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The relative importance of topography and vegetation in shaping local soil invertebrate communities is poorly understood. We investigated changes in the species and trait compositions of Collembola communities associated with different topography and forest types. Our study sites encompassed various topographies with different slope aspects and forest types; specifically, natural deciduous broad-leaved forests and deciduous coniferous plantations in Japan. The body length and edaphic adaptation score, which are representative of a certain life-form, were used as Collembola traits. Across a topographic gradient from south to north-facing slopes, the total biomass of Collembola was higher in dry plots than in humid plots, and the forest types did not affect either abundance, biomass, or species diversity. Variation partitioning analysis revealed that topography played an important role in determining species composition and functional traits, whereas forest types and spatial distance had minor roles. The responses of community weighted mean traits (CWMs) to environmental change were more pronounced than those of species composition. The CWM and functional diversity of body length were higher in the dry plots than in the humid plots. These changes in traits might be explained by food supply or habitat structure increasing large individuals via topography, rather than by physical stress filtering out intolerant small individuals. These results suggest that slope topography is an important determinant of the Collembola community structure in cool-temperate forest ecosystems.

Tropical trees account for most emissions of isoprene, a reactive biogenic volatile organic compound into the atmosphere. The Guenther 1993 (G-93) model is the most widely used algorithm for predicting isoprene emissions by leaves of terrestrial plants. Several studies have reported on the poor performance of the G-93 model in predicting emissions from tropical tree species. To improve the performance of the G-93 model in tropical regions, we carried out diurnal leaf-scale observations of tropical trees Casuarina equisetifolia and Ficus septica outdoors. We developed an iterative method that uses mutual and repetitive step-by-step optimization of the G-93 parameters for temperature (CT) and light (CL) variables using best fit practices, named “Ping-Pong” optimization. Using temperate tree species (Poplar) for comparison, we show that emissions from C. equisetifolia and F. septica had a diversion from predictions of the G-93 formula, especially at high temperatures and high light intensities during mid-day, whilst emissions from temperate tree species were fairly captured by G-93. Results demonstrate that our optimized formulas greatly improved capturing of high light and temperature responses of emission profiles from tropical trees whilst it also performed well for poplar species. Parameterization of the G-93 formula greatly improved its performance on predicting diurnal isoprene emission from tropical trees C. equisetifolia and F. septica; explaining 81–96% of variation up from 73 to 77% explained by default G-93. We propose that there is a need to optimize the G-93 model to more accurately predict regional emissions from tropical ecosystems.

Isoprene emission from plants is very sensitive to environmental temperature both at short-term and long-term scales. Our previous study demonstrated suppression of isoprene emission by cold temperatures in a high emitting tropical tree Ficus septica and revealed a strong correlation of emission to isoprene synthase (IspS) protein levels. When challenged with decreasing daily temperatures from 30 to 12 °C, F. septica completely stopped isoprene emission at 12 °C, only to recover on the second day after re-exposure to 30 °C. Here, we explored this regulation of isoprene emission in response to environmental temperature by a comprehensive analysis of transcriptome data, gene expressions and metabolite pools of the 2-C-methyl-D-erythritol 4-phosphate (MEP) pathway. MEP pathway genes and metabolites dynamics did not support substrate-level limitations as major control over observed basal emission, but transcriptome data, network inferences and putative regulatory elements on IspS promoter suggested transcriptional regulation of IspS gene through circadian rhythm and phytohormone signalling processes. Expression levels of 29 genes involved in these pathways were examined by quantitative real-time PCR. We propose that temperature controls over basal isoprene emission at a time-scale of hours to few days are regulated by phytohormone-mediated transcriptional modulation of IspS gene under synchronization by the circadian clock.

Isoprene is the most abundant type of nonmethane, biogenic volatile organic compound in the atmosphere, and it is produced mainly by terrestrial plants. The tropical tree species Ficus septica Burm. F. (Rosales: Moraceae) has been shown to cease isoprene emissions when exposed to temperatures of 12 °C or lower and to re-induce isoprene synthesis upon subsequent exposure to temperatures of 30 °C or higher for 24 h. To elucidate the regulation of genes underlying the disabling and then induction of isoprene emission during acclimatization to ambient temperature, we conducted gene expression analyses of F. septica plants under changing temperature using quantitative real-time polymerase chain reaction and western blotting. Transcription levels were analyzed for 17 genes that are involved in metabolic pathways potentially associated with isoprene biosynthesis, including isoprene synthase (ispS). The protein levels of ispS were also measured. Changes in transcription and protein levels of the ispS gene, but not in the other assessed genes, showed identical temporal patterns to isoprene emission capacity under the changing temperature regime. The ispS protein levels strongly and positively correlated with isoprene emission capacity (R(2) = 0.92). These results suggest that transcriptional regulation of ispS gave rise to the temporal variation in isoprene emission capacity in response to changing temperature.

