtained from these samples were consistent with R. eximius and R. incultus identification. To our knowledge, this is the first report of R. incultus in Tunisia. Consequently, all these data suggest that spiral nematode species of the genus Rotylenchus are predominant in olive as previously reported in other Mediterranean areas (Ali et al., 2014). [ABSTRACT FROM AUTHOR]
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- 2016
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21. First Report of Longidorus kuiperi and Rotylenchus eximius from Coastal Sand Dunes in Crete, Greece.
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TZORTZAKAKIS, EMMANUEL A., CANTALAPIEDRA-NAVARRETE, CAROLINA, ARCHIDONA-YUSTE, ANTONIO, PALOMARES-RIUS, JUAN E., and CASTILLO, PABLO
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NEMATODES ,SAND dunes ,RHIZOSPHERE ,ROTYLENCHUS ,SOILS - Abstract
Plant-parasitic nematode species have been reported on several occasions from coastal sand dunes, including Longidorus and Rotylenchus species (Vovlas et al., 2008; De Luca et al., 2009; Mateille et al., 2014). In April 2016, 10 soil samples of 3 to 4 kg from the rhizosphere of Tamarix smyrnensis with different vegetation around (viz. Elymus farctus, Lycium schweinfurthii, Crithmum maritimum, and Arthrocnemum sp.) were collected for diagnosis of plant-parasitic nematodes. The area of sampling was a coastal sand dune near the archeological site of Komos, southwest of Crete, Greece. Low soil populations of a needle and a spiral nematode were detected (3 and 8 individuals/1,000 cm³ of soil, respectively), which prompted us to undertake a detailed morphological and molecular com- parative study with previous reported data. Nematodes were extracted from soil with the wet sieving and decanting method (Cobb, 1918). Morphological and molecular analyses of females identified these species as Longidorus kuiperi Brinkman, Loof and Barbez, 1987, and Rotylenchus eximius Siddiqi, 1964. Themorphology of L. kuiperi females (six specimens studied) was characterized by having a slender body; very broad lip region (27 ± 1.5 [25 to 30] mm in width); short, hemispherical tail; body length of (7.1 ± 0.8 [6.5 to 8.5] mm); vulva position at 47% to 55% of body length; odontostyle length of (105 ± 6.5 [90 to 115] mm); males very common (but less frequent than females [45% vs 55%]); tail region with 15 to 20 supplements and bulged terminal cuticle. The morphology of R. eximius females (four specimens studied) was characterized by having a hemispherical lip region clearly set off; with four annuli; body without longitudinal striations; lateral fields areolated in the pharyngeal region only; stylet 36 to 38 mm; and broadly rounded tail. The morphology of the isolated nematodes agreed with previous descriptions of L. kuiperi (Brinkman et al., 1987; De Luca et al., 2009), and R. eximius (Siddiqi, 1964; Castillo and Vovlas, 2005). A single individual was used for DNA extraction. Primers and polymerase chain reaction conditions used in this research were specified in Cantalapiedra et al. (2013) and Archidona-Yuste et al. (2016), and a single amplicon of 800 and 1,100 bp was obtained and sequenced, respectively. D2-D3 (KX398055-KX398056) and ITS sequence alignments (751 and 648 bp, respectively) from L. kuiperi (KX398057) showed 98% to 99% similarity, differing in 4, and from ± to 12 nucleotides, respectively, to other sequences of L. kuiperi deposited in GenBank from Italy and Spain (AM911623, AM905267-AM905270, respectively), with a query coverage of 99%. Similarly, D2-D3 sequence alignment fromR. eximius (KX398058) showed 100% to 99% to another sequence of R. eximius deposited in GenBank from Italy and Spain (EU280794, DQ328741), differing in zero to three nucleotides, respectively, and a query coverage of 81%. Phylogenetic analyses using Bayesian Inference placed L. kuiperi in a highly supported (100%) clade that included all L. kuiperi sequences deposited in GenBank (Archidona-Yuste et al., 2016), and R. eximius in a highly supported (100%) clade that included all R. eximius sequences deposited in GenBank (Cantalapiedra-Navarrete et al., 2013). All identification methods were consistent with L. kuiperi and R. eximius. To our knowledge, this is the first report of L. kuiperi and R. eximius in Greece. Consequently, all these data suggest that coastal sand dunes in Europe constitute environmental conditions optimal for colonization and development of L. kuiperi, as previously reported (De Luca et al., 2009). Similarly, R. eximius has been reported in several Mediterranean countries, including Italy, Morocco, Spain, and Tunisia (Castillo and Vovlas, 2005), and this report extend the geographical distribution of this species. [ABSTRACT FROM AUTHOR]
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- 2016
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22. First Report of the Carolina Spiral Nematode, Scutellonema brachyurus, from Soil of a Garden in Crete, Greece.
