Morphological and molecular characterisation of some Hemicriconemoides species (Nematoda: Criconematidae) together with a phylogeny of the genus

Nematology ◽  
2014 ◽  
Vol 16 (5) ◽  
pp. 519-553 ◽  
Author(s):  
Esther Van den Berg ◽  
Louwrens R. Tiedt ◽  
Renato N. Inserra ◽  
Jason D. Stanley ◽  
Nicola Vovlas ◽  
...  
Keyword(s):  
Fruit Trees ◽  
Valid Species ◽  
Rrna Gene ◽  
28S Rrna ◽  
Molecular Characterisation ◽  
Electron Microscopic ◽  
Specific Primers ◽  
Scanning Electron Microscopic ◽  
The Usa ◽  
Species Specific

Sheathoid nematodes of the genusHemicriconemoidesare migratory root-ectoparasites of many plants including various agricultural crops and fruit trees. They are generally found inhabiting warm areas of the world and presently consist of 52 valid species. In this study we provide morphological and molecular characterisation of 12 species of this genusviz.:H. alexis,H. brachyurus,H. californianus,H. chitwoodi,H. macrodorus,H. minutus,H. ortonwilliamsi,H. promissus,H. silvaticus,H. strictathecatus,H. wessoniandHemicriconemoidessp. originating from China, Greece, Japan, Myanmar, Spain, South Africa and the USA. Morphological descriptions, measurements, light and scanning electron microscopic observations and drawings are given for several species. Phylogenetic relationships withinHemicriconemoides, as inferred from the analyses of the D2-D3 of 28S rRNA and ITS-rRNA gene sequences, resulted in trees with three major clades that corresponded with species groupings based on morphology of the lip pattern and vulval flap. PCR with species-specific primers were developed forH. californianus,H. chitwoodiandH. strictathecatus.

Morphological and molecular characterisation of one new and several known species of the reniform nematode, Rotylenchulus Linford & Oliveira, 1940 (Hoplolaimidae: Rotylenchulinae), and a phylogeny of the genus

Nematology ◽  
2016 ◽  
Vol 18 (1) ◽  
pp. 67-107 ◽  
Author(s):  
Esther Van Den Berg ◽  
Juan E. Palomares-Rius ◽  
Nicola Vovlas ◽  
Louwrens R. Tiedt ◽  
Pablo Castillo ◽  
...  
Keyword(s):  
South Africa ◽  
Valid Species ◽  
Rrna Gene ◽  
28S Rrna ◽  
Molecular Characterisation ◽  
Rapid Diagnostics ◽  
Sister Relationship ◽  
Species Specific ◽  
Reniform Nematodes ◽  
A New Species

The reniform nematodes of the genusRotylenchulusare semi-endoparasites of numerous herbaceous and woody plant roots and are mainly distributed in tropical and subtropical regions. In this study, we provide morphological and molecular characterisation of six out of ten presently known valid species ofRotylenchulus:R. clavicaudatus,R. leptus,R. macrodoratus,R. macrosoma,R. reniformisandR. saccharifrom South Africa, USA, Italy and Spain.Rotylenchulus parvuswas only studied morphologically. A new species,R. macrosomoidessp. n., isolated from soil and roots of sugarcane in South Africa, is described. The phylogeny ofRotylenchulus, as inferred from the analyses of D2-D3 of 28S rRNA, ITS rRNA,coxImtDNA andhsp90gene sequences, is presented. The study revealed thatR. reniformisandR. macrosomahave a sister relationship, but that relationships between otherRotylenchulusspecies remain unresolved. The phylogenetic analysis also confirmed the hypothesis that this genus originated from the Afrotropical zoogeographical region. Our study revealed thatR. reniformisandR. macrosomoidessp. n. have two distinct rRNA gene types andR. macrosomahave three rRNA gene types in their genomes. PCR with species-specific primers was developed for rapid diagnostics ofR. reniformis.