Alimentary canals of higher termites are equipped with a complex structural segment consisting of mesenteric and proctodeal epithelial tissues, which is termed the mixed segment. To assess metabolic functions occurring in the mixed segment, Nasutitermes takasagoensis (Shiraki) transcriptomes were obtained from the mixed segment and adjacent gut regions (midgut and first proctodeal segment) and were annotated using the Kyoto Encyclopedia of Genes and Genomes (KEGG) database. The predominant metabolic function detected in the mixed segment was energy production via oxidative phosphorylation. As KEGG orthologs involved in lysosomal and phagosomal systems were also characteristic of this segment, cellular components are probably recycled rather than newly produced, allowing higher energy production. The predominant adenosine triphosphate (ATP) consumer in the mixed segment was vacuolar-type H+-ATPase (V-ATPase), known to be involved in K+ transport in many insects. Based on our data, V-ATPases appear to electrogenically drive NHE6 or NHE7 (NHE6/7) K+/H+ antiporters to stimulate K+ secretion, increasing gut lumen pH. The mixed segment also expressed higher levels than did the adjacent gut regions of carbonic anhydrase, another contributor to gut alkalization through HCO3 − production. Thus, metabolic pathways in the mixed segment might promote lignocellulose solubilization in higher termites’ gut, contributing to their success.

Aim Drug resistance has been a major issue in cancer chemotherapy. It is therefore important to screen more potent natural anti-cancer agents for use against the drug-resistant cancer phenotype. The present study investigated the cytotoxicity of the traditional and natural medicinal agent nitidine (NTD) against the drug-resistant cancer phenotype of A549 human lung adenocarcinoma cells. Methods The camptothecin (CPT) tolerant phenotype was selected after A549 cells were exposed to CPT for 1 week. The cytotoxicity of NTD in these CPT-resistant cells (CRC) was studied in vitro in relation to its accumulation and expression of the ATP binding cassette (ABC) transporter gene. Results CRC expressing higher levels of ABCC1, ABCC2, ABCC3, and ABCA1 were more resistant to CPT compared with normal A549 cells. CRC were, however, more susceptible to NTD compared with normal A549 cells with preferential accumulation of NTD. A statistically significant difference in protein level between normal A549 and CRC was noted only for ABCC1 and ABCC2. Furthermore, gene expression of ABCC2 and ABCC3 had a positive correlation with NTD accumulation, and negative correlation with the effective dose that inhibited 50% of cell growth (D50) of this agent in nine cancer cell panels. Conclusion CRC were found to be more susceptible to NTD compared with the normal phenotype. The cytotoxicity of NTD was associated with its accumulation and the expression of ABCC2, suggesting the involvement of this transporter in the selective incorporation of NTD into cancer cells, resulting in enhanced cytotoxicity.

Whole-genome sequencing has emerged as one of the most effective means to elucidate the biological roles and molecular features of obligate intracellular symbionts (endosymbionts). However, the de novo assembly of an endosymbiont genome remains a challenge when host and/or mitochondrial DNA sequences are present in a dataset and hinder the assembly of the genome. By focusing on the traits of genome evolution in endosymbionts, we herein developed and investigated a genome-assembly strategy that consisted of two consecutive procedures: the selection of endosymbiont contigs from an output obtained from a de novo assembly performed using a TBLASTX search against a reference genome, named TBLASTX Contig Selection and Filtering (TCSF), and the iterative reassembling of the genome from reads mapped on the selected contigs, named Iterative Mapping and ReAssembling (IMRA), to merge the contigs. In order to validate this approach, we sequenced two strains of the cockroach endosymbiont Blattabacterium cuenoti and applied this strategy to the datasets. TCSF was determined to be highly accurate and sensitive in contig selection even when the genome of a distantly related free-living bacterium was used as a reference genome. Furthermore, the use of IMRA markedly improved sequence assemblies: the genomic sequence of an endosymbiont was almost completed from a dataset containing only 3% of the sequences of the endosymbiont's genome. The efficiency of our strategy may facilitate further studies on endosymbionts.

