Descriptions of Trichodorus lownsberyi sp. n., Nanidorus minor (Colbran, 1956) Siddiqi, 1974 (Nematoda: Trichodoridae) and Longidorus quercus sp. n. (Nematoda: Longidoridae) from Mexico

Summary A new species of stubby root nematode, Trichodorus lownsberyi sp. n., collected from soil around Buxus sempervirens at Montecillo Campus, Colegio de Postgraduados, Mexico, is described and illustrated. Trichodorus lownsberyi sp. n. is characterised by the abundant males with a characteristic spicule shape, the narrower part about mid-blade having visible bristles in most specimens, and the presence and position of the three ventromedian cervical papillae, which are all anterior to the secretory-excretory pore. Females possess a rhomboid-shaped vagina and oval to rounded triangular vaginal sclerotised pieces with a pore-like vulva. The phylogenetic relationships of T. lownsberyi sp. n. with related species were constructed using the ITS2 rRNA and the D2-D3 expansion segments of 28S rRNA gene sequences. Trichodorus lownsberyi sp. n. was a sister species to T. viruliferus in the phylogenetic trees. A Mexican population of Nanidorus minor from a peach orchard is also described and illustrated. The new needle nematode, Longidorus quercus sp. n., was recovered from soil around roots of oak, Quercus crassipes, from Cerro Jusda ‘El Diablo’ in Mexico State. Longidorus quercus sp. n. females are characterised by the C-shaped posterior end of the body after fixation, L = 4.9 (3.9-5.6) mm, a = 71 (60.8-93.2), lip region rounded, sometimes slightly flattened, marked by depression, odontostylet 170 (144-206) μm long, hemispherical to bluntly conoid tail and no males. The new species was characterised using the D2-D3 of 28S rRNA and COI gene sequences. The phylogenetic relationships of L. quercus sp. n. with other Longidorus species were reconstructed using the D2-D3 of 28S rRNA gene sequences.

Assessment of Factors Associated with Molecular Quantification of Stubby Root Nematode Paratrichodorus allius from Field Soil DNA

Factors relating to SYBR Green-based quantitative real-time PCR (qPCR) quantification of stubby root nematode Paratrichodorus allius using soil DNA were evaluated in this study. Soils used were loamy sand from potato fields in North Dakota and Idaho. Results showed that the largest nematode individuals (body length >720 µm) produced significant lower Cq values than the smallest individuals (<359 µm), indicating more total DNA amount in the largest nematodes. Soil pre-treatments showed that autoclaved field soil had significantly reduced DNA amount and quality. The air- or oven-dried soil yielded a lower amount of DNA with similar purity, compared with natural field soil. PCR inhibitors were detected in soil DNA substrates targeting pBluescript II SK(+)-plasmid DNA. Al(NH4)(SO4)2 treatment during DNA preparation significantly reduced the inhibitors compared with post-treatment of soil DNA with polyvinylpolypyrrolidone column. The effect of PCR inhibitors on qPCR was suppressed by bovine serum albumin. Quantification results did not significantly change when increasing the number of DNA extractions from three to six per soil sample when soil grinding and grid sampling strategies were used. Two standard curves, generated from serial dilutions of plasmid DNA containing P. allius ITS1 rDNA and soil DNA containing known nematode numbers, produced similar correlations between Cq values and amount of targets. The targets in soil DNA quantified by qPCR using either standard curve correlated well with microscopic observations using both artificially and naturally infested field soils. This is the first study for assessing various factors that may affect qPCR quantification of stubby root nematodes. Results will be useful during the setup or optimization of qPCR-based quantification of plant-parasitic nematodes from soil DNA.

Molecular characterisation of some stubby root nematodes (Nematoda: Trichodoridae) from the USA and other countries

Summary The stubby root nematodes are world-wide distributed polyphagous root ectoparasites and can cause damage to a wide range of crops and natural vegetation. In this study, 22 valid and putative species of stubby root nematodes were identified in 37 samples collected in Malaysia, Mexico, Russia, and the USA (California, Florida, Minnesota and Nevada). The analysis of 58 new sequences of the D2-D3 expansion fragments of 28S rRNA gene revealed the following species: Nanidorus minor from California, Florida and Mexico, N. renifer from Florida, two unidentified Nanidorus species from California and Malaysia, respectively, Paratrichodorus allius from Florida and Minnesota, P. pachydermus and Paratrichodorus sp. from Russia, Trichodorus californicus, T. intermedius, and T. obscurus from California, T. obtusus from Florida, eight unidentified Trichodorus species from California, one unidentified Trichodorus species from Nevada, and two unidentified species of stubby root nematodes, one of Trichodorus and another undetermined species from Mexico. Molecular characterisation of T. californicus, T. intermedius and T. obscurus is given for the first time. The phylogenetic tree reconstructed from the analysis of 108 D2-D3 of 28S rRNA gene sequences of 58 valid and putative species of the stubby root nematodes contained five major clades: i) Trichodorus from Europe, Asia and North America; ii) Nanidorus and Trichodorus from Asia; iii) Trichodorus from California; iv) Paratrichodorus from several distant geographical regions; and v) Monotrichodorus from Central and South America. It has been hypothesised that the California Floristic Province is one of the centres of origin and diversification of stubby root nematodes. There is no information on the economic importance of the stubby root nematode species found in California and Mexico. The confirmation of the occurrence of N. renifer and P. allius in Florida should be of concern for the blueberry and potato industries in the state.

Stubby-Root Nematode, Trichodorus obtusus Cobb (Nematoda: Adenophorea: Triplonchida: Diphtherophorina: Trichodoridea: Trichodoridae)

Nematodes in the family Trichodoridae (Thorne, 1935) Siddiqi, 1961, are commonly called "stubby-root" nematodes, because feeding by these nematodes can cause a stunted or "stubby" appearing root system. Trichodorus obtusus is one of the most damaging nematodes on turfgrasses, but also may cause damage to other crops. This document is EENY-340, one of a series of Featured Creatures from the Entomology and Nematology Department, Florida Cooperative Extension Service, Institute of Food and Agricultural Sciences, University of Florida. Published: November 2004. EENY340/IN617: Stubby-Root Nematode, Trichodorus obtusus Cobb (syn T. proximus) (Nematoda: Adenophorea: Triplonchida: Diphtherophorina: Trichodoridea: Trichodoridae) (ufl.edu)

Stubby-Root Nematode, Paratrichodorus minor (Colbran) Siddiqi (Nematoda: Adenophorea: Triplonchida: Diphtherophorina: Trichodoridae: Trichodoridae)

Nematodes in the family Trichodoridae (Thorne, 1935) Siddiqi, 1961, are commonly called "stubby-root" nematodes, because feeding by these nematodes can cause a stunted or "stubby" appearing root system. Paratrichodorus minor is the most common species of stubby-root nematode in Florida, and in tropical and sub-tropical regions worldwide. Paratrichodorus minor is important because of the direct damage it causes to plant roots, and also because it can transmit certain plant viruses. This document is EENY-339, one of a series of Featured Creatures from the Entomology and Nematology Department, Florida Cooperative Extension Service, Institute of Food and Agricultural Sciences, University of Florida. Published: November 2004. EENY339/IN616: Stubby-Root Nematode, Nanidorus minor (Colbran) Siddiqi (syn. Paratrichodorus minor, P. christiei, Trichodorus minor, T. christiei) (Nematoda: Adenophorea: Triplonchida: Diphtherophorina: Trichodoridea: Trichodoridae) (ufl.edu)