Promise for plant pest control: root-associated pseudomonads with insecticidal activities.

Insects are an important and probably the most challenging pest to control in agriculture, in particular when they feed on belowground parts of plants. The application of synthetic pesticides is problematic owing to side effects on the environment, concerns for public health and the rapid development of resistance. Entomopathogenic bacteria, notably Bacillus thuringiensis and Photorhabdus/Xenorhabdus species, are promising alternatives to chemical insecticides, for they are able to efficiently kill insects and are considered to be environmentally sound and harmless to mammals. However, they have the handicap of showing limited environmental persistence or of depending on a nematode vector for insect infection. Intriguingly, certain strains of plant root-colonizing Pseudomonas bacteria display insect pathogenicity and thus could be formulated to extend the present range of bioinsecticides for protection of plants against root-feeding insects. These entomopathogenic pseudomonads belong to a group of plant-beneficial rhizobacteria that have the remarkable ability to suppress soil-borne plant pathogens, promote plant growth, and induce systemic plant defenses. Here we review for the first time the current knowledge about the occurrence and the molecular basis of insecticidal activity in pseudomonads with an emphasis on plant-beneficial and prominent pathogenic species. We discuss how this fascinating Pseudomonas trait may be exploited for novel root-based approaches to insect control in an integrated pest management framework.

Direct and indirect root defences of milkweed (Asclepias syriaca): trophic cascades, trade-offs and novel methods for studying subterranean herbivory

P>1. Entomopathogenic nematodes can function as indirect defence for plants that are attacked by root herbivores. By releasing volatile organic compounds (VOCs), plants signal the presence of host insects and thereby attract nematodes.2. Nonetheless, how roots deploy indirect defences, how indirect defences relate to direct defences, and the ecological consequences of root defence allocation for herbivores and plant biomass are essentially unknown.3. We investigate a natural below-ground tritrophic system, involving common milkweed, a specialist root-boring beetle and entomopathogenic nematodes, and asked whether there is a negative genetic correlation between direct defences (root cardenolides) and indirect defences (emission of volatiles in the roots and nematode attraction), and between constitutive and inducible defences.4. Volatiles of roots were analysed using two distinct sampling methods. First, we collected emissions from living Asclepias syriaca roots by dynamic headspace sampling. This method showed that attacked A. syriaca plants emit five times higher levels of volatiles than control plants. Secondly, we used a solid phase micro-extraction (SPME) method to sample the full pool of volatiles in roots for genetic correlations of volatile biosynthesis.5. Field experiments showed that entomopathogenic nematodes prevent the loss of biomass to root herbivory. Additionally, suppression of root herbivores was mediated directly by cardenolides and indirectly by the attraction of nematodes. Genetic families of plants with high cardenolides benefited less from nematodes compared to low-cardenolide families, suggesting that direct and indirect defences may be redundant. Although constitutive and induced root defences traded off within each strategy (for both direct and indirect defence, cardenolides and VOCs, respectively), we found no trade-off between the two strategies.6. Synthesis. Constitutive expression and inducibility of defences may trade off because of resource limitation or because they are redundant. Direct and indirect defences do not trade off, likely because they may not share a limiting resource and because independently they may promote defence across the patchiness of herbivore attack and nematode presence in the field. Indeed, some redundancy in strategies may be necessary to increase effective defence, but for each strategy, an economy of deployment reduces overall costs.

Weights in the balance: jasmonic acid and salicylic acid signaling in root-biotroph interactions.

Work on the interaction of aerial plant parts with pathogens has identified the signaling molecules jasmonic acid (JA) and salicylic acid (SA) as important players in induced defense of the plant against invading organisms. Much less is known about the role of JA and SA signaling in root infection. Recent progress has been made in research on plant interactions with biotrophic mutualists and parasites that exclusively associate with roots, namely arbuscular mycorrhizal and rhizobial symbioses on one hand and nematode and parasitic plant interactions on the other hand. Here, we review these recent advances relating JA and SA signaling to specific stages of root colonization and discuss how both signaling molecules contribute to a balance between compatibility and defense in mutualistic as well as parasitic biotroph-root interactions.

