Bacteriocinogenesis in cells of Xenorhabdus nematophilus and Photorhabdus luminescens: Enterobacteriaceae associated with entomopathogenic nematodes

Summary— Xenorhabdus nematophilus FI strain and Photorhabdus luminescens NC19 strain produced bacteriocins after mitomycin C treatment and under natural conditions respectively. The ultrastructure of these two strains was described and compared to the ultrastructure of untreated or normal cells. After image processing of purified bacteriocins we found morphological homology in infected cells with protoplasmic rods in longitudinal section and hexagonal aggregates in transversal section. We concluded that these particular structures, so-called ‘lattice structures’ and previously interpreted as ‘photosomes’, are in fact the early stages of in situ production of bacteriocins in these two bacterial genera. Natural occurrence of Photorhabdus spp bacteriocinogenesis was observed in other strains, while other lysogenic strains of Xenorhabdus spp are lysed after a mitomycin C treatment.     

Arabidopsis thaliana as a new model host for plant-parasitic nematodes

We have established culture conditions for successful infection and development of several economically important cyst-forming and root-knot nematodes on Arabidopsis thaliana under monoxenic conditions. Complete life cycles were obtained with the sedentary cyst nematodes Heterodera schachtii, H. trifolii, H. cajani and the root-knot nematodes Meloidogyne incognita and M. arenariaas well as with the migratory nematode Pratylenchus penetrans. In contrast, H. goettingiana and Globodera rostochiensis were unable to develop on Arabidopsis roots. Tissue-culture quality agar and medium conditions optimized for hydroponic root culture were essential for successful infections. Detailed in-vivo observations were made inside Arabidopsis roots during the early infection stages of M. incognita and during complete development of H. schachtii. Seventy-four different ecotypes of Arabidopsis were screened for their susceptibility towards H. schachtii resulting in a range of infection rates. None of the ecotypes tested showed complete resistance in vitro. The use of Arabidopsis as a host for plant-parasitic nematodes will provide a new model system for the molecular genetic analysis of this interaction.     

Caenorhabditis elegans as a model for parasitic nematodes: a focus on the cuticle

The phylum Nematoda consists of over half a million species of worms that inhabit astoundingly diverse environments. Nematodes can live as obligatory parasites of plants and animals, or alternate a parasitic with a free-living life style. The fact that the vast majority of species are strictly free living often surprises parasitology students, for obviously the highest research priorities in this field have involved parasites of medical, veterinary and agricultural importance. Here Samuel Politz and Mario Philipp contend that some basic questions concerning the biology of the parasite cuticle can be investigated more easily and in greater depth in the free-living nematode Caenorhabditis elegans than in the parasites themselves.     

Hydrophobe-water interactions: methane as a model

Recent molecular-dynamics simulations have demonstrated that the use of an empirical hydrophobic potential displaying two minima, i.e., one for hydrophobes in close contact and one for hydrophobes separated by a hydration layer, leads to a marked improvement in protein structure prediction. This potential is supported by experimental data and simulations, but its physical origin and mathematical formulation have not been derived as yet. Here we show that water-mediated attraction (the “wetting regime”) between two hydrophobic molecules originates in the interaction between the dipoles induced at the surface of the hydrophobes by the surrounding structured water. As an example, we derive the effective hydrophobic potential that describes the interaction between two methane molecules, a classical model of a double-well energy function. We found an excellent agreement with published results from all-atom, explicit solvent molecular-dynamics simulations of this interaction. The approach presented here provides the theoretical basis for implementing an adequate representation of the wetting regime of the hydrophobic interactions in force fields used for structure prediction. The results are useful for modeling both protein folding and binding.     

Photorhabdus: a model for the analysis of pathogenicity and mutualism

Photorhabdus are entomopathogenic members of the family Enterobacteriaceae. In addition to killing insects Photorhabdus also have a mutualistic association with nematodes from the family Heterorhabditidiae. Therefore, the bacteria have a complex life cycle that involves temporally separated pathogenic and mutualistic associations with two different invertebrate hosts. This tripartite Photorhabdus-insect-nematode association provides researchers with a unique opportunity to characterize the prokaryotic contribution to two different symbioses, i.e. pathogenicity and mutualism while also studying the role of the host in determining the outcome of association with the bacteria. In this review I will outline the life cycle of Photorhabdus and describe recent important advances in our understanding of the symbiology of Photorhabdus. Finally, the contribution made by this model to our understanding of the nature of symbiotic associations will be discussed.     