Morphological and molecular characterisation of several Paratylenchus Micoletzky, 1922 (Tylenchida: Paratylenchidae) species from South Africa and USA, together with some taxonomic notes

Nematology ◽  
2014 ◽  
Vol 16 (3) ◽  
pp. 323-358 ◽  
Author(s):  
Esther Van den Berg ◽  
Esther Van den Berg ◽  
Louwrens R. Tiedt ◽  
Esther Van den Berg ◽  
Louwrens R. Tiedt ◽  
...  
Keyword(s):  
South Africa ◽  
Rrna Genes ◽  
Morphological Identification ◽  
Rrna Gene ◽  
28S Rrna ◽  
Molecular Characterisation ◽  
Electron Microscopic ◽  
Wide Range ◽  
Its Rrna Gene ◽  
Its Rrna

Pin nematodes of the genus Paratylenchus are widely distributed across the world and associated with many plant species. Morphological identification of Paratylenchus species is a difficult task because it relies on many characters with a wide range of intraspecific variation. In this study we provide morphological and molecular characterisation of several pin nematodes: Paratylenchus aquaticus, P. dianthus, P. hamatus, P. nanus and P. straeleni, collected in different states of the USA and South Africa. Paratylenchus aquaticus is reported from South Africa and Hawaii and P. nanus is found from South Africa for the first time. Morphological descriptions, morphometrics, light and scanning electron microscopic photos and drawings are given for these species. Molecular characterisation of nematodes using the D2-D3 of 28S rRNA and ITS rRNA gene sequence revealed that samples morphologically identified as P. aquaticus, P. hamatus and P. nanus indeed represent species complexes containing several species. Sequences of the rRNA genes are also provided for several unidentified Paratylenchus. Phylogenetic relationships within the genus Paratylenchus are given as inferred from the analyses of the D2-D3 of 28S rRNA and ITS rRNA gene sequences. We present here the most complete phylogenetic analysis of the genus.

Morphological and molecular characterisation of Helicotylenchus pseudorobustus (Steiner, 1914) Golden, 1956 and related species (Tylenchida: Hoplolaimidae) with a phylogeny of the genus

Nematology ◽  
2015 ◽  
Vol 17 (1) ◽  
pp. 27-52 ◽  
Author(s):  
Sergei A. Subbotin ◽  
Nicola Vovlas ◽  
Gregor W. Yeates ◽  
Johannes Hallmann ◽  
Sebastian Kiewnick ◽  
...  
Keyword(s):  
New Zealand ◽  
Related Species ◽  
Phylogenetic Relationships ◽  
Valid Species ◽  
Molecular Characteristics ◽  
Morphological Identification ◽  
Rrna Gene ◽  
28S Rrna ◽  
Molecular Characterisation ◽  
Its Rrna Gene

Morphological identification of spiral nematodes of the genus Helicotylenchus is a difficult task because most characters used for their diagnosis vary within species. In this paper we provide morphological and molecular characterisations of several spiral nematodes, H. broadbalkiensis, H. digonicus, H. dihystera, H. microlobus, H. paxilli and H. pseudorobustus, collected in different geographical areas of USA, Switzerland, Italy, New Zealand, Spain, UK, South Korea and Russia. We suggest that H. microlobus and H. pseudorobustus are valid species separated from each other morphologically and molecularly. Seven species with distinct molecular characteristics are also distinguished, but are not ascribed morphologically to any specific taxon because of the low number of specimens available. Phylogenetic relationships of H. pseudorobustus with other Helicotylenchus species are given as inferred from the analyses of 154 sequences of the D2-D3 of 28S rRNA gene and 37 sequences of ITS rRNA gene.

Molecular characterisation and phylogenetic relationships of cystoid nematodes of the family Heteroderidae (Nematoda: Tylenchida)

Nematology ◽  
2017 ◽  
Vol 19 (9) ◽  
pp. 1065-1081 ◽  
Author(s):  
Sergei A. Subbotin ◽  
Jerry Akanwari ◽  
Chau N. Nguyen ◽  
Ignacio Cid del Prado Vera ◽  
John J. Chitambar ◽  
...  
Keyword(s):  
New Species ◽  
North America ◽  
Phylogenetic Relationships ◽  
Valid Species ◽  
28S Rrna ◽  
Molecular Characterisation ◽  
The Family ◽  
The Usa ◽  
Unidentified Species ◽  
Its Rrna

