@article{GuptaKupperRatzkaetal.2015, author = {Gupta, Shishir K. and Kupper, Maria and Ratzka, Carolin and Feldhaar, Heike and Vilcinskas, Andreas and Gross, Roy and Dandekar, Thomas and F{\"o}rster, Frank}, title = {Scrutinizing the immune defence inventory of Camponotus floridanus applying total transcriptome sequencing}, series = {BMC Genomics}, volume = {16}, journal = {BMC Genomics}, number = {540}, doi = {10.1186/s12864-015-1748-1}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-125279}, year = {2015}, abstract = {Background Defence mechanisms of organisms are shaped by their lifestyle, environment and pathogen pressure. Carpenter ants are social insects which live in huge colonies comprising genetically closely related individuals in high densities within nests. This lifestyle potentially facilitates the rapid spread of pathogens between individuals. In concert with their innate immune system, social insects may apply external immune defences to manipulate the microbial community among individuals and within nests. Additionally, carpenter ants carry a mutualistic intracellular and obligate endosymbiotic bacterium, possibly maintained and regulated by the innate immune system. Thus, different selective forces could shape internal immune defences of Camponotus floridanus. Results The immune gene repertoire of C. floridanus was investigated by re-evaluating its genome sequence combined with a full transcriptome analysis of immune challenged and control animals using Illumina sequencing. The genome was re-annotated by mapping transcriptome reads and masking repeats. A total of 978 protein sequences were characterised further by annotating functional domains, leading to a change in their original annotation regarding function and domain composition in about 8 \% of all proteins. Based on homology analysis with key components of major immune pathways of insects, the C. floridanus immune-related genes were compared to those of Drosophila melanogaster, Apis mellifera, and other hymenoptera. This analysis revealed that overall the immune system of carpenter ants comprises many components found in these insects. In addition, several C. floridanus specific genes of yet unknown functions but which are strongly induced after immune challenge were discovered. In contrast to solitary insects like Drosophila or the hymenopteran Nasonia vitripennis, the number of genes encoding pattern recognition receptors specific for bacterial peptidoglycan (PGN) and a variety of known antimicrobial peptide (AMP) genes is lower in C. floridanus. The comparative analysis of gene expression post immune-challenge in different developmental stages of C. floridanus suggests a stronger induction of immune gene expression in larvae in comparison to adults. Conclusions The comparison of the immune system of C. floridanus with that of other insects revealed the presence of a broad immune repertoire. However, the relatively low number of PGN recognition proteins and AMPs, the identification of Camponotus specific putative immune genes, and stage specific differences in immune gene regulation reflects Camponotus specific evolution including adaptations to its lifestyle.}, language = {en} } @article{RatzkaFoersterLiangetal.2012, author = {Ratzka, Carolin and F{\"o}rster, Frank and Liang, Chunguang and Kupper, Maria and Dandekar, Thomas and Feldhaar, Heike and Gross, Roy}, title = {Molecular characterization of antimicrobial peptide genes of the carpenter ant Camponotus floridanus}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-75985}, year = {2012}, abstract = {The production of antimicrobial peptides (AMPs) is a major defense mechanism against pathogen infestation and of particular importance for insects relying exclusively on an innate immune system. Here, we report on the characterization of three AMPs from the carpenter ant Camponotus floridanus. Due to sequence similarities and amino acid composition these peptides can be classified into the cysteine-rich (e.g. defensin) and glycine-rich (e.g. hymenoptaecin) AMP groups, respectively. The gene and cDNA sequences of these AMPs were established and their expression was shown to be induced by microbial challenge. We characterized two different defensin genes. The defensin-2 gene has a single intron, whereas the defensin-1 gene has two introns. The deduced amino acid sequence of the C. floridanus defensins is very similar to other known ant defensins with the exception of a short C-terminal extension of defensin-1. The hymenoptaecin gene has a single intron and a very peculiar domain structure. The corresponding precursor protein consists of a signal- and a pro-sequence followed by a hymenoptaecin-like domain and six directly repeated hymenoptaecin domains. Each of the hymenoptaecin domains is flanked by an EAEP-spacer sequence and a RR-site known to be a proteolytic processing site. Thus, proteolytic processing of the multipeptide precursor may generate several mature AMPs leading to an amplification of the immune response. Bioinformatical analyses revealed the presence of hymenoptaecin genes with similar multipeptide precursor structure in genomes of other ant species suggesting an evolutionary conserved important role of this gene in ant immunity.