Oesophagostomum dentatum: potential as a model for genomic studies of strongylid nematodes, with biotechnological prospects

There are substantial gaps in the knowledge of the molecular processes of development and reproduction in parasitic nematodes, despite the fact that understanding such processes could lead to novel ways of treating and controlling parasitic diseases, through blocking or disrupting key biological pathways. Biotechnological advances through large-scale sequencing projects, approaches for the analysis of differential gene and protein expression and functional genomics (e.g., double-stranded RNA interference) now provide opportunities to investigate the molecular basis of developmental processes in some parasitic nematodes. The porcine nodule worm, Oesophagostomum dentatum (order Strongylida), may provide a platform for testing the function of genes from this and related nematodes, given that this species can be grown and maintained in culture in vitro for periods longer than other nematodes of the same order. In this article, we review relevant biological, biochemical and molecular biological and genomic information about O. dentatum and propose that the O. dentatum - pig system provides an attractive model for exploring molecular developmental and reproductive processes in strongylid nematodes, leading toward new intervention methods and biotechnological outcomes.     

Spider-web kleptoparasites as a model for studying producer-consumer interactions

In this study, we documented that the kleptoparasitic spidersArgyroes elvatus consume and assimilate web material from thehost spider Nephila clavipes. We also demonstrated quantitativelythat the amount of web material consumed by the kleptopa asiteis equivalent to the amount of insect material comsumed whenhost vigilance is low, as expected when foraging conditionsare very good. Argyrodes vary in their impact on their hosts,as they may steal large prey, small prey, or silk. This host-kieptoparasite interaction is therefore an ideal system for experimentallycramming a variable producer-consumer interaction. We compareour experimental results to published experiments showing thatthe impact of Arg on a Nephila host can be deleterious whenforaging conditions are poor.     

Nicotiana benthamiana: its history and future as a model for plant-pathogen interactions

Nicotiana benthamiana is the most widely used experimental host in plant virology, due mainly to the large number of diverse plant viruses that can successfully infect it. Additionally, N. benthamiana is susceptible to a wide variety of other plant-pathogenic agents (such as bacteria, oomycetes, fungi, and so on), making this species a cornerstone of host-pathogen research, particularly in the context of innate immunity and defense signaling. Moreover, because it can be genetically transformed and regenerated with good efficiency and is amenable to facile methods for virus-induced gene silencing or transient protein expression, N. benthamiana is rapidly gaining popularity in plant biology, particularly in studies requiring protein localization, interaction, or plant-based systems for protein expression and purification. Paradoxically, despite being an indispensable research model, little is known about the origins, genetic variation, or ecology of the N. benthamiana accessions currently used by the research community. In addition to addressing these latter topics, the purpose of this review is to provide information regarding sources for tools and reagents that can be used to support research in N. benthamiana. Finally, we propose that N. benthamiana is well situated to become a premier plant cell biology model, particularly for the virology community, who as a group were the first to recognize the potential of this unique Australian native.     

Type 1 fimbriae of insecticidal bacterium Xenorhabdus nematophila is necessary for growth and colonization of its symbiotic host nematode Steinernema carpocapsiae

Xenorhabdus nematophila produces type 1 fimbriae on the surface of Phase I cells. Fimbriae mediate recognition and adhesion of the bacteria to its target cell. To investigate the role of fimbriae in the biology of X. nematophila , we have produced a fimbrial mutant strain by insertional inactivation of the mrx A gene, encoding the structural subunit of type 1 fimbriae. Phenotypic characterization of the mutant revealed loss of fimbriae on the cell surface. Cell surface characteristics like dye absorption, biofilm formation, red blood cell agglutination remained unaltered. The mrx A mutant was defective in swarming on soft agar, although swimming motility was not affected. Flagellar expression was suppressed in the mrxA strain under swarming conditions, but not swimming conditions. Agglutination and cytotoxicity of the mutant to larval haemocytes was also reduced. When the mutant cells were injected in the haemocoel of the fourth instar larvae of Helicoverpa armigera , an increase in the LT₅₀ of 9–12 h was observed relative to the wild-type strain. The nematode growth was slow on the lawn of the fimbrial mutant. The mrxA negative strain was unable to colonize the nematode gut efficiently. This study demonstrates importance of type 1 fimbriae in establishment of bacteria-nematode symbiosis, a key to successful pest management program.     

