Growth, death, and resource competition in sessile organisms.

Population-level scaling in ecological systems arises from individual growth and death with competitive constraints. We build on a minimal dynamical model of metabolic growth where the tension between individual growth and mortality determines population size distribution. We then separately include resource competition based on shared capture area. By varying rates of growth, death, and competitive attrition, we connect regular and random spatial patterns across sessile organisms from forests to ants, termites, and fairy circles. Then, we consider transient temporal dynamics in the context of asymmetric competition, such as canopy shading or large colony dominance, whose effects primarily weaken the smaller of two competitors. When such competition couples slow timescales of growth to fast competitive death, it generates population shocks and demographic oscillations similar to those observed in forest data. Our minimal quantitative theory unifies spatiotemporal patterns across sessile organisms through local competition mediated by the laws of metabolic growth, which in turn, are the result of long-term evolutionary dynamics.

TGFß signaling related genes are involved in hormonal mediation during termite soldier differentiation.

A working knowledge of the proximate factors intrinsic to sterile caste differentiation is necessary to understand the evolution of eusocial insects. Genomic and transcriptomic analyses in social hymenopteran insects have resulted in the hypothesis that sterile castes are generated by the novel function of co-opted or recruited universal gene networks found in solitary ancestors. However, transcriptome analysis during caste differentiation has not been tested in termites, and evolutionary processes associated with acquiring the caste are still unknown. Termites possess the soldier caste, which is regarded as the first acquired permanently sterile caste in the taxon. In this study, we performed a comparative transcriptome analysis in termite heads during 3 molting processes, i.e., worker, presoldier and soldier molts, under natural conditions in an incipient colony of the damp-wood termite Zootermopsis nevadensis. Although similar expression patterns were observed during each molting process, more than 50 genes were shown to be highly expressed before the presoldier (intermediate stage of soldier) molt. We then performed RNA interference (RNAi) of the candidate 13 genes, including transcription factors and uncharacterized protein genes, during presoldier differentiation induced by juvenile hormone (JH) analog treatment. Presoldiers induced after RNAi of two genes related to TGFß (Transforming growth factor beta) signaling were extremely unusual and possessed soldier-like phenotypes. These individuals also displayed aggressive behaviors similar to natural soldiers when confronted with Formica ants as hypothetical enemies. These presoldiers never molted into the next instar, presumably due to the decreased expression levels of the molting hormone (20-hydroxyecdysone; 20E) signaling genes. These results suggest that TGFß signaling was acquired for the novel function of regulating between JH and 20E signaling during soldier differentiation in termites.

The making of the defensive caste: Physiology, development, and evolution of the soldier differentiation in termites.

Termites (Blattodea, Termitoidea, or Isoptera) constitute one of the major lineages of eusocial insects. In termite societies, multiple types of functional individuals, that is, castes, perform divisions of labors to coordinate social behaviors. Among other castes, the soldier caste is distinctive since it is sterile and exclusively specialized into defensive behavior with largely modified morphological features. Therefore, many of the previous studies have been focused on soldiers, in terms of ecology, behavior, and evolution as well as developmental and physiological mechanisms. This article overviews the accumulation of studies especially focusing on the developmental and physiological mechanisms underlying the soldier differentiation in termites. Furthermore, the evolutionary trajectories that have led the acquisition of soldier caste and have diversified the soldier characteristics in association with the social evolution are discussed.

Unique Morphogenesis in the Damp-Wood Termite: Abscission of the Stylus during Female Reproductive Caste Differentiation.

