Drosophila melanogaster is one of the most widely used model systems in biology. However, little is known about its associated bacterial community. As a first step towards understanding these communities, we compared bacterial 16S rRNA gene sequence libraries recovered from 11 natural populations of adult D. melanogaster. Bacteria from these sequence libraries were grouped into 74 distinct taxa, spanning the phyla Proteobacteria, Bacteroidetes, and Firmicutes, which were unevenly spread across host populations. Summed across populations, the distribution of abundance of genera was closely fit by a power law. We observed differences among host population locations both in bacterial community richness and in composition. Despite this significant spatial variation, no relationship was observed between species richness and a variety of abiotic factors, such as temperature and latitude. Overall, bacterial communities associated with adult D. melanogaster hosts are diverse and differ across host populations.Insects harbor diverse microbial communities (11,29,34,65), and interactions between hosts and their microbes can range from mutualistic, such as the interaction between termites and their gut microbes (8,65), to parasitic, such as the interaction of the bacterium Paenibacillus larvae (American foulbrood) in honeybees (62). Some of these interactions are relatively well characterized, owing to their economic importance or because of their remarkable biology. However, the exact nature of many other potentially interesting and experimentally tractable insect-microbe interactions, specifically those between microbes and the major insect model systems, remains poorly understood.In addition to the immediate association between insect hosts and the bacterial communities they harbor, the bacteria that insects carry can also associate with and affect the fitness of other hosts through vector transmission. The most common vector-borne zoonotic inflammatory disease in the United States, Lyme disease (caused by Borrelia burgdorferi), is transmitted by the deer tick, Ixodes scapularis, and infected more than 23,000 people in 2002 (26). In addition, Erwinia carotova, responsible for soft rot in many species of plants and for significant economic losses, can be vector transmitted by a variety of insects, including Drosophila melanogaster (38). Clearly, vector-borne bacterial infections can have large economic and health impacts and are important determinants of fitness for a variety of potential hosts.It is estimated that approximately 99% of the bacteria in nature are unculturable (3). With the advent of molecular techniques, such as PCR and genome sequencing, and metagenomic approaches, researchers have uncovered an astonishing level of microbial diversity in natural habitats, ranging from soil (7, 54, 69) and marine environments (23,64,69,70) to the human gut (25). The same techniques are currently being applied to understanding the microbiota of a range of insects (11,29,34,47,53,55,56,65). For example, using such sequence-b...

The developmental cycle of the myxobactetium Myxococcus xanthus consists of three partially overlapping morphological stages referred to as rippling, fruiting body formation, and sporulation, all of which are absent in csgA null mutants. The CsgA gene product is an extracellular protein, referred to as the C signal, which is essential for developmental cell-cell interactions. csgA expression increases throughout development, reaching its peak during sporulation. CsgA was made limiting for development by constructing nested deletions upstream from the csgA gene, which resulted in reduced csgA expression. Successively larger deletions resulted in termination of development at earlier and earlier stages, with rippling requiring -20% maximum csgA expression, fruiting body formation requiring -30% expression, and sporulation requiring 82% expression. Conversely, artificial induction of csgA also induced development provided nutrients were limiting. These results suggest that steady increases in CsgA over the course of development entrain the natural sequence of morphological events. The csgA upstream region appears to process information concerning the levels of nutrients, peptidoglycan components, and the B signal. In the absence of nutrients, a region extending 400 bp upstream from the start site of transcription was necessary for development and maximal csgA expression. In the presence of low levels of nutrients, a region extending -930 bp upstream was essential for the same tasks. It appears that the upstream region extending from -400 to -930 stimulates csgA expression in the presence of excess carbon, nitrogen, and phosphate, thereby allowing development to go to completion.

Myxococcus xanthus social (S) gliding motility has been previously reported by us to require the chemotaxis homologues encoded by the dif genes. In addition, two cell surface structures, type IV pili and extracellular matrix fibrils, are also critical to M. xanthus S motility. We have demonstrated here that M. xanthus dif genes are required for the biogenesis of fibrils but not for that of type IV pili. Furthermore, the developmental defects of dif mutants can be partially rescued by the addition of isolated fibril materials. Along with the chemotaxis genes of various swarming bacteria and the pilGHIJ genes of the twitching bacterium Pseudomonas aeruginosa, the M. xanthus dif genes belong to a unique class of bacterial chemotaxis genes or homologues implicated in the biogenesis of structures required for bacterial surface locomotion. Genetic studies indicate that the dif genes are linked to the M. xanthus dsp region, a locus known to be crucial for M. xanthus fibril biogenesis and S gliding.