The draft genome sequence of Lactobacillus paracasei strain LC-Ikematsu, isolated from a pineapple in Okinawa, was determined. The total length of the 87 contigs was 3.08 Mb with a G+C content of 46.2% and 2,946 coding sequences. The genome analysis revealed its biosynthetic ability of 11 amino acids.

Influence of living roots on the community composition of Collembola was investigated in a coniferous forest of Chamaecyparis obtusa. We conducted a buried pot experiment that constructed two different systems for carbon (C) availability. We used two types of pots with or without a C. obtusa seedling. The former pots were used to make a system with soil including living roots (i.e. a system based on root-derived and litter-derived C), while the latter was equivalent to soil system without living roots (i.e. a system based on litter-derived C). After 8 months, we harvested the pots and examined the collembolan community and environmental factors. The presence of living roots affected collembolan abundance and species-specific responses. These changes could be explained in terms of leaf biomass of the seedlings, indicating a possible linkage between above-ground productivity and collembolan community. Given the possibility that root-derived C is associated with aboveground plant mass, we speculate that collembolan community responds to root-derived C. The three dominant species, which are widespread in Japanese temperate forests, were more abundant in the presence of living roots. Moreover they were positively correlated with leaf biomass in the system with a seedling, again suggesting the fundamental importance of living roots for the organisation of the collembolan community.

Eight microsatellite markers for the population genetics and evolutionary ecology of Ischnura senegalensis (Rambur 1842), which shows body color polymorphism in females, were developed using a streptavidin-bead enrichment library and pyrosequence by a next generation sequencer. The number of alleles per locus and effective number of alleles ranged from 3 to 11 and from 1.24 to 5.51, respectively. Observed and expected heterozygosities were 0.18–0.75 and 0.19–0.77, respectively. No linkage disequilibrium between loci was detected. One locus, IsenAC75, deviated significantly from the Hardy–Weinberg equilibrium, and the locus and additional two loci, IsenAC40 and IsenAC8, were suspected for the presence of null alleles. Altogether, these eight microsatellite loci are considered to be useful for population genetic analyses because of the high polymorphic status and independency.

We investigated soil microarthropod communities in two physically dissimilar inorganic soil materials and in a mixture of these two materials to examine the effect of the structural complexity of a habitat on microarthropod abundance and communities, teasing it out from that of nutritional factors. Mesh boxes were filled with perlite (a highly porous material), similar size of granite gravels (no pores inside), or their mixture, and placed on a forest floor. The boxes were collected after 8 or 20 months, and the microarthropods were extracted and identified to the species level, with a focus on Collembola. We also evaluated fine-root biomass and the amount of organic matter in the boxes. It was found that the mixture of perlite and granite enhanced microarthropod abundance and root development. A partial redundancy analysis revealed that collembolan communities developed differently among the substrate materials. We also found that variation in the collembolan communities was related to fine-root development and the abundance of other microarthropods, implying that habitat structural complexity affects collembolan communities indirectly by affecting soil food webs.

Bacteria of the genus Rhodococcus (Actinomycetes) have the ability to catabolize various organic compounds and are therefore considered potential genetic resources for applications such as bioremediation. We investigated a next-generation sequencing-based procedure to rapidly identify candidate functional gene(s) from rhodococci on the basis of their frequent genome recombination. The Rhodococcus rhodochrous strain B-276 and its alkene monooxygenase (AMO) gene cluster were the focus of our investigation. Firstly, 2 types of cultures of the R. rhodochrous strain B-276 were prepared, one of which was supplied with propene, which requires AMO genes for its assimilation, whereas the other was supplied with glucose as the sole energy source. The latter culture was anticipated to have a lower gene frequency of AMO genes because of their deletion during cultivation. We then conducted whole genome shotgun sequencing of the genomic DNA extracted from both cultures. Next, all sequence data were pooled and assembled into contiguous sequences (contigs). Finally, the abundance of each contig was quantified in order to detect contigs that were highly biased between the 2 cultures. We identified contigs that were overrepresented by 2 orders of magnitude in the AMO-required culture and successfully identified an AMO gene cluster among these contigs. We propose this procedure as an efficient method for the rapid detection and sequencing of deleted region, which contributes to identification of functional genes in rhodococci.

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A free-living ciliate, Trimyema compressum , found in anoxic freshwater environments harbors methanogenic archaea and a bacterial symbiont named TC1 in its cytoplasm. Here, we report the complete genome sequence of the TC1 symbiont, consisting of a 1.59-Mb chromosome and a 35.8-kb plasmid, which was determined using the PacBio RSII sequencer.

