Description of Globodera agulhasensis n. sp. (Nematoda: Heteroderidae) from South Africa

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A new cyst nematode, herein described as Globodera agulhasensis n. sp., was found parasitising Senecio burchelli 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.

Description of Globodera agulhasensis n. sp. (Nematoda: Heteroderidae) from South Africa

in Nematology

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References

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Figures

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    The type locality of Globodera agulhasensis n. sp. in the Cape Floristic Region of South Africa. The sampling location is indicated by an X.

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    Globodera agulhasensis n. sp. A: Whole cysts; B: Cyst vulval basin; C: Female anterior region; D: Male anterior region; E: Male tail region; F: J2 anterior region; G: J2 tail region.

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    Photomicrographs of females and cysts of Globodera agulhasensis n. sp. A: Female; B: Female anterior region showing median bulb; C: Female anterior region showing stylet and lip annuli; D: Female vulval basin showing cuticular ornamentation; E: Female vulval basin showing tubercles; F: Female vulval basin from the inside, showing vulval slit; G: Cyst vulval basin showing cuticular ornamentation; H: Cyst vulval basin showing V-shaped subsurface mark in cuticle; I: Females on roots. Abbreviations: a = anus; c = cuticular ridges; f = vulval fenestra; k = stylet knobs; s = stylet; mb = median bulb; t = tubercles; v = vulval slit. Scale bars: A, I = 100 μm; B-H = 10 μm. This figure is published in colour in the online edition of this journal, which can be accessed via http://booksandjournals.brillonline.com/content/journals/15685411.

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    Photomicrographs of second-stage juveniles and males of Globodera agulhasensis n. sp. A-C: Anterior region of second-stage juveniles; D-F: Tail region of second-stage juveniles; G, H: Anterior region of male; I: Tail region of male. Abbreviations: a = anus; g = gubernaculum; h = hyaline region; s = stylet; sp = spicule; k = knobs; mb = median bulb. Scale bars = 10 μm. This figure is published in colour in the online edition of this journal, which can be accessed via http://booksandjournals.brillonline.com/content/journals/15685411.

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    Cysts of Globodera agulhasensis n. sp. (SEM). A: Detail of anus; B: Cuticular ornamentation in vicinity of anus; C: Punctations on cyst wall; D: Internal morphology of cyst wall; E: External morphology of cyst wall; F: Cross section of cyst wall. Abbreviations: a = anus; c = cuticular ridges; f = vulval fenestra; p = punctations.

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    Second-stage juveniles of Globodera agulhasensis n. sp. (SEM). A, B: Head (A: Lateral view; B: Frontal view); C: Mid-body, showing lateral lines; D-F: Tail (D: Lateral view of entire tail; E: Ventral view of tail area showing anus (arrow); F: Lateral view of ending of lateral lines showing phasmid (arrow)).

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    RsaI digestion products of the amplified ITS1 region of Globodera spp. A: 100 bp marker (Fermentas); B: G. rostochiensis; C: G. capensis; D: G. agulhasensis n. sp.; E: WK1.

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    Phylogenetic relationships of Globodera species as inferred from ITS-rRNA sequences by using the neighbour-joining method. The optimal tree with the sum of branch length = 0.66840077 is shown. Tree drawn to scale with branch lengths in same units as those of evolutionary distances used to infer phylogenetic tree. The percentage of replicate trees in which associated taxa clustered together in the bootstrap test (1000 replicates) is shown next to the branches.

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    Phylogenetic relationships of Globodera species as inferred from ITS-rRNA sequences by using the maximum likelihood method. The tree with the highest log likelihood (−4197.4930) is shown. The tree is drawn to scale, with branch lengths measured in the number of substitutions per site. The percentage of trees in which the associated taxa clustered together in the bootstrap test (1000 replicates) is shown next to the branches.

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    Phylogenetic relationships of Globodera species as inferred from ITS-rRNA sequences by using the maximum parsimony method. The bootstrap consensus tree inferred from 1000 replicates is taken to represent the evolutionary history of the taxa analysed. The percentage of replicate trees in which the associated taxa clustered together in the bootstrap test is shown next to the branches.

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