Managing Soybean Cyst Nematode, 2007

Soybean cyst nematode (SCN) causes the greatest yield loss of any single pathogen of soybean in Iowa. An estimated 50 million bushels were lost in Iowa to SCN in 2004. Damage from SCN is not limited to yield loss from root feeding; SCN also makes other diseases like sudden death syndrome, iron deficiency chlorosis, Pythium, Phytophthora root and stem rot and brown stem rot worse. Once established in a field, SCN cannot be eradicated. However, the use of multiple management tactics can help minimize yield loss.

Predicting root defence against herbivores during succession

P>1. Root herbivores and pathogens interfere with basic below-ground plant function, and can thereby affect plant fitness and spatial and temporal patterns in natural plant communities. However, there has been little development of concepts and theories on below-ground plant defence, a deficit that is in contrast to the abundance of theorizing for above-ground plant parts.2. A review of the past 10 years of research on below-ground plant-herbivore interactions has revealed that, similar to above-ground tissues, root defences can be expressed constitutively or induced upon herbivore attack, and can be classified into direct and indirect traits, tolerance, and escape. Indeed, it has been shown that roots tolerate herbivory by outgrowing or re-growing lost tissues, or resist it by producing secondary metabolites that are toxic to herbivores or attract natural enemies of herbivores.3. We propose that, similar to above-ground plant-herbivore theories, the partition of abiotic and biotic factors over ecological succession can serve as the basis for predicting investment in defence strategies below-ground.4. Investigation of herbivore pressure and root responses along primary and secondary successional gradients suggests that: (i) roots are often fast growing, thinner and softer in early compared to later succession. (ii) Insect and nematode herbivore pressure increases until mid-succession and later decreases. (iii) Mycorrhizal abundance increases with succession, and the composition of fungal species changes through succession, often shifting from arbuscular mycorrhizae to ecto-mycorrhizae.5. Based on these findings, and on classical (above-ground) plant defence theory, we suggest the following set of testable hypotheses for below-ground plant defence: (i) During succession, early plants invest most of their resources in growth and less in defences (associated with a general lack of herbivores and pathogens, and with limited availability of resources in the system), therefore relying more on re-growth (tolerance) strategies. (ii) During mid-succession, a buildup of herbivore pressure facilitates replacement by plant species that exhibit greater direct and indirect defence strategies. (iii) Constitutive and inducible levels of defences may trade-off, and early successional plants should rely more on induction of defences after herbivore attack, whereas late successional plants will increasingly rely on constitutively produced levels of physical and chemical defence. (iv) Successional changes in microbial associations have consequences for root defence by improving plant nutrition and defence expression as well as directly competing for root space; however, toxic or impenetrable root defences may also limit association with root symbionts, and so may constrain the expression of root defence.

Nematode community, trophic structure and population fluctuation in soybean fields

Temporal (monthly in three fields for 12 months) and spatial (once in 23 fields during March-April) samplings were conducted in the major soybean (Glycine max)-growing region of the Brazilian Federal District. Fifty-three nematode genera were found in both samplings, but 13 were detected only by the temporal sampling, and one only by the spatial sampling. Fifty-three percent were plant-parasites, 35% were bacterivores, and about 12% were fungivores, predators and omnivores constituted the community that was dominated by the genera Helicotylenchus (40% of total abundance), Acrobeles (15%), Cephalobus (7.6%), Meloidogyne(5.6%) and Pratylenchus (4.9%). Heterodera glycines was not found in this study. There were no differences in ten ecological measurements [Ds, H', Es, T, FF/BF, (FF+BF)/PP, MI, PPI, mMI, and Dorylaimida (%)] between the two sampling types, but differences in indexes d and J'. Plant parasite populations dropped at the end of the crop cycle, remained at low levels during the dry season and the seedling period, then increased again in the crop-growing season. Fungivores maintained their low populations throughout the year, increasing only in June and July, the post-harvest period, when soil fungi decomposed root tissue. The population of bacterivores slightly declined during the dry season and the initial rainy season, but peaked in the middle of the rainy season, apparently associated with soil humidity. In the five most abundant nematodes, those of Acrobeles and Pratylenchus were more populous in wet soils, Cephalobus and Meloidogyne adapted well in dry soils, but Helicotylenchus survived abundantly in a wide range of soil moisture.