}, subject = {Biologie}, language = {en} } @phdthesis{Ratzka2012, author = {Ratzka, Carolin}, title = {Immune responses of the ant Camponotus floridanus towards pathogens and its obligate mutualistic endosymbiont Blochmannia floridanus}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-69350}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2012}, abstract = {Ants of the species Camponotus floridanus live in huge colonies composed of genetically identical or closely related animals, which should predispose them to an increased vulnerability towards infection by pathogens (Cremer et al. 2007). Therefore the question is how ants (or social insects in general) can nevertheless efficiently combat infections. In order to investigate the immune response of the ant C. floridanus, the present study initially focused on the identification of possible immune factors, encoded by the ant´s genome. By using the method "suppression subtractive hybridization" as well as by Illumnia sequencing technology, several immune-related genes could be identified. Among these were genes encoding proteins involved in pathogen recognition, signal transduction, antimicrobial activity, or general stress response. In accordance with the ant´s genome sequence (Bonasio et al. 2010), only three antimicrobial peptide (AMP) genes could be identified in C. floridanus. The gene and cDNA sequences of these AMPs were established and their expression was shown to be induced by microbial challenge. Two different defensin genes (type 1 and 2) were characterized. A detailed characterization of the mRNA and gene sequence of the other AMP, a hymenoptaecin, revealed a special repeat structure. The C. floridanus hymenoptaecin has a signal and a pro-sequence followed by a hymenoptaecin-like domain and six directly repeated hymenoptaecin domains (HDs). Since each HD is flanked by two known processing sites, proteolytic processing of the precursor protein may generate several mature AMPs. Bioinformatical analyses revealed the presence of hymenoptaecin genes with similar multipeptide precursor structure in genomes of other ant species suggesting an evolutionary conserved important role of this gene in ant immunity. C. floridanus ants harbor the obligate intracellular bacterium, Blochmannia floridanus, in specialized cells (so-called bacteriocytes), which are intercalated between midgut cells as well as in ovaries of females (Blochmann 1882; Sauer et al. 2002; Schr{\"o}der et al. 1996). Ant hosts face the problem that on the one hand they have to maintain the beneficial symbiotic bacteria and on the other hand they need to raise an immune response against harmful pathogenic bacteria during an infection. It was investigated, if endosymbionts are actually detected by the host immune system. Injection of B. floridanus induced an immune response of its host C. floridanus, which was comparable to the one towards pathogens. This means that, despite the evolutionary established cooperation of the endosymbionts and their hosts, these bacteria are still recognized as „non-self" by the host immune system. This finding led to the question, if the ant immune system might be involved in regulation of the endosymbiont number in the midgut tissue in order to avoid their uncontrolled replication. During the holometabolous life cycle of the ant hosts the distribution of bacteriocytes and of Blochmannia endosymbionts is remarkably dynamic and peaks in late pupal stages, in which the entire midgut is transformed into a symbiotic organ (Stoll et al. 2010). It was hypothesized that hosts could regulate the number of endosymbionts present in their tissues via the innate immune system. A quantitative gene expression analysis of assumed symbiosis-relevant candidate genes revealed distinct expression patterns of some genes according to developmental stage and tissue. Moreover, the immune gene expression in response to bacterial challenge was investigated in the pupal stage. By an artificial immune-challenge of pupae it was confirmed that in fact the immune response of the endosymbiont-bearing midgut tissue differs from that of other body parts. The data support a key role for amidase peptidoglycan recognition proteins (PGRPs), especially PGRP-LB, in endosymbiont tolerance and suggest an involvement of the lysosomal system in control of Blochmannia endosymbionts. In sum, this thesis provides a first description of the immune response of the ant C. floridanus. A comprehensive set of immune-relevant genes was determined. Especially, the identification and molecular characterization of the hymenoptaecin gene delivered new insights into the immune competence of ants in general. Moreover, first indications could be gathered for the involvement of the immune system in controlling the endosymbiont B. floridanus.}, subject = {Humorale Immunit{\"a}t}, language = {en} }