Eusocial insects organize their colonies based on division of labor and caste differentiation, in which caste-specific morphologies are sculpted during postembryonic development. In the differentiation between reproductive and sterile castes, characteristics related to mating and reproduction are developed in reproductives, and degenerated in neuters, although little is known about the developmental regulations during the differentiation. In some species of termites, a sensory protrusion at the posterior end of the abdomen, the stylus, is known to disappear in female reproductives. In the present study, we performed anatomical and histological analyses in the damp-wood termite Hodotermopsis sjostedti to elucidate the developmental process underlying the disappearance of the stylus during neotenic and alate differentiation. Although it was first hypothesized that styli were hidden beneath the enlarged seventh sternite, our observation results found out that the styli were completely lost in reproductive females. Further histological observations revealed that the stylus disappearance was not accomplished by degeneration process; rather, styli were separated from the abdomen and discarded with the exoskeleton (exuviae) during the molt into the reproductive caste. This phenomenon in which live tissues are discarded through developmental processes is suggested to be a case of abscission, as seen in plant leaves.

Mitochondrial Phylogenomics Resolves the Global Spread of Higher Termites, Ecosystem Engineers of the Tropics.

The higher termites (Termitidae) are keystone species and ecosystem engineers. They have exceptional biomass and play important roles in decomposition of dead plant matter, in soil manipulation, and as the primary food for many animals, especially in the tropics. Higher termites are most diverse in rainforests, with estimated origins in the late Eocene (∼54 Ma), postdating the breakup of Pangaea and Gondwana when most continents became separated. Since termites are poor fliers, their origin and spread across the globe requires alternative explanation. Here, we show that higher termites originated 42-54 Ma in Africa and subsequently underwent at least 24 dispersal events between the continents in two main periods. Using phylogenetic analyses of mitochondrial genomes from 415 species, including all higher termite taxonomic and feeding groups, we inferred 10 dispersal events to South America and Asia 35-23 Ma, coinciding with the sharp decrease in global temperature, sea level, and rainforest cover in the Oligocene. After global temperatures increased, 23-5 Ma, there was only one more dispersal to South America but 11 to Asia and Australia, and one dispersal back to Africa. Most of these dispersal events were transoceanic and might have occurred via floating logs. The spread of higher termites across oceans was helped by the novel ecological opportunities brought about by environmental and ecosystem change, and led termites to become one of the few insect groups with specialized mammal predators. This has parallels with modern invasive species that have been able to thrive in human-impacted ecosystems.

A lipocalin protein, Neural Lazarillo, is key to social interactions that promote termite soldier differentiation.

Social communication among castes is a crucial component of insect societies. However, the genes involved in soldier determination through the regulation of inter-individual interactions are largely unknown. In an incipient colony of the damp-wood termite Zootermopsis nevadensis, the first larva to develop into a third instar always differentiates into a soldier via frequent trophallactic feeding from the reproductives. Here, by performing RNA-seq analysis of third instar larvae, a homologue of Neural Lazarillo (named ZnNLaz1) was found to be the most differentially expressed gene in these soldier-destined larvae, compared with worker-destined larvae. This gene encodes a lipocalin protein related to the transport of small hydrophobic molecules. RNAi-induced knockdown of ZnNLaz1 significantly inhibited trophallactic interactions with the queen and decreased the soldier differentiation rates. This protein is localized in the gut, particularly in the internal wall, of soldier-destined larvae, suggesting that it is involved in the integration of social signals from the queen through frequent trophallactic behaviours. Based on molecular phylogenetic analysis, we suggest that a novel function of termite NLaz1 has contributed to social evolution from the cockroach ancestors of termites. These results indicated that a high larval NLaz1 expression is crucial for soldier determination through social communication in termites.

Evidence from the gut microbiota of swarming alates of a vertical transmission of the bacterial symbionts in Nasutitermes arborum (Termitidae, Nasutitermitinae).

