Root-knot nematodes (Meloidogyne spp.) are the most common and destructive plant-parasitic nematode group worldwide and adversely influence both crop quality and yield. In this study, a total of 51 root-knot nematode populations from turfgrasses were tested, of which 44 were from North Carolina, 6 from South Carolina and 1 from Virginia. Molecular characterisation was performed on these samples by DNA sequencing on the ribosomal DNA 18S, ITS and 28S D2/D3. Species-specific primers were developed to identify turfgrass root-knot nematode through simplex or duplex PCR. Four species were identified, including M. marylandi Jepson & Golden in Jepson, 1987, M. graminis (Sledge & Golden, 1964) Whitehead, 1968, M. incognita (Kofoid & White, 1919) Chitwood, 1949 and M. naasi Franklin, 1965 through a combined analysis of DNA sequencing and PCR by species-specific primers. M. marylandi has been reported from North Carolina and South Carolina for the first time. Molecular diagnosis using PCR by species-specific primers provides a rapid and cheap species identification approach for turfgrass root-knot nematodes.

Oscheius carolinensis n. sp. (Rhabditidae) was recovered through the Galleria bait method from vermicompost produced in Raleigh, NC, USA. Morphological studies with light microscopy and scanning electron microscopy, as well as molecular analyses of the near-full-length small subunit rDNA gene (SSU), D2/D3 expansion segments of the large subunit rDNA gene (LSU) and internal transcribed spacer (ITS), revealed this as a new species, described herein as Oscheius carolinensis n. sp. The new species is characterised by a combination of characters including its unique DNA sequences, amphimictic reproduction, six separate lips with bristle-like sensilla, lateral field with four lines, leptoderan and open male bursa, arrangement of bursal papillae 1 + 1 + 1/3 + 3 + ph, evenly spaced first, second and third papillae, and separate spicules that are distally shaped like a crochet needle. Oscheius carolinensis n. sp. belongs to the Insectivorus-group and is closest to O. colombianus, O. chongmingensis n. comb., O. insectivorus and O. lucianii. A Bacillus-like bacterium appears to be associated with this nematode, based on our microscopic and SEM observations. Exposed Galleria larvae were killed within 5 days and numerous nematodes were recovered from the cadavers within 48 h. Preliminary tests revealed that this nematode is capable of infecting at least two other insect species (Pieris rapae and Tenebrio molitor) under laboratory conditions and therefore has potential as a biological control agent. The status of Heterorhabditidoides chongmingensis is discussed, the genus is proposed as a junior synonym of Oscheius, and O. chongmingensis n. comb. is proposed.

Root-knot nematodes (Meloidogyne spp.) are the most common major pathogens of many crops throughout the world, impacting both the quantity and quality of marketable yields. In this study, a total of 244 root-knot nematode populations from various hosts from 39 counties in Arkansas were tested to determine the species diversity. Molecular characterization was performed on these populations by DNA sequencing of the ribosomal DNA 18S-ITS-5.8S, 28S D2/D3 and a mitochondrial DNA fragment flanking cytochrome oxidase gene subunit II - the intergenic spacer. Five species were identified, including M. incognita (Kofoid & White, 1919) Chitwood, 1949 from soybean, cotton, corn and various vegetables (232 samples); M. hapla Chitwood, 1949 from rose (1 sample); M. haplanaria Eisenback, Bernard, Starr, Lee & Tomaszewski, 2003 from okra, tomato, peanut, Indian hawthorn, ash, willow and elm trees (7 samples); M. marylandi Jepson & Golden in Jepson, 1987 from grasses (3 samples); and M. partityla Kleynhans, 1986 from pecan (1 sample) through a combined analysis of DNA sequencing and PCR by species-specific primers. Meloidogyne incognita is the most abundant species that was identified in 95% samples and was the only species in field crops including soybean and cotton, except for one population of M. haplanaria from soybean in Logan County (TK201). Species-specific primers were used to verify M. incognita through PCR by species-specific primers. Unlike historical data, M. arenaria, M. javanica and M. graminis were not detected from any of the samples collected during this study. This result is essential for effective and sustainable management strategies against root-knot nematodes in Arkansas.

Acrostichus puri n. sp., isolated from the body of a sweat bee, Augochlora pura mosieri, is described and figured. The new species is morphologically, molecularly and biologically very similar to A. halicti, i.e., these two species share a very complicated gubernaculum structure, distally hooked spicules with a triangular flap-like rostrum, stomatal polymorphism and synchronised associations with halcitid bees. The new species can be distinguished from A. halicti by its spicule morphology, distinctive squared manubrium and sharply bent (= ca 90° or L-shaped) distal tip vs an indistinctive rounded manubrium and a strongly recurved (= ca 180° or U-shaped) distal tip; gubernaculum with narrower, longer, anterior part and more complex posterior part than A. halicti, and tail of both sexes, with a long and filiform spike on the male or female tail vs a short spike (male) or conical tail (female). These two species also differ from each other by significant molecular sequence differences in SSU (2.0%), D2/D3 LSU (4.5%) and mtCOI (9.2%), and are reproductively incompatible in mating tests.

Taxonomy and morphology of plant-pa rasitic nematodes associated with turfgrasses in North and South Carolina, USA

In a search for an entomopathogenic nematode to control cranberry insect pests, three Oscheius populations (Rhabditidae) were recovered through the Galleria-bait method from one sample taken in a wild cranberry marsh in Jackson County, Wisconsin, USA. Morphological studies with light microscopy and scanning electron microscopy, as well as molecular analyses of the near-full-length small subunit rDNA gene, D2/D3 expansion segments of the large subunit rDNA gene, internal transcribed spacer, and mitochondrial cytochrome oxidase subunit 1 (CoxI) genes revealed this as Oscheius onirici, a species recently described from a karst cave soil of central Italy. The species belongs to the dolichura-group and is characterized by its DNA sequences; hermaphroditic reproduction; and males not found. A Bacillus-like bacterium appears to be associated with this nematode based on our microscopic and SEM observations; however its identity and persistent association with the nematode has not been confirmed. Nonetheless, this nematode is capable of infecting and killing the sparganothis fruitworm Sparganothis sulfureana Clemens (Lepidoptera: Tortricidae), the brown-banded cockroach Supella longipalpa Fabricius (Blattodea: Ectobiidae), and the cranberry fruitworm Acrobasis vaccinii Riley (Lepidoptera: Pyralidae), under laboratory conditions, and each in less than 72 hr. The mealworm Tenebrio molitor Linnaeus (Coleoptera: Tenebrionidae) and the greater wax moth Galleria mellonella Linnaeus (Lepidoptera: Pyralidae), are also susceptible, but take 3.5 and 5.2 days to die, respectively. This species is a new potential bio-control agent on insects.