A new cyst nematode, herein described as Globodera capensis n. sp., was found on several farms in the Swartland and Sandveld areas in South Africa. As the host plant of G. capensis n. sp. is currently unknown, the species is represented by cysts and second-stage juveniles (J2) only. The cysts have a spherical form, are yellow to dark brown in colour and have bullae (vulval bodies) and subcuticular punctations. Six to 20 cuticular ridges/lines are present between the anus and vulval basin, Granek’s ratio is 0.7-4.0, the vulval basin diam. is 16-28.5 μm and vulval basin to anus distance is 19-60 μm. The J2 is 430-528 μm long and the stylet is 23-28 μm long with anteriorly flattened to rounded stylet knobs. These stages are morphologically and morphometrically very near to, or indistinguishable from PCN, G. artemisiae, G. millefolii and G. tabacum tabacum. It can be distinguished by the molecular characteristics of the ITS-region of the ribosomal DNA. Pairwise distances between closely related Globodera species show that G. capensis n. sp. differs the least in number of base differences per sequence from G. millefolii (63-64 bp) and most from an undescribed Globodera from Chile (72-73 bp). Phylogenetic relationships of G. capensis n. sp. with selected species of Punctoderinae, inferred from ITS-rRNA sequences by using the Maximum Likelihood method, indicate that G. capensis n. sp. occupies a basal position within a lineage of Globodera species from Europe, Asia and New Zealand that parasitises non-solanaceous plants. For diagnostic purposes, digestion of the ITS1 with AluI will distinguish G. capensis n. sp. from G. mexicana, G. pallida, G. rostochiensis and G. tabacum, but not from G. artemisiae, G. millefolii and G. zelandica, whilst FauI will distinguish G. capensis n. sp. from all the other Globodera species tested.
During a survey for entomopathogenic nematodes in citrus orchards throughout South Africa, a new species of Steinernema was isolated from a citrus orchard on Rietkloof farm, near the town of Piketberg in the Western Cape Province, South Africa. The nematode was isolated from soil using the Galleria-baiting technique. Steinernema citrae n. sp. is characterised by the following morphological characters: third-stage infective juvenile with a body length of 754 (623-849) μm, distance from head to excretory pore of 56 (49-64) μm, tail length of 71 (63-81) μm, and ratio E value of 110 (85-132). The lateral pattern for the new species is 2, 7, 8, 6, 4, 2 and is not typical for the genus. Steinernema citrae n. sp. is closely related the feltiae-group. The body length of the IJ is close to that of S. texanum and S. weiseri, though it differs in body diam., the length of the pharynx and E%. The male of S. citrae n. sp. differs from S. feltiae in the length and shape of the spicule and body diam. Steinernema citrae n. sp. differs from all species in the feltiae-group in the morphology of the vulva, as it has a single flapped, low, epiptygma. It also differs from the most closely related species, S. feltiae, as there is no interbreeding between the two species. In addition, the new nematode differs from other species of the feltiae-group by characteristics of the ITS and D2D3 regions of its rDNA.
Steinernema nguyeni n. sp. was recovered by baiting from beneath an Olea africana tree in South Africa. The combination of morphological and molecular features suggests that S. nguyeni n. sp. is a member of the feltiae-kraussei-oregonense group, clustering with members of this group in Clade III. The new species is morphologically characterised by the infective juvenile body length of 737 (673-796) μm and the number of ridges in the infective juvenile lateral field is 2, 8, 2. The male of the first generation can be recognised by the spicule length of 66 (58-75) μm and a gubernaculum length of 43 (30-55) μm. The first generation female can be recognised by the vulval lips only slightly protruding and the presence of low, double-flapped epiptygmata. Analysis of the ITS and D2-D3 regions of the ribosomal DNA confirms that S. nguyeni n. sp. differs from all other known Steinernema species.
