research-article | 06-June-2019
process as well as to delineate more phylogenetically distant species, which share the same morphology (cryptic species), the support of a molecular approach is needed.
To date, only about 20% of Helicotylenchus species has been molecularly characterized using mostly ribosomal DNA fragments (18S, ITS, 28S rDNA; GenBank resources). Mitochondrial cytochrome c oxidase subunit I (mtCOI) sequences were reported for one species, the recently described H. oleae (Palomares-Rius et al., 2018), and a cytochrome
K. Rybarczyk-Mydłowska,
E. Dmowska,
K. Kowalewska
Journal of Nematology, Volume 51 , 1–17
research-article | 30-November-2019
, measurements and pictures were taken using Carl Zeiss Axio Lab. A1 light microscope equipped with a Zeiss Axiocam ERc5s digital camera. For molecular characterization, the D2-D3 region of 28S rDNA and COI mtDNA gene were amplified using D2A/D3B (5′–ACAAGTACCGTGGGGAAAGTTG–3′/5′–TCGGAAGGAACCAGCTACTA–3′) (Subbotin et al., 2006) and JB3/JB4 (5′-TTTTTTGGGCATCCTGAGGTTTAT-3′/5′-TAAAGAAAGAACATAATGAAAATG-3′) (Nguyen et al., 2019b) primers. Forward and reverse sequences were assembled using Geneious R11
Thi Mai Linh Le,
Huu Tien Nguyen,
Thi Duyen Nguyen,
Quang Phap Trinh
Journal of Nematology, Volume 52 , 1–4
research-article | 30-November-2019
. A1 light microscope equipped with a Zeiss Axiocam ERc5s digital camera. For molecular characterization, the 5′-end region of 28S rDNA was amplified using DP391/501 primers (5′-AGCGGAGGAAAAGAAACTAA-3′/5′-TCGGAAGGAACCAGCTACTA-3′) following Nguyen et al. (2019b). Forward and reverse sequences were assembled and analyzed using Geneious R11 (Nguyen et al., 2019b, 2019c). The best fit model was chosen using Mega 7 following Nguyen et al. (2019b).
Results and discussion
Measurements
Eight females: L
Huu Tien Nguyen,
Thi Duyen Nguyen,
Thi Mai Linh Le,
Quang Phap Trinh
Journal of Nematology, Volume 52 , 1–4
research-article | 30-November-2020
Tanha Maafi et al. (2003), and used as template for polymerase chain reaction (PCR). The D2-D3 expansion segments of 28S rDNA were amplified using the forward D2A (5′-ACAAGTACCGTGAGGGAAAGTTG-3′) and reverse D3B (5′-TCGGAAGGAACCAGCTACTA-3′) primers (Nunn, 1992). Each PCR reaction mixure with a final volume of 30 μl, contained: 15 μl Taq DNA Polymerase 2x Master Mix RED (Ampliqon, Denmark), 1 μl (10 pmol μl−1) of each forward and reverse primers, 2 μl of DNA template and 11 μl deionised water
Manouchehr Hosseinvand,
Ali Eskandari,
Joaquín Abolafia,
Reza Ghaderi
journal of Nematology, Volume 53 , 1–10
Article | 21-July-2017
sequences of 28S rDNA D2/D3 and internal transcribed spacer 1 (ITS1) fragments. Compared to Enchodorus neodolichurus, it has basic differences in tail characters and spicule lengths. Molecular phylogenetic studies using partial sequences of 28S rDNA D2/D3 fragment of the new species and available sequences of Nordiidae members and several other dorylaim species/genera, revealed E. yeatsi n. sp. and E. dolichurus forming a clade with 0.81 Bayesian posterior probability (BPP). This
MAJID PEDRAM
Journal of Nematology, Volume 49 , ISSUE 1, 21–26
Research Article | 17-October-2018
duct, offset spermatheca filled with small spheroid sperm cells, 106 to 127 µm long elongate-conoid tail with filiform distal region and finely rounded tip. Molecular phylogenetic analyses were performed using a near-full length fragment of the 18S rDNA and the D2–D3 expansion segments of the 28S rDNA using Bayesian inference and maximum likelihood methods. In the inferred phylogenetic tree with 18S rDNA, the new species has a close affinity with several isolates of the type species, Labrys
Yousef Panahandeh,