We developed a novel protocol with superior quantitative analysis results for DNA metabarcoding of Collembola, a major soil microarthropod order. Degenerate PCR primers were designed for conserved regions in the mitochondrial cytochrome c oxidase subunit I (mtCOI) and 16S ribosomal RNA (mt16S) genes based on published collembolan mitogenomes. The best primer pair was selected based on its ability to amplify each gene, irrespective of the species. DNA was extracted from 10 natural communities sampled in a temperate forest (with typically 25–30 collembolan species per 10 soil samples) and 10 mock communities (with seven cultured collembolan species). The two gene regions were then amplified using the selected primers, ligated with adapters for 454 technology, and sequenced. Examination of the natural community samples showed that 32 and 36 operational taxonomic units (defined at a 90% sequence similarity threshold) were recovered from the mtCOI and mt16S data, respectively, which were comparable to the results of the microscopic identification of 25 morphospecies. Further, sequence abundances for each collembolan species from the mtCOI and mt16S data of the mock communities, after normalization by using a species as the internal control, showed good correlation with the number of individuals in the samples (R = 0.91–0.99), although relative species abundances within a mock community sample estimated from sequences were skewed from community composition in terms of the number of individuals or biomass of the species. Thus, this protocol enables the comparison of collembolan communities in a quantitative manner by metabarcoding.

Isoprene emission from plants is very sensitive to environmental temperature both at short-term and long-term scales. Our previous study demonstrated suppression of isoprene emission by cold temperatures in a high emitting tropical tree Ficus septica and revealed a strong correlation of emission to isoprene synthase (IspS) protein levels. When challenged with decreasing daily temperatures from 30 to 12 °C, F. septica completely stopped isoprene emission at 12 °C, only to recover on the second day after re-exposure to 30 °C. Here, we explored this regulation of isoprene emission in response to environmental temperature by a comprehensive analysis of transcriptome data, gene expressions and metabolite pools of the 2-C-methyl-D-erythritol 4-phosphate (MEP) pathway. MEP pathway genes and metabolites dynamics did not support substrate-level limitations as major control over observed basal emission, but transcriptome data, network inferences and putative regulatory elements on IspS promoter suggested transcriptional regulation of IspS gene through circadian rhythm and phytohormone signalling processes. Expression levels of 29 genes involved in these pathways were examined by quantitative real-time PCR. We propose that temperature controls over basal isoprene emission at a time-scale of hours to few days are regulated by phytohormone-mediated transcriptional modulation of IspS gene under synchronization by the circadian clock.

Nasutitermes takasagoensis soldiers defend their colonies using characteristic diterpenes. Diterpenes are thought to be synthesized in the frontal gland cells surrounding the gland reservoir. To identify the genes involved in diterpene synthesis, a cDNA library was prepared from the frontal gland cells and exhaustively sequenced using a 454 pyrosequencer (GS Junior; Roche, Branford, CT, USA). A total of 50,290 clean sequences were assembled into 1111 contigs, which were grouped into 774 genes (isogroups). Based on sequence similarity with known proteins, we identified seven genes encoding the following four enzymes associated with diterpene synthesis: 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) synthase (HMGS), HMG-CoA reductase (HMGR), farnesyl diphosphate synthase, and geranylgeranyl diphosphate synthases. The expression levels of two enzymes, HMGS and HMGR, involved in the mevalonate pathway were examined, assuming that the site of the defensive terpenoid synthesis strongly activates the mevalonate pathway, which produces a precursor of terpenoids. Real-time quantitative reverse-transcriptase PCR confirmed significantly higher expression of HMGS and HMGR in the heads of soldiers. We then divided the head into three parts and found that the expression levels of HMGS and HMGR were significantly higher in the part containing class 1 secretory cells of the frontal gland. Overall, the results suggested that the mevalonate pathway for diterpene synthesis occurs in class 1 cells around the frontal gland reservoir.

We measured the natural stable carbon (C) and nitrogen (N) isotope ratio patterns of collembola and the organic substrates of their habitats and potential food sources in a warm temperate coniferous forest. Based on previous studies, we classified collembola into successional classes along litter decomposition gradients: early colonizers, late colonizers, and dominants-throughout. The stable C and N isotope ratios of late colonizers exceeded those of early colonizers, and those of the dominants-throughout were intermediate between early and late colonizers, which is consistent with previous studies on two macro-invertebrates, earthworms and termites. The C and N isotopic signature differences in collembola may reflect food resource partitioning along decomposition gradients.