The cystoid sedentary nematodes of the Heteroderidae include more than 30 recognised species belonging to from nine to 11 genera, depending on the opinion of the authority. In the present study, we collected nematode populations of species belonging to some of these genera, as well as other cyst-forming species from different locations in the USA, Vietnam, Germany and Russia. The populations of cystoid nematodes represent seven valid species, Atalodera carolynae, Cryphodera sinensis, Meloidodera astonei, M. floridensis, M. mexicana, M. sikhotealiniensis, Rhizonemella sequoiae, two unidentified species of Atalodera, six unidentified species of Cryphodera, and three putative new species of Rhizonemella. We also obtained samples of cyst-forming nematodes that include an unidentified species of Betulodera from California, and Heterodera guangdongensis from Vietnam. A population of Rotylenchulus sp. from Arizona, intercepted in Florida, and a population of an unidentified species of Verutus from Germany were also added to this study. All of these populations were characterised using sequences of the D2-D3 of 28S rRNA, ITS rRNA or mitochondrial COI genes. A total of 89 new sequences were obtained from these analyses. Phylogenetic relationships within the family Heteroderidae were reconstructed based on the D2-D3 of 28S rRNA, ITS rRNA and mitochondrial COI gene sequences. Results revealed that the subfamily Ataloderinae was paraphyletic. Ekphymatodera thomasoni, a non-cyst-forming species, clustered with the cyst-forming nematodes. Representatives of the subfamily Verutinae formed clades within Heteroderidae. The genus Meloidodera was non-monophyletic and distributed within two clades: i) M. sikhotealiniensis and Cryphodera spp. from Asia and Europe; and ii) M. astonei, M. floridensis and M. mexicana from North America. Based on comparative molecular analysis Meloidodera alni syn. n. is proposed as a junior synonym of M. sikhotealiniensis. Problems of taxonomy and phylogeography of cystoid nematodes are also discussed.

Description of Tylenchulus musicola sp. n. (Nematoda: Tylenchulidae) from banana in Iran with molecular phylogeny and characterisation of species of Tylenchulus Cobb, 1913

Nematology ◽  
2012 ◽  
Vol 14 (3) ◽  
pp. 353-369 ◽  
Author(s):  
Zahra Tanha Maafi ◽  
Majid Amani ◽  
Jason D. Stanley ◽  
Renato N. Inserra ◽  
Esther Van den Berg ◽  
...  
Keyword(s):  
New Species ◽  
Rapid Identification ◽  
Rrna Genes ◽  
Rrna Gene ◽  
28S Rrna ◽  
Specific Primers ◽  
Diagnostic Pcr ◽  
Second Stage ◽  
Species Specific ◽  
A New Species

During a survey conducted on banana plantations in Sistan and Blouchestan province, south-east Iran, a new species of Tylenchulus was extracted from the soil and roots of banana plants. This species, named Tylenchulus musicola sp. n., is characterised by mature females having a swollen, hook-shaped body with a conical and elongate post-vulval portion ending in a round terminus, males having a weak stylet and a cylindrical and thick tail ending in a bluntly rounded and smooth terminus, and by second-stage juveniles having a slender body and a posterior body portion ending in a finely pointed or mucronate terminus. The results of glasshouse host tests indicated that the new species does not parasitise sugarcane ratoons or sour orange seedlings. Tylenchulus musicola sp. n. is distinguished from other known Tylenchulus species by the sequences of D2-D3 expansion segments of 28S rRNA and ITS rRNA genes. Phylogenetic relationships within Tylenchulus were reconstructed based on rRNA gene sequences using Bayesian inference. Diagnostic PCR-ITS-RFLP profiles are presented for T. musicola sp. n., T. furcus, T. graminis, T. palustris, T. semipenetrans and Trophotylenchulus floridensis. PCR with species-specific primers and genus-specific primer are tested and developed for rapid identification of five Tylenchulus species. An identification key to Tylenchulus species is provided.

Molecular characterisation of some stubby root nematodes (Nematoda: Trichodoridae) from the USA and other countries

Nematology ◽  
2019 ◽  
Vol 22 (1) ◽  
pp. 39-52 ◽  
Author(s):  
Sergei A. Subbotin ◽  
Ignacio Cid Del Prado Vera ◽  
Renato N. Inserra ◽  
Vladimir N. Chizhov ◽  
Wilfrida Decraemer
Keyword(s):  
Nematode Species ◽  
Rrna Gene ◽  
28S Rrna ◽  
Molecular Characterisation ◽  
28S Rrna Gene ◽  
Putative Species ◽  
Geographical Regions ◽  
Wide Range ◽  
The Usa ◽  
Stubby Root Nematode

