Asexual queen succession in the subterranean termite Reticulitermes virginicus

Termite colonies are founded by a pair of primary reproductives. In many species, including subterranean termites (family Rhinotermitidae), the primary king and queen can be succeeded by neotenic reproductives that are produced from workers or nymphs within the colony. It is generally believed that these neotenics inbreed within the colony, sometimes for many generations. Here, we show that primary queens of the North American subterranean termite, Reticulitermes virginicus, are replaced by numerous parthenogenetically produced female neotenics. We collected functional female neotenics from five colonies of R. virginicus in North Carolina and Texas, USA. Genetic analysis at eight microsatellite loci showed that 91-100% of the neotenics present within a colony were homozygous at all loci, indicating that they were produced through automictic parthenogenesis with terminal fusion. In contrast, workers, soldiers and alates were almost exclusively sexually produced by mating between the female neotenics and a single king. This is the second termite species shown to undergo asexual queen succession, a system first described in the Japanese species, Reticulitermes speratus. Thus, the conditional use of sexual and asexual reproduction to produce members of different castes may be widespread within Reticulitermes and possibly other subterranean termites.     

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.     

Workers do not mediate the inhibitory power of queens in a termite, <Emphasis Type="Italic">Reticulitermes speratus</Emphasis> (Isoptera,Rhinotermitidae)

In social insects, the caste systems are based on reproductive division of labor; queens specialize in reproduction and workers primarily maintain the colony. Recently, a volatile pheromone containing n-butyl-n-butyrate and 2-methyl-1-butanol was identified as a termite queen pheromone that inhibits the differentiation of female neotenic reproductives (secondary queens). Although this volatile inhibitory pheromone regulates caste differentiation directly, the method by which it reaches members without direct contact with the queen in large colonies is not well understood. Therefore, additional mechanisms of indirect communication must exist, such as worker-mediated queen signal transport. We found that workers exposed to female reproductives did not mediate queens’ inhibitory signal in a termite Reticulitermes speratus. The experiment assessed worker transfer from direct to indirect contact groups and determined that the differentiation of new female reproductives in the indirect contact groups was not influenced by the direct contact groups, whereas direct contact with functional female reproductives and artificial queen pheromone did suppress neotenic differentiation. This suggests that worker transfer of the queen signal is unlikely and that for colony-wide inhibition direct contact by the majority of infertile members with reproductives or eggs, which emit the same volatiles as female reproductives, is necessary within a certain time interval.     

Head-butting as an Early Indicator of Reproductive Disinhibition in the Termite Zootermopsis nevadensis

In lower termites, functionally sterile larval helpers are totipotent—capable of becoming reproductively active with the loss of their colony’s king or queen. Full reproductive development may take several weeks, but initiation of this developmental response most likely occurs shortly after colony members detect when a reproductive-specific signal is missing. We investigated the early response of termite helpers to the removal of their king and queen in the basal termite species Zootermopsis nevadensis. Within 6–12 h after reproductives were removed, helpers displayed an increase in head-butting—a behavior associated with dominance in other termite species as well as in closely related roaches. The loss of just one reproductive, either the king or queen, was also sufficient to cause an increase in head-butting. We did not find evidence, however, that this response was sex-specific: males and females were equally likely to increase head-butting independent of the sex of the reproductive that was removed. Finally, we discovered that reproductive-specific compounds present on the cuticle of king and queen termites were also present in their feces, but the presence of the feces did not seem sufficient to inhibit the increased head-butting after the reproductives were removed. Collectively, these results indicate that termite workers readily detect the loss of reproductives in their colony and that they at least initially respond in a non sex-specific manner.     

Behavioural differences between male and female replacement reproductives in Kalotermes flavicollis (Isoptera, Kalotermitidae)

Summary: An ethogram type study was performed on representative colonies of Kalotermes flavicollis to investigate the behavioural repertoires of replacement king and queen. Our observations suggested a sex-based behavioural specialisation in the two reproductives, affecting colony organisation and equilibrium. Interesting differences were detected in overall activity of the two reproductives, with the male showing the greatest frequency of movements in the nest, vibratory acts, and the highest rate and range of social contacts. In addition to his importance for periodical insemination of the queen, the king might also play a basic role in the social structure and dynamic development of the colony, as a "mediator" in social interactions between the queen and the other colony members.     