Long-term response of the nematode community to elevated atmospheric CO2 in a temperate dry grassland soil

Long-term effects of the elevated atmospheric CO2 on biosphere have been in focus of research since the last few decades. In this experiment undisturbed soil monoliths of loess grassland were exposed to an elevated CO2 environment (two-times the ambient CO2 level) for a period of six years with the aid of the open top chamber method. Control without a chamber and CO2 elevation was applied as well. Elevated CO2 level had very little impact oil soil food web. It did not influence either root and microbial biomass or microbial and nematode community structure. The only significant response was that density of the bacterial feeder genus Heterocephalobus increased in the chamber with elevated CO2 concentration. Application of the open top chambers initiated more changes on nematodes than the elevated CO2 level. Open top chamber (OTC) method decreased nematode density (total and plant feeder as well) to less than half of the original level. Negative effect was found on the genus level in the case of fungal feeder Aphelenchoides, plant feeder Helicotylenchus and Paratylenchus. It is very likely that the significantly lower belowground root biomass and partly its decreased quality reflected by the increased C/N ratio are the main responsible factors for the lower density of the plant feeder nematodes in the plots of chambers. According to diversity profiles, MI and MI(2-15) parameters, nematode communities in the open top chambers (both on ambient and elevated CO2 level) seem to be more structured than those under normal circumstances six years after start of the experiment.

Susceptibility of winter crops to Pratylenchus brachyurus and effect on the nematode population in the maize crop

The aim of this study was to evaluate the susceptibility of winter crops to Pratylenchus brachyurus and their effect on the population of phytonematodes in the maize. To study the effect of the plants on nematodes, an experiment was set up in sandy, naturally-infested soil. The area was divided into strips, consisting of six blocks of 16 treatments, with eight winter treatments, subdivided on the basis of the fertilizer used (organic: bird litter, and synthetic: NPK). The initial nematode population was determined by sampling the soil (100 cm(3)) and weeds (10 g of root). The winter treatments put in place (bristle oats, chickpea, vetch bean, common bean, oilseed radish, wheat, intercropped bristle oats + oilseed radish and fallow), and the nematode population determined 100 days after sowing. Subsequently, two maize crops (summer and short season) were planted, and the nematode population in the soil and roots determined during crop full bloom. To evaluate the susceptibility of winter crops to nematodes, an experiment was conducted under controlled conditions, determining the nematode reproduction factor (RF) in the treatments described above. Both in the field and under controlled conditions, it was observed that the bristle oats, oilseed radish and intercropped oats + oilseed radish exhibited lower reproduction rates for P. brachyurus. In the field, lower population of nematodes was observed with the application of bird litter. Under controlled conditions, the highest RF were observed in the fallow plot and under common bean and chickpea, in that order.

Resistance of rose rootstocks to Meloidogyne hapla nematode

Roses are widely used in landscaping. One of the most important fitossanitary problems of this ornamental plant in gardens is the fitonematodes, especially Meloidogyne hapla, which is the most important one in colder climate regions. This work had the objective of study the resistance of nine rose rootstocks (Rosa multiflora 'Paulista', R. multiflora 'Japanese', R. multiflora 'Iowa', R. multiflora 'Kopman's', R. indica × multiflora, R. indica 'Mayor', R. sp. 'Natual Brier', R. manetti and R. canina 'Inermis') to Meloidogyne hapla nematode. Ten replications of each rootstock were used. The roots, collected in a commercial area of cut roses, visually had galls there were isolated and identified. The inoculum of M. hapla was previously produced in rose and tomatoes seedlings under greenhouse conditions. The plants were harvested ninety days after inoculation. The roots were washed and the number of eggs and juveniles recuperated in the root system of each plant was estimated by the Final Population (FP), and the Reproductive Factor (RF) was calculated. The rootstocks with RF<1 values were considered resistant, and the ones with RF>1, susceptible. Rosa manetti and Rosa sp. 'Natual Brier' rootstocks showed the lowest values for RF, indicating that, although they are susceptible, they provide the lowest nematode reproduction. Based on the RF values, it was concluded that all the rose rootstocks evaluated were susceptible to M. hapla nematode.