Summary The stubby root nematodes are world-wide distributed polyphagous root ectoparasites and can cause damage to a wide range of crops and natural vegetation. In this study, 22 valid and putative species of stubby root nematodes were identified in 37 samples collected in Malaysia, Mexico, Russia, and the USA (California, Florida, Minnesota and Nevada). The analysis of 58 new sequences of the D2-D3 expansion fragments of 28S rRNA gene revealed the following species: Nanidorus minor from California, Florida and Mexico, N. renifer from Florida, two unidentified Nanidorus species from California and Malaysia, respectively, Paratrichodorus allius from Florida and Minnesota, P. pachydermus and Paratrichodorus sp. from Russia, Trichodorus californicus, T. intermedius, and T. obscurus from California, T. obtusus from Florida, eight unidentified Trichodorus species from California, one unidentified Trichodorus species from Nevada, and two unidentified species of stubby root nematodes, one of Trichodorus and another undetermined species from Mexico. Molecular characterisation of T. californicus, T. intermedius and T. obscurus is given for the first time. The phylogenetic tree reconstructed from the analysis of 108 D2-D3 of 28S rRNA gene sequences of 58 valid and putative species of the stubby root nematodes contained five major clades: i) Trichodorus from Europe, Asia and North America; ii) Nanidorus and Trichodorus from Asia; iii) Trichodorus from California; iv) Paratrichodorus from several distant geographical regions; and v) Monotrichodorus from Central and South America. It has been hypothesised that the California Floristic Province is one of the centres of origin and diversification of stubby root nematodes. There is no information on the economic importance of the stubby root nematode species found in California and Mexico. The confirmation of the occurrence of N. renifer and P. allius in Florida should be of concern for the blueberry and potato industries in the state.

Pratylenchus speijeri n. sp. (Nematoda: Pratylenchidae), a new root-lesion nematode pest of plantain in West Africa

Nematology ◽  
2012 ◽  
Vol 14 (8) ◽  
pp. 987-1004 ◽  
Author(s):  
Francesca De Luca ◽  
Alberto Troccoli ◽  
Larry W. Duncan ◽  
Sergei A. Subbotin ◽  
Lieven Waeyenberge ◽  
...  
Keyword(s):  
New Species ◽  
West Africa ◽  
Species Complex ◽  
Rrna Gene ◽  
28S Rrna ◽  
New Taxon ◽  
Root Lesion Nematode ◽  
Specific Primers ◽  
Lesion Nematode ◽  
Species Specific

A new root-lesion nematode, particularly pathogenic to Musa spp. and causing important plantain losses in Ghana, is described and named Pratylenchus speijeri n. sp. The cryptic status of this species within the P. coffeae species complex has been assessed and confirmed in this study. An extensive comparison of the morphological and molecular characteristics of this new species with those of P. coffeae and other related amphimictic species did not result in an unambiguous separation of this species from P. coffeae because only a few morphological features of diagnostic value were found. Sequence and phylogenetic analyses of the D2-D3 expansion segments of the 28S rRNA gene, the ITS rRNA gene and a portion of the hsp90 gene of P. speijeri n. sp. and P. coffeae species complex populations from different sources generated majority consensus BI trees with three major clades: P. speijeri n. sp. from Musa spp. roots in Ghana; unidentified or putative new Pratylenchus sp. C1 from Colocasia esculenta roots in Japan and P. coffeae with non-homogeneous relationships from different hosts and distant geographical areas. These results confirmed the validity of P. speijeri n. sp. as a new taxon and indicated that P. coffeae populations from Colocasia in Japan also need to be considered as a new species. Sequence differences in the ITS were used to design group- and species-specific primers to detect P. speijeri n. sp. and other species of P. coffeae species complex. The use of these species-specific primers for the separation of P. speijeri n. sp., Pratylenchus sp. C1 and P. coffeae has important practical application in breeding programmes for agriculture in West Africa.

Morphological and molecular characterisation and diagnostics of some species of Scutellonema Andrássy, 1958 (Tylenchida: Hoplolaimidae) with a molecular phylogeny of the genus

Nematology ◽  
2013 ◽  
Vol 15 (6) ◽  
pp. 719-745 ◽  
Author(s):  
Esther van den Berg ◽  
Esther van den Berg ◽  
Louwrens R. Tiedt ◽  
Esther van den Berg ◽  
Louwrens R. Tiedt ◽  
...  
Keyword(s):  
South Africa ◽  
Type A ◽  
28S Rrna ◽  
Molecular Characterisation ◽  
Type B ◽  
Gene Sequences ◽  
Electron Microscopic ◽  
Coi Mtdna ◽  
The Usa ◽  
Its Rrna