Benefits and costs of secondary polygyny in the dampwood termite Zootermopsis angusticollis

Newly molted female neotenic reproductives of the dampwood termite Zootermopsis angusticollis Hagen were allowed to mature in the presence of a neotenic male, a fixed number of larval helpers, and varying numbers of sibling neotenic queens to assess the impact of secondary polygyny to the individual and colony. Under monogyne conditions, neotenics developed more ovarioles per ovary and had higher individual fecundities after 60 d compared with females under polygyne conditions. Queens in groups of three females were able to gain more body mass than those in groups of five. Although the division of resources provided by helpers reduced individual female development and fecundity under polygyne conditions, it resulted in an overall increase in colony fecundity. In addition, neotenic females in polygynous colonies did not differ significantly in reproductive competence. There was no evidence that neotenics were attacked or injured by other reproductives or larval helpers, suggesting little if any reproductive competition among sibling queens. The physiological responses of neotenics to the increasing queen/worker ratio may have the benefit of enhancing the colony growth at the cost of the fecundity of individual queens.     

Queen pheromone regulates egg production in a termite

In social insects, resource allocation is a key factor that influences colony survival and growth. Optimal allocation to queens and brood is essential for maximum colony productivity, requiring colony members to have information on the total reproductive power in colonies. However, the mechanisms regulating egg production relative to the current labour force for brood care remain poorly known. Recently, a volatile chemical was identified as a termite queen pheromone that inhibits the differentiation of new neotenic reproductives (secondary reproductives developed from nymphs or workers) in Reticulitermes speratus. The same volatile chemical is also emitted by eggs. This queen pheromone would therefore be expected to act as an honest message of the reproductive power about queens. In this study, we examined how the queen pheromone influences the reproductive rate of queens in R. speratus. We compared the number of eggs produced by each queen between groups with and without exposure to artificial queen pheromone. Exposure to the pheromone resulted in a significant decrease in egg production in both single-queen and multiple-queen groups. This is the first report supporting the role of queen pheromones as a signal regulating colony-level egg production, using synthetically derived compounds in a termite.     

尖唇散白蚁工蚁、兵蚁和繁殖蚁精子发生的比较研究

为探讨白蚁非生殖品级和生殖品级生殖细胞发育差异,采用组织学染色技术对尖唇散白蚁Reticulitermes aculabialis Tsaiet Hwang繁殖蚁、工蚁和兵蚁的精巢发育以及精子发生进行了显微观察和比较研究。结果发现3个品级间精巢发育的程度差异很大,三者精巢切面面积相对大小之比为:繁殖蚁∶工蚁=1.7∶1;繁殖蚁∶兵蚁=29.3∶1;工蚁∶兵蚁=17.1∶1。繁殖蚁和工蚁精巢管内有精子的形成,工蚁和繁殖蚁精子的发生都经历了精原细胞、初级精母细胞、次级精母细胞、精细胞和精子时期,但工蚁有大量次级精母细胞呈细胞凋亡状态。兵蚁生殖细胞发育仅有精原细胞、初级精母细胞、次级精母细胞,没有精细胞和精子产生。工蚁的生殖细胞显著小于同一时期繁殖蚁的生殖细胞,兵蚁的各时期生殖细胞均极显著小于繁殖蚁同一时期的生殖细胞。研究表明各品级之间生殖功能分化与生殖细胞发育有直接关系,工蚁有转化为补充繁殖蚁和兵蚁的能力;而兵蚁由于精巢极度退化不能产生精细胞和精子,因此是非生殖品级分化的终极形式,不具有转化成补充繁殖蚁或其它品级的能力。     

Clinal variation in colony breeding structure and level of inbreeding in the subterranean termites Reticulitermes flavipes and R. grassei