The mixed segment is a unique part of the gut present only in the most apical lineage of termites and consists of a complex of overlapping mesenteric and proctodeal epithelia. In spite of its unique structure, the physiological functions of the mixed segment have been poorly studied. We performed transcriptome analysis to identify functional enzymes acting in the mixed segment of the wood-feeding higher termite Nasutitermes takasagoensis. We sequenced the transcripts (4563 isotigs) of the mixed segment and compared them with those of the midgut (4813 isotigs) and the first proctodeal segment (3629 isotigs). We found that vacuolar H+-ATPase (V-ATPase) subunits were predominant in the mixed segment, which was confirmed by RT-qPCR analysis. The V-ATPase activity in these three tissues was in a good agreement with the expression patterns, suggesting that V-ATPase is a prevalent enzyme in the mixed segment of the termites. The results confirmed the proposed role of the mixed segment as a transporting epithelium.

The Ryukyu ground geckos, which comprise Goniurosaurus kuroiwae (sensu stricto) and four close relatives in the Ryukyu Archipelago, face conservation threats, and are listed as endangered in the Red List of both IUCN and Japan. We developed 24 microsatellite markers in G. kuroiwae and cross-amplified for another species, G. orientalis. All 24 and 22 loci were successfully amplified and genotyped in G. kuroiwae and G. orientalis, respectively. The number of alleles per locus and expected heterozygosity at polymorphic loci ranged from 1 to 11 and 0.050–0.891, respectively, in G. kuroiwae, and ranged from 1 to 15 and 0.050–0.932, respectively, in G. orientalis.

The paper concerns two species of Collembola having a strong spine armature at the end of abdomen. Sahacanthella saoriae sp.nov. (Hokkaido, Japan), the second species of the genus, is described. Octodonthophora ornata (Magadan district, Russia) is redescribed with remarks on its taxonomical position. A key to the world genera of Anurophorinae with anal spines is given. The genera having four anal spines on the fifth abdominal segment are illustrated. DNA sequences of genes for mitochondrial cytochrome c oxidase subunit I, 16S ribosomal RNA, and nucler 28S ribosomal RNA were sequenced as DNA barcodes for S. saoriae sp.nov. and several taxa of Anurophorinae.

The unicellular green alga Chlamydomonas reinhardtii has a haplontic life cycle, and forms diploid zygotes for reproduction. The zygospore, a sporulating zygote, begins germination in response to light signals, generating haploid progenies and inducing several cell-biological events; e.g., DNA synthesis and meiotic division, successively. Their regulatory mechanisms remain largely unknown, so we focused on the early stages of germination and analyzed the dynamics of gene expression associated with the germination process. The gene expression levels of zygospores at 1 and 6h after light exposure were analyzed by a next-generation sequencing platform, the 454 GS Junior. At 6h, the photosynthesis pathway, including its antenna proteins and two methionine metabolism-related genes (methionine synthase and sulfite reductase), were up-regulated compared to 1h after light exposure. Meanwhile, three uncharacterized genes that contained an antibiotic biosynthesis monooxygenase domain and an HSP20/alpha crystallin family protein were specifically expressed at 1h after light exposure. These gene expressions were also verified by quantitative real-time PCR analysis. These results suggest that the photosynthesis and methionine synthesis pathways, both of which occur in the chloroplast, are activated in zygospores at around 6h after light exposure, and that some polyketides and/or a small heat shock protein may be related to the initiation of zygospore germination.

We sequenced a complete mitochondrial genome of the marine sponge, Cacospongia mycofijiensis, which is the first report for the family Thorectidae in the class Demospongiae. The mitogenome was obtained from a de novo assembly of shotgun genome sequencing using Illumina Miseq technology, which reconstructed a circular genome with 97 × of sequence coverage. The assembled mitochondrial genome consisting of 16,227 bp includes 14 protein-coding genes, 2 rRNAs and 2 tRNAs. This complete mitogenome sequence will be useful especially for the phylogenic studies of Demospongiae.

