Beyond Bt: New Bacterial Resources for Insect Biocontrol

The Gram-positive bacterium Bacillus thuringiensis (Bt) is the economically most important entomopathogen for insect biocontrol, and for excellent reason a large part of the effort invested in the development of microbial insecticides has concentrated on Bt strains and their cry genes. Hundreds of cry gene sequences have been determined from nature, and molecular modeling has been used to create artificial recombinant Cry proteins with potentially new properties. However, alternative insect biocontrol agents are increasingly solicited. More recently, research efforts have concentrated upon bacterial insect-pathogens as potential biocontrol resources as, e.g., non-cry toxins of Bt and other bacteria of the family Bacillaceae as well as the bacterium Saccharopolyspora spinosa (Actinobacteria). Moreover, several different γ-proteobacterial entomopathogens belonging to the species Serratia entomophila, Yersinia entomophaga or Pseudomonas entomophila as well as to the genera Providencia or Rickettsiella are currently being evaluated for their biocontrol potential. The present literature review gives a brief update on these entomopathogens and toxins

Pathogenicity of Pythium species to maize

AbstractPythium isolates from diseased and dead bait plants of maize and cress grown in compost or various soils (maize fields, parkland under deciduous trees, grassland) were characterised and tested for pathogenicity to maize (Zea mays L.). In pot tests performed under controlled conditions, pathogenicity of the isolates to maize was apparent by reduction of root and shoot growth, whereas damping-off of maize seedlings was less frequent. Contrarily, pea seedlings were killed by pathogenic Pythium isolates. Pythium isolates from diseased maize seedlings and pathogenic strains from other gramineous plants (P. phragmitis, P. aff.phragmitis, P. catenulatum) were not necessarily more virulent to maize compared to isolates originating from dicotyledonous plants (cress). The most virulent isolates originated from compost and caused a reduction of maize shoot growth of up to 60%. Phylogenetic analysis revealed that they were very closely related to P. ultimum var. ultimum and P. arrhenomanes, respectively. Isolates originating from maize fields, grassland and parkland under deciduous trees, a reference culture of P. arrhenomanes and strains of P. phragmitis, P. aff. phragmitis and P. catenulatum with known pathogenicity on reed were non-pathogenic on maize. Isolates from compost, and from maize fields generally had a higher temperature optimum for mycelial growth (30 °C) and a faster growth rate (1.5–2.0 mm h−1) compared to the isolates from parkland under deciduous trees and grassland soil (20–25 °C, ~1.0 mm h−1), respectively. This study indicates a potential impact of pathogenic Pythium on maize plants even in the absence of visible symptoms

A Rickettsiella Bacterium from the Hard Tick, Ixodes woodi: Molecular Taxonomy Combining Multilocus Sequence Typing (MLST) with Significance Testing

Hard ticks (Acari: Ixodidae) are known to harbour intracellular bacteria from several phylogenetic groups that can develop both mutualistic and pathogenic relationships to the host. This is of particular importance for public health as tick derived bacteria can potentially be transmitted to mammals, including humans, where e.g. Rickettsia or Coxiella act as severe pathogens. Exact molecular taxonomic identification of tick associated prokaryotes is a necessary prerequisite of the investigation of their relationship to both the tick and possible vertebrate hosts. Previously, an intracellular bacterium had been isolated from a monosexual, parthenogenetically reproducing laboratory colony of females of the hard tick, Ixodes woodi Bishopp, and had preliminarily been characterized as a “Rickettsiella-related bacterium”. In the present molecular taxonomic study that is based on phylogenetic reconstruction from both 16 S ribosomal RNA and protein-encoding marker sequences complemented with likelihood-based significance testing, the bacterium from I. woodi has been identified as a strain of the taxonomic species Rickettsiella grylli. It is the first time that a multilocus sequence typing (MLST) approach based on a four genes comprising MLST scheme has been implemented in order to classify a Rickettsiella-like bacterium to this species. The study demonstrated that MLST holds potential for a better resolution of phylogenetic relationships within the genus Rickettsiella, but requires sequence determination from further Rickettsiella-like bacteria in order to complete the current still fragmentary picture of Rickettsiella systematics

Control of Bemisia tabaci by entomopathogenic fungi isolated from arid soils in Argentina

