Termite queens have disproportionately more DNA in their fat body cells: reproductive division of labor and endoreduplication

Increases in DNA content caused by endoreduplication are widely observed in the metabolically active tissues of plants and animals. During egg production, insect females synthesize very large amounts of vitellogenin in their fat bodies, and female fat bodies of some insects become polyploid to accelerate vitellogenin production. Social insects have developed reproductive division of labor, wherein queens lay most of the eggs while other individuals have reduced fertility and undertake tasks required for maintaining the colony. Therefore, only queens are engaged in vitellogenin synthesis for egg production in social insects. Here, we show that termite queens have disproportionately more DNA in their fat body cells. Our DNA content analysis using flow cytometry demonstrated that more cells contained 4C‐DNA than 2C‐DNA in the fat bodies of Reticulitermes speratus queens. This high level of endoreduplication was not found in the fat body cells of other castes. This caste‐dependent doubling of DNA content in fat body cells suggests that termites exploit endoreduplication to boost egg production, in conjunction with the development of reproductive division of labor. This study highlights nuclear polyploidization as an adaptive strategy in social insects.     

Ultrastructure and formation of the physogastric termite queen cuticle

The physogastric termite queen is the most striking example in insects of growth in size without cuticular moulting. This phenomenon has been studied with electron microscopy and histochemical tests in two species of higher termites, Cubitermes fungifaber and Macrotermes bellicosus. The abdominal hypertrophy (physogastry) is allowed by growth of the arthrodial membranes of the swarming imago. The growth is slow (over several years) but important: the cuticular dry weight is multiplied by 20 in C. fungifaber, by 100-150 in M. bellicosus. The termite queen cuticle arises from the transformation of the cuticle of the swarming imago or imaginal cuticle (unfolding and growing of the epicuticle, stretching of the endocuticle, resorption of the subcuticle) and from the secretion of a new endocuticle or royal endocuticle. The termite queen is the first example known in insects of epicuticular growth. In the physogastric queen, three cuticular types are observed: the rigid cuticle of the sclerites, the soft cuticle of the arthrodial membranes and the partially rigid cuticle of special structures, the neosclerites, which show both rigidity and growth. The fibrillar architecture varies according to the abdominal zones and the position within the cuticle. It appears to be determined by the forces arising from the musculature and the anisometric abdominal growth. The king does not become physogastric, although its cuticle is also modified.     

On the relationship between queen number and fecundity in polygyne colonies of the fire ant Solenopsis invicta

ABSTRACT. The reproductive status of individual queens in relation to the number of queens in polygyne colonies of Solenopsis invicta Buren was investigated under both field and laboratory conditions. In field colonies, the weight of individual queens is inversely related to the number of resident queens present. The weight of a queen is positively correlated with fecundity, as measured by the number of eggs laid in 5 h in isolation. Thus increasing queen number has a strong negative effect on the fecundity of individual queens in these colonies. A similar relationship was established in standardized laboratory colonies using queens from a single experimental population. Colony size by itself had no significant effect on queen weight in either the field or laboratory colonies, suggesting that the inverse relationship between queen number and fecundity results from mutual inhibition among queens, possibly involving pheromones, rather than reduced nutrition through lower worker/queen ratios. While uninseminated queens can comprise up to 50% of the functional (i.e. egglaying) queens in polygyne colonies of this ant, they tend to be less fecund than their inseminated nestmates; uninseminated queens within a given colony generally weigh 5–25% less.     

Experimental increase in fecundity causes upregulation of fecundity and body maintenance genes in the fat body of ant queens

In most organisms, fecundity and longevity are negatively associated and the molecular regulation of these two life-history traits is highly interconnected. In addition, nutrient intake often has opposing effects on lifespan and reproduction. In contrast to solitary insects, the main reproductive individual of social hymenopterans, the queen, is also the most long-lived. During development, queen larvae are well-nourished, but we are only beginning to understand the impact of nutrition on the queens'' adult life and the molecular regulation and connectivity of fecundity and longevity. Here, we used two experimental manipulations to alter queen fecundity in the ant Temnothorax rugatulus and investigated associated changes in fat body gene expression. Egg removal triggered a fecundity increase, leading to expression changes in genes with functions in fecundity such as oogenesis and body maintenance. Dietary restriction lowered the egg production of queens and altered the expression of genes linked to autophagy, Toll signalling, cellular homeostasis and immunity. Our study reveals that an experimental increase in fecundity causes the co-activation of reproduction and body maintenance mechanisms, shedding light on the molecular regulation of the link between longevity and fecundity in social insects.     