This paper describes the existence of temperature threshold in isoprene emission from tropical trees for the first time. Isoprene emission capacity of Ficus virgata leaves kept outdoors were measured over a period of 20 d in the wintertime. When the plants experienced the low temperature below 12 °C in the daytime, isoprene emission was completely suppressed, and re-activated by elevated temperature above the threshold of 12 °C. Photosynthesis is also decreased similarly, but too much smaller extent compared to the reduction observed for isoprene emission. The cut off level of accumulated temperature to shut off the isoprene emission was roughly estimated to be 300 degree/d. The shut off in the isoprene emission by the low temperature was confirmed experimentally by use of phytotron with tropical trees of F. virgata and F. septica. Isoprene emission of both species ceased at 12 °C, and re-activated by raising the temperature above the threshold level. Photosynthesis was lowered slightly as was the case for the field observation, and no close association between photosynthesis and isoprene emission was noted. The expression of isoprene synthase gene showed much greater variation after exposure to the cold temperature compared to those involved in photosynthesis. These observations therefore suggested the existence of putative direct connection between perception of temperature and isoprene emission in tropical trees, which may merit further investigation to estimate the net output of isoprene from subtropical or tropical forest.

Research on the soft coral genus Sarcophyton extends over a wide range of fields, including marine natural products and the isolation of a number of cembranoid diterpenes. However, it is still unknown how soft corals produce this diverse array of metabolites, and the relationship between soft coral diversity and cembranoid diterpene production is not clear. In order to understand this relationship, we examined Sarcophyton specimens from Okinawa, Japan, by utilizing three methods: morphological examination of sclerites, chemotype identification, and phylogenetic examination of both Sarcophyton (utilizing mitochondrial protein-coding genes MutS homolog: msh1) and their endosymbiotic Symbiodinium spp. (utilizing nuclear internal transcribed spacer of ribosomal DNA: ITS- rDNA). Chemotypes, molecular phylogenetic clades, and sclerites of Sarcophyton trocheliophorum specimens formed a clear and distinct group, but the relationships between chemotypes, molecular phylogenetic clade types and sclerites of the most common species, Sarcophyton glaucum, was not clear. S. glaucum was divided into four clades. A characteristic chemotype was observed within one phylogenetic clade of S. glaucum. Identities of symbiotic algae Symbiodinium spp. had no apparent relation to chemotypes of Sarcophyton spp. This study demonstrates that the complex results observed for S. glaucum are due to the incomplete and complex taxonomy of this species group. Our novel method of identification should help contribute to classification and taxonomic reassessment of this diverse soft coral genus.

The effectiveness of cancer chemotherapy is often limited by the toxicity to other tissues in the body. Therefore, the identification of non-toxic chemotherapeutics from herbal medicines remains to be an attractive goal to advance cancer treatments. This study evaluated the cytotoxicity profiles of 364 herbal plant extracts, using various cancer and normal cell lines. The screening found occurrence of A549 (human lung adenocarcinoma) specific cytotoxicity in nine species of herbal plants, especially in the extract of Arctium lappa L. Moreover, purification of the selective cytotoxicity in the extract of Arctium lappa L. resulted in the identification of arctigenin as tumor specific agent that showed cytotoxicity to lung cancer (A549), liver cancer (HepG2) and stomach cancer (KATO III) cells, while no cytotoxicity to several normal cell lines. Arctigenin specifically inhibited the proliferation of cancer cells, which might consequently lead to the induction of apoptosis. In conclusion, this study found that arctigenin was one of cancer specific phytochemicals, and in part responsible for the tumor selective cytotoxicity of the herbal medicine.

In addition to harbouring intestinal symbionts, some animal species also possess intracellular symbiotic microbes. The relative contributions of gut-resident and intracellular symbionts to host metabolism, and how they coevolve are not well understood. Cockroaches and the termite Mastotermes darwiniensis present a unique opportunity to examine the evolution of spatially separated symbionts, as they harbour gut symbionts and the intracellular symbiont Blattabacterium cuenoti . The genomes of B. cuenoti from M. darwiniensis and the social wood-feeding cockroach Cryptocercus punctulatus are each missing most of the pathways for the synthesis of essential amino acids found in the genomes of relatives from non-wood-feeding hosts. Hypotheses to explain this pathway degradation include: (i) feeding on microbes present in rotting wood by ancestral hosts; (ii) the evolution of high-fidelity transfer of gut microbes via social behaviour. To test these hypotheses, we sequenced the B. cuenoti genome of a third wood-feeding species, the phylogenetically distant and non-social Panesthia angustipennis . We show that host wood-feeding does not necessarily lead to degradation of essential amino acid synthesis pathways in B. cuenoti , and argue that ancestral high-fidelity transfer of gut microbes best explains their loss in strains from M. darwiniensis and C. punctulatus .