The product(s) of the Myxococcus xanthus spoC locus is required for two multicellular activities in fruiting body development, rippling and sporulation. Ripples, which are formed early in development, are, spatialy separated ridges of cells that move synchronously. Myxospores are heat-resistant resting cells that are formed near the end of the developmental process. To investigate the function of spoC, it was cloned in an Escherichia coli.plasmid, then transferred to M. xanthus by specialized transduction with.coliphage P1. The plasmid, which cannot.replicate in M. xanthus, integrated into the M. xanthus chromosome, producing two copies of the spoC locus in tandem. Cells containing one copy of a mutant allele and one copy of the wild-type allele displayed the wild-type phenotype. Cells containing two different mutant alleles failed to ripple or sporulate, implying that all.four independent.spoC mutations. are in the same gene or unit of transcription. Homozygous mutant duplications arose from constructions in which DNA from a spo' donor was transduced into a spoC recipient, or vice versa, at an average frequency of 14%, indicating that gene conversion was a frequent event.

No abstract

The myxobacterium Myxococcus xanthus has a life cycle that is dominated by social behavior. During vegetative growth, cells prey on other bacteria in large groups that have been likened to wolf packs. When faced with starvation, cells form a macroscopic fruiting body containing thousands of spores. The social systems that guide fruiting body development have been examined through the isolation of conditional developmental mutants that can be stimulated to develop in the presence of wild-type cells. Extracellular complementation is due to the transfer of soluble and cell contact-dependent intercellular signals. This review describes the current state of knowledge concerning cell-cell signaling during development.

An agglutination assay was used to study cell cohesion in the myxobacterium Myxococcus xanthus. Vegetative cells agglutinated in the presence of the divalent cations Mg2+ and Ca2+. Agglutination was blocked by energy poisons that inhibit electron transport, uncouple oxidative phosphorylation, or inhibit the membrane-bound ATPase. However, energy was not required for the maintenance of cells in the multicellular aggregate. Cyanide, a strong inhibitor of agglutination, did not cause cells to dissociate from the aggregate even when shear forces were applied. While gliding motility was not necessary for agglutination, some gliding mutants exhibited aberrant agglutination that was generally correlated with cell behavior. Cells with an intact social motility system were cohesive and glided in large multicellular swarms. Cells with a mutation in their social motility system were 5- to 10-fold less cohesive and tended to glide as single cells. One group of social motility mutants, known as Dsp, did not agglutinate.

The gliding behavior of Myxococcus xanthus cells is controlled by two multigene systems, A and S, which encode information for adventurous and social behaviors, respectively. The S system can be genetically disrupted through mutation, such as a dsp mutation, or phenotypically disrupted by treating cells with the diazo dye Congo red (Arnold and Shimkets, J. Bacteriol. 170:5765-5770, 1988). One of the functions controlled by the S system is cell agglutination. Immediately after the induction of agglutination, wild-type cells begin to form aggregates, and within 30 min the cells are packed side-to-side in clumps containing thousands of cells. Changes in the cohesive properties of S+ cells are correlated with changes in the topology of the cell surface observed by electron microscopy. Two types of cell-associated appendages were observed on wild-type cells: thin filaments (ca. 5 nm in diameter), which have been called fimbriae or pili, at one cell pole, and thick, flaccid ifiaments (ca. 50 nm in diameter), referred to as fibrils, at both the sides and tips of cells. Cohesion was correlated with the secretion of the thick fibrils, which coat the cell surface and form an extracellular matrix in which the cells are interconnected. Several lines of evidence suggest that these thick fibrils are involved in cohesion. First, Dsp cells were unable to agglutinate or secrete this extracellular material. Second, wild-type cells which were treated with Congo red neither agglutinated nor secreted the extracellular fibrils. Finally, removal of the Congo red from wild-type cells restored cohesion and also restored production of the thick fibrils. Attempts to estimate the efficiency with which two cells cohered following collision suggested that under optimal conditions, one in three collisions resulted in stable contact. The collision efficiency decreased linearly as the cell density increased, suggesting a cell density-dependent regulation of cohesion. Some aspects of gliding behavior can be explained in terms of an inducer and an inhibitor of S motility.