Joaquín Abolafia,
Ebrahim Pourjam,
Robin M. Giblin-Davis,
Farahnaz Jahanshahi Afshar,
Majid Pedram
Journal of Nematology, Volume 50 , ISSUE 3, 343–354
research-article | 30-November-2020
adding 15 μl TE buffer. The DNA sample was stored at −20°C. Primers for 28S rDNA D2-D3 amplification/sequencing were forward primer D2A (5´-ACAAGTACCGTGAGGGAAAGT-3´) and reverse primer D3B (5´-TCGGAAGGAACCAGCTACTA-3´) (Nunn, 1992). The PCR cycles and sequencing of amplified fragments were according to Jahanshahi Afshar et al. (2019) and sequenced directly for both strands using the same primers with an ABI 3730XL sequencer (Bioneer Corporation, South Korea). The newly generated sequence for the new
Fariba Heydari,
Mohammad Reza Atighi,
Ebrahim Pourjam,
Majid Pedram
Journal of Nematology, Volume 53 , 1–11
Research Article | 03-September-2018
molecular analysis of many D. weischeri specimens from Canada is presented. Individuals from 41C. arvense or yellow pea grain samples with seeds of C. arvense from the Prairie Provinces were sequenced for the internal transcribed spacer (ITS rDNA), large subunit (LSU) D2D3 28S rDNA, partial segment of small subunit (SSU) 18S rDNA, and the heat shock protein Hsp90 gene. The analysis also included D. weischeri individuals from C. arvense from Russia and garlic with D. dipsaci from the Provinces of Ontario
Mehrdad Madani,
Mario Tenuta
Journal of Nematology, Volume 50 , ISSUE 2, 163–182
research-article | 30-November-2019
microscopy (SEM)
Specimens preserved in glycerine were selected for observation under SEM according to the methods of Abolafia (2015). They were hydrated in distilled water, dehydrated in a graded ethanol-acetone series, critical point dried, coated with gold, and observed with a Zeiss Merlin microscope (5 kV) (Zeiss, Oberkochen, Germany).
Phylogenetic analyses
For phylogenetic relationships, analyses were based on 18S and 28S rDNA gene sequences available in GenBank. The sequences were aligned using
Joaquín Abolafia,
Reyes Peña-Santiago
Journal of Nematology, Volume 52 , 1–20
research-article | 30-November-2018
Applied Biosystems Hitachi 3500 Genetic Analyzer. The sequences obtained were submitted to the GenBank database.
Phylogenetic analyses
For phylogenetic relationships, analyses were based on 18S and 28S rDNA. The newly obtained sequences were manually edited using BioEdit 7.2.6 (Hall, 1999) and aligned with another 18S or 28S rRNA gene sequences available in GenBank using Muscle alignment tool implemented in the MEGA7 (Kumar et al., 2016). The ambiguously aligned parts and divergent regions were
Joaquín Abolafia,
Reyes Peña-Santiago
journal of nematology, Volume 51 , 1–21
research-article | 14-December-2020
′ (reverse) (Vrain et al., 1992). The fragment containing the D2/D3 regions of the 28S rDNA gene was amplified using primers D2F: 5′-CCTTAGTAACGGCGAGTGAAA-3′ (forward) and 536: 5′-CAGCTATCCTGAGGAAAC-3′ (reverse) (Nadler et al., 2006). The 18S rDNA was amplified using primers NEM18SF: 5′-CGCGAATRGCTCATTACAACAGC-3′ (forward) and NEM18SR: 5′-GGGCGGTATCTGATCGCC-3′ (reverse) (Floyd et al., 2005). The Polymerase Chain Reaction (PCR) protocol for ITS, 18S, and D2/D3 rDNA gene amplification followed was
Aashaq Hussain Bhat,
Shreyansh Srivastava,
Aasha Rana,
Ashok Kumar Chaubey,
Ricardo A. R. Machado,
Joaquín Abolafia
Journal of Nematology, Volume 52 , 1–23
research-article | 01-October-2021
, Progen Scientific, London, UK). The bands were stained with 1.25 µl RedSafe (20,000x) previously added to the agarose gel solution (25 ml). The sequencing reactions of the PCR products were performed at Sistemas Genómicos (Paterna, Valencia, Spain) according the Sanger et al. (1977) method. The DNA sequences obtained for P. pycnus (MZ656001 for the 18S rDNA and MZ656000 for the 28S rDNA) and Tarantobelus arachnicida Abolafia and Peña-Santiago, 2018 (MZ655998–MZ655999 for the 18S rDNA and MZ656002
