Proximate factors influencing soldier development in the basal termite <Emphasis Type="Italic">Cryptotermes secundus</Emphasis> (Hill)

Summary Regulation of soldier development in termites at the endocrinological and social scale was intensively studied in the 60s and 70s, but conclusive results are still scanty. In termites, the presence of soldiers inhibits further soldier differentiation, but the mechanism is not well understood. Secretions from the frontal glands seem to be important, but dry- and dampwood termites do not have frontal glands. We investigated the influence of precocene I, a JH-antagonist, and of soldier head extracts on the development of soldiers in the drywood termite, Cryptotermes secundus (Hill). Soldierless colonies produced fewer soldiers when treated with precocene I or soldier head extract than control colonies. Furthermore, colonies treated with precocene I had produced worker-nymph intercastes, last stage nymphs and alates by the end of the experiment (after six months) which were not present in control colonies. Hence, both treatments inhibited soldier development while precocene I simultaneously promoted the development of adult traits, probably by reducing (directly or indirectly) the JH level. The inhibition effect of the soldier head extract indicates that at least in termites without a frontal gland, other sources of material in the head have a regulatory function.     

Preliminary studies on the control of caste formation in the harvester termiteHodotermes mossambicus (Hagen)

The control of caste formation in the harvester termiteHodotermes mossambicus (Hagen) was investigated.Normal laboratory colonies contained a primary reproductive pair, between 6.59 and 24.98% workers, 0.39 and 3.45% soldiers and 74.64 and 92.06% larvae. Supplementary reproductives did not develop in colonies deprived of their king and queen. Continuous removal of soldiers stimulated soldier production. We have suggested that a pheromone produced by the soldiers plays a major role in controlling the soldier population of a colony. A similar phermone system appreared to control the production of workers.Evidence was also obtained that the primary reproductive pair influenced the production of workers and soldiers.     

Reproductive schedule and factors affecting soldier production in the eusocial bamboo aphid Pseudoregma bambucicola (Homoptera, Aphididae)

Summary: The reproductive characteristics of the soldier-producing aphid Pseudoregma bambucicola were studied in Kagoshima, Southern Japan, to know the factors affecting soldier production of eusocial aphids. The soldier proportion in aphid colonies was highest from October to November. In some large colonies, soldiers were observed in all seasons except in July when colony size was relatively small. Multiple regression analysis showed that the colony size was a principal factor affecting soldier proportion throughout a year. Other social or environmental factors such as aphid composition, host plant conditions and predator abundance were not always significant. Rearing experiments revealed that large colonies (̿,000 individuals) produced soldiers in almost all seasons while small colonies (<1,000) never produced any soldiers. The caste-production schedule of adult females was examined in the field. When solitary females produced both castes, they usually produced normal nymphs first and then soldiers. Females from large colonies tended to produce more soldiers in the earlier period of their lifetime, whereas females from newly established small colonies produced no or only a few soldiers at later times. The average number of soldiers and normal nymphs produced consecutively by a single female was >10 and >20, respectively. Because they have a small number of ovarioles (<15 on average), females should alter caste production within the same ovarioles according to changes in environmental conditions. Artificial removal or introduction of predators and reduction of colony size did not affect soldier production over two successive generations, revealing maternal effects on soldier production. Females cannot shift caste production quickly in response to changes in predator abundance and colony size. This is probably due to early developmental determination of castes within the mother's body.     

截头堆砂白蚁性比的研究

为了解截头堆砂白蚁Cryptotermes domesticus（Haviland）群体中不同品级、不同虫态的雌雄性比,本文研究了该种白蚁自行配对室温条件下饲养4年巢龄的实验群体与采集的自然群体的各个品级和不同虫态的雌雄性比。结果表明：自行配对室温饲养4年巢龄群体与自然环境群体中原始繁殖蚁、幼蚁、若蚁、兵蚁都有雌雄之分,其中自行配对室温饲养群体的原始繁殖蚁雌雄比例为1：1,兵蚁雌雄比例为1：1．24,幼蚁和若蚁雌雄比例为1：1．46;自然环境群体原始繁殖蚁雌雄比例为1：1,兵蚁雌雄比例为1：1．15,幼蚁和若蚁雌雄比例为1：1．43。     

Soldiers are differentiated from male larval stages in incipient colonies of <Emphasis Type="Italic">Nasutitermes takasagoensis</Emphasis> (Isoptera: Termitidae)