Emergence of mutualism: Application of a differential model to endosymbiosis

A population dynamics approach based on a system of differential equations allows us to establish conditions for the emergence of mutualism for cases such as coelenterates-algae symbionts. A central assumption of the model is that a host organism is able to discriminate, via some molecular recognition mechanisms, among different invading organisms and preferentially rejectparasites rather thanbona fide symbionts. Large differential rejection rates allow the emergence of mutualism. Different attractors of the population dynamics correspond to the emergence of mutualism, predominance of “selfish” species, or coexistence of many species.     

A female nervous system is necessary for normal sperm storage in Drosophila melanogaster: a masculinized nervous system is as good as none

A male Drosophila melanogaster deposits many more sperm in a female''s bursa copulatrix than are stored in her ventral receptacle or paired spermathecae soon after copula has ended. The remaining sperm are expelled by the female. These observations suggest a sexual conflict over the processes involved in sperm storage. We used genetically manipulated flies to study the role of the central nervous system in sperm storage. Flies with female bodies but masculinized nervous systems, or isolated female abdomens, stored significantly fewer sperm than did control females. Furthermore, compared with control flies, there were relatively more sperm in the ventral receptacle and relatively fewer in the spermathecae. These results suggest that the female nervous input counteracts the male''s attempts to force sperm into the ventral receptacle during copula and promotes active transport of sperm to the spermathecae during and after copula. The female is clearly a very active partner in influencing processes involved in sperm competition, especially as only stored sperm can be used later to fertilize eggs. To our knowledge, this is the first study to show directly the involvement of the female nervous system in sperm storage.     

Tricellulin is a tight-junction protein necessary for hearing

The inner ear has fluid-filled compartments of different ionic compositions, including the endolymphatic and perilymphatic spaces of the organ of Corti; the separation from one another by epithelial barriers is required for normal hearing. TRIC encodes tricellulin, a recently discovered tight-junction (TJ) protein that contributes to the structure and function of tricellular contacts of neighboring cells in many epithelial tissues. We show that, in humans, four different recessive mutations of TRIC cause nonsyndromic deafness (DFNB49), a surprisingly limited phenotype, given the widespread tissue distribution of tricellulin in epithelial cells. In the inner ear, tricellulin is concentrated at the tricellular TJs in cochlear and vestibular epithelia, including the structurally complex and extensive junctions between supporting and hair cells. We also demonstrate that there are multiple alternatively spliced isoforms of TRIC in various tissues and that mutations of TRIC associated with hearing loss remove all or most of a conserved region in the cytosolic domain that binds to the cytosolic scaffolding protein ZO-1. A wild-type isoform of tricellulin, which lacks this conserved region, is unaffected by the mutant alleles and is hypothesized to be sufficient for structural and functional integrity of epithelial barriers outside the inner ear.     

Potential role of parasitism in the evolution of mutualism in astigmatid mites: Hemisarcoptes cooremani as a model

Phoresy is a symbiotic interaction that results in dispersal, benefiting the relocated organism without negatively impacting the phoretic host. It has long been considered that phoresy among astigmatid mites is somehow an intermediate precursor to the evolution of parasitism within the group. In astigmatid mites, only the heteromorphic deutonymph (hypopode) participates in phoretic dispersal, and the plesiomorphic hypopode may be the key to understanding the dynamics of the evolution of that parasitism. Hypopodes of Hemisarcoptes cooremani (Acari: Acariformes) and their phoretic beetle host Chilocorus cacti (Coleoptera: Coccinellidae) have become the experimental focus for studies concerned with the potential forces that influence the transition of a free-living life style into various coevolved relationships. Previous radiolabeling studies applied to H. cooremani and C. cacti determined that hypopodes of H. cooremani acquired resources from adults of C. cacti while in transit, negating the paradigm that the heteromorphy was purely phoretic. To further probe this relationship, we tested whether materials could be passed from the mites to their hosts. We report here a study using a tritium radiolabel, which indicated that beetles also acquire resources from the hypopodes. These results have implications for understanding the complex relationship between H. cooremani and C. cacti. We propose that this relationship should now correctly be defined as mutualistic (not phoretic) and develop a general model for the potential role of parasitism in the evolution of mutualism among the Astigmata.