Joaquín Abolafia,
Matteo Vecchi
Journal of Nematology, Volume 53 , 1–20
research-article | 30-November-2020
phylogenetic relationships, analysis was based on 18S and 28S rDNA. The newly obtained sequences were manually edited using BioEdit 7.2.6 (Hall, 1999) and aligned with other 28S rRNA gene sequences available in GenBank using ClustalW (Thompson et al., 1994) alignment tool implemented in the MEGA7 (Kumar et al., 2016). Alignments ends were trimmed using MEGA7. The best-fit model of nucleotide substitution used for the phylogenetic analysis was statistically selected using jModelTest 2.1.10 (Darriba et al
Joaquín Abolafia,
Manouchehr Hosseinvand,
Ali Eskandari
Journal of Nematology, Volume 53 , 1–16
Research Article | 03-September-2018
. sturhani. The morphological differences of the new species with the aforementioned species are discussed. For all the aforementioned species (except L. protae, currently lacking molecular data) the differences of the new species was also confirmed with differences in molecular sequences of D2-D3 expansion domains of 28S rDNA and the corresponding phylogenetic analyses. The partial sequence of the internal transcribed spacer 1 (ITS1) of the new species was also used in phylogenetic analyses. In partial
Farshad Gharibzadeh,
Ebrahim Pourjam,
Majid Pedram
Journal of Nematology, Volume 50 , ISSUE 2, 207–218
research-article | 16-April-2019
phylogenetic tree (Fig. 5).
Phylogenetic analyses
The newly obtained sequences were aligned using MEGA6 (Tamura et al., 2013) and compared with other Xiphinema D2–D3 expansion segment of 28S rDNA gene sequences available in GenBank using the Nblast homology search program. Longidorus helveticus (Lamberti et al., 2001) (AY601566) was chosen as out group. The best-fitted model of DNA evolution was obtained using MrModeltest 2.3 (Nylander, 2004) with the Akaike Information Criterion (AIC). Phylogenetic
Nasir Vazifeh,
Gholamreza Niknam,
Habibeh Jabbari,
Arezoo Naghavi
Journal of Nematology, Volume 51 , 1–17
research-article | 30-November-2020
″) (De Ley et al., 1999), were used in the PCR reactions for partial amplification of the 18S and 28S rDNA region, respectively. PCR was conducted with 8 μl of the DNA template, 12.5 μl of 2X PCR Master Mix Red (Promega, USA) for the Botswanan specimens, 1 μl of each primer (10 pmol μl−1), and ddH2O for a final volume of 30 μl. The amplification was processed using an Eppendorf master cycler gradient (Eppendorf, Hamburg, Germany), with the following program: initial denaturation for 3 min at 94°C, 37
Ebrahim Shokoohi
Journal of Nematology, Volume 53 , 1–5
research-article | 30-November-2018
Zeiss Axio Lab.A1 light microscope. Measurements and pictures were taken using a ZEN lite software on ZEISS Axiocam ERc5s digital camera (Nguyen et al., 2017).
For molecular studies, Primers D2A (5′-ACAAGTACCGTGGGGAAA GTTG-3′) and D3B (5′-TCGGAAGGAACCAGCTAC TA-3′) were used to amplify D2D3 of 28S rDNA region (Nguyen et al., 2017). Obtained sequence was used for a Blast search in GenBank (Altschul et al., 1997). The data set was analyzed using maximum likelihood (ML) method in MEGA 6 program with
Thi Duyen Nguyen,
Huu Tien Nguyen,
Thi Mai Linh Le,
Thi Tuyet Thu Tran,
Neriza Nobleza,
Quang Phap Trinh
Journal of Nematology, Volume 51 , 1–4
research-article | 30-November-2019
were prepared according to Taylor and Netscher (1974). The determination of the esterase profile was made according to Carneiro and Almeida (2001), using 20 female.