In phylogenetically ancestral taxa of termites (the so-called lower termites), at least one soldier emerges and is maintained longitudinally in each incipient colony. However, in apical taxa (the so-called higher termites), the developmental pathway and regulation of soldiers in incipient colonies currently remain unknown. We therefore examined soldier and worker development in incipient colonies of higher termites (Nasutitermes takasagoensis Shiraki). Developmental stages and castes were successfully discriminated by head width in incipient colonies 4 months after colony foundation. Furthermore, differences were observed in the number of bristles on antennae between first- and second-instar larvae. In N. takasagoensis, there was more than one soldier in each incipient colony 4 months after its foundation. Presoldiers in the incipient colonies were differentiated from an earlier instar (male second-instar larvae), whereas, in mature colonies, they were differentiated from male third instars (= minor workers). The developmental period of the former (7 days) was markedly shorter than that of the latter (14 days). All female second-instar larvae molted into workers. The developmental processes shown here are useful for obtaining a clearer understanding of the mechanisms of soldier/worker differentiation in higher termites.     

Morphometric changes during soldier differentiation of the damp-wood termite Hodotermopsis japonica (Isoptera, Termopsidae)

Summary. To clarify the allometric development of body parts accompanying soldier differentiation in termites, we measured 16 body parts of soldiers, presoldiers, pseudergates (workers), nymphs and larvae of the damp-wood termite Hodotermopsis japonica. Principal component analysis (PCA) was performed using these parameters, which revealed that differentiation into soldiers differed distinctly from development into adult (reproductive) individuals. In particular, the anterior body parts enlarged during development of soldiers. Similarly, elongation of the apical portion of both mandibles was noted during soldier differentiation. X-ray analysis of mandibles revealed sclerotization of the soldier mandibles after differentiation into terminal soldiers. These morphological changes during soldier differentiation are associated with changes in their roles within the colony. Through soldier differentiation, the morphology of this caste of termite becomes functionally suited for attacking predators, and unsuitable for feeding on wood using their mandibles. Based on these data, we suggest that there must be some morphogenetic factors leading caste specific morphology such as soldier mandibles.     

Selection on defensive traits in a sterile caste – caste evolution: a mechanism to overcome life‐history trade‐offs?

SUMMARY During development and evolution individuals generally face a trade-off between the development of weapons and gonads. In termites, characterized by reproductive division of labor, a caste evolved—the soldiers—which is completely sterile and which might be released from developmental trade-offs between weapons and testes. These soldiers are exclusively dedicated to defense. First, we investigated whether defensive traits are under selection in sterile termite soldiers using allometric analyses. In soldiers of the genus Cryptotermes phragmotic traits such as a sculptured and foreshortened head evolve rapidly but were also lost twice. Second, we compared the scaling relationships of these weapons with those in solitary insects facing a trade-off between weapons and gonads. Defensive traits consistently had lower slopes than nondefensive traits which supports the existence of stabilizing selection on soldier phragmotic traits in order to plug galleries. Moreover, soldier head widths were colony specific and correlated with the minimum gallery diameter of a colony. This can proximately be explained by soldiers developing from different instars. The scaling relationships of these termite soldiers contrast strikingly with those of weapons of solitary insects, which are generally exaggerated (i.e., overscaling) male traits. These differences may provide important insights into trait evolution. Trade-offs constraining the development of individuals may have been uncoupled in termites by evolving different castes, each specialized for one function. When individuals in social insect are "released" from developmental constraints through the evolution of castes, this certainly contributed to the ecological and evolutionary success of social insects.     

X‐ray computed tomography reveals that intraspecific competition promotes soldier differentiation in a one‐piece nesting termite

Investment in soldier production in eusocial lineages involves a trade‐off between maintenance costs and defense benefits. Termites are eusocial insects that live in colonies organized into three castes: primary reproductives, soldiers, and workers or pseudergates. Neotermes chilensis (Blanchard) (Isoptera: Kalotermitidae) is a one‐piece nesting termite that nests and forages in a single piece of wood. Two scenarios may be of importance in a defense context of one‐piece nesting termites: during swarms, when colonies may be invaded by winged termites (alates) in search of a place to found a new colony, and when colonies of conspecifics are present within the same substrate. It was hypothesized that the ratio of soldiers to non‐soldiers would be higher at the onset of the swarming period and in substrates bearing more than one termite colony. A method based on X‐ray computed tomography (CT) was developed to study gallery connectivity in colonies of N. chilensis and caste composition within colonies. Computed tomography allowed the digital reconstruction of the galleries within the substrate, even when they belonged to different colonies, and was effective in distinguishing termites from substrate, and soldiers from reproductives and pseudergates. Using CT, the ratio of soldiers to non‐soldiers was shown to be highest in colonies within multicolonial scapes (i.e., neighboring colonies were present in the same substrate) during the swarming season, thus supporting our initial hypotheses. These results constitute a unique example of induced defenses arising from intraspecific interactions in termites.     