Aggressiveness of pratylenchus brachyurus to sugarcane, compared with key nematode p. zeae

Pratylenchus zeae, Meloidogyne javanica and M. incognita are considered key species of nematodes in sugarcane in Brazil, but P. brachyurus is also frequently found. This study was conducted to determine the aggressiveness of P. brachyurus compared with P. zeae to sugarcane. Plants were grown in pots (100 L) in an open area with initial inoculation of 10, 100, 1,000, 10,000 and 100,000/plant for P. brachyurus and P. zeae. The nematode inocula were from in vitro, carrot-cylinder cultures. Sampling was performed every 60 days until 300 days after inoculation. At harvest, we evaluated the population dynamics of the nematodes and plant growth characteristics. The population for the initial levels of 10 and 100,000 specimens/plant, for P. brachyurus and P. zeae at 300 days after inoculation were similar. This fact shows that, upon detection of nematodes in a certain place during the planting of sugarcane, the ratoon on this area should be treated so as to control populations of P. brachyurus and/or P. zeae. The damage caused by the initial population of 10 specimens of P. brachyurus was similar to those of 10.000 specimens of P. zeae. The variety CTC 2 was classified as susceptible to P. zeae and intolerant to P. brachyurus. Compared to the control, the losses as measured by the volume and fresh weight of shoots by the nematode species were 29.82% and 40.34%, respectively. Pratylenchus brachyurus was more aggressive than P. zeae to the CTC 2 sugarcane variety.

Systemic root signalling in a belowground, volatile-mediated tritrophic interaction

Plants attacked by leaf herbivores release volatile organic compounds (VOCs) both locally from the wounded site and systemically from non-attacked tissues. These volatiles serve as attractants for predators and parasitoids. This phenomenon is well described for plant leaves, but systemic induction of VOCs in the roots has remained unstudied. We assessed the spatial and temporal activation of the synthesis and release of (E)-β-caryophyllene (EβC) in maize roots upon feeding by larvae of Diabrotica virgifera virgifera, as well as the importance of systemically produced EβC for the attraction of the entomopathogenic nematode Heterorhabditis megidis. The production of EβC was found to be significantly stronger at the site of attack than in non-attacked tissues. A weak, but significant, increase in transcriptional activity of the EβC synthase gene tps23 and a corresponding increase in EβC content were observed in the roots above the feeding site and in adjacent roots, demonstrating for the first time that herbivory triggers systemic production of a volatile within root systems. In belowground olfactometers, the nematodes were significantly more attracted towards local feeding sites than systemically induced roots. The possible advantages and disadvantages of systemic volatile signalling in roots are discussed.

Effects of Seed Treatments and a Soil-applied Nematicide on Corn Yields and Nematode Population Densities

Plant-parasitic nematodes are microscopic worms that feed on plants. Almost every nematode that feeds on corn is capable of feeding on many other plants. These nematode parasites are thought to be native to most Iowa soils and to have fed upon native plants before corn was grown as a cultivated crop. Population densities (numbers) of most species of plant-parasitic nematodes that feed on corn have to increase to damaging levels (called damage thresholds) before yield loss occurs. Products that are currently available to manage plant-parasitic nematodes on corn in the state include the soil-applied insecticide/nematicide Counter® and two relatively new protectant seed treatments, Avicta® and Votivo®. Counter® is a contact and systematic nematicide with the active ingredient terbufos. Avicta® is a contact nematicide (active ingredient abamectin) that moves on the surface of the root, and Votivo® is a special strain of the natural soil bacterium Bacillus firmus that grows on the root. Counter® is available from AMVAC, Avicta® from Syngenta Seedcare, and Votivo® from Bayer CropScience. The objective of this experiment was to assess and compare the nematode population densities and yields of corn growing in plots with and without the seed-treatment nematode protectants and the soil-applied nematicide Counter®.