For molecular identification, the D2 to D3 region of 28S rDNA segment was amplified and sequenced using the primers D2A (5′-ACAAGTACCGTGAGGGAAAGTTG-3′) and D3B (5′-TCGGAAGGAACCAGCTACTA-3′) (De Ley et al., 1999) and ITS primers with VRAIN2F (5′-CTTTGTACACACCGCCCGTCGCT-3′) and VRAIN2R (5′-TTTCACTCGCCGTTACTAAGGGAATC-3′) (Vrain et al., 1992
Francisco Jorge Carlos Souza Junior,
Mayara Castro Assunção
Journal of Nematology, Volume 52 , 1–4
research-article | 30-November-2020
identification of specimens from the population of Pratylenchus was carried out by amplifying and sequencing the regions ITS primers with VRAIN2F (5´-CTTTGTACACACCGCCCGTCGCT-3´) and VRAIN2R (5´-TTTCACTCGCCGTTACTAAGGGAATC-3´) (Vrain et al., 1992) and D2-D3 of 28S rDNA segment with the primers D2A (5´-ACAAGTACCGTGAGGGAAAGTTG-3´) and D3B (5´-TCGGAAGGAACCAGCTACTA-3´) (De Ley et al., 1999).
The consensus sequences were formed from the forward and reverse sequences, using the Staden package (Staden et al., 1998
Francisco Jorge Carlos Souza Junior,
Mayara Castro Assunção
Journal of Nematology, Volume 53 , 1–5
research-article | 30-November-2018
(Majd Taheri et al., 2013). Those species have been studied by morphological characters except for two unknowns which have been studied by morphological and molecular DNA barcoding using 28S rDNA (Majd Taheri et al., 2013). Root-lesion nematodes, Pratylenchus (Filipjev, 1936), are after root-knot and cyst nematodes listed as the third economically most important genus that adversely affects crop production worldwide (Castillo and Vovlas, 2007; Jones et al., 2013). Pratylenchus hippeastri, the
Ebrahim Shokoohi,
Joaquín Abolafia,
Phatu William Mashela,
Nafiseh Divsalar
Journal of Nematology, Volume 51 , 1–26
research-article | 30-November-2020
Cristiano Bellé,
Paulo Sergio dos Santos,
Tiago Edu Kaspary
Journal of Nematology, Volume 53 , 1–4
Article | 24-July-2017
ALI YAGHOUBI,
EBRAHIM POURJAM,
SERGIO A LVAREZ-ORTEGA,
GRACIA LIE´BANAS,
MOHAMMAD REZA ATIGHI,
MAJID PEDRAM
Journal of Nematology, Volume 48 , ISSUE 3, 214–221
Research Article | 26-September-2018
SOLOMIA SUSULOVSKA,
PABLO CASTILLO,
ANTONIO ARCHIDONA-YUSTE
Journal of Nematology, Volume 49 , ISSUE 4, 396–402
research-article | 30-March-2020
sequences that overlapped with a 540 bp region from JF741961 previously described from the TN population of V. zeaphila (Bernard et al., 2010). Except for a few ambiguous positions in the latter sequence (possible sequencing artifacts), the ITS sequences from both populations were identical. 28S rDNA sequences obtained from four J2 were assembled into a 762 bp alignment along with JF741960 previously described from the TN population of V. zeaphila, differing at 0 to 2 bp from each other and from the TN
Andrea M. Skantar,
Zafar A. Handoo,
Mihail R. Kantor,
Lynn K. Carta,
Jamal Faghihi,
Virginia Ferris
Journal of Nematology, Volume 52 , 1–8
research-article | 30-November-2020
Abbas Abdolkhani,
Sedighe Azimi
Journal of Nematology, Volume 53 , 1–10
research-article | 30-November-2019
using CLUSTAL W (Thompson et al., 1994). The length of each alignment was 946 and 1186 bp for ITS rDNA and 28S rDNA, respectively. Bayesian inference was used to reconstruct the phylogeny, with Bayesian trees generated using the Bayesian inference method as implemented in the program MrBayes 3.1.2 (Ronquist and Huelsenbeck, 2003). The GTR + I + G model was selected using jModeltest 2.1.10 (Guindon and Gascuel, 2003; Darriba et al., 2012). Analysis using the GTR + I + G model was initiated with a
Ebrahim Shokoohi,
Phatu W. Mashela
Journal of Nematology, Volume 52 , 1–5
Article | 21-July-2017
phylogenetic studies based on partial sequences of 28S rDNA D2/D3 fragments, all species formed a clade with high Bayesian posterior probability in Bayesian inference, indicating the monophyly of the genus. The clade of Coslenchus spp. formed a highly supported monophyletic group, a sister clade to two species of the genus Aglenchus.