Social insects exhibit remarkable variation in their colony breeding structures, both within and among species. Ecological factors are believed to be important in shaping reproductive traits of social insect colonies, yet there is little information linking specific environmental variables with differences in breeding structure. Subterranean termites (Rhinotermitidae) show exceptional variation in colony breeding structure, differing in the number of reproductives and degree of inbreeding; colonies can be simple families headed by a single pair of monogamous reproductives (king and queen) or they can be extended families headed by multiple inbreeding neotenic reproductives (wingless individuals). Using microsatellite markers, we characterized colony breeding structure and levels of inbreeding in populations over large parts of the range of the subterranean termites Reticulitermes flavipes in the USA and R. grassei in Europe. Combining these new data with previous results on populations of both species, we found that latitude had a strong effect on the proportion of extended‐family colonies in R. flavipes and on levels of inbreeding in both species. We examined the effect of several environmental variables that vary latitudinally; while the degree of inbreeding was greatest in cool, moist habitats in both species, seasonality affected the species differently. Inbreeding in R. flavipes was most strongly associated with climatic variables (mean annual temperature and seasonality), whereas nonclimatic variables, including the availability of wood substrate and soil composition, were important predictors of inbreeding in R. grassei. These results are the first showing that termite breeding structure is shaped by local environmental factors and that species can vary in their responses to these factors.     

圆唇散白蚁雌性补充生殖蚁不同发育阶段表皮长链碳氢化合物的变化

【目的】确定圆唇散白蚁 Reticulitermes labralis 补充生殖蚁不同发育阶段表皮长链碳氢化合物(cuticular long-chained hydrocarbons, CHCs) 的变化。【方法】人工隔离巢群饲养圆唇散白蚁 R. labralis,观测其翅芽型补充生殖蚁的分化时间和补充生殖蚁数量趋于稳定的时间,并采用气质联用仪(GC-MS)对补充生殖蚁不同发育阶段的表皮长链碳氢化合物变化趋势进行分析。【结果】随着补充生殖蚁的出现,每一巢群会逐渐形成稳定数量的具有生殖能力的生殖蚁。若蚁在分化为补充生殖蚁的第8天时,43.3%的组补充生殖蚁数量趋于稳定,第16天时83.3%的组趋于稳定,第25天时93.3%的组趋于稳定,36 d后100%的组趋于稳定。通过补充生殖蚁不同发育阶段中CHCs各峰面积与若蚁阶段相应峰峰面积的比值随发育阶段的变化趋势分析发现,在卵中没有检测到的保留时间为38.52 min的化合物在补充生殖蚁产7~8粒卵时峰面积比值是若蚁阶段的14.14倍;在卵中检测到的保留时间为35.50和39.21 min的峰面积比值在补充生殖蚁开始产卵后显著升高。【结论】CHCs不能用来区分有无生殖能力的补充生殖蚁个体,但可以很好地用来区分补充生殖蚁个体与尚未转化为生殖蚁的若蚁个体;保留时间为38.52 min的化合物是补充生殖蚁的特有物质;保留时间为35.50和39.21 min的化合物可作为补充生殖蚁已经产卵的标志。     

The influence of group size on ovarian development in adult and neotenic reproductives of the termite Zootermopsis angusticollis (Hagen) (Hodotermitidae)

Termite imagines of Zootermopsis angusticollis (Hagen), while still in the colony as winged individuals, do not show ovary development, although vitellogenin is present in the blood at low levels. When removed from the colony, artificially dealated, and paired, vitellogenin levels rise: subsequently ovaries develop resulting in oviposition approximately 15–16 days after removal from the colony. Imagines removed from the colony, dealated and held together in a heterosexual group, however, show little or no ovary development after 25 days, although vitellogenin is present in the haemolymph in low levels. Newly developed neotenic reproductives, on the other hand, removed from the colonly 1–2 days after the neotenic moult, and isolated in heterosexual pairs for 60 days, had ovaries which did not become larger than 0.02 mm³, and no eggs were laid. This is in contrast to the normal ovary development of neotenic reproductives produced in groups of larvae, as described earlier. Vitellogenin levels in isolated pairs of neotenics were low until 25 days after isolation, while vitellogenin levels increased 7–9 days after the moult in female neotenics kept in a colony with larvae. The results of this study on ovarian development and vitellogenesis in neotenic and adult reproductives reflect the normal biology of the two reproductive types where the latter leave the colony to found new colonies, while neotenics become sexually mature within the colony in which they developed.     