A new cyst nematode, herein described as Globoderaagulhasensis n. sp., was found parasitising Senecioburchelli in the Western Cape Province, South Africa. Second-stage juveniles are characterised by a well developed stylet of 23.5 (22.5-24.8) μm with rounded to anteriorly flattened knobs. The dorsal pharyngeal gland outlet is 4.4 (3.5-6.5) μm posterior to the stylet knobs. The tail is 56 (49-64) μm long and the length of the hyaline region is 25 (19-29) μm. The cysts are characterised by their ovate to spherical shape, short neck, the presence of subcuticular punctations over the entire body and the absence of bullae or vulval bodies. Six to 12 cuticular ridges/lines are present on the outer surface of the cyst between the anus and vulval basin. Granek’s ratio is 1.7 (1.0-3.0), the vulval basin diam. 17.6 (11.7-26.1) μm and the distance between vulval basin and anus is 28.6 (19.1-47.0) μm. Males have a stylet length of 26.1 (24.4-27.7) μm and spicule length of 30.3 (27.2-33.8) μm with a rounded tip. Females have a stylet length of 22.1 (19.0-24.4) μm, a large median bulb almost filling the body diam., and a short vulval slit 4.2 (3.2-6.6) μm long. Phylogenetic relationships of G. agulhasensis n. sp. with other species of the genus, inferred from ITS-rRNA sequences by using the neighbour-joining (NJ), maximum likelihood (ML) and maximum parsimony method (MP), indicate that G.agulhasensis n. sp. is included in the clade of Globodera sp. that parasitise non-solanaceous plants, forming a monophyletic group with unidentified Globodera spp. from Portugal, G. millefolii and G. artemisiae. For diagnostic purposes, three restriction enzymes, Hpy8I, RsaI and XceI were selected as being able to discriminate between G. agulhasensis n. sp. and other Globodera spp. present in South Africa.
A new cyst nematode, herein described as Globoderasandveldensis n. sp., was found in the Sandveld, Western Cape Province, South Africa. As the host plant of G. sandveldensis n. sp. is currently unknown, the species is represented by cysts and second-stage juveniles (J2) only. The J2 are characterised by a well developed stylet of 26.4 (24.8-28.5) μm with rounded to anteriorly flattened knobs. The distance from the dorsal pharyngeal gland outlet to the stylet knobs is 4.5 (2.4-7.2) μm. The tail is 64 (56-77) μm long and the length of the hyaline region is 33 (22-39) μm. The cysts are characterised by their ovate to spherical shape, short neck, and the presence of subcuticular punctations and bullae or vulval bodies. Six to 14 cuticular ridges/lines are present on the outer surface of the cyst between the anus and vulval basin. Granek’s ratio is 1.7 (0.9-3.0), the vulval basin diam. is 20.4 (13.1-34.6) μm and the distance between vulval basin and anus is 33.5 (19.7-54.0) μm. Phylogenetic relationships with other species of the genus, inferred from ITS-rDNA sequences, indicate that G. sandveldensis n. sp. is included in the clade of Globodera sp. that parasitise non-solanaceous plants and is closely related to unidentified Globodera spp. from Portugal, G. millefolii, and G. artemisiae.
A new species of entomopathogenic nematode, Steinernema sacchari n. sp., was isolated by trapping with the sugar cane borer, Eldana saccharina, from soil of a sugar cane field in the KwaZulu-Natal province of South Africa. The new species is morphologically characterised by the length of the infective juvenile (IJ) of 680 (630-722) μm, tail length of 64 (51-74) μm, ratio a = 19 (14-23), H% = 49 (43-57) and E% = 82 (70-109). The pattern of the lateral field of the IJ of the new species is 2, 5, 2 ridges (3, 6, 3 lines or incisures). The male of the first generation can be recognised by the long spicule of 83 (73-89) μm, gubernaculum of 61 (50-68) μm, D% = 67 (54-88) and GS% = 73 (66-81). The first generation male lacks a mucron, while the second generation male always has one. The first generation female can be recognised by the vulval lips not being raised, the possession of long double-flapped epiptygmata and the lack of a postanal swelling. Analysis of the ITS and D2D3 regions showed S. sacchari n. sp. to differ from all other Steinernema species and to belong to a new monophyletic group, the ‘Cameroonian’ clade, consisting of S. cameroonense, S. nyetense and S. sacchari n. sp. This group is closely related to the feltiae-kraussei-oregonense Clade III.