Soldier diterpene patterns in relation with aggressive behaviour,spatial distribution and reproduction of colonies in Nasutitermes princeps

The territorial organization of the colonies of N. princeps (Desneux) (Isoptera: Temitidae) was studied by four methods: maps of nest systems, tests of aggressiveness, comparison of soldier diterpene patterns, and determination of the reproductive status of the nests. The diterpene patterns are rather variable among sympatric colonies, and allow the tracing of soldiers from their respective colonies throughout their foraging range. Soldiers from neighbouring nests display in some instances closely resembling diterpene patterns, suggesting close genetic relatedness and polycalic colonies, i.e. composed of several linked nests spread over large territories. Most likely, N, princeps colonies reproduce by budding. The number and type of reproductives found in the nests are consistent with this hypothesis. Tests of aggressiveness supported the above conclusions, but absence of aggression did not always indicate that the termites belonged to the same colony.     

Social structure and the defensive role of soldiers in a eusocial bamboo aphid,Pseudoregma bambucicola (Homoptera: Aphididae): A test of the defence-optimization hypothesis

A complete understanding of the evolution of sociality in aphids requires a detailed knowledge of the patterns of soldier investment in their ecology. The eusocial bamboo aphidPseudoregma bambucicola has a morphologically specialized first-instar soldier caste. The proportion of soldiers was positively correlated with colony size. Within a colony, soldiers were evenly distributed among subcolonies; within each subcolony, however, their distribution was biased toward peripheries which were exposed to many predators. Field experiments introducing natural enemies such asEupeodes confrater (Diptera: Syrphidae) andSynonycha grandis (Coleoptera: Coccinellidae) revealed that the survival rate of these predators was negatively correlated with the density of soldiers, suggesting that soldiers can more or less defend their colonies by killing or removing a range of natural enemies. Observations suggest that large mature colonies attract more predators than newly established small colonies and that, within a colony, the predators attack each subcolony regardless of its position on bamboo shoots. This implies the presence of a positive correlation between colony size and predation risk. Thus, the investment in soldiers seems to reflect the attacking pattern of predators within a colony. These results agree with the defence-optimization hypothesis in soldier investment ofP. bambucicola colonies.     

八种白蚁的兵蚁和工蚁消化道形态比较研究(等翅目)

为探讨白蚁消化道形态品级差异及其系统学意义,对4个科8种白蚁兵蚁和工蚁的消化道进行比较形态学研究.结果表明:低等白蚁的兵蚁和(拟)工蚁形态较为相似,高等白蚁的兵蚁与工蚁局部特征存在明显差异.由低等到高等兵蚁和工蚁消化道各部分差异呈增多趋势.前、中、后肠占消化道的百分比在山林原白蚁Hodotermopsis sjoest...     

Impact of a juvenile hormone analogue on the anatomy and the frontal gland secretion of Prorhinotermes simplex (Isoptera: Rhinotermitidae)

In termites, juvenile hormone plays a key role in soldier differentiation. To better understand the evolutionary origin of the soldiers, we studied the external and inner morphology of pseudergate-soldier intercastes and neotenic-soldier intercastes formed artificially by the application of juvenile hormone analogue in Prorhinotermes simplex. A majority of these intercastes had a soldier phenotype, whereas the inner anatomy had an intermediary form between two castes or a form specific to intercastes. Our experiments showed that traits of neotenics and soldiers can be shared by the same individuals, although such individuals do not exist naturally in P. simplex, and they have not been reported in other species but in some Termopsidae. Our results reinforce the hypothesis that soldiers may have emerged from soldier neotenics during the evolution of termites.     

Group size effect on worker juvenile hormone titers and soldier differentiation in Formosan subterranean termite

In a finite environment, population growth can lead to crowding, increased densities and stress. Termites live in highly organized societies and densities can increase astronomically as colonies grow. However, little is known about juvenile hormone (JH) changes in these insects as numbers increase, despite the fact that JH is a critical caste regulator in this insect. Using Formosan subterranean termites as a model, we evaluated minimum group size requirements for soldier differentiation and the effect of density (50, 100, 500, and 1000 individuals/experimental unit) on worker JH titers either with or without initial soldiers being present. The minimum group size investigation indicated that groups initiated with 5 workers were insufficient for soldier differentiation. Soldiers were produced in groups initiated with 10 or more workers. As density increased from 50 to 1000 individuals per experimental unit, worker JH levels were elevated. Presence of soldiers lessened the effect of density on rising worker JH titers, indicating that soldiers have the ability to down-regulate worker JH. The study provides direct evidence of a population density effect on JH in eusocial insects and sheds light on understanding of the regulatory mechanisms associated with termite soldier caste differentiation.     