Entomopathogenic Hypocreales were isolated from arid soils in Argentina using Tenebrio molitor as bait and tested for their biological performance at 30°C and 45–65% RH. Conidial germination was tested in three vegetable oils (sunflower, olive and maize) at two concentrations (1% and 10%) to evaluate their compatibility for further liquid formulations. According to radial growth and germination results, we selected four isolates to test their pathogenicity against second instar B. tabaci nymphs with the selected oil formulations at 30°C. CEP381 and CEP401 showed the highest radial growth. Isolates CEP381, CEP401, CEP413 and CEP409 (Metarhizium spp.) had similar germination percentages as compared with water control when germinated on either sunflower, olive or maize oils at 10% v/v. The highest mortality of B. tabaci was observed for the isolates CEP381 in sunflower oil and CEP401 in olive oil. Molecular identification of isolates was performed using ITS4–5 primers. All isolates belong to the Metarhizium core group. Tested isolates could grow and infect B. tabaci nymphs at 30°C in some of the vegetable oils as carriers, providing new possibilities for integrated pest management of Bemisia tabaci.Fil: Aguilera Sammaritano, Juan Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de San Juan. Facultad de Ingeniería. Instituto de Biotecnología; ArgentinaFil: Lopez Lastra, Claudia Cristina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Estudios Parasitológicos y de Vectores. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Centro de Estudios Parasitológicos y de Vectores; ArgentinaFil: Leclerque, Andreas. Hochschule Geisenheim University; AlemaniaFil: Vazquez, Fabio. Universidad Nacional de San Juan. Facultad de Ingeniería. Instituto de Biotecnología; ArgentinaFil: Toro, Maria Eugenia. Universidad Nacional de San Juan. Facultad de Ingeniería. Instituto de Biotecnología; ArgentinaFil: D´Alessandro, Celeste Paola. Universidade de Sao Paulo; BrasilFil: Cuthbertson, Andrew G.S.. Independent Science Advisor; Reino UnidoFil: Lechner, Bernardo Ernesto. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Micología y Botánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Micología y Botánica; Argentin

Entomopathogenic fungi: Are polisporic isolates more pathogenic than monosporic strains?

Currently only monosporic strains from several entomopathogenic fungi have been selected and used for mass production and bioinsecticide manufacturing worldwide. The main reasons for the use of single spore instead of multispore strains are the attenuated virulence and the contaminants of the same species. In this study, different polisporic isolates and their monosporic combinations were tested against Tenebrio molitor (Linnaeus) larvae as an insect model. Isolates were obtained from arid soils. Four Metarhizium sp. (Metschn.) multisporic isolates (CEP413, CEP589, CEP590 and CEP591) were selected for bioassays. Trials were performed to evaluate mortality on three treatments, Full Polisporic (FP), Partial Polisporic (PP) and Pure Monosporic (PM). Cumulative mortality was measured at day 4 post infection. Sporulation percentage was assessed at day 6 post infection. The highest mortality was found at FP treatment (94%), the lowest mortality at day 4 was found at PM-CEP413 (32%). At day 6 the sporulation percentage was higher on FP (94%) and it was different from the rest of the treatments. To elucidate different polisporic and monosporic combinations to improve their effectiveness, may help to expand the use of bioinsecticides based on entomopathogenic fungi.Centro de Estudios Parasitológicos y de Vectore

Entomopathogenic fungi: Are polisporic isolates more pathogenic than monosporic strains?

Currently only monosporic strains from several entomopathogenic fungi have been selected and used for mass production and bioinsecticide manufacturing worldwide. The main reasons for the use of single spore instead of multispore strains are the attenuated virulence and the contaminants of the same species. In this study, different polisporic isolates and their monosporic combinations were tested against Tenebrio molitor (Linnaeus) larvae as an insect model. Isolates were obtained from arid soils. Four Metarhizium sp. (Metschn.) multisporic isolates (CEP413, CEP589, CEP590 and CEP591) were selected for bioassays. Trials were performed to evaluate mortality on three treatments, Full Polisporic (FP), Partial Polisporic (PP) and Pure Monosporic (PM). Cumulative mortality was measured at day 4 post infection. Sporulation percentage was assessed at day 6 post infection. The highest mortality was found at FP treatment (94%), the lowest mortality at day 4 was found at PM-CEP413 (32%). At day 6 the sporulation percentage was higher on FP (94%) and it was different from the rest of the treatments. To elucidate different polisporic and monosporic combinations to improve their effectiveness, may help to expand the use of bioinsecticides based on entomopathogenic fungi.Centro de Estudios Parasitológicos y de Vectore