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

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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.

Nematology

International Journal of Fundamental and Applied Nematological Research

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Figures

  • Phylogenetic relationships within populations and species of the Pratylenchus coffeae species complex as inferred from Bayesian analysis using the D2-D3 of 28S rRNA gene sequence dataset with the GTR + I + G model. Posterior probability more than 70% is given for appropriate clades. Newly obtained sequences are indicated in bold. *Sequences from Nguyen (2010); **only D3 segment of a sample originally identified as P. musicola.

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  • Phylogenetic relationships within populations and species of the Pratylenchus coffeae species complex as inferred from Bayesian analysis using the ITS rRNA gene sequence dataset with the GTR + I + G model. Posterior probability more than 70% is given for appropriate clades. Newly obtained sequences are indicated in bold. *Sequence from Cheng et al. (2009) incorrectly annotated as Caenorhabditis elegans; **sequence from Uehara et al. (1998).

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  • Phylogenetic relationships within populations and species of the Pratylenchus coffeae species complex as inferred from Bayesian analysis using the hsp90 gene sequence dataset with the GTR + I + G model. Posterior probability more than 70% is given for appropriate clades. Newly obtained sequences are indicated in bold.

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  • PCR-D2-D3-RFLP for the Pratylenchus coffeae species complex. A: Cfr42I; B: Bpu1102I; Lanes: M: 100 bp DNA ladder (Promega); 1, 2: P. speijeri n. sp. Kade, Ghana; 3, 4: Pratylenchus sp. C1 (Japan, Colocasia esculenta, CA80 and CA96); 5: P. coffeae (USA, FL, Aglaonema sp., CD820); 6: P. coffeae (Viet Nam, Musa sp.); 7: P. coffeae (Haiti, Musa sp.).

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  • Duplex PCR with the Pratylenchus coffeae group-specific and P. speijeri n. sp. species-specific primers. Lanes: M: 100 bp DNA marker (Promega); 1, 2: P. speijeri n. sp. Kade, Ghana; 3, 4: Pratylenchus sp. C1 (Japan, Colocasia esculenta, CA80 and CA96); 5: P. coffeae (USA, FL, Aglaonema sp., CD820); 6: P. neglectus; 7: P. brachyurus; 8: control without DNA. Arrows indicate a group-specific band for Pratylenchus coffeae species complex (417-425 bp) and a species-specific band for P. speijeri n. sp. (102 bp).

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  • Pratylenchus speijeri n. sp. A, B: Female and male entire body; C: Female pharyngeal region; D: Male pharyngeal region; E: Female anterior region; F: Schematic view of undivided face pattern of female; G: Female vulval region with spermatheca; H: Female posterior region; I, J: Male tail; K-O: Female tail.

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  • Light micrographs of Pratylenchus speijeri n. sp. A, B: Female pharyngeal region showing hemizonid and excretory pore; C: Male pharyngeal region; D: Female anterior end; E: Female vulval region; F, H: Female tail; G: Female lateral field; I: Female posterior body portion; J: Male tail. (Scale bars = 20 μm.)

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  • Common tail shapes observed in specimens of Pratylenchus speijeri n. sp. Note tail terminus indentation (F, J), a common feature of this root-lesion nematode.

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  • SEM morphology of Pratylenchus speijeri n. sp. Left and right: Undivided face pattern of female with all labial sectors fused together and with oral disc; Centre: Female face lateral view showing second lip annulus higher than first.

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  • Comparative line drawings of morphological features of the closely related Pratylenchus auracensis (P.a.), P. coffeae (P.c.), P. floridensis (P.f.), P. jaehni (P.j.), P. loosi (P.l.), P. parafloridensis (P.p.) and P. speijeri n. sp. (P.s.). (5, 18, 25 after Seinhorst, 1977, and 7, 20, 27 after Múnera et al., 2009, modified.)

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