Some isolates of the nematophagous fungus Pochonia chlamydosporia promote root growth and reduce flowering time of tomato

The fungal parasite of nematode eggs Pochonia chlamydosporia is also a root endophyte known to promote growth of some plants. In this study, we analysed the effect of nine P. chlamydosporia isolates from worldwide origin on tomato growth. Experiments were performed at different scales (Petri dish, growth chamber and greenhouse conditions) and developmental stages (seedlings, plantlets and plants). Seven P. chlamydosporia isolates significantly (P < 0.05) increased the number of secondary roots and six of those increased total weight of tomato seedlings. Six P. chlamydosporia isolates also increased root weight of tomato plantlets. Root colonisation varied between different isolates of this fungus. Again P. chlamydosporia significantly increased root growth of tomato plants under greenhouse conditions and reduced flowering and fruiting times (up to 5 and 12 days, respectively) versus uninoculated tomato plants. P. chlamydosporia increased mature fruit weight in tomato plants. The basis of the mechanisms for growth, flowering and yield promotion in tomato by the fungus are unknown. However, we found that P. chlamydosporia can produce Indole-3-acetic acid and solubilise mineral phosphate. These results suggest that plant hormones or nutrient ability could play an important role. Our results put forward the agronomic importance of P. chlamydosporia as biocontrol agent of plant parasitic nematodes with tomato growth promoting capabilities.

Interspecific competition between the potato cyst nematode species Globodera pallida and G. rostochiensis

The two potato cyst nematode species, Globodera pallida and G. rostochiensis, are among the most important pests of potato. PCN are difficult to manage, while the two species respond differently to the main control methods. An increase in the incidence of G. pallida had been reported and is generally attributed to greater effectiveness of control measures against G. rostochiensis. The status of PCN in Ireland was studied using PCR. The results demonstrated qPCR to be an efficient means of high-throughput PCN sampling, being able to accurately identify both species in mixed-species populations. Species discrimination using qPCR revealed an increase in the incidence of G. pallida in Ireland in the absence of G. pallida-selective control measures. The population dynamics of G. pallida and G. rostochiensis in Ireland were studied in mixed- and single-species competition assays in vivo. G. pallida proved to be the more successful species, with greater multiplication in mixed- than single-species populations, with G. rostochiensis showing the opposite. This effect was similarly observed in staggered inoculation trials and population proportion trials. It was hypothesised that the greater G. pallida competitiveness could be attributed to its later hatch. G. pallida exhibited a later peak in hatching activity and more prolonged hatch, relative to G. rostochiensis. G. rostochiensis hatch was significantly reduced in mixedspecies hatching assays. G. pallida hatch was significantly higher when hatch was induced in potato root leachates containing G. rostochiensis-specific compounds, indicating that G. pallida hatch is stimulated upon perception of G. rostochiensis–derived compounds. Rhizotron studies revealed that root damage, caused by feeding of the early-hatching G. rostochiensis, resulted in increased lateral root proliferation and significantly increased G. pallida multiplication. Split-root trials indicated a significant G. pallida-induced ISR effect. G. rostochiensis multiplication was significantly reduced in split-root rhizotrons when G. pallida colonised roots before or after G. rostochiensis infection.

Evaluation Of The Radiopacity Of Esthetic Root Canal Posts.

The radiopacity of esthetic root canal posts may impair the assessment of their fit to the root canal when using radiographic images. This study determined in vitro the radiographic density of esthetic root canal posts using digital images. Thirty-six roots of human maxillary canines were assigned to six groups (N=6 per group): Reforpost (RP); Aestheti-Plus (AP); Reforpost MIX (RPM); D.T. Light Post (LP); Reforpost Radiopaque (RPR); and White Post DC (WP). Standardized digital images of the posts were obtained in different conditions: outside the root canal, inside the canal before and after cementation using luting material, and with a tissue simulator. Analysis of variance was used to compare the radiopacity mean values among the posts outside the root canal and among the posts under the other conditions, and the t unpaired test to compare the radiopacity between the posts and the dentin, and between the posts and the root canal space. There was no statistically significant difference in radiopacity between RP and RPM, and LP and WP. AP posts showed radiopacity values significantly lower than those for dentin. No statistically significant difference was found between posts (RP and AP) and the root canal space. A statistically significant difference was observed between the luted and non-luted posts; additionally, luted posts with and without tissue simulator showed no significant differences. Most of the cement-luted posts analyzed in this study were distinguishable from the density of adjacent dentin surfaces, allowing radiographic confirmation of the fit of the post in the canal. The success of using esthetic root canal posts depends mainly on the fit of the post within the canal.[1] The radiopacity of a post allows for radiographic imaging to be used to determine the fit, an important factor in a clinical perspective.