Studies on termite symbiosis have revealed that significant symbiont lineages are maintained across generations. However, most studies have focused only on the worker caste. Little is known about the gut microbiota of reproductives, the most probable vectors for transmitting these lineages to offspring. Using 16S rRNA gene-based Illumina MiSeq sequencing, we compared the gut microbiota of swarming alates of the higher termite Nasutitermes arborum with those of their nestmates from the parental colony. The OTU-based alpha diversity indices showed that the gut microbiota of the alates was at least as diverse as those of non-reproductive adults. It was largely dominated by Spirochaetes mostly of the Treponema I cluster (63.1% of reads), the same dominant taxa found in soldiers and workers of this species and in workers of closely related Nasutitermes species. The termite-specific lineages also included other representative taxa such as several clusters of Bacteroidetes and Fibrobacteres-TG3 group. The microbiota of alates was dominated by a core set of host-specific lineages (87% of reads, 77.6% of OTUs), which were always present across all castes/stages. This first comprehensive survey of the microbiota of the founding reproductives of these xylophagous higher termites shows that the bulk of the host endogenous symbionts, mostly taxa that cannot thrive outside the gut, are brought from the parent colony. The royal pair therefore seems to be a key player in the transmission of symbionts across generations and thereby in host-symbiont codiversification. The high proportion of fiber-degrading lineages in their gut suggests a wood-rich diet unlike the larval stages.

Secondary queens in the parthenogenetic termite Cavitermes tuberosus develop through a transitional helper stage.

In termite species with asexual queen succession (AQS), parthenogenetically produced immatures mostly differentiate into secondary queens, called "neotenics." In order to elucidate the ontogenetic origin of neotenics in Cavitermes tuberosus (Termitidae: Termitinae), a neotropical termite with AQS, we investigated developmental pathways of offspring according to their sex and genetic origin using both morphometric and genetic tools. The caste system of C. tuberosus follows the classical pathway of Termitidae. After the first larval instar, there is a bifurcation between two developmental lines. The apterous line is composed of a second larval instar, several worker instars, presoldiers, and soldiers. Workers display a consistent male bias and soldiers are female-only. The nymphal line is composed of five nymphal instars and the imago stage. We highlight that neotenic queens derive from third and fourth instar nymphs displaying peculiar morphological traits, here termed "aspirants," most of which are produced by parthenogenesis. Aspirants are present in all nests and perform worker tasks while waiting for the queen's death to differentiate into neotenic queens. Aspirants can successfully be used to demonstrate the occurrence of parthenogenesis in termite species whose reproductive cores are difficult to access.

The transition path from female workers to neotenic reproductives in the termite Reticulitermes labralis.

Termite workers are characterized by unique flexibility in that a worker can develop in one of three ways: remain a worker, become a soldier within two successive moults, or become a neotenic reproductive (NR) within a single moult. However, is it true that workers can transform into NRs within a single moult? Actually, the developmental pathways of workers turning into NRs remains unclear. In this study, we show for the first time that the female workers of Reticulitermes labralis develop into NRs after a pre-NRs stage. We found that a female worker became a NR after two successive moults, whereas the male workers copulated directly with queens without undergoing any moults. After the first moult led the female workers into the pre-NR stage, the length of their abdomens, seventh sternites and ovaries significantly increased. After the second moult from the pre-NRs stage into NRs, the follicle cells returned to normal, and a few oocytes and follicle cells underwent apoptosis. These results demonstrated that the female pre-NR type was a transitional type during the development of female workers to the NR caste, and the starting point for oogenesis resumption was the NR stage. We confirmed there were fundamental differences in the reproductive pathway of the male and female workers. Therefore, we determined that the transformation process of the female NRs from workers may be a very complex process, and the reproductive biology of the workers has great potential to provide important and spectacular insights into the evolution strategy of termites.

An American termite in Paris: temporal colony dynamics.

Termites of the genus Reticulitermes are widespread invaders, particularly in urban habitats. Their cryptic and subterranean lifestyle makes them difficult to detect, and we know little about their colony dynamics over time. In this study we examined the persistence of Reticulitermes flavipes (Kollar) colonies in the city of Paris over a period of 15 years. The aim was (1) to define the boundaries of colonies sampled within the same four areas over two sampling periods, (2) to determine whether the colonies identified during the first sampling period persisted to the second sampling period, and (3) to compare the results obtained when colonies were delineated using a standard population genetic approach versus a Bayesian clustering method that combined both spatial and genetic information. Herein, colony delineations were inferred from genetic differences at nine microsatellite loci and one mitochondrial locus. Four of the 18 identified colonies did not show significant differences in their genotype distributions between the two sampling periods. While allelic richness was low, making it hard to reliably distinguish colony family type, most colonies appeared to retain the same breeding structure over time. These large and expansive colonies showed an important ability to fuse (39% were mixed-family colonies), contained hundreds of reproductives and displayed evidence of isolation-by-distance, suggesting budding dispersal. These traits, which favor colony persistence over time, present a challenge for pest control efforts, which apply treatment locally. The other colonies showed significant differences, but we cannot exclude the possibility that their genotype distributions simply changed over time.

