scholarly journals Comparative Genomics and Pathogenicity Analysis of Two Bacterial Symbionts of Entomopathogenic Nematodes: The Role of The GroEL Protein in Virulence

Author(s):  
Abraham Rivera-Ramírez ◽  
Rosalba Salgado-Morales ◽  
Alfredo Jiménez-Pérez ◽  
Rebeca Pérez-Martínez ◽  
Blanca Inés García-Gómez ◽  
...  

Abstract Bacteria of the genera Xenorhabdus and Photorhabdus are symbionts of entomopathogenic nematodes. Despite their close phylogenetic relationship, they show differences in their pathogenicity and virulence mechanisms in target insects. These differences can be explored by the analysis of the pangenome, as it provides a framework for characterizing and defining the gene repertoire. Here, we report the genome of strain SC 0516. In addition, we performed the first pangenome analysis of 91 strains of Xenorhabdus and Photorhabdus, obtaining a total of 23,603 gene clusters and a core genome of 348 genes. Phylogenetic analysis performed with the core genome showed that our strain belonged to the X. nematophila group. Biological tests showed that whole cells of X. nematophila SC 0516 were more virulent than those of P. luminescens HIM3 when both were injected into Galleria mellonella larvae. In addition, we cloned and expressed the GroEL proteins of both bacteria, as this protein has been previously indicated to show insecticidal activity in the genus Xenorhabdus. Cpn60-Xn was found to be the most toxic at all concentrations tested, with an LC50 value of 102.34 ng/larva. Sequence analysis suggested that the Cpn60-Xn toxin was homologous to Cpn60-Pl; however, Cpn60-Xn contained thirty-five differentially substituted amino acid residues that could be responsible for its insecticidal activity.

1991 ◽  
Vol 56 (9) ◽  
pp. 1963-1970 ◽  
Author(s):  
Jan Hlaváček ◽  
Václav Čeřovský ◽  
Jana Pírková ◽  
Pavel Majer ◽  
Lenka Maletínská ◽  
...  

In a series of analogues of the cholecystokinin octapeptide (CCK-8) the amino acid residues were gradually modified by substituting Gly by Pro in position 4, Trp by His in position 5, Met by Cle in position 6, or the Gly residue was inserted between Tyr and Met in positions 2 and 3 of the peptide chain, and in the case of the cholecystokinin heptapeptide (CCK-7) the Met residues were substituted by Nle or Aib. These peptides were investigated from the point of view of their biological potency in the peripheral and central region. From the results of the biological tests it follows that the modifications carried out in these analogues and in their Nα-Boc derivatives mean a suppression of the investigated biological activities by 2-3 orders of magnitude (at a maximum dose of the tested substance of 2 . 10-2 mg per animal).This means that a disturbance of the assumed biologically active conformation of CCK-8, connected with a considerable decrease of the biological potency of the molecule, takes place not only after introduction of the side chain into its centre (substitution of Gly4), but also after the modification of the side chains of the amino acids or by extension of the backbone in further positions around this central amino acid.


2021 ◽  
Vol 9 (4) ◽  
pp. 768
Author(s):  
Karel Kopejtka ◽  
Yonghui Zeng ◽  
David Kaftan ◽  
Vadim Selyanin ◽  
Zdenko Gardian ◽  
...  

An aerobic, yellow-pigmented, bacteriochlorophyll a-producing strain, designated AAP5 (=DSM 111157=CCUG 74776), was isolated from the alpine lake Gossenköllesee located in the Tyrolean Alps, Austria. Here, we report its description and polyphasic characterization. Phylogenetic analysis of the 16S rRNA gene showed that strain AAP5 belongs to the bacterial genus Sphingomonas and has the highest pairwise 16S rRNA gene sequence similarity with Sphingomonas glacialis (98.3%), Sphingomonas psychrolutea (96.8%), and Sphingomonas melonis (96.5%). Its genomic DNA G + C content is 65.9%. Further, in silico DNA-DNA hybridization and calculation of the average nucleotide identity speaks for the close phylogenetic relationship of AAP5 and Sphingomonas glacialis. The high percentage (76.2%) of shared orthologous gene clusters between strain AAP5 and Sphingomonas paucimobilis NCTC 11030T, the type species of the genus, supports the classification of the two strains into the same genus. Strain AAP5 was found to contain C18:1ω7c (64.6%) as a predominant fatty acid (>10%) and the polar lipid profile contained phosphatidylglycerol, diphosphatidylglycerol, phosphatidylethanolamine, sphingoglycolipid, six unidentified glycolipids, one unidentified phospholipid, and two unidentified lipids. The main respiratory quinone was ubiquinone-10. Strain AAP5 is a facultative photoheterotroph containing type-2 photosynthetic reaction centers and, in addition, contains a xathorhodopsin gene. No CO2-fixation pathways were found.


