Design and evaluation of nematode 18S rDNA primers for PCR and denaturing gradient gel electrophoresis (DGGE) of soil community DNA (original) (raw)

Metabarcoding of soil nematodes: the importance of taxonomic coverage and availability of reference sequences in choosing suitable marker(s)

Metabarcoding and Metagenomics, 2019

For many organisms, there is agreement on the specific genomic region used for developing barcode markers. With nematodes, however, it has been found that the COI region designated for most animals lacks the taxonomic coverage (ability to amplify a diverse group of taxa) required of a metabarcoding marker. For that reason, studies on metabarcoding of nematodes thus far have utilized primarily regions within the highly conserved 18S ribosomal DNA. Two popular markers within this region are the ones flanked by the primer pairs NF1-18Sr2b and SSUF04-SSUR22. The NF1-18Sr2b primer pair, especially, has been critiqued as not being specific enough for nematodes leading to suggestions for other candidate markers while the SSUF04-SSUR22 region has hardly been tested on soil nematodes. The current study aimed to evaluate these two markers against other alternative ones within the 28S rDNA and the COI region for their suitability for nematode metabarcoding. The results showed that the NF1-18Sr...

Characterization of soil nematode communities in three cropping systems through morphological and DNA metabarcoding approaches

Scientific reports, 2018

We used complementary morphological and DNA metabarcoding approaches to characterize soil nematode communities in three cropping systems, conventional till (CT), no-till (NT) and organic (ORG), from a long-term field experiment. We hypothesized that organic inputs to the ORG system would promote a more abundant nematode community, and that the NT system would show a more structured trophic system (higher Bongers MI) than CT due to decreased soil disturbance. The abundance of Tylenchidae and Cephalobidae both showed positive correlations to soil organic carbon and nitrogen, which were highest in the ORG system. The density of omnivore-predator and bacterial-feeding nematodes was reduced in NT soils compared to CT, while some plant-parasitic taxa increased. NT soils had similar Bongers MI values to CT, suggesting they contained nematode communities associated with soils experiencing comparable levels of disturbance. Metabarcoding revealed within-family differences in nematode diversit...

Distribution and diversity of cyst nematode (Nematoda: Heteroderidae) populations in the Republic of Azerbaijan, and their molecular characterization using ITS-rDNA analysis

Nematropica, 2019

Cereal cyst nematodes (Heterodera spp.) are a significant threat to global cereal production systems and choosing the correct management strategy requires knowledge and identification of various species. We conducted a survey across the main cereal-growing regions of the Republic of Azerbaijan in 2017. Cyst-forming nematodes were detected in 34 samples (44.7%), of which 28 were identified as H. filipjevi or H. avenae using internal transcribed spacer (ITS)-rDNA sequencing. Six populations were unidentifiable to species level and were recorded as belonging to the H. avenae group. H. filipjevi was the dominant species, found in 19 samples (25%) from the provinces of Qobustan, Ismailli, Oguz, Sheki, Barda, and Kurdamir. H. avenae was detected in 9 samples (11.8%) across the provinces of Ismailli, Oguz, and Sheki. ITS-rDNA phylogenetic analyses showed that populations of H. filipjevi clustered in one group with two subgroups, all supported by high bootstrap values. Populations of H. ave...

Nematode 28 S rDNA sequences for selected taxa in Figure 3 . Species Isolate Locality Accession

2018

Millipedes may cause unexpected damage when they are introduced to new locations, becoming invaders that leave behind their old parasites and predators. Therefore, it was interesting to find numerous rhabditid nematodes within the gut of the invasive phytophagous millipede Chamberlinius hualienensis Wang, 1956 (Diplopoda, Paradoxosomatidae) from Hachijojima (Japan) in November, 2014. This millipede originated in Taiwan but was discovered in Japan in 1986. The nematodes were identified as juvenile Oscheius rugaoensis (Zhang et al., 2012) Darsouei et al., 2014 (Rhabditidae), and juvenile and adult Mononchoides sp. (Diplogastridae) based on images, morphometrics, and sequences of 18S and 28S rDNA. A novel short 28S sequence of a separate population of Oscheius necromenus SB218 from Australian millipedes was also included in a phylogenetic comparison of what can now be characterized as a species complex of millipede-associated Oscheius. The only other nematode associates of millipedes b...