Soldier caste influences on candidate primer pheromone levels and juvenile hormone-dependent caste differentiation in workers of the termite Reticulitermes flavipes

Caste systems and the division of labor they make possible are common underlying features of all social insects. Multiple extrinsic factors have been shown to impact caste composition in social insect colonies. Primer pheromones are one type of extrinsic caste-regulatory factor; they are chemical signaling molecules produced by certain colony members to impact developmental physiology of recipient nestmates. However, only limited evidence exists regarding primer pheromones and their actions in eusocial termites. In previous research we identified two soldier-produced terpenes, γ-cadinene (CAD) and γ-cadinenal (ALD), as candidate primer pheromones of the lower termite Reticulitermes flavipes. In the present study we tested hypotheses related to CAD and ALD action in recipient individuals. We examined the influences of terminally developed soldier termites on (1) CAD and ALD levels and (2) caste differentiation in developmentally totipotent workers. Our findings show CAD and ALD (respectively) are caste stimulatory and inhibitory components of chemical blends present in soldier heads, ALD levels increase significantly (10.9×) in workers only in the presence of soldiers, and soldiers can reduce developmental-hormone response thresholds of workers, presumably via ALD action. These findings provide novel evidence supporting that CAD and ALD are authentic caste-regulatory primer pheromones in Reticulitermes.     

Male or female soldiers? An evaluation of several factors which may influence soldier sex ratio in lower termites

In termites, the soldiers’ sex ratio is often biased toward one sex. Unlike in the Hymenoptera, this bias cannot easily be explained by relatedness asymmetries because termites are diploid. Matsuura proposed that when large body size is adaptive for colony defence (e.g. in case of phragmotic defence) then the larger sex (given sexual size dimorphism exists) should be more likely to reach a threshold size and develop into soldiers. This would explain biased sex ratios. Matsuura validated his hypothesis for four Reticulitermes species. Here, we tested his hypothesis for two species of Cryptotermes with phragmotic defence. These drywood termites have a life type that is thought to be ancestral in termite’s evolution, thus giving us potential insights into the evolution of the soldier caste. In one of these species, the sex ratio of soldiers was highly female biased, but we could not support Matsuura’s hypothesis. Both species lacked sexual size dimorphism in all castes. Additionally, in both species, the sex ratio of helpers and sexuals did not deviate from a 1:1 ratio, and hence can also not account for the bias observed in soldiers. However, this study showed that there were behavioural differences between the sexes in both species, which could shed some light on biased sex ratio in soldiers. Our findings also indicate that the developmental pathway taken by individuals reflects a ‘decision’ at the colony level. The discovery of behavioural differences between sexes in termites should open the way to similar studies in other taxa with helpers/ workers of both sexes, as it might reveal more task partitioning in colonies than previously thought and it raises questions concerning the selective pressures that acted on caste evolution in termites. Received 30 October 2007; revised 17 January and 27 February; accepted 4 March 2008.     

Asexual queen succession in the higher termite Embiratermes neotenicus

Asexual queen succession (AQS), in which workers, soldiers and dispersing reproductives are produced sexually while numerous non-dispersing queens arise through thelytokous parthenogenesis, has recently been described in three species of lower termites of the genus Reticulitermes. Here, we show that AQS is not an oddity restricted to a single genus of lower termites, but a more widespread strategy occurring also in the most advanced termite group, the higher termites (Termitidae). We analysed the genetic structure in 10 colonies of the Neotropical higher termite Embiratermes neotenicus (Syntermitinae) using five newly developed polymorphic microsatellite loci. The colonies contained one primary king accompanied either by a single primary queen or by up to almost 200 neotenic queens. While the workers, the soldiers and most future dispersing reproductives were produced sexually, the non-dispersing neotenic queens originated through thelytokous parthenogenesis of the founding primary queen. Surprisingly, the mode of thelytoky observed in E. neotenicus is most probably automixis with central fusion, contrasting with the automixis with terminal fusion documented in Reticulitermes. The occurrence of AQS based on different mechanisms of ploidy restoration raises the hypothesis of an independent evolutionary origin of this unique reproductive strategy in individual lineages of lower and higher termites.     