YOUSEF PANAHANDEH,
EBRAHIM POURJAM,
MAJID PEDRAM
Journal of Nematology, Volume 48 , ISSUE 4, 268–279
Original Paper | 10-December-2018
of the following rDNA regions: ITS1, ITS2, 5.8S, 28S rDNA, LSU and β-tubulin were carried out for the isolates studied. A high level of similarity was found between sequences from certain rDNA regions, i.e. ITS1-5.8S-ITS2 and LSU, what confirmed the classification of the isolates to the species A. penicillioides. The classification of our isolates to A. penicillioides species was confirmed also by the phylogenetic analysis.
EULALIA MACHOWICZ-MATEJKO,
AGNIESZKA FURMAŃCZYK,
EWA DOROTA ZALEWSKA
Polish Journal of Microbiology, Volume 67 , ISSUE 4, 407–416
Article | 03-December-2017
The taxonomy and the systematics of the genus Makatinus are discussed by means of the characterization of its morphological pattern and the first molecular (D2–D3 expansion segments of 28S rDNA) analysis of a representative of this taxon, Makatinus crassiformis from Costa Rica. The presence of two or more pairs of male ad-cloacal genital papillae is the most characteristic autapomorphy of the genus, but the status of its species on this concern differ among them. Both morphological and
REYES PENA-SANTIAGO,
INGRID VARELA
Journal of Nematology, Volume 49 , ISSUE 3, 245–253
Research Article | 03-December-2018
juvenile Oscheius rugaoensis (Zhang et al., 2012) Darsouei et al., 2014 (Rhabditidae), and juvenile and adult Mononchoides sp. (Diplogastridae) based on images, morphometrics, and sequences of 18S and 28S rDNA. A novel short 28S sequence of a separate population of Oscheius necromenus SB218 from Australian millipedes was also included in a phylogenetic comparison of what can now be characterized as a species complex of millipede-associated Oscheius. The only other nematode associates of millipedes
L. K. Carta,
W. K. Thomas,
V. B. Meyer-Rochow
Journal of Nematology, Volume 50 , ISSUE 4, 479–486
research-article | 30-November-2018
M. L. Inácio,
L. C. Rusinque,
M. J. Camacho,
F. Nóbrega
Journal of Nematology, Volume 51 , 1–6
research-article | 30-November-2019
, the mixture was incubated at room temperature (23 ± 2°C) for 10 min and then at 95°C for 3 min. Finally, 20 µL of neutralization solution was added to the tube and vortexed briefly. This DNA extract was stored at −20°C and used as DNA template for PCR reactions in 2 µL aliquots. Three primer pairs targeting 18S rDNA (360F/932R), 28S rDNA (D2A/D3B) and ITS1 rDNA (BL18/5818) (Riga et al., 2007; Duarte et al., 2010; Ye et al., 2015) were used in singleplex PCR (Hajihassani et al., 2018a). The
Ganpati B. Jagdale,
Fereidoun Forghani,
Katherine Martin,
Abolfazl Hajihassani,
Alfredo Dick Martinez-Espinoza
Journal of Nematology, Volume 52 , 1–3
research-article | 24-November-2020
diagnosis is gaining more reliability for precise and accurate identification of cyst-forming nematodes (Peng et al., 2003). The internal transcribed spacer region of the ribosomal DNA (ITS-rDNA), the D2 and D3 expansion fragments of the 28S ribosomal DNA genes (D2-D3 of 28S-rDNA), and mitochondrial DNA (COI gene) units are good candidate genes for molecular taxonomic and phylogenetic studies (Subbotin et al., 2001; Subbotin et al., 2006; Madani et al., 2004; Vovlas et al., 2017). Based on
Wenhao Li,
Huixia Li,
Chunhui Ni,
Deliang Peng,
Yonggang Liu,
Ning Luo,
Xuefen Xu
Journal of Nematology, Volume 52 , 1–16
Article | 04-December-2017
monodelphic-prodelphic reproductive system, 15 to 19 mm long conical tail with broad rounded tip, and males absent. The new species is compared with two known species of the genus, Anguillonema poligraphi and A. crenati. Molecular phylogenetic studies of the new species using partial sequences of small subunit (SSU) rDNA revealed that it forms a clade with an unidentified nematode species and two species of the genus Howardula. In phylogenetic analyses using partial sequences of the 28S rDNA (D2-D3
