Morphological and molecular characterisation of Heterodera filipjevi (Madzhidov, 1981) from the Slovak Republic

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Cysts, males and juveniles from a population of Heterodera filipjevi found on sports-ground turf in Trnava, Slovak Republic, in 2016 are described based on morphology and morphometrics. The identity of species of juveniles extracted from cysts was subsequently determined and confirmed by PCR-RFLP and sequencing. Morphologically, cysts, juveniles and males from the Slovak Republic are similar to paratypes from Tajikistan. The results of the phylogenetic analysis of the ITS rRNA gene sequences confirmed the species identification and phylogenetic relationship of H. filipjevi with other Heterodera species. Of interest, this analysis showed the close similarity between the Slovakian (KY349106) and Chinese (KU896216) samples, which differed by a single nucleotide and clustered together. To our knowledge, this is also the first report of H. filipjevi from the Slovak Republic, thus increasing the total number of cyst nematode species of the Heterodera genus known to occur in the country to a total of 15 species.

Morphological and molecular characterisation of Heterodera filipjevi (Madzhidov, 1981) from the Slovak Republic

in Nematology

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References

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Figures

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    Light microscope photos of cysts and juveniles of Heterodera filipjevi. A: Cysts; B-E: Vulval area of cysts; F: Entire body of second-stage juvenile (J2); G: Head region and stylet of J2; H-I: Tail of J2. (Scale bars: A = 500 μm; B-E, G-I = 10 μm; F = 100 μm.)

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    Light microscope photos of males of Heterodera filipjevi. A: Entire body of male; B, C: Head region; D: Excretory pore; E: Sperm; F: Tail region with spicule. (Scale bars: A = 100 μm; B-F = 10 μm.)

  • View in gallery

    PCR-ITS-RFLP profile for the cyst nematode Heterodera filipjevi (Madzhidov, 1981): M = 100 bp DNA ladder (CEB), 1: AluI; 2: AvaI; 3: Bsh1236I; 4: HaeIII; 5: CfoI; 6: MvaI; 7: PstI; 8: RsaI.

  • View in gallery

    Phylogenetic tree resulting from alignment of ITS-rRNA sequences. The tree was inferred by Maximum Likelihood method under the GTR + G model. Bootstrap values were obtained from 100 replicates. Values higher than 50% are presented. Original sequence is indicated by bold font.

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