Scutellonema spp. are widely distributed across tropical and subtropical regions of the world and are associated with numerous agricultural and horticultural crops. Identification of many Scutellonema species is not always reliable, in part because many species share very similar diagnostic characters. In this study, we provide morphological and molecular characterisation of S. brachyurus from the USA and South Africa, S. bradys from Nigeria and three unidentified species from California, USA, New Zealand and Burkina Faso. Morphological descriptions, measurements, light and scanning electron microscopic photos and drawings are given for S. brachyurus. Females of S. brachyurus from the USA (type A) and South Africa (type B) showed a significant variation in the number of sectors and blocks on the lip annuli, ranging from about 4-12 and from 8-20, respectively. Molecular analysis using the D2-D3 of 28S rRNA, ITS rRNA and the COI mtDNA gene sequences revealed two distinct genotypes within S. brachyurus samples: type A (samples from USA, Italy, Korea, Taiwan) and type B (South Africa). Multivariate analyses determined that S. brachyurus from the USA and Taiwan (type A) differed from that from South Africa (type B) mainly in body, tail and DGO lengths, and ratios b′, c′, c and V. Phylogenetic relationships within Scutellonema are given as inferred from the analyses of the D2-D3 of 28S rRNA, ITS rRNA and the COI mtDNA gene sequences. PCR-RFLP diagnostic profiles and PCR with species-specific primers are developed for the studied Scutellonema species.

Species-specific duplex PCR for the detection of Pratylenchus penetrans

Nematology ◽  
2009 ◽  
Vol 11 (6) ◽  
pp. 847-857 ◽  
Author(s):  
Lieven Waeyenberge ◽  
Nicole Viaene ◽  
Maurice Moens
Keyword(s):  
Internal Control ◽  
Dna Sequences ◽  
Rrna Gene ◽  
Duplex Pcr ◽  
Specific Primers ◽  
5.8S Rrna Gene ◽  
5.8S Rrna ◽  
Wide Range ◽  
Pcr Products ◽  
Species Specific

Abstract ITS1, the 5.8S rRNA gene and ITS2 of the rDNA region were sequenced from 20 different Pratylenchus species. Additionally, the same region was sequenced from seven populations of P. penetrans. After purifying, cloning and sequencing the PCR products, all sequences were aligned in order to find unique sites suitable for the design of species-specific primers for P. penetrans. Since ITS regions showed variability between and even within populations of P. penetrans, only three small DNA sequences were suitable for the construction of three potentially useful species-specific primers. New species-specific primers were paired with existing universal ITS primers and tested in all possible primer combinations. The best performing primer set, supplemented with a universal 28S rDNA primer set that served as an internal control, was tested in duplex PCR. The ideal annealing temperature, Mg2+ concentration and primer ratios were then determined for the most promising primer set. The optimised duplex PCR was subsequently tested on a wide range of different Pratylenchus spp. and 25 P. penetrans populations originating from all over the world. To test the sensitivity, the duplex PCR was conducted on DNA extracted from a single P. penetrans nematode mixed with varying amounts of nematodes belonging to another Pratylenchus species. Results showed that a reliable and sensitive P. penetrans species-specific duplex PCR was constructed.

scholarly journals Multiplex PCR with 16S rRNA Gene-Targeted Primers of Bifidobacterium spp. To Identify Sources of Fecal Pollution

2004 ◽  
Vol 70 (5) ◽  
pp. 3171-3175 ◽  
Author(s):  
X. Bonjoch ◽  
E. Ballesté ◽  
A. R. Blanch
Keyword(s):  
16S Rrna ◽  
Multiplex Pcr ◽  
Fecal Pollution ◽  
Sensitivity Threshold ◽  
Rrna Gene ◽  
Specific Primers ◽  
Animal Wastewater ◽  
Wastewater Samples ◽  
Species Specific ◽  
Different Origins

ABSTRACT Bifidobacteria are one of the most common bacterial types found in the intestines of humans and other animals and may be used as indicators of human fecal pollution. The presence of nine human-related Bifidobacterium species was analyzed in human and animal wastewater samples of different origins by using species-specific primers based on 16S rRNA sequences. Only B. adolescentis and B. dentium were found exclusively in human sewage. A multiplex PCR approach with strain-specific primers was developed. The method showed a sensitivity threshold of 10 cells/ml. This new molecular method could provide useful information for the characterization of fecal pollution sources.

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