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  • Author or Editor: Renato N. Inserra x

Renato N. Inserra, Jason D. Stanley, Alberto Troccoli, John Chitambar and Sergei A. Subbotin

Hemicaloosia vagisclera n. sp. is described from Bermuda grass (Cynodon dactylon) in Florida. This new species is characterised by females with body slightly ventrally arcuate in death, face with a prominent elliptical oral disc, large amphids, slender stylet with mean length 64 μm, lateral field consisting of a single longitudinal line marked by continuous and discontinuous transverse striae, oval and full spermatheca, sclerotised vagina vera and tail annuli width greater than that of remaining body annuli. Diagnostic characters for the males are a C-shaped body, head with 4-5 faint lip annuli and pronounced oval oral disc, lateral field with two longitudinal lines intersected by transverse striae, tail digitate in the distal portion posterior to bursa, distinctly annulated and with a round terminus. Molecular characterisation of H. vagisclera n. sp. using the D2-D3 domain of 28S rRNA, partial 18S rRNA and ITS rRNA gene sequences is also provided. The phylogenetic relationships of this species with other representatives of the suborder Criconematina are presented and indicate that H. vagisclera n. sp. has sister relationships with Caloosia longicaudata supporting the classification of Caloosia together with Hemicaloosia as separate genera in the family Caloosiidae. A diagnostic PCR-ITS-RFLP profile for H. vagisclera n. sp. is also given together with an identification key for seven known species of Hemicaloosia.

Esther Van den Berg, Louwrens R. Tiedt, Jason D. Stanley, Renato N. Inserra and Sergei A. Subbotin

The genus Scutellonema contains more than 40 species of spiral nematodes with enlarged phasmids called scutella. In this study, we provide morphological and molecular characterisation of S. clavicaudatum sp. n., S. brachyurus, S. bradys, S. cavenessi, S. transvaalense, S. truncatum and Scutellonema sp. A. from North and Central America, and Africa. The new species, S. clavicaudatum sp. n., was found on sugarcane in South Africa and is characterised by a lack of lip annuli as in S. africanum, S. siamense and S. truncatum. The lip region, in both males and females, is conical and marked by six large rectangular blocks separated or fused with the submedian and lateral lip sectors, which surround a round and distinct labial disc. Females of this new species also have large vaginal glands, a functional spermatheca, the lateral field posterior to the scutellum ending in a bluntly pointed shape and a clavate tail. Morphological descriptions, measurements, light and scanning electron microscopic photos and drawings are also given for S. bradys, S. cavenessi, S. transvaalense and S. truncatum. The study of spiral nematode samples from Florida, USA, confirmed the presence of a morphologically and genetically atypical populations of S. bradys. The morphology of the S. bradys population from Bermuda grass in pasture land from central Florida fits that of type specimens of this species, but differs in having a truncate tail terminus rather than round and also a prominent spermatheca filled with flagellate spermatozoa. Other Florida Scutellonema samples analysed in this study belonged to S. cavenessi, a species native to West Africa. This is the first report of S. cavenessi in Florida, where it parasitises the ornamental plant Sansevieria trifasciata. Our study showed a high level of intraspecific variation for Scutellonema rRNA and mtDNA genes, which can reach 5.6% for the D2-D3 of 28S rRNA, 12.9% for the ITS rRNA genes and 14.4% for the COI gene. 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.

Alberto Troccoli, Sergei A. Subbotin, Jason D. Stanley, Brian Alford, Nicola Vovlas and Renato N. Inserra

Morphological and molecular analyses of three populations of Meloidoderita whittoni (Sledge & Christie, 1962) comb. n. (syn. Sphaeronema whittoni; Tumiota whittoni) collected in Florida from sweet gum (Liquidambar styraciflua L.) indicated that this species is a representative of Meloidoderita rather than Sphaeronema, where it was included in the original description, or Tumiota, to where it was subsequently transferred. However, this species can be considered an atypical representative of the genus, because it differs from the other species of this genus in having females retaining the eggs inside a thin-walled uterus, which remains encased inside their body. After egg deposition inside the uterus, the female dies and its body is transformed into a persistent tanned sac with a thick cuticle, devoid of ornamentations, which protects the eggs like a heteroderid cyst. The female secretes from the vulva a gelatinous matrix, which becomes hardened in time and encases its body for protection against predation and parasitism by biological antagonists as has been suggested for other tylenchuloid nematodes. No egg deposition outside the female body was observed. Second-stage juveniles of this species have a semi-endoparasitic, rather than endoparasitic, habit as in other known Meloidoderita species. This is the first report of a sphaeronematid having a cyst stage fitting the definition of a heteroderid cyst. Phylogenetic relationships between some species of Tylenchuloidea and Criconematina are analysed using the partial 18S rRNA, the D2-D3 of 28S rRNA and the ITS rRNA gene sequences.

Rodrigo Rodríguez-Kábana, Rosa H. Manzanilla-López, Renato N. Inserra and Nahum Marbán-Mendoza

Zahra Tanha Maafi, Majid Amani, Jason D. Stanley, Renato N. Inserra, Esther Van den Berg and Sergei A. Subbotin

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.

Sergei A. Subbotin, Jerry Akanwari, Chau N. Nguyen, Ignacio Cid del Prado Vera, John J. Chitambar, Renato N. Inserra and Vladimir N. Chizhov

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: iM. sikhotealiniensis and Cryphodera spp. from Asia and Europe; and iiM. 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.

Sergei A. Subbotin, Nicola Vovlas, Gregor W. Yeates, Johannes Hallmann, Sebastian Kiewnick, Vladimir N. Chizhov, Rosa H. Manzanilla-López, Renato N. Inserra and Pablo Castillo

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.

Sergei A. Subbotin, Ignacio Cid Del Prado Vera, Renato N. Inserra, Vladimir N. Chizhov and Wilfrida Decraemer

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.

Esther Van den Berg, Louwrens R. Tiedt, Renato N. Inserra, Jason D. Stanley, Nicola Vovlas, Juan E. Palomares Rius, Pablo Castillo and Sergei A. Subbotin

Sheathoid nematodes of the genus Hemicriconemoides are 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 genus viz.: H. alexis, H. brachyurus, H. californianus, H. chitwoodi, H. macrodorus, H. minutus, H. ortonwilliamsi, H. promissus, H. silvaticus, H. strictathecatus, H. wessoni and Hemicriconemoides sp. 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 within Hemicriconemoides, 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 for H. californianus, H. chitwoodi and H. strictathecatus.

Francesca De Luca, Alberto Troccoli, Larry W. Duncan, Sergei A. Subbotin, Lieven Waeyenberge, Daniel L. Coyne, Francis C. Brentu and Renato N. Inserra

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.