Nematology ◽  
1999 ◽  
Vol 1 (7) ◽  
pp. 735-743 ◽  
Author(s):  
Parwinder S. Grewal ◽  
Edwin E. Lewis ◽  
Sudha Venkatachari

Abstract A possible mechanism of suppression of a plant-parasitic nematode Meloidogyne incognita by entomopathogenic nematodes is described. Heat-killed entomopathogenic nematodes Steinernema feltiae and S. riobrave temporarily suppressed penetration of the root-knot nematode M. incognita into tomato roots, but live nematodes had no effect. Infective juvenile M. incognita were repelled from all entomopathogenic nematode treatments that included their symbiotic bacteria. They were repelled by Galleria mellonella cadavers infected with S. carpocapsae, S. feltiae, and S. riobrave and from cell-free culture filtrates of the symbiotic bacteria Xenorhabdus nematophilus, X. bovienii, and Xenorhabdus sp. "R" from the three nematode species, respectively. Cell-free filtrates from all three Xenorhabdus spp. were toxic to M. incognita infective juveniles causing 98-100% mortality at 15% concentration. Cell-free filtrate of Xenorhabdus sp. "R" also reduced the hatch of M. incognita eggs. Application of formulated bacterial cell-free filtrates temporarily suppressed M. incognita penetration into tomato roots in a greenhouse trial. The short-term effects of cell-free bacterial filtrates, namely toxicity and repellency, were almost entirely due to ammonium. These results demonstrate allelopathic interactions between plant-parasitic nematodes, entomopathogenic nematodes and their symbiotic bacteria. The likely role of allelopathy in the suppression of plant-parasitic nematodes by innundative applications of entomopathogenic nematodes is discussed. Allelopathie: Ein moglicher Mechanismus zur Unterdruckung pflanzenparasitarer Nematoden durch insektenpathogene Nematoden - Es wird ein moglicher Mechanismus zur Unterdruckung des pflanzenparasitaren Nematoden Meloidogyne incognita durch insektenpathogene Nematoden beschrieben. Durch Hitze abgetotete insektenpathogene Nematoden Steinernema feltiae und S. riobrave underdruckten das Eindringen des Wurzelgallenalchens M. incognita in Tomatenwurzeln, lebende Nematoden hatten keine Wirkung. Infektionsjuvenile von M. incognita wurden von allen Behandlungen mit insektenpathogenen Nematoden abgestossen, die auch die symbiontischen Bakterien einschlossen. Sie wurden durch die Kadaver von Galleria mellonella abgestossen, die mit S. carpocapsae, S. feltiae und S. riobrave infiziert waren sowie durch zellfreie Kultursubstrate der symbiontischen Bakterien Xenorhabdus nematophilus, X. bovienii und Xenorhabdus sp. "R" aus den drei genannten Nematodenarten. Zellfreie Kultursubstrate von allen drei Xenorhabdus spp. waren giftig fur die Infektionsjuvenilen von M. incognita und verursachten in einer Konzentration von 15% Abtotungsraten von 98-100%. Zellfreie Kultursubstrate von Xenorhabdus sp. "R" vermiderten ausserdem das Schlupfen von M. incognita-Eiern. In einem Gewachshausversuch unterdruckten formulierte zellfreie Bakterienfiltrate vorubergehend das Eindringen von M. incognita in Tomatenwurzeln. Die Kurzzeitwirkungen von zellfreien Bakterien filtraten, namentlich Giftigkeit und Abstossung, waren nahezu ganz bedingt durch Ammoniak. Diese Ergebnisse zeigen das Vorhandensein von allelopathischen Wechselwirkungen zwischen pflanzenparasitaren Nematoden, insektenpathogenen Nematoden und deren symbiontischen Bakterien. Die wahrscheinliche Rolle von Allelopathie bei der Unterdruckung pflanzenparasitarer Nematoden durch eine Massenanwendung insektenpathogener Nematoden wird diskutiert.