对蜜蜂性别决定和蜂王级型确立理论的新诠释

从全新视角概貌性解读蜜蜂Apis诸多生殖机制,依据的就是蜂王或产卵工蜂的卵子发生、性别决定假说、性位点研究以及蜂王级型确立机制。这种不同于单倍-二倍性性别决定机制的新诠释,适用于明确性别决定机制、简化定向育种方法、减少比较性实验的难度和时间、实施亲子鉴定和抽象并量化种群的概念。     

Development of physogastry in the queen of the fungus-growing termite Macrotermes michaelseni (Isoptera: Macrotermitinae)

Female reproductives of the fungus-growing termite Macrotermes michaelseni never showed signs of physogastry if they were kept in petri dishes to establish an incipient colony. During the 200 days of observation, their corpora allata volume, juvenile hormone titre and number of active ovarioles remained on a more or less constant level, and their gut was filled with soil. In contrast, these parameters are enhanced in partially and especially in fully physogastric queens, and these females had only a transparent liquid in the gut.Application of the juvenile hormone mimic ZR 515 to young females accelerated yolk incorporation, increased the number of active ovarioles and brought about the release of the soil from the gut. The same changes were induced by adding workers to increase the population of incipient colonies from about 45 to 400. This also resulted in an increase of the corpus allatum volume and of the juvenile hormone titre in young females. Their intestine was in this case filled with transparent liquid associated with the increased population of workers. It was, therefore, concluded that the development of physogastry depends on a positive feedback: the more workers are with the young female, the more nutrients are available and the more juvenile hormone the female produces, the more eggs are laid and the more workers can again develop to nurture the queen.     

Sex ratio conflicts, mating frequency, and queen fitness in the ant Formica truncorum

We examined the effect of facultative sex allocation by workerson queen fitness in a Furnish population of the ant Formicatruncorum. Workers rear female-biased broods in colonies headedby a singly mated queen and male-biased broods in colonies headedby a multiply mated queen. As a result, multiply mated queenshave a 37% fitness advantage over singly mated queens. Neitherreproductive output nor worker population of colonies variedwith queen mating frequency. We suggest that singly mated queenspersist in the population because fitness benefits to multiplymated queens via sex allocation are balanced by costs of additionalmatings. Alternatively, singly mated queens may persist simplybecause some queens lack opportunities to mate multiply or becausemale control sometimes prevents additional matings by queens.     

Gyne development in the Argentine ant Iridomyrmex humilis: role of overwintering and queen control

Abstract A possible stimulatory effect of overwintering on gyne development in Iridomyrmex humilis (Mayr) was investigated. Although gyne-potent larvae are present in the nest throughout the year, small queenless colony fragments composed of freshly overwintered ants (collected in late winter or early spring) produced 3–8 times more gynes than comparable fragments made up of non-freshly overwintered ants (collected at other times of the year). Apparently, this stimulatory effect of overwintering acts on both the developmental potential of larvae and the tendency of workers to rear sexually competent larvae as gynes; queenless colony fragments in which either the brood or workers were freshly overwintered produced more gynes than fragments composed of non-freshly overwintered workers or brood but fewer than fragments containing both brood and workers freshly overwintered. This increased sexualization potential of larvae due to overwintering is enough to overcome weak pheromonal inhibition of queens associated with low queen number; a single queen in a large freshly overwintered colony fragment is insufficient to inhibit gyne development, whereas ten queens are totally inhibitory. In non-freshly overwintered colony fragments one queen is completely inhibitory. Variability in egg developmental potential according to queen age does not appear to play a major role in the seasonal production of gynes, because at least some eggs of very young queens (less than 3 weeks old) are capable of gyne development. In the field this stimulatory effect of overwintering is superimposed on a seasonal fluctuation in the combined strength of pheromonal queen control. In southern France, gynes are produced only in spring where they arise primarily from overwintered larvae just after a sharp drop in queen number, and presumably the total level of inhibitory queen pheromone, due to the massive execution of queens by workers.     

