Project description:The objective of this work was to determine the effectiveness of cross-hybridization of gDNA from five native soil nematodes to an Affymetrix Caenorhabditis elegans tiling array. Cross-hybridization experiments using C. briggsae, for which genome information is available, allowed hybridisation intensities to be correlated with known sequence differences. Initial analysis of data by conventional array-based Comparative Genomic Hybridization (aCGH) techniques at the chip level lead to misleading results due to an artefact from the combination of scaling, bandwidth smoothing, and differential GC content in exon and intron regions. To circumvent this artefact, individual probes were instead normalized and centered by adjusting for probe-specific thermodynamic binding affinity. However, cross-hybridization of C. briggsae DNA revealed that the resultant probe intensities alone were still uncorrelated to sequence similarity below 90% identity. Below 90% similarity, all probes hybridize uniformly poorly, and above 90% similarity the hybridization differences are not large enough to detect over background, therefore, no 'threshold' ratio of hybridization intensity was successful at identifying probes with similarity to the heterologous genome. In light of the observations described here, we suggest that the criteria for replication and verification of gene expression profiles generated from cross-species microarray hybridizations be more stringent than typically adopted for con-specific hybridizations.
Project description:The objective of this work was to determine the effectiveness of cross-hybridization of gDNA from five native soil nematodes to an Affymetrix Caenorhabditis elegans tiling array. Cross-hybridization experiments using C. briggsae, for which genome information is available, allowed hybridisation intensities to be correlated with known sequence differences. Initial analysis of data by conventional array-based Comparative Genomic Hybridization (aCGH) techniques at the chip level lead to misleading results due to an artefact from the combination of scaling, bandwidth smoothing, and differential GC content in exon and intron regions. To circumvent this artefact, individual probes were instead normalized and centered by adjusting for probe-specific thermodynamic binding affinity. However, cross-hybridization of C. briggsae DNA revealed that the resultant probe intensities alone were still uncorrelated to sequence similarity below 90% identity. Below 90% similarity, all probes hybridize uniformly poorly, and above 90% similarity the hybridization differences are not large enough to detect over background, therefore, no 'threshold' ratio of hybridization intensity was successful at identifying probes with similarity to the heterologous genome. In light of the observations described here, we suggest that the criteria for replication and verification of gene expression profiles generated from cross-species microarray hybridizations be more stringent than typically adopted for con-specific hybridizations. Genomic DNA from Caenorhabditis elegans N2 (Bristol), C. elegans CB4856 (Hawaiian), C. briggsae AF16, Oscheius tipulae KS585, Oscheius FVV-2 KS555, Mesorhabditis sp. KS587, Acrobeloides sp. KS586, and Chiloplacus sp. KS584 were hybridized onto C. elegans Affymetrix tiling array (two replicate chips were performed for each species).
Project description:Two new species, Neotobrilus nicsmolae n. sp. and Chronogaster carolinensis n. sp. are described from a small, acidic, temperate, natural lake in North Carolina. N. nicsmolae n. sp. comes close to three members of the genus reported from North America, N. filipjevi, N. longus, and N. hopei. However, N. nicsmolae is unique with in the genus in having a combination of characters: size smaller than 1,700 μm, shorter outer labial and cephalic setae, tail shorter than 250 μm, last ventromedian supplement close (about 5 μm) to cloacal opening, spicule length of 61 to 85 μm, flagelloid sperm, and possession of subterminal setae. Assessment of relationships among clades within the Triplonchida using DNA sequences of the D2D3 expansion segment of the LSU rDNA showed that the family Trichodoridae and the genus Tripyla were recovered as monophyletic. The genus Tobrilus was recovered as monophyletic in the neighbor-joining and maximum likelihood trees, but that was not so in the maximum-parsimony tree. The separation among genera of the Trichodoridae, i.e., Trichodorus and Paratrichodorus, was not clear-cut in all phylograms. Chronogaster carolinensis n. sp. in having one ventral mucro with no spine and vacuolated lateral glandular bodies comes close to C. typica and C. ethiopica but differs from all hitherto known species in a combination of characteristics: in having long cephalic setae, long stoma, crystalloid bodies, vacuolated lateral glandular bodies, and a tail terminus with blunt ventral mucro, and its lack of lateral line.
Project description:Sectonema caobangense sp. n. from evergreen forest soil in Vietnam is described, including scanning electron micrograph (SEM) observations and D2-D3 LSU rDNA analysis. The new species is characterized by its 3.12 to 5.80 mm long body, lip region offset by deep constriction and 21 to 23 μm broad, mural tooth 13 to 14 μm long at its ventral side, 940 to 1,112 μm long neck, pharyngeal expansion occupying 61% to 69% of total neck length, uterus a long simple tube-like structure 292 to 363 μm long or 2.7 to 2.9 times the corresponding body diameter, pars refringens vaginae well developed, V = 48 to 56, short (36-51 μm, c = 77-132, c' = 0.5-0.8) and rounded tail, 87 to 99 μm long spicules, and four or five irregularly spaced ventromedian supplements bearing hiatus. Sectonema caobangense sp. n. differs from the typical pattern of Sectonema in the nature of the stomatal protrusible structure, bearing a mural tooth attached to the ventral side of the stoma. Molecular data obtained and the derived evolutionary trees support a close phylogenetic relationship with other Sectonema species.
Project description:A new species of the genus Geocenamus was isolated from soil and root samples from the rhizosphere of Casuarina equisetifolia (Casuarinaceae) in Quang Nam province, Vietnam. This species is characterized by a round-to-hexagonal labial disc, the presence of a labial region, which is continuous or slightly offset from the body with six sectors, lateral sectors of first labial annulus being smaller than the submedian sectors, the presence of six to seven labial annules; the absence of deirids; stylet length 24 to 28???m long, body length 776 to 979???m long; lateral field with six to eight lateral lines, without areolation at mid-body and with areolation in outer bands at the tail region and a pointed tail terminus. Geocenamus vietnamensis n. sp. most closely resembles G. boghiae in having a non-sclerotized head framework and lacking a bursa in the males. It can be clearly distinguished from all other species of the genus Geocenamus by these characteristics. The combination of morphology, morphometric features, and phylogenetic trees, based on D2-D3 of 28S and ITS rDNA sequences, showed that this new species can be clearly separated from all other sequenced species. This record is the first for Geocenamus in Vietnam.A new species of the genus Geocenamus was isolated from soil and root samples from the rhizosphere of Casuarina equisetifolia (Casuarinaceae) in Quang Nam province, Vietnam. This species is characterized by a round-to-hexagonal labial disc, the presence of a labial region, which is continuous or slightly offset from the body with six sectors, lateral sectors of first labial annulus being smaller than the submedian sectors, the presence of six to seven labial annules; the absence of deirids; stylet length 24 to 28???m long, body length 776 to 979???m long; lateral field with six to eight lateral lines, without areolation at mid-body and with areolation in outer bands at the tail region and a pointed tail terminus. Geocenamus vietnamensis n. sp. most closely resembles G. boghiae in having a non-sclerotized head framework and lacking a bursa in the males. It can be clearly distinguished from all other species of the genus Geocenamus by these characteristics. The combination of morphology, morphometric features, and phylogenetic trees, based on D2–D3 of 28S and ITS rDNA sequences, showed that this new species can be clearly separated from all other sequenced species. This record is the first for Geocenamus in Vietnam.