MAHYAR MOBASSERI,
MAJID PEDRAM,
EBRAHIM POURJAM
Journal of Nematology, Volume 49 , ISSUE 3, 286–294
Research Article | 03-December-2018
only 95% similarity with PWN B. xylophilus. Compared to the previously described Portuguese population of B. antoniae, the sequences generated for the MA population were 98.3% similar in the ITS1, 2 rDNA and 99.9% similar for 28S rDNA. There was 99.2% similarity between the COI sequences of the US and Portuguese isolates of B. antoniae. This population has morphology consistent with that of Penas et al., 2006; however, the female tail on this MA pine population is mucronate and more attenuated than
Lynn K. Carta,
R. L. Wick
Journal of Nematology, Volume 50 , ISSUE 4, 473–478
research-article | 28-April-2020
transferred to an Eppendorf tube containing 25.65 μl ddH2O, 2.85 μl 10 × PCR buffer and 1.5 μl proteinase K (600 μg/ml) (Promega, Benelux, the Netherlands). The tubes were incubated at −80°C (1 h), 65°C (1 h) and 95°C (15 min). The extracted DNA was stored at −20°C until use. The D2-D3 domains of the 28S rDNA were amplified with forward primer D2A (5´-ACAAGTACCGTGAGGGAAAGTTG-3´) and reverse primer D3B (5´-TCGGAAGGAACCAGCTACTA-3´) (Nunn, 1992). In total, 25 μl PCR reaction mixture was prepared constituting
Nasir Vazifeh,
Gholamreza Niknam,
Habibeh Jabbari,
Reyes Peña-Santiago
Journal of Nematology, Volume 52 , 1–12
research-article | 30-November-2020
length of stylet, tail and hyaline tail in second-stage juvenile, and the surface differentiation in eggs (Subbotin et al., 2010). However, traditional identification of cyst forming nematode based on morphology is imprecise and time-consuming to separate the related species. During the past 30 years, molecular data, including ITS-rDNA, D2-D3 region of 28S-rDNA, are more accurate tool for identification of cyst-forming nematode species. Sequence analysis of the ITS-rDNA and the D2-D3 region of 28S
Wenhao Li,
Huixia Li,
Chunhui Ni,
Mingming Shi,
Xuejuan Wei,
Yonggang Liu,
Yiwen Zhang,
Deliang Peng
Journal of Nematology, Volume 53 , 1–15
Research Article | 26-September-2018
rRNA gene, D2–D3 expansion domains of the 28S rDNA, the ITS region, and the partial mitochondrial COI were carried out. Sequences of the 18S rRNA gene, the D2–D3 domains, and the ITS were analyzed using several methods for inferring phylogeny to reconstruct the relationships among Sheraphelenchus and Bursaphelenchus species. The bacterial feeder Panagrellus sp. was characterized at the molecular level only. The D2–D3 expansion domains and ITS sequences of this Italian panagrolaimid were determined
ELENA FANELLI,
ALBERTO TROCCOLI,
NICOLA VOVLAS,
GIANLUCA SCARCIA,
ANNAMARIA MINCUZZI,
SIMONA M. SANZANI,
ANTONIO IPPOLITO,
FRANCESCA DE LUCA
Journal of Nematology, Volume 49 , ISSUE 4, 418–426
research-article | 02-April-2019
28s rDNA gene (Nunn, 1992). The amplification of near full length 18s rDNA gene was attempted by primers SSU_F_07 (AAAGATTAAGCCATGCATG) and SSU_R_81 (TGATCCWKCYGCAGGTTCAC) (Gutiérrez-Gutiérrez et al., 2012). Each PCR reaction comprised of 1 unit of Platinum Taq polymerase (Invitrogen Carlsbad, CA, USA), 1x Taq polymerase buffer, 0.2 mM dNTPs, 1.5 mM MgCl2, 0.4 µM forward and reverse primers and 6 µl of template DNA. The thermocycling conditions for ITS were – Initial denaturation at 95 °C for 5
Puneet Kumar,
Wajih Jamal,
Vishal S. Somvanshi,
Khushbu Chauhan,
Sabia Mumtaz
Journal of Nematology, Volume 51 , 1–11
research-article | 30-November-2020
illustrations were edited by using Adobe Photoshop CC 2018.
