Influences of space, soil, nematodes and plants on microbial community composition of chalk grassland soils

Microbial communities respond to a variety of environmental factors related to resources (e.g. plant and soil organic matter), habitat (e.g. soil characteristics) and predation (e.g. nematodes, protozoa and viruses). However, the relative contribution of these factors on microbial community composition is poorly understood. Here, we sampled soils from 30 chalk grassland fields located in three different chalk hill ridges of Southern England, using a spatially explicit sampling scheme. We assessed microbial communities via phospholipid fatty acid (PLFA) analyses and PCR-denaturing gradient gel electrophoresis (DGGE) and measured soil characteristics, as well as nematode and plant community composition. The relative influences of space, soil, vegetation and nematodes on soil microorganisms were contrasted using variation partitioning and path analysis. Results indicate that soil characteristics and plant community composition, representing habitat and resources, shape soil microbial community composition, whereas the influence of nematodes, a potential predation factor, appears to be relatively small. Spatial variation in microbial community structure was detected at broad (between fields) and fine (within fields) scales, suggesting that microbial communities exhibit biogeographic patterns at different scales. Although our analysis included several relevant explanatory data sets, a large part of the variation in microbial communities remained unexplained (up to 92% in some analyses). However, in several analyses, significant parts of the variation in microbial community structure could be explained. The results of this study contribute to our understanding of the relative importance of different environmental and spatial factors in driving the composition of soil-borne microbial communities.

The combined effect of the application of a biocontrol agent Paecilomyces lilacinus, with various practices for the control of root-knot nematodes

The effectiveness of a formulated product containing spores of the naturally occurring fungus Paecilomyces lilacinus, strain 251, was evaluated against root-knot nematodes in pot and greenhouse experiments. Decrease of second-stage juveniles hatching from eggs was recorded by using the bio-nematicide at a dose of 4 kg ha(-1), while a further decrease was recorded by doubling the dose. However, the mortality rate decreased by increasing the inoculum level. Application of P. lilacinus and Bacillus firmus, singly or together in pot experiments, provided effective control of second-stage juveniles, eggs or egg masses of root-knot nematodes. In a greenhouse experiment, the bio-nematicide was evaluated for its potential to control root-knot nematodes either as a stand-alone method or in combination with soil solarization. Soil was solarized for 15 d and the bio-nematicide was applied just after the removal of the plastic sheet. Soil solarization for 15 d either alone or combined with the use of P. lilacinus did not provide satisfactory control of root-knot nematodes. The use of oxamyl, which was applied 2 weeks before and during transplanting, gave results similar to the commercial product containing P. lilacinus but superior to soil solarization. (C) 2007 Elsevier Ltd. All rights reserved.

Effects of a non-chemical nematicide combined with soil solarization for the control of root-knot nematodes

The effectiveness of a formulated bio-nematicide product containing lyophilized bacteria spores of Bacillus firmus was evaluated against root-knot nematodes (RKN) in greenhouse and field experiments. A decrease of second stage juveniles hatching from eggs was recorded by using the bio-nematicide at a dose of 0.9 g kg(-1) of soil while further a decrease was recorded by doubling the dose. However, the mortality rate decreased as the inoculurn level increased. Exposure of either second stage juveniles or egg masses to temperatures of 35-40 degrees C for 1-4 weeks had a marked effect on their survival. In a field experiment, the bio-nematicide was evaluated for its potential to control RKN either as a stand-alone method or in combination with soil solarization. The latter was tested for 15-30 days and the bionematicide was applied just before soil coverage with the plastic sheet or just after its removal. Soil solarization either for 15-30 days provided satisfactory control of RKN. The combination of soil solarization with the bio-nematicide improved nematode control and gave results similar to the chemical treatment. (c) 2007 Elsevier Ltd. All rights reserved.

