Photorhabdus antumapuensis sp. nov., a novel symbiotic bacterial species associated with Heterorhabditis atacamensis entomopathogenic nematodes.

One motile, Gram-negative, non-spore-forming and rod-shaped symbiotic bacterium, strain UCH-936T, was isolated from Heterorhabditis atacamensis nematodes. Results of biochemical, physiological, molecular and genomic analyses suggest that it represents a new species, which we propose to name Photorhabdus antumapuensis sp. nov. Digital DNA-DNA hybridization shows that strain UCH-936T is more closely related to Photorhabdus kleinii DSM 23513T, but shares solely 50.5 % similarity, which is below the 70% cut-off value that delimits species boundaries in bacteria. Phylogenetic reconstructions using whole-genome sequences show that strain UCH-936T forms a unique clade, suggesting its novel and distinct taxonomic status again. Similarly, comparative genomic analyses shows that the virulence factor flagella-related gene fleR, the type IV pili-related gene pilL and the vibriobactin-related gene vibE are present in the genome of strain UCH-936T but absent in the genomes of its closest relatives. Biochemically and physiologically, UCH-936T differs also from all closely related Photorhabdus species. Therefore, Photorhabdus antumapuensis sp. nov. is proposed as a new species with the type strain UCH-936T (CCCT 21.06T=CCM 9188T=CCOS 1991T).

Xenorhabdus lircayensis sp. nov., the symbiotic bacterium associated with the entomopathogenic nematode Steinernema unicornum.

Xenorhabdus is a symbiotic group of bacteria associated with entomopathogenic nematodes of the family Steinernematidae. Although the described Steirnernema species list is extensive, not all their symbiotic bacteria have been identified. One single motile, Gram-negative and non-spore-forming rod-shaped symbiotic bacterium, strain VLST, was isolated from the entomopathogenic nematode Steinernema unicornum. Analyses of the 16S rRNA gene determined that the VLST isolate belongs to the genus Xenorhabdus, and its closest related species is Xenorhabdus szentirmaii DSM 16338T (98.2 %). Deeper analyses using the whole genome for phylogenetic reconstruction indicate that VLST exhibits a unique clade in the genus. Genomic comparisons considering digital DNA-DNA hybridization (dDDH) values confirms this result, showing that the VLST values are distant enough from the 70 % threshold suggested for new species, sharing 30.7, 30.5 and 30.3 % dDDH with Xenorhabdus khoisanae MCB, Xenorhabdus koppenhoeferi DSM 18168T and Xenorhabdus miraniensis DSM 18168T, respectively, as the closest species. Detailed physiological, biochemical and chemotaxonomic tests of the VLST isolate reveal consistent differences from previously described Xenorhabdus species. Phylogenetic, physiological, biochemical and chemotaxonomic approaches show that VLST represents a new species of the genus Xenorhabdus, for which the name Xenorhabdus lircayensis sp. nov. (type strain VLST=CCCT 20.04T=DSM 111583T) is proposed.

Morphological, molecular and ecological characterization of a native isolate of Steinernema feltiae (Rhabditida: Steinernematidae) from southern Chile.

BACKGROUND: Steinernema feltiae is an entomopathogenic nematode used in biological control programs with a global distribution. Populations of this species show phenotypic plasticity derived from local adaptation and vary in different traits, such as location and host penetration. The aim of this work was to describe a Chilean isolate of this nematode species, using integrative approaches. METHODS: Nematode morphological and morphometric studies were conducted along with molecular analysis of nuclear genes. The symbiotic bacterium was also identified by sequencing the 16S rRNA gene. Some ecological characteristics were described, including the temperature requirements for the nematode life cycle and the effect of soil water content for optimal reproduction. RESULTS: Morphometric characterization revealed a large intra-specific variability. The isolate identity was also corroborated with the analysis of nuclear genes. Based on the 16S gene, its symbiont bacteria, Xenorhabdus bovienii, was identified. The lowest, optimal and highest temperatures found to limit the infestation and reproduction on Galleria mellonella were 10, 20 and 30 °C, respectively; the emergence from the host larvae occurred approximately 10 days after inoculation. Differences were observed in offspring, and 120 infective juveniles (IJ)/larva was the most prolific dose at 20 °C. The soil water content did not affect the number of IJ invaders, penetration efficacy and IJ emergence time or offspring per larva, but it caused a delay in achieving full mortality at the permanent wilting point with respect to saturation and field capacity. CONCLUSIONS: For the first time, a Chilean isolate of S. feltiae is described in detail considering morphological, molecular and ecological aspects. The isolate was shown to be efficient in soil containing water, with optimal temperatures ranging from 15 to 25 °C for host infestation and production of an abundant offspring; these characteristics would allow its potential use as control agents in a wide geographical area of the country.

Soil texture, infective juvenile concentration, and soil organic matter influence the efficacy of Steinernema feltiae isolate Lican Ray.

The influence of infective juveniles (IJs) concentration, soil texture, IJ-host distance and organic matter (OM) content, at different decomposition degree, on the activity of the nematode Steinernema feltiae isolate Lican Ray (LR) was examined using Galleria mellonella larvae. Bioassays were conducted in tubes of varied length, filled with soil of different textures, placed either vertically or horizontally. In the concentration assay, highest IJ concentrations caused maximum larval mortality in all soil types (440, 2,200 and 4,400 IJs in clay, loam and sandy loam). In the second assay, soil texture (loam, clay or sandy loam) interacted significantly with IJ-host distance (10, 20, 30, 40 cm, horizontally; 30, 50, 70 cm, vertically), and distances of 30 cm or more affected IJ effectiveness on the control of G. mellonella. The effect was stronger in clay and sandy loam than in loam soils, where IJ moved up to 40 cm horizontally and 70 cm vertically. In the third assay, OM content (0, 2, 4, 6 and 8%) and its decomposition degree (initial, medium and advanced) did not interact to influence IJ movement in all treatments that contained any percentage of OM (2-8%). Only in the soil with no OM, IJ did not cause death of larvae at all. These results show the potential of S. feltiae LR to be used in different soil textures, as long as the content of soil OM allows its dispersal and host infection, in order to optimize the pest-control activity of the nematode.The influence of infective juveniles (IJs) concentration, soil texture, IJ-host distance and organic matter (OM) content, at different decomposition degree, on the activity of the nematode Steinernema feltiae isolate Lican Ray (LR) was examined using Galleria mellonella larvae. Bioassays were conducted in tubes of varied length, filled with soil of different textures, placed either vertically or horizontally. In the concentration assay, highest IJ concentrations caused maximum larval mortality in all soil types (440, 2,200 and 4,400 IJs in clay, loam and sandy loam). In the second assay, soil texture (loam, clay or sandy loam) interacted significantly with IJ-host distance (10, 20, 30, 40 cm, horizontally; 30, 50, 70 cm, vertically), and distances of 30 cm or more affected IJ effectiveness on the control of G. mellonella. The effect was stronger in clay and sandy loam than in loam soils, where IJ moved up to 40 cm horizontally and 70 cm vertically. In the third assay, OM content (0, 2, 4, 6 and 8%) and its decomposition degree (initial, medium and advanced) did not interact to influence IJ movement in all treatments that contained any percentage of OM (2­8%). Only in the soil with no OM, IJ did not cause death of larvae at all. These results show the potential of S. feltiae LR to be used in different soil textures, as long as the content of soil OM allows its dispersal and host infection, in order to optimize the pest-control activity of the nematode.