DNA extraction, polymerase chain reaction (PCR), and sequencing
Nematode DNA was extracted from a single individual as described by Holterman et al. (2006) and DNA extracts were stored at –20° until used as PCR template. The D2-D3 expansion segment 28S rDNA and 18S were amplified using the forward D2A (5′–ACAAGTACCGTGGGGAAAGTTG–3′) and reverse D3B (5′–TCGG AAGGAACCAGCTACTA–3′) primers (Subbotin et al., 2006) and primers 18S (18F : 5
Tam T. T. Vu,
Thi Mai Linh Le,
Thi Duyen Nguyen
Journal of Nematology, Volume 53 , 1–22
research-article | 24-April-2019
morphology and morphometrics along with molecular characteristics and phylogeny of the D2-D3 expansion segment of 28S rDNA, ITS rDNA, and COI mtDNA sequences.
Materials and methods
Sampling and nematode extraction
The soil and root samples were collected around the rhizosphere of banana (Musa basjoo Siebold & Zucc. ex Iinuma) (GPS coordinates N: 51°2′6.8″, E: 3°43′22.7″) and Yam (Dioscorea tokoro) (GPS coordinates: N: 51°2′6.9″, E: 3°43′22.6″) at the Botanical garden of Ghent University. The nematodes
Huu Tien Nguyen,
Quang Phap Trinh,
Marjolein Couvreur,
Phougeishangbam Rolish Singh,
Wilfrida Decraemer,
Wim Bert
Journal of Nematology, Volume 51 , 1–20
research-article | 30-November-2020
extract) was used either immediately as a DNA template for a PCR reaction or stored at ‒ 20°C for future use.
Nearly full length 18S rDNA was amplified from the analyzed Parkellus individuals in two overlapping fragments, using the following primer combinations: 988F combined with 1912R and 1813F combined with 2646R (Holterman et al., 2006). The 28S rDNA sequence was also amplified in two parts using the 61F primer (Holterman et al., 2006) combined with the MCB1R (Dobosz et al., 2013) and the D2A
Tam T.T. VU,
Katarzyna Rybarczyk-Mydłowska,
Andrij Susulovsky,
Magdalena Kubicz,
Łukasz Flis,
Thi Mai Linh Le,
Grażyna Winiszewska
Journal of Nematology, Volume 53 , 1–22
Research Article | 03-September-2018
Gary Phillips,
Robert J. Pivar,
Xiocaun Sun,
John K. Moulton,
Ernest C. Bernard
Journal of Nematology, Volume 50 , ISSUE 2, 133–146
research-article | 03-June-2019
Eukaryotic nuclear ribosomal DNA (rDNA) is arranged in tandem repeat arrays in the genome. Each repeat unit consists of one copy of small subunit (SSU) 18S, internal transcribed spacers (ITS1 and ITS2), 5.8S, and large subunit (LSU) 28S rDNA, and is separated by an external transcribed spacer (EST) and an intergenic spacer (IGS) (Hillis and Dixon, 1991). The copy number of the repeats within most eukaryotic genomes is high, which provide large quantities of template DNA for PCR. In
L. K. Carta,
S. Li
Journal of Nematology, Volume 51 , 1–8
research-article | 28-April-2020
between present strains of Acrobeloides and already described species.
S. No.
28S rDNA
Country
1
2
3
4
5
6
7
8
9
10
11
12
13
14
1
MK935147 A. saeedi KMW
India
1
1
5
5
5
38
51
59
67
74
75
76
76
2
MN101167 A. saeedi DH1
India
100
0
5
5
5
38
51
59
68
74
75
77
77
3
KY914573 A. saeedi
Iran
100
100
10
10
10
38
56
64
68
79
80
82
82
4
MF325168 A. sexlineatus
Germany
98.9
98.9
98.0
0
0
5
8
0
11
0
0
0
0
5
Aasha Rana,
Aashaq Hussain Bhat,
Suman Bhargava,
Ashok Kumar Chaubey,
Joaquín Abolafia
Journal of Nematology, Volume 52 , 1–21