圆唇散白蚁补充生殖蚁的类型与建群能力

摘要: 【目的】为了探讨圆唇散白蚁Reticulitermes labralis补充生殖蚁对巢群稳定和发展的作用。【方法】对野外巢群进行调查研究, 及对婚飞成虫通过雌雄配对与补充生殖蚁隔离巢群进行人工饲养的对比研究。【结果】在野外巢群只发现1对原始蚁王蚁后, 而补充生殖蚁的数量最多可达到689头/巢。圆唇散白蚁有3种类型的补充生殖蚁, 即由工蚁转化来的无翅型补充生殖蚁、若蚁转化来的翅芽型补充生殖蚁和末龄若蚁羽化来的拟成虫型补充生殖蚁。实验室条件下婚飞配对群体和隔离群体建群1个月后的存活率分别为64%和96%。建巢初期婚飞配对群体的子代数目增长缓慢, 2个月时的群体数量为6.3±1.54, 10个月时的群体数量也仅为8.4±1.47; 而隔离建群补充生殖蚁2个月时的群体数量为52.4±6.44, 10个月时的群体数量为164.3±20.85, 都高于婚飞配对群体。 此外, 野外巢群的补充蚁后跟原始蚁后一样都具有发达的卵巢。【结论】在圆唇散白蚁中补充生殖蚁是白蚁巢群主要的繁殖力量, 也是建立新巢群的重要繁殖品级。     

Morphology of the male reproductive systems of two Indopacific blenniid fishes,Salarias fasciatus and Ecsenius bicolor (Blenniidae,Teleostei)1

The testes and associated accessory organs of two blenniid fishes are described. The testicular organs of Salarias fasciatus consist of a testicular gland adjacent to the testis, chambered seminal vesicles serving as a reservoir for spermatozoa and testicular blind ouches. Ecsenirts bicolor has no testicular gland, has elongated chambered seminal vesicles whii are not used as a reservoir for spermatozoa, and has testicular blind pouches.     

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.     

The origins and relatedness of multiple reproductives in colonies of the termite Nasutitermes corniger

Colonies of the termite Nasutitermes corniger often contain multiple reproductive queens and kings. We used double-strand conformation polymorphism (DSCP) analysis of mitochondrial DNA (mtDNA) to determine the probable origins of co-occurring reproductives. Colonies differed in queen and king number, in the number of nests containing reproductives, and in the genetic relationships among reproductives. Most of the 44 colonies contained a single pair of maternally unrelated reproductives. In the two single-nest colonies with a pair of queens, the two queens differed in mtDNA haplotype, suggesting nest-founding by unrelated queens. In the seven single-nest colonies with larger numbers of reproductives (11–49), all reproductives shared the same haplotype, a pattern consistent with replacement of a single pair by several offspring. As predicted by theory, the number of coexisting queens was greater for replacement reproductives than for co-foundresses. Several complex colonies contained multiple queens of two or more haplotypes distributed among several interconnected nests. This indicates that several matrilines can persist within a colony through one or more generations of budding and replacement, a hypothesis confirmed by orphaning experiments. The various modes of termite colony formation rival the diversity seen in ant species and demonstrate the remarkable convergence of behaviours between the two groups.     

Adaptation of a radiochemical assay for juvenile hormone biosynthesis to study caste differentiation in a primitive termite

A radiochemical assay measuring juvenile hormone synthesis by corpora allata incubated in vitro was adapted for use with the termite Zootermopsis angusticollis. Corpora allata from 3–4-day old virgin female neotenic reproductives were used in these studies because this caste showed the highest rates of juvenile hormone synthesis (0.6 pmol h^?1 per pair corpora allata). Juvenile hormone-III synthesis was linear for up to 6 h over the range of concentrations of labelled l-methionine from 27–280 μM. Rates of juvenile hormone synthesis were stimulated up to 10-fold in a dose-dependent manner by the addition of farnesoic acid to the incubation medium. However, the relatively high concentration of 120 μM farnesoic acid reduced the rates of juvenile hormone synthesis. The radiochemical assay was used to determine rates of juvenile hormone synthesis in vitro by corpora allata from larvae with a queen and king vs orphaned larvae. The presence of reproductives resulted in a suppression of larval corpus allatum activity relative to orphaned controls.     