Susceptibility of different insect pupae to the bacterial symbiont, Xenorhabdus nematophila, isolated from the entomopathogenic nematode Steinernema carpocapsae

Cells of the bacterial symbiont Xenorhabdus nematophila from the entomopathogenic nematode, Steinernema carpocapsae entered the pupae of Plutella xylostella after 15 minutes treatment with suspensions containing the bacterial cells. Secretions of Xenorhabdus nematophila, in either broth or water, were found lethal to the pupae of P. xylostella when applied in moist sand. The bacterial symbiont Xenorhabdus nematophila was found lethal to the pupae of greater wax moth (Galleria mellonella), beet armyworm (Spodoptera exigua), diamondback moth (Plutella xylostella) and black vine weevil (Otiorhynchus sulcatus) in the absence of the nematode vector and the cells of X. nematophila entered the haemocoele of the pupae.

The potential of combining Pasteuria penetrans and neem (Azadirachta indica) formulations as a management system for root-knot nematodes on tomato

Initial applications of 10(4) spores g(-1) of Pasteuria penetrans, and dried neem cake and leaves at 3 and 2% w:w, respectively, were applied to soil in pots. Juveniles of Meloidogyne javanica were added immediately to the pots (500, 5,000 or 10,000) before planting 6-week-old tomato seedlings. The tomatoes were sampled after 64 days; subsequently a second crop was grown for 59 days and a third crop for 67 days without further applications of P. penetrans and neem. There was significantly less root-galling in the P. penetrans combined with neem cake treatment at the end of the third crop and this treatment also had the greatest effect on the growth of the tomato plants. At the end of the third crop, 30% of the females were infected with P. penetrans in those treatments where spores had been applied at the start of the experiment. The effects of neem leaves and neem cake on the nematode population did not persist through the crop sequences but the potential for combining the amendments with a biological control agent such as P. penetrans is worthy of further evaluation.

Efficacy of neem (Azadirachta indica) formulations on biology of root-knot nematodes (Meloidogyne javanica) on tomato

Second stage juveniles of Meloidogyne javanica were exposed to aqueous extracts of neem crude formulations (leaves and cake) at 10%, 5%, and 2.5% w/v and a refined product, Aza at 0.1% w/v. The 10% extracts of neem leaf and cake caused 83% and 85% immobility and 35% and 28% mortality, respectively. Aza caused neither immobility or mortality of juveniles. When egg masses were placed in extracts of these formulations, hatching did not occur at all the concentrations (10%, 5%, 2.5% and 1.25% w/v) of the crude formulations. When the treated egg masses were returned to water, the eggs resumed hatching. Aza did not affect the nematode hatching. In glasshouse experiments, soil application of neem formulations significantly reduced the invasion of tomato roots by root-knot nematodes but once the nematodes managed to invade them, no effect detected on their development. Soil applications of Aza at 0.05% and 0.1% w/v significantly reduced the invasion and delayed development of nematodes within tomato roots whereas 0.025% did not. There were significantly fewer egg masses on tomato roots exposed to single egg mass in neem amended soil as compared to control. (C) 2007 Elsevier Ltd. All rights reserved.

Systemic and persistent effect of neem (Azadirachta indica) formulations against root-knot nematodes, Meloidogyne javanica and their storage life

Neem leaves, neem cake (a by-product left after the extraction of oil from neem seed) and a commercially refined product aza (azadirachtin) extracted from seed were evaluated. Aqueous extracts of crude neem formulations used as a seedling dip treatment significantly reduced the number of females and egg masses in roots whereas the refined one did not. A split-root technique was used to demonstrate the translocation of active compounds within a plant and their subsequent effect on the development of nematodes. When applied to the root portion all formulations significantly reduced the number of egg masses and eggs per egg mass. Whereas on the untreated root portion, neem cake at 3% w/w and aza at 0.1% w/w significantly reduced the number of egg masses as compared with neem leaves at 3% w/w, aza at 0.05% and control. All the neern formulations significantly reduced the number of eggs per egg mass on' the untreated root portion. The effect of neem leaves and cake on the development of root-knot nematodes was tested at 2, 4, 6, 8, and 16 weeks after their application to soil. Even after 16 weeks all the treatments significantly reduced the galling index and number of egg masses but their effectiveness declined over time. After storing neem leaves, cake and aza for 8 months under ambient conditions the efficacy of neem leaves and aza, against root-knot nematodes, remained stable whereas that of cake declined. (c) 2006 Elsevier Ltd. All rights reserved.