During a survey in the Mpumalanga province of South Africa, a Steinernema species was isolated from a soil sample taken from a litchi orchard. Steinernema litchii n. sp. can be separated from other, closely related, species in the glaseri-group by morphological, morphometric and molecular analyses. The infective third-stage juvenile of the new species has a body length of 1054 (953-1146) μm, distance from head to excretory pore of 78 (64-86) μm, as well as eight ridges (i.e., nine lines) in the mid-body region. The c-ratio of 10 (9-13) is low and the tail is long at 95 (73-105) μm. First generation males have a spicule length of 86 (76-96) μm and a gubernaculum length of 65 (59-72) μm. The tail of the first generation male lacks a mucron; that of the second generation always bears one. The genital papillae total 23 and consist of 11 pairs and an unpaired precloacal papilla. The vulva of S. litchii n. sp. has a slightly asymmetrical protuberance and short, double-flapped epiptygmata. The female has a slightly protuberant postanal swelling. Phylogenetic analysis of the internal transcribed spacer (ITS) and of the 28S (D2-D3) regions of the ribosomal DNA (rDNA) confirmed the close relationship of S. litchii n. sp. to the Karii-clade. Both morphological and molecular evidence support the species status of S. litchii n. sp.
Aphelenchoides arachidis is reported for the first time from South Africa and for the fourth time outside Nigeria. The A. arachidis-infested pods from South Africa showed the following symptoms: small seeds with the testa wrinkled and darker in colour than that of non-infested seeds; the pods showed dark lesions and some seeds within the pods showed early germination. Differences between the two South African and the Nigerian populations of A. arachidis include more lateral lines in some specimens (2-4 vs 2) and, on average, longer post-uterine sac length (extending for 74 (41-96) and 62 (33-82) vs about 50% of vulva to anus distance). Scanning electron micrographs of this species are presented for the first time. The ITS regions of ribosomal DNA were amplified, sequenced, aligned and compared with other sequences of Aphelenchoides species. Two pathogenic fungi, Thielaviopsis basicola and Neocosmospora vasinfecta, were also isolated from this material.
Globodera presently contains 13 valid and three as yet undescribed species. Three species, G. rostochiensis, G. pallida and G. ellingtonae, the potato cyst nematodes (PCN), cause significant economic losses on potatoes around the world. In our study we provide comprehensive phylogenetic analyses of 455 ITS rRNA, 219 COI and 164 cytb gene sequences of 11 valid and two undescribed species of Globodera using Bayesian inference, maximum likelihood and statistical parsimony. New 205 COI, 116 cytb and 21 ITS rRNA gene sequences were obtained from 148 populations of these species collected from 23 countries. The phylogenetic analysis revealed that Globodera displayed two main clades in the trees: i) Globodera from South and North America parasitising plants from Solanaceae; and ii) Globodera from Africa, Europe, Asia and New Zealand parasitising plants from Asteraceae and other families. Based on the results of phylogeographical analysis and age estimation of clades with a molecular clock approach, it is hypothesised that Globodera species originated and diversified from several centres of speciation located in mountain regions and then dispersed across the world from these regions during the Pleistocene. High genetic diversity of Bolivian populations of G. rostochiensis was observed for both mtDNA genes. Analysis of phylogenetic relationships of G. pallida and G. rostochiensis populations revealed incongruence in topology between networks inferred from mtDNA genes, which might be an indication of possible recombination and selective introgression events through gene flow between previously isolated populations. This puts some limitations on the use of the mtDNA marker as universal DNA barcoding identifier for PCN. Globodera bravoae syn. n. is proposed as a junior synonym of G. mexicana.