The isolation of a single spore isolate of Pasteuria penetrans and its pathogenicity on Meloidogyne javanica

Trotter, James R.; Darban, D.A.; Gowen, S. R.; Bishop, Alistair H.; Pembroke, Barbara

doi:10.1163/1568541042665250

Cited by 5 publications

(7 citation statements)

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“…Trotter et al . () developed a method to generate single spore lines of Pasteuria by incubating spore suspension and J2 for a few minutes. Although this procedure generated a few single spore lines (SSP), the process had to be repeated several times and each J2 examined microscopically to ensure that only a single spore was attached.…”

Section: Introductionmentioning

confidence: 99%

Genotyping of single spore isolates of aPasteuria penetranspopulation occurring in Florida using SNP-based markers

et al. 2016

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Aims To generate single spore lines of a population of bacterial parasite of root‐knot nematode (RKN), Pasteuria penetrans, isolated from Florida and examine genotypic variation and virulence characteristics exist within the population. Methods and Results Six single spore lines (SSP), 16SSP, 17SSP, 18SSP, 25SSP, 26SSP and 30SSP were generated. Genetic variability was evaluated by comparing single‐nucleotide polymorphisms (SNPs) in six protein‐coding genes and the 16S rRNA gene. An average of one SNP was observed for every 69 bp in the 16S rRNA, whereas no SNPs were observed in the protein‐coding sequences. Hierarchical cluster analysis of 16S rRNA sequences placed the clones into three distinct clades. Bio‐efficacy analysis revealed significant heterogeneity in the level virulence and host specificity between the individual clones. Conclusions The SNP markers developed to the 5′ hypervariable region of the 16S rRNA gene may be useful in biotype differentiation within a population of P. penetrans. Significance and Impact of the Study This study demonstrates an efficient method for generating single spore lines of P. penetrans and gives a deep insight into genetic heterogeneity and varying level of virulence exists within a population parasitizing a specific Meloidogyne sp. host. The results also suggest that the application of generalist spore lines in nematode management may achieve broad RKN control.

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Section: Introductionmentioning

confidence: 99%

Genotyping of single spore isolates of aPasteuria penetranspopulation occurring in Florida using SNP-based markers

et al. 2016

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“…While direct PCR of individual and infected females gave no amplification products, the correct-sized products were obtained for two genes in half of the females tested after an initial MDA treatment. Sequencing of the MDA-amplified DNA uncovered single nucleotide polymorphisms (SNPs) in the genes analysed from a single female; this demonstrates that infection by more than one spore had occurred and also illustrates the genetic heterogeneity of the "populations" of P. penetrans commonly used (Trotter et al 2004). …”

Section: S Rrna Gene Fragmentsmentioning

confidence: 86%

“…This obviously makes interpretation of experiments difficult because the population of spores used cannot be expected to be homogenous. Trotter et al (2004) finally resolved this problem by producing a clone resulting from a single spore that had been allowed to attach to J2 of Meloidogyne incognita. A single spore cannot be guaranteed to result in infection of a nematode so this process had to be repeated several times and each J2 examined microscopically to ensure that only one spore was attached.…”