Genotypic differences in the growth of bananas (Musa spp.) infected with migratory endoparasitic nematodes. 1. Roots

The effects of nematodes on root morphology and the association of root characteristics with resistance to nematodes of seven banana varieties were investigated in two experiments. Banana plants were grown in controlled conditions within polytunnels and harvested on three occasions for the measurement of root morpholopy, and biomass. Varieties differed in their resistance to nematodes, from resistant (Yg Km5, FHIA 17, FHIA 03) and partly resistant (FHIA 01, FHIA 25) to not resistant ((FHIA 23, Williams). Nematodes reduced the root dry weight of FHIA 01, FHIA 17 and FHIA 23 at some harvests. Primary root number was on average 9.5% lower in nematode-infected plants than controls, with no differences among the varieties. Thus, there was no simple association between the resistance of these varieties and their tolerance to nematodes. Varieties differed in root morphology. Root dry weight was greatest for resistant varieties Yg Km5 and FHIA 03, and least for non-resistant varieties FHIA 23 and Williams. Thus, resistance to nematodes was associated with varieties with greater root mass and more and larger primary roots.

Genotypic differences in the growth of bananas (Musa spp.) infected with migratory endoparasitic nematodes. 2. Shoots

In order to identify the effect of burrowing nematodes on the shoots (pseudostem and leaves) of banana plants and to determine whether or not shoot characteristics are associated with plant resistance to nematodes two experiments were conducted in controlled conditions within polytunnels. The banana plants were harvested on three occasions for the measurement of root morphology and biomass. Varieties differed in their resistance to nematodes from resistant (Yg Km5, FHIA 17, FHIA 03) and partly resistant (FHIA 01, FHIA 25) to not resistant (FHIA 23, Williams). Nematodes reduced total plant dry weight at the first harvest in Experiment 1 and by an average of 8.8% in Experiment 2, but did not affect leaf area in either experiment. The ratio of above-ground Weight to total plant weight was reduced from 75% to 72% in nematode-infected plants compared with the control plants for all varieties tested in Experiment 1, but was only reduced in FHIA 25 and FHIA 23 in Experiment 2. Varieties differed in above-ground growth. The FHIA varieties had greater shoot weights and leaf area than YgKm5 and Williams. Overall, resistance to nematodes was associated with the partitioning of a greater proportion of biomass to the roots than to above-ground parts.

Use of entomopathogenic bacterium Pseudomonas putida (Enterobacteriaceae) and it secretions against the Greater Wax Moth Galleria mellonella pupae

The bacterium from Pseudomonas putida from Steinernema abbasi and its metabolic secretions caused the mortality of the Galleria mellonella pupae. Experiments were conducted in sand and filter paper on time exposure, temperature, moisture, dose and time of penetration of bacterium in pupae and tested stored or dried toxic metabolites using G. mellonella pupae as a test target organism. Death of pupae was probably due to the toxic metabolites. Pseudomonas putida cells were recovered from the haemocoele when bacterial cells were applied to the G. mellonella pupae indicating that bacterial cells can enter the haemocoele in the absence of nematode vector. Penetration of bacterium was found rapidly after application on G. mellonella pupae. Pseudomonas putida or its toxic secretions can be used as a microbial control for insect control. The experimental results indicate that there is possibility of using P. putida and its toxic secretions as a biopesticide and can contribute in the development of new microbial and biological control against insect pests.