Postecdysis Sclerotization of Mouthparts of the Formosan Subterranean Termites (Isoptera: Rhinotermitidae).

In termites, it is challenging to recognize the incidence of molting in workers because of their successive stationary molt, asynchronous molting among individuals, cryptic behavior, a soft and poorly sclerotized cuticle, and they immediately consume the shed exuvia of nestmates. This study describes a method in which the degree of sclerotization of the mouthparts in newly molted workers of the Formosan subterranean termite can be quantified and used to determine if an individual has recently molted, within a 36-h time frame. Changes in the tanning of mouthparts over time were used as a measure of the index of sclerotization in workers postmolting. Upon ecdysis, the primary point of articulation of the mandible already initiated sclerotization, which may allow the movement of the mandibles during the shedding of the exuvium. The sclerotization of the secondary point of articulation and the mandibular teeth, and the width of sclerotization of the mandibles, progressively increased until reaching a plateau around 36-h postecdysis, which imply that workers can regain some level of activity as early as 2 d after ecdysis. Our observations allowed for the determination of variables for the sclerotization of the mouthparts to easily identify workers that recently molted, and this method will be useful in future studies that focus on the molting activity of workers over time and space within a termite colony, in the scope of improving current control strategies for termite pests.

High juvenile hormone titre and abdominal activation of JH signalling may induce reproduction of termite neotenics.

Termite castes are a key example of polyphenism, in which reproductive division of labour is clearly seen in colonies. The reproductive castes in termites include primary and neotenic reproductives; primary reproductives found a new colony whereas neotenics succeed them in the reproductive role when the primary reproductives die or become senescent. Neotenics usually differentiate from nymphs or workers by developing functional gonads while retaining juvenile characteristics; however, the developmental mechanism during neotenic differentiation remains poorly understood. Juvenile hormone (JH) mediates a number of aspects of developmental regulation in caste differentiation in termites. In the present study we quantified JH titres in neotenic reproductives of Reticulitermes speratus, and compared these with other developmental stages. In addition, expression changes in JH signalling gene homologues (Methoprene-tolerant [Met], Krüppel-homolog1, Broad-Complex) in the head, thorax and abdomen were investigated during neotenic differentiation. Finally, we examined the function of Met in reproduction of neotenics by RNA interference (RNAi). Our results showed that the JH titres of neotenics were significantly higher than those of nymphs and workers. JH signalling genes were highly expressed in neotenic abdomens, compared with those in workers and nymphs. Met RNAi resulted in the inhibition of vitellogenin gene expression in newly moulted neotenics. These results suggest that the fertility of neotenics might be controlled by a large increase of JH titres and body-part-specific activation of JH signalling pathways.

Expansion of presoldier cuticle contributes to head elongation during soldier differentiation in termites.

In termites, the soldier caste possesses morphological features suitable for colony defence, despite some exceptions. Soldiers are differentiated via two moultings through a presoldier stage with dramatic morphogenesis. While a number of morphological modifications are known to occur during the presoldier moult, growth and morphogenesis seem to continue even after the moult. The present study, using the damp-wood termite Hodotermopsis sjostedti, carried out morphological and histological investigations on the developmental processes during the presoldier stage that is artificially induced by the application of a juvenile hormone analogue. Measurements of five body parameters indicated that head length significantly increased during the 14-day period after the presoldier moult, while it did not increase subsequently to the stationary moult (pseudergate moult as control). Histological observations also showed that the cuticular development played a role in the presoldier head elongation, suggesting that the soft and flexible presoldier cuticle contributed to the soldier morphogenesis in termites.