Extreme genetic mixing within colonies of the wood-dwelling termite Kalotermes flavicollis (Isoptera,Kalotermitidae)

The existence of altruism in social insects is commonly attributed to altruistic individuals gaining indirect fitness through kin selection. However, recent studies suggest that such individuals might also gain direct fitness through reproduction. Experimental studies on primitive wood-dwelling termites revealed that colony fusion often causes the death of primary reproductives (queen and king), allowing opportunities for workers to inherit the nest by developing into replacement reproductives (neotenics). Therefore, colony fusion has been proposed as an important factor that may have favoured sociality in termites. However, whether colony fusion occurs frequently in natural populations of wood-dwelling termites remains an open question. We analysed eleven colonies of the wood-dwelling termite Kalotermes flavicollis (Kalotermitidae), using two mitochondrial and five nuclear microsatellite markers. Nine of eleven colonies (82%) were mixed families, with offspring of three or more primary reproductives. To our knowledge, this result represents the highest frequency of mixed-family colonies ever reported in termites. Moreover, genetic mixing of colonies appeared extreme in two ways. First, the number of haplotypes per colony was exceptionally high (up to nine), indicating that colonies were composed of multiple queens' offspring. Second, some mixed-family colonies included individuals belonging to two highly divergent genetic lineages. F-statistics and relatedness values suggest that mixed-family colonies most likely result from colony fusion, giving support to the accelerated nest inheritance theory. These findings raise important questions about the mode of foundation of mixed-family colonies and the evolutionary forces that maintain them within populations.     

Asexual queen succession mediates an accelerated colony life cycle in the termite Silvestritermes minutus

Mixed modes of reproduction, combining sexual processes with thelytokous parthenogenesis, occur in all major clades of social insects. In several species of termites, queens maximize their genetic input into nondispersing replacement queens through parthenogenesis, while maintaining genetically diverse sterile offspring and dispersing reproductives via sexual reproduction. This so‐called asexual queen succession (AQS) has multiple independent origins and its presumed advantages are diverse as well, ranging from multiplication of colony reproductive potential to extension of its lifespan beyond that of the foundress. However, how AQS shapes colony life cycles under natural conditions remains poorly known. The neotropical termite Silvestritermes minutus inhabits small but conspicuous nests, offering a unique opportunity to investigate the impact of AQS on life history. We report on its breeding system, life cycle and sex allocation using social structure census in 137 nests and genotyping of 12 colonies at 12 microsatellite loci. We show that colonies are established by an outbred pair of primary reproductives. In less than 2 years, the foundress is replaced by multiple neotenic queens, arising mostly through automixis with central fusion. Sterile castes, male and most (93%) female dispersers are produced sexually. Colony reproduction is usually restricted to a single dispersal of alates with unbiased sex ratio, taking place after 3 years. We conclude that S. minutus benefits from AQS to maximize colony growth rate and alate production within a very short life cycle rather than to extend colony lifespan. This highlights the versatile role of AQS in different cases of its polyphyletic origin.     

Aberrant spermatozoa and spermatids,and observations on phagocytosis,in Sparganophilus tamesis (Sparganophilidae,Oligochaeta, Annelida): an ultrastructural study

In Sparganophilus tamesis replication, misalignment and misplacement phenomena occur in aberrant spermatids and spermatozoa, and giant nuclei are recorded in some sperm.

Sperm components replicate independently of each other, but midpiece replication was not observed. Aberrancy was commonest and showed greatest variation in the acrosomal region, and it is suggested that double acrosome tubes may share a common precursor primary vesicle.

Phagocytosis by coelomocytes and by cytophores occurs in the seminal vesicles. The spermathecae were empty and the possibility that the population is parthenogenetic is discussed.