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TZORTZAKAKIS, EMMANUEL A., CANTALAPIEDRA-NAVARRETE, CAROLINA, ARCHIDONA-YUSTE, ANTONIO, PALOMARES-RIUS, JUAN E., and CASTILLO, PABLO
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NEMATODE infections ,HOOKWORM disease ,CUCUMBERS ,RADICCHIO ,SPINACH - Abstract
Members of the genus Scutellonema can cause substantial crop losses to ornamental and cultivated plants directly by feeding ectoparasitically on plant roots (Bridge et al., 2005; Coyne et al., 2006). In May 2015, a soil sample from a house garden from Heraklion city in Crete, Greece, was sent for diagnosis of plant-parasitic nematodes. In this place, there had been cactus (Opuntia sp.) plants (probably imported), which were uprooted 3 to 4 years earlier. After that, the area was cropped with cucumber (Cucumis sativus L.) in spring-summer and leaf vegetables such as spinach (Spinacia oleracea L.) and chicory (Cichorium intybus L.) in autumn-winter. The soil was collected 1 mon after the end of chicory crop. A population density (ca. 30 individuals/100 cm³ of soil) of spiral nematodes (Scutellonema sp.) was found by extracting soil with the wet sieving and decanting method (Cobb, 1918). Morphological and molecular analyses of females identified the species as Scutellonema brachyurus (Steiner, 1938) Andrássy, 1958. The morphology of females was characterized by a hemispherical lip region with four to six annuli, morphometric data for 12 females were L, 640 to 760 µm; a, 24.6 to 30.6; b, 5.8 to 7.4; c, 69.1 to 99.3; c'0.5 to 0.6; stylet, 24.5 to 27.5 µm with anterior part shorter than posterior; and spermatheca nonfunctional and male absent. The morphology agreed with the description of S. brachyurus (van den Berg et al., 2013). Alignment indicated that the D2-D3 and ITS sequences (KU059494 and KU059495, respectively) showed 99% and 100% to 99% similarity, respectively, to other sequences of S. brachyurus (type A) deposited in GenBank from the United States, Italy, and Korea (JX472037-JX472046, DQ328753, FJ485643; and JX472069, JX472070, JX472071, respectively), differing from one to six nucleotides. Phylogenetic analyses using Bayesian inference of these sequences placed the Scutellonema sp. in a highly supported (100%) clade that included all S. brachyurus (type A) sequences deposited in GenBank (van den Berg et al., 2013). All identification methods were consistent with S. brachyurus. To our knowledge, this is the first report of S. brachyurus for Greece. As the cucumbers and the leaf vegetables cultivated in the area were seed planted, we consider that the nematode originated most probably from the cactus plants which had been previously root ball planted. Scutellonema brachyurus may represent a threat for ornamental and cultivated plants production in Crete, Greece. The nematode has been already reported, mainly in greenhouses of six European countries (CABI and EPPO, 2006). In most of these cases, it is hypothesized that the nematode was introduced by imported plant material. [ABSTRACT FROM AUTHOR]
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- 2016
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