Association between caste and genotype in the termite Mastotermes darwiniensis Froggatt (Isoptera: Mastotermitidae)

Abstract Termite workers and soldiers differ markedly in their morphology and behaviour. We sought evidence for genetic influences on caste determination in the giant northern termite, Mastotermes darwiniensis , by investigating if workers and soldiers from the same colony differed genetically. The genotypes of 795 termites from 11 distinct colonies were assayed at six polymorphic microsatellite loci. We found that the multilocus genotypes of workers and soldiers from 8 of the 11 colonies did not differ significantly. Thus, the majority of the data provided no evidence for a genetic association with caste in workers and soldiers of M. darwiniensis . However, the genotype frequencies of workers and soldiers from three colonies differed, suggesting that genotype is occasionally associated with caste in this species. The genetic differentiation of castes within these colonies could reflect differences in the propensities of termites with distinct genotypes to develop into particular castes and provide a selective advantage to colonies headed by multiple reproductives.     

Density triggers soldier production in a social aphid

For evolution and maintenance of the social system of insect colonies, investment in the sterile caste should be adequately controlled in response to environmental cues. Recent developments using artificial diet rearing techniques have revealed an underlying mechanism of caste control in a gall-forming aphid, Tuberaphis styraci, which has a soldier caste in the second instar. Statistical analyses of field-collected galls detected a significant positive correlation between aphid density and soldier proportion in the natural colonies of T. styraci. Artificial diet experiments showed that soldiers are produced under crowded conditions. Detailed experiments demonstrated that soldiers are produced in a density-dependent manner rather than in a colony size-dependent manner. From these results, it was concluded that aphid density is the crucial cue that triggers soldier production in T. styraci. This study provides, to our knowledge, the first experimental demonstration of an environmental factor involved in aphid soldier differentiation.     

The evolution of soldier reproduction in social thrips

We estimated the degree of reproductive differentiation betweenfoundresses and soldiers in multiple populations of five speciesof haplodiploid Australian gall-forming thrips using microsatellitedata, ovarian dissections, and census data. Microsatellite-basedspecies estimates of average per capita reproductive outputof soldiers relative to the foundresses ranged from 0.005 to0.64, and dissection and census-based estimates ranged from0.17 to 1.1. Mapping of these estimates onto a phylogeny showedthat levels of soldier reproduction were apparently higherin three basal lineages than in two more derived lineages.We infer from this phylogenetic pattern that soldier morphologyand behavior of thrips evolved in the presence of substantial levels of soldier reproduction. This pattern of evolutionarychange is similar to that proposed for the origin of soldiersin aphids and termites, but it differs from the scenario proposedfor the origin of workers in Hymenoptera, within which helpingand strong reproductive division of labor apparently evolvedbefore morphological differentiation. We suggest that this difference in evolutionary routes to eusociality between taxa with soldiersand taxa with foraging workers was driven by a weaker trade-offbetween helping and reproducing, and a greater ability of thehelpers to withstand reproductive domination, in taxa withsoldiers. This is the first study to analyze the social-evolutionarytrajectories of reproductive, behavioral, and morphologicaldifferentiation in the context of a species-level phylogeny.     

Soldier-specific modification of the mandibular motor neurons in termites

Social insects exhibit a variety of caste-specific behavioral tendencies that constitute the basis of division of labor within the colony. In termites, the soldier caste display distinctive defense behaviors, such as aggressively attacking enemies with well-developed mandibles, while the other castes retreat into the colony without exhibiting any aggressive response. It is thus likely that some form of soldier-specific neuronal modification exists in termites. In this study, the authors compared the brain (cerebral ganglion) and the suboesophageal ganglion (SOG) of soldiers and pseudergates (workers) in the damp-wood termite, Hodotermopsis sjostedti. The size of the SOG was significantly larger in soldiers than in pseudergates, but no difference in brain size was apparent between castes. Furthermore, mandibular nerves were thicker in soldiers than in pseudergates. Retrograde staining revealed that the somata sizes of the mandibular motor neurons (MdMNs) in soldiers were more than twice as large as those of pseudergates. The enlargement of MdMNs was also observed in individuals treated with a juvenile hormone analogue (JHA), indicating that MdMNs become enlarged in response to juvenile hormone (JH) action during soldier differentiation. This enlargement is likely to have two functions: a behavioral function in which soldier termites will be able to defend more effectively through relatively faster and stronger mandibular movements, and a developmental function that associates with the development of soldier-specific mandibular muscle morphogenesis in termite head. The soldier-specific enlargement of mandibular motor neurons was observed in all examined species in five termite families that have different mechanisms of defense, suggesting that such neuronal modification was already present in the common ancestor of termites and is significant for soldier function.