Section: Genetic Heterogeneity Of Pasteuria "Strains"mentioning

confidence: 99%

Pasteuria penetrans and Its Parasitic Interaction with Plant Parasitic Nematodes

Bishop

2011

Soil Biology

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Until quite recently, bacteriology has been the study of culturable organisms. The proportion of unculturable bacteria in the soil is commonly put at over 99% (Sharma et al. 2005). The ecology of many of these will be very difficult to discover (Nannipieri et al. 2003;Riesenfeld et al. 2004) but this is not the case for one extraordinary group of non-culturable, aerobic, spore-forming bacteria: members of the genus Pasteuria. The composite species are exclusively obligate parasites of either water fleas or plant parasitic nematodes. These represent the two lineages of the different species of Pasteuria. This chapter predominantly focuses on the nematode parasites because of their residence in soil.The first time that the characteristic spores of this genus were represented in publication were as drawings by Metchnikoff (1888) of what is now called Pasteuria ramosa (Ebert et al. 1996), the parasite of the water fleas Daphnia magna and D. pulex. This is now the type species of the genus Pasteuria and was recognized by Metchnikoff (1888) as an endospore-forming bacterium. A Pasteuria sp. was first reported in a nematode (Dorylaimus bulbiferous), by Cobb (1906), and the suggestion was made that it should be classified as a protozoan. In keeping with this suggestion the nematode parasite was later named Duboscqia penetrans by Thorne (1940) and placed in the family Microsporidiae.Until quite recently, developments in taxonomy were based solely upon observations and measurements made from electron micrographs. Mankau (1975) noted that the parasite of the root knot nematode (RKN) Meloidogyne javanica divided with the formation of cross walls reminiscent of bacteria, and that it formed spores similar to those of known endospore-formers. He suggested the adoption of a position within the eubacteria with the name Bacillus penetrans n. comb. In spite of the outlandish appearance of the highly differentiated vegetative phase (Fig. 9.1)

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“…In addition, the few studies on this topic reveal contradictory results (Samšiňáková & Kálalová, 1983;Trotter et al, 2004). Therefore, to improve the biocontrol potential of several EF worldwide, it is interesting to know if polisporic isolates may enhance the efficacy of monosporic isolates in controlling insect pest populations.…”

Section: Discussionmentioning

confidence: 99%

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

Sammaritano¹,

Leclerque²,

Deymié³

et al. 2017

Rev. Soc. Entomol. Arg.

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https://doi.org/10.25085/rsea.763406Hongos entomopatógenos: ¿Son los aislamientos polispóricos más patógenos que las cepas monospóricas? RESUMEN. Actualmente existen varias cepas de hongos entomopatógenos utilizadas para la fabricación de bio-insecticidas comerciales. Sin embargo, la selección de éstas sigue algunas pautas como la obtención y evaluación de cepas monospóricas (cultivo en masa de microorganismos que provienen de una sola espora) Las principales razones para el uso de cultivos monospóricos en lugar de cultivos polispóricos son: la virulencia atenuada y los posibles "contaminantes" de la misma especie. En este estudio, diferentes aislamientos polispóricos y sus combinaciones monospóricas se pusieron a prueba para evaluar su eficacia respecto a larvas de Tenebrio molitor (L.) como insecto modelo. Los aislamientos polispóricos de hongos entomopatógenos fueron obtenidos de muestras de suelo de regiones agrícolas áridas. Se seleccionaron cuatro aislamientos polispóricos de Metarhizium sp. (Metschn.) (CEP413, CEP589, CEP590 y CEP591) para los bioensayos. Se realizaron pruebas de patogenicidad para evaluar la mortalidad en tres tratamientos, Polispórico completo (FP), Polispórico parcial (PP) y Monospórico puro (PM). La mortalidad acumulada se midió al día 4 después de la infección. El porcentaje de esporulación se evaluó al día 6 después de la infección. La mayor mortalidad se encontró en el tratamiento PF (94%), la menor mortalidad en el día 4 se encontró en PM-CEP413 (32%). Al día 6 el porcentaje de esporulación fue mayor en FP (94%) y fue diferente del resto de tratamientos. Elucidar diferentes combinaciones polispóricas y monospóricas para aumentar la eficacia de las aplicaciones es crucial para ayudar a expandir el uso de bio-insecticidas basados en hongos entomopatógenos. ABSTRACT. 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 (L.) 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 b...

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The isolation of a single spore isolate of Pasteuria penetrans and its pathogenicity on Meloidogyne javanica

Cited by 5 publications

References 0 publications

Genotyping of single spore isolates of aPasteuria penetranspopulation occurring in Florida using SNP-based markers

Genotyping of single spore isolates of aPasteuria penetranspopulation occurring in Florida using SNP-based markers

Pasteuria penetrans and Its Parasitic Interaction with Plant Parasitic Nematodes

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

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