Nematology ◽  
2000 ◽  
Vol 2 (5) ◽  
pp. 515-521 ◽  
Author(s):  
Hara Menti ◽  
Denis Wright ◽  
Roland Perry

AbstractThe infectivity of populations of the entomopathogenic nematodes Steinernema feltiae and Heterorhabditis megidis from Greece (GR) and the UK was compared using Galleria mellonella larvae as hosts. Dose-response tests showed that the two Steinernema populations did not differ in their establishment rates but they were more infective than H. megidis UK 211. The temperature range for infectivity was greater than that for development. However, the optimal temperature for infection and development for all populations was 23°C. Infectivity of Steinernema populations was not affected by storage for 12 weeks. However, 12 week-old H. megidis UK 211 infective juveniles (IJ) were less infective than fresh IJ. H. megidis GR showed very low establishment rates at all the doses and temperatures tested, before and after storage. The results are discussed in relation to the nematodes' climatic origin and lipid content. Pouvoir infestant de populations des nématodes entomopathogènes Steinernema feltiae et Heterorhabditis megidis suivant la température, l'âge et le contenu lipidique - Le pouvoir infestant de populations des nématodes entomopathogènes Steinernema feltiae et Heterorhabditis megidis provenant de Grèce et du Royaume Uni a été comparée, utilisant comme hôte Galleria mellonella. Les tests de dose/réaction ont montré que les taux d'établissement des deux populations ne diffèrent pas mais que leur pouvoir infestant était plus élevée que celle de H. megidis UK211. La plage des températures permettant l'infestation était plus étendue que celle relative au développement. Cependant, les températures optimales pour l'infestation et pour le développement étaient l'une et l'autre de 23°C pour toutes les populations. L'infestivité des populations de Steinernema n'a pas été affectée par un stockage de 12 semaines. Les juvéniles infestants de H. megidis UK211 âgés de 12 semaines montraient toutefois une infestivité plus faible que celle d'individus frais. Les specimens de H. megidis provenant de Grèce présentaient - que ce soit avant ou après le stockage - des taux d'établissement très faibles pour toutes les doses et les températures testées. Ces résultats sont discutés en relation avec l'origine climatique et le contenu lipidique des nématodes.


2021 ◽  
Vol 9 (8) ◽  
pp. 1761
Author(s):  
Gaurav Agarwal ◽  
Ronald D. Gitaitis ◽  
Bhabesh Dutta

Pantoea stewartii subsp. indologenes (Psi) is a causative agent of leafspot on foxtail millet and pearl millet; however, novel strains were recently identified that are pathogenic on onions. Our recent host range evaluation study identified two pathovars; P. stewartii subsp. indologenes pv. cepacicola pv. nov. and P. stewartii subsp. indologenes pv. setariae pv. nov. that are pathogenic on onions and millets or on millets only, respectively. In the current study, we developed a pan-genome using the whole genome sequencing of newly identified/classified Psi strains from both pathovars [pv. cepacicola (n = 4) and pv. setariae (n = 13)]. The full spectrum of the pan-genome contained 7030 genes. Among these, 3546 (present in genomes of all 17 strains) were the core genes that were a subset of 3682 soft-core genes (present in ≥16 strains). The accessory genome included 1308 shell genes and 2040 cloud genes (present in ≤2 strains). The pan-genome showed a clear linear progression with >6000 genes, suggesting that the pan-genome of Psi is open. Comparative phylogenetic analysis showed differences in phylogenetic clustering of Pantoea spp. using PAVs/wgMLST approach in comparison with core genome SNPs-based phylogeny. Further, we conducted a horizontal gene transfer (HGT) study using Psi strains from both pathovars along with strains from other Pantoea species, namely, P. stewartii subsp. stewartii LMG 2715T, P. ananatis LMG 2665T, P. agglomerans LMG L15, and P. allii LMG 24248T. A total of 317 HGT events among four Pantoea species were identified with most gene transfer events occurring between Psi pv. cepacicola and Psi pv. setariae. Pan-GWAS analysis predicted a total of 154 genes, including seven gene-clusters, which were associated with the pathogenicity phenotype (necrosis on seedling) on onions. One of the gene-clusters contained 11 genes with known functions and was found to be chromosomally located.


2008 ◽  
Vol 65 (4) ◽  
pp. 433-437 ◽  
Author(s):  
Juan Carlos Lara ◽  
Cláudia Dolinski ◽  
Elias Fernandes de Sousa ◽  
Rogério Figueiredo Daher