Seasonal Changes in the Caste Distribution of Foraging Populations of Formosan Subterranean Termite in New Orleans, Louisiana.

This study examined the relationship between temperature, precipitation, soil composition, levels of feeding damage, and the caste distribution (workers, soldiers, nymphs) of the Formosan subterranean termite, Coptotermes formosanus Shiraki, collected in underground monitoring stations over a 12 mo period. Because nymphs are the caste that develops into alates, the seasonal abundance of nymphs was examined over a 5 yr period. Numbers of workers, soldiers, and soldier/worker ratio were significantly affected by month. Recruitment and retention of foraging termites in stations was significantly affected by the level of feeding damage. The number of nymphs collected in monitoring stations was highly variable. In the 12 mo test, there was a significant correlation between numbers of nymphs and level of feeding damage, temperature, precipitation, and soil composition. Over a 5 yr period, significantly more nymphs were collected in 2011 than in 2007 and 2008. Peak nymph collections varied from year to year. Overall, peak nymph collections were more likely to occur in Mar., Sept., and Oct. Increasing our knowledge of the environmental factors that influence recruitment and retention of foraging termites in monitoring stations could influence termite bait placement and improve baiting strategies for termite control. Identifying the key factors that cause aggregations of nymphs in underground stations could increase our ability to predict the intensity and location of alate swarms.

The development of adultoid reproductives and brachypterous neotenic reproductives from the last instar nymphs in Reticulitermes labralis (Isoptera: Rhinotermitidae): a comparative study.

Secondary reproductives develop primarily from nymphs. However, they have been rarely studied; in particular, the development of adultoid reproductives (AR) with floppy wings is still unclear. In this study, the change in juvenile hormone (JH) levels, vitellogenin gene expression, and oogenesis during the development of AR and brachypterous neotenic reproductives (BN) from the last instar nymphs of Reticulitermes labralis are investigated and compared. The results showed that the AR derived from the last instar nymphs by molting, and they were more similar to neotenic reproductives in morphology. In addition, the paired AR were not able to survive in the absence of workers. In R. labralis, the process of the last instar nymphs developing into AR and BN took an increase in JH level as a starting point. The JH level of the last instar nymphs molting into BN was approximately 1.5-fold higher than that of the AR. Additionally, The JHIII level of BN peaked on day 5, and that of AR peaked on day 10, which induced the onset of vitellogenesis in BN and AR, respectively. After molting, the vitellogenin gene expression levels of both BN and AR initially increased and then declined, and the expression levels in the BN were significantly higher than those in the AR. In addition, the oocytes of BN matured earlier than those of the AR, and the number of eggs laid by the BN was higher than the number laid by the AR. Our results demonstrate that, in R. labralis, the last instar nymphs can develop into AR, which are significantly different from BN in their development.

Intracolonial genetic variation affects reproductive skew and colony productivity during colony foundation in a parthenogenetic termite.

BACKGROUND: In insect societies, intracolonial genetic variation is predicted to affect both colony efficiency and reproductive skew. However, because the effects of genetic variation on these two colony characteristics have been tested independently, it remains unclear whether they are affected by genetic variation independently or in a related manner. Here we test the effect of genetic variation on colony efficiency and reproductive skew in a rhinotermitid termite, Reticulitermes speratus, a species in which female-female pairs can facultatively found colonies. We established colonies using two types of female-female pairs: colonies founded by sisters (i.e., sister-pair colonies) and those founded by females from different colonies (i.e., unrelated-pair colonies). Colony growth and reproductive skew were then compared between the two types of incipient colonies. RESULTS: At 15 months after colony foundation, unrelated-pair colonies were larger than sister-pair colonies, although the caste ratio between workers and nymphs, which were alternatively differentiated from young larvae, did not differ significantly. Microsatellite DNA analyses of both founders and their parthenogenetically produced offspring indicated that, in both sister-pair and unrelated-pair colonies, there was no significant skew in the production of eggs, larvae, workers and soldiers. Nymph production, however, was significantly more skewed in the sister-pair colonies than in unrelated-pair colonies. Because nymphs can develop into winged adults (alates) or nymphoid reproductives, they have a higher chance of direct reproduction than workers in this species. CONCLUSIONS: Our results support the idea that higher genetic variation among colony members could provide an increase in colony productivity, as shown in hymenopteran social insects. Moreover, this study suggests that low genetic variation (high relatedness) between founding females increases reproductive skew via one female preferentially channeling her relatives along the reproductive track. This study thus demonstrated that, in social insects, intracolonial genetic variation can simultaneously affect both colony efficiency and reproductive skew.

