Molecular and morphological characterisation of Sphaeronema alni Turkina & Chizhov, 1986 (Nematoda: Sphaeronematidae) from Spain compared with a topotype population from Russia

The occurrence of a male-less population of Sphaeronema alni parasitising chestnut (Castanea sativa) roots and inducing a stelar syncytium is reported for the first time in Pola de Somiedo (Oviedo province), Spain.Morphometric and molecular characters of the Spanish population matched those of a topotype population from Russia. SEM observations showed swollen females having the first lip annulus wider than the second and appearing as a cap-like, circumoral elevation. The second-stage juveniles, having a single band in the lateral fields, were characterised by a non-annulated dome-shaped lip region derived from the fusion of the oral disc with all the lip sectors and lip annuli, and showing slit-like amphidial apertures and an oval prestoma. The sequences of the D2-D3 expansion segments of 28S rRNA, partial 18S rRNA and ITS rRNA gene for the Spanish and topotype populations of S. alni were congruent and matched those deposited in GenBank for another population from Germany, thereby confirming their conspecificity. A PCR-RFLP profile of D2-D3 of 28S rRNA for identification of this species was also provided. The phylogenetic relationships between S. alni populations and representatives of the suborder Criconematina, as inferred from analysis of partial 18S rRNA and D2-D3 of 28S gene sequences obtained in this and previous studies, indicated that S. alni formed a basal clade on the majority consensus Bayesian phylogenetic trees, standing together with Meloidoderita sp. or alone. These findings provide additional evidence of the need to clarify the position of Sphaeronema within Criconematina and its relationships with representatives of Tylenchulinae.