Entomopathogenic nematodes (EPNs) are currently being used as successful biological control agents of soil-dwelling insect pests. Previous field and greenhouse studies demonstrated that application techniques and non-biotic factors (temperature and pressure) have a significant effect on EPNs efficacy. The objective of this study was to evaluate the influence of an irrigation spray application system on the viability, infectivity and host search capability of Heterorhabditis baujardi LPP7 (Nematoda: Heterorhabditidae) infective juveniles (IJ). Two assays were proposed. Their viability was evaluated under the microscope after the IJ passed through the irrigation system. Infectivity on Galleria mellonella larvae, and host search capability, as evidenced by larval mortality, were evaluated in containers (Experiment 1). In the field (Experiment 2), mortality of G. mellonella larvae was evaluated under different nematode concentrations (0, 100,000, 300,000 and 500,000 IJ per tree). No differences were recorded on the viability, infectivity and host search capability of the IJ in Experiment 1. In Experiment 2, differences were recorded among the different concentrations used (p < 0.05), and a higher mortality was observed at the highest nematode concentration (28.3% and 37% in each one of the two experiment repetitions). This irrigation system did not affected adversely the viability, infectivity and host search capability of H. baujardi LPP7.


Nematology ◽  
2015 ◽  
Vol 17 (9) ◽  
pp. 1057-1069 ◽  
Author(s):  
Hugues Baimey ◽  
Lionel Zadji ◽  
Leonard Afouda ◽  
Maurice Moens ◽  
Wilfrida Decraemer

The influence of three pesticides on the viability and infectivity of four Beninese isolates of entomopathogenic nematodes (EPN), Heterorhabditis indica Ayogbe1, H. sonorensis Azohoue2, H. sonorensis Ze3, and Steinernema sp. Bembereke, was determined. The impact of both soil temperature and soil moisture on the virulence of these EPN to Trinervitermes occidentalis was investigated in laboratory assays. The effect of EPN-infected Galleria mellonella larvae on underground populations of Macrotermes bellicosus was also examined. All tested Heterorhabditis species were more tolerant to glyphosate and fipronil than the Steinernema species. Heterorhabditis sonorensis Azohoue2, showed the best results with 63.2% termite mortality at a soil temperature of 35°C. The increase of soil moisture to 20% (w/w) did not negatively influence the virulence of tested EPN. The underground populations of 71% or 60% treated nests were controlled by H. sonorensis Azohoue2- or H. indica Ayogbe1-infected G. mellonella larvae, respectively.


2020 ◽  
Vol 86 (13) ◽  
Author(s):  
M. Veronica Fumero ◽  
Alessandra Villani ◽  
Antonia Susca ◽  
Miriam Haidukowski ◽  
Maria T. Cimmarusti ◽  
...  

ABSTRACT Fusarium subglutinans and Fusarium temperatum are common maize pathogens that produce mycotoxins and cause plant disease. The ability of these species to produce beauvericin and fumonisin mycotoxins is not settled, as reports of toxin production are not concordant. Our objective was to clarify this situation by determining both the chemotypes and genotypes for strains from both species. We analyzed 25 strains from Argentina, 13 F. subglutinans and 12 F. temperatum strains, for toxin production by ultraperformance liquid chromatography mass spectrometry (UPLC-MS). We used new genome sequences from two strains of F. subglutinans and one strain of F. temperatum, plus genomes of other Fusarium species, to determine the presence of functional gene clusters for the synthesis of these toxins. None of the strains examined from either species produced fumonisins. These strains also lack Fum biosynthetic genes but retain homologs of some genes that flank the Fum cluster in Fusarium verticillioides. None of the F. subglutinans strains we examined produced beauvericin although 9 of 12 F. temperatum strains did. A complete beauvericin (Bea) gene cluster was present in all three new genome sequences. The Bea1 gene was presumably functional in F. temperatum but was not functional in F. subglutinans due to a large insertion and multiple mutations that resulted in premature stop codons. The accumulation of only a few mutations expected to disrupt Bea1 suggests that the process of its inactivation is relatively recent. Thus, none of the strains of F. subglutinans or F. temperatum we examined produce fumonisins, and the strains of F. subglutinans examined also cannot produce beauvericin. Variation in the ability of strains of F. temperatum to produce beauvericin requires further study and could reflect the recent shared ancestry of these two species. IMPORTANCE Fusarium subglutinans and F. temperatum are sister species and maize pathogens commonly isolated worldwide that can produce several mycotoxins and cause seedling disease, stalk rot, and ear rot. The ability of these species to produce beauvericin and fumonisin mycotoxins is not settled, as reports of toxin production are not concordant at the species level. Our results are consistent with previous reports that strains of F. subglutinans produce neither fumonisins nor beauvericin. The status of toxin production by F. temperatum needs further work. Our strains of F. temperatum did not produce fumonisins, while some strains produced beauvericin and others did not. These results enable more accurate risk assessments of potential mycotoxin contamination if strains of these species are present. The nature of the genetic inactivation of BEA1 is consistent with its relatively recent occurrence and the close phylogenetic relationship of the two sister species.


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