Body part-specific development in termite caste differentiation: crosstalk between hormonal actions and developmental toolkit genes.

In social insects, interactions among colony members trigger caste differentiation with morphological modifications. During caste differentiation in termites, body parts and caste-specific morphologies are modified during postembryonic development under endocrine controls such as juvenile hormone (JH) and ecdysone. In addition to endocrine factors, developmental toolkit genes such as Hox- and appendage-patterning genes also contribute to the caste-specific body part modifications. These toolkits are thought to provide spatial information for specific morphogenesis. During social evolution, the complex crosstalks between physiological and developmental mechanisms should be established, leading to the sophisticated caste systems. This article reviews recent studies on these mechanisms underlying the termite caste differentiation and addresses implications for the evolution of caste systems in termites.

Phylogenetic analyses of termite post-embryonic sequences illuminate caste and developmental pathway evolution.

Termites are highly eusocial insects with a caste polyphenism (i.e., discontinuous morphological differences between castes) and elaborated behaviors. While the developmental pathways leading to caste occurrence are well-known in many species, the evolutionary origin of these pathways is still obscure. Recent molecular phylogenetic studies suggest multiple independent origins of sterile castes in termites, reviving a 30 years old debate. We demonstrate here that diploid sterile castes ("true" workers) evolved several times independently in this group and that this caste was lost at least once in a lineage with developmentally more flexible workers called pseudergates or "false" workers. We also infer that flexibility in post-embryonic development was acquired multiple times independently during termite evolution. We suggest that focusing on detailed developmental pathways in phylogenetic analyses is essential for elucidating the origin of caste polyphenism in termites.

Soldier morphogenesis in the damp-wood termite is regulated by the insulin signaling pathway.

Eusocial insects exhibit various morphological castes associated with the division of labor within a colony. Termite soldiers possess defensive traits including mandibles that are greatly exaggerated and enlarged, as compared to termite reproductives and workers. The enlarged mandibles of soldiers are known to result from dynamic morphogenesis during soldier differentiation that can be induced by juvenile hormone and its analogs. However, the detailed developmental mechanisms still remain unresolved. Because the insulin/insulin-like growth factor signaling (IIS) pathway has been shown to regulate the relative sizes of organs (i.e., allometry) in other insects, we examined the expression profiles of major IIS factors in the damp-wood termite Hodotermopsis sjostedti, during soldier differentiation. The relative expression patterns of orthologs for termite InR (HsjInR), PKB/Akt (HsjPKB/Akt), and FOXO (HsjFOXO) suggest that HsjInR and HsjPKB/Akt were up-regulated in the period of elongation of mandibles during soldier development. In situ hybridization showed that HsjInR was strongly expressed in the mandibular epithelial tissues, and RNA interference (RNAi) for HsjInR disrupted soldier-specific morphogenesis including mandibular elongation. These results suggest that signaling through the IIS pathway is required for soldier-specific morphogenesis. In addition, up-regulation of the IIS pathway in other body tissues occurred at earlier stages of development, indicating that there is tissue-specific IIS regulation. Because the IIS pathway is generally thought to act upstream of JH in insects, our results suggest the damp-wood termite may have evolved a novel feedback loop between JH and IIS that enables social interactions, rather than nutrition, to regulate caste determination.

Spatial dynamics and ecosystem functioning.

Ecosystem functioning is dependent upon the way species become distributed across space.