Facultative use of thelytokous parthenogenesis for queen production in the polyandrous ant Cataglyphis cursor

The evolutionary paradox of sex remains one of the major debates in evolutionary biology. The study of species capable of both sexual and asexual reproduction can elucidate factors important in the evolution of sex. One such species is the ant Cataglyphis cursor, where the queen maximizes the transmission of her genes by producing new queens (gynes) asexually while simultaneously maintaining a genetically diverse workforce via the sexual production of workers. We show that the queen can also produce gynes sexually and may do so to offset the costs of asexual reproduction. We genotyped 235 gynes from 18 colonies and found that half were sexually produced. A few colonies contained both sexually and asexually produced gynes. Although workers in this species can also use thelytoky, we found no evidence of worker production of gynes based on genotypes of 471 workers from the six colonies producing sexual gynes. Gynes are thus mainly, and potentially exclusively, produced by the queen. Simulations of gynes inbreeding level following one to ten generations of automictic thelytoky suggest that the queen switches between or combines thelytoky and sex, which may reduce the costs of inbreeding. This is supported by the relatively small size of inbred gynes in one colony, although we found no relationship between the level of inbreeding and immune parameters. Such facultative use of sex and thelytoky by individual queens contrasts with other known forms of parthenogenesis in ants, which are typically characterized by distinct lineages specializing in one strategy or the other.     

Evolution of eusociality and the soldier caste in termites: a validation of the intrinsic benefit hypothesis

In termites the evolution of reproductive altruism is not based on a particularly high relatedness between nestmates. For the evolution and maintenance of the ancestral sterile soldier caste, the benefits generated by the soldiers' presence must compensate the loss of the soldiers' reproductive potential. To study the impact of soldiers on colony's fitness, we manipulated the proportion of soldiers to nonsoldiers in colonies of the dry-wood termite Cryptotermes secundus.'Soldier-less' colonies were obtained by removing soldiers and inhibiting their development with an extract of soldier heads. The colonies were set up for 1 year in experimental nests in the field. 'Soldier-less' colonies produced fewer soldiers. The reduction of soldiers neither affected colony survival nor helper growth, but fewer dispersing sexuals were produced in 'soldier-less' than in control colonies. This confirms what was only supposed so far, that in termites soldiers are maintained for their intrinsic benefit to cost ratio.     

白蚁的分子生物学研究进展

白蚁是危险性社会昆虫,建立安全有效的白蚁防治方法有赖多学科的参与,分子生物学已经成为白蚁研究的重要工具。目前,DNA序列分析方法已应用于白蚁鉴定和分类,其中线粒体基因是最通用的分子标记;基因工程技术成功构建了可用于白蚁防治的白蚁肠道工程菌;Hexamerin、COX Ⅲ、纤维素酶等白蚁功能基因以及白蚁品级分化相关的若干蛋白相继得到了分离和鉴定。文章从白蚁分类、防治、功能基因、品级分化4个方面综述白蚁分子生物学的研究进展,为白蚁的防治提供新的方法和思路。     

Brood care and social evolution in termites

Cooperative brood care is assumed to be the common driving factor leading to sociality. While this seems to be true for social Hymenoptera and many cooperatively breeding vertebrates, the importance of brood care for the evolution of eusociality in termites is unclear. A first step in elucidating this problem is an assessment of the ancestral condition in termites. We investigated this by determining the overall level of brood care behaviour across four termite species that cover the phylogenetic diversity of the lower termites. Brood care was low in the three species (all from different families) that had an ancestral wood-dwelling lifestyle of living in a single piece of wood that serves as food and shelter. In the fourth species, a lower termite that evolved outside foraging, brood care was more common. Together with data for higher termites, this suggests that brood care in termites only becomes important when switching from a wood-dwelling to a foraging lifestyle. These results imply that early social evolution in termites was driven by benefits of increased defence, while eusociality in Hymenoptera and cooperative breeding in birds and mammals are primarily based on brood care.     

Can endopolyploidy explain body size variation within and between castes in ants?

Endoreduplication is the process by which the nuclear genome is repeatedly replicated without mitotic cell division, resulting in nuclei that contain numerous additional genome copies. Endoreduplication occurs widely throughout Eucarya and is particularly common in angiosperms and insects. Although endoreduplication is an important process in the terminal differentiation of some specialized cell types, and often increases cell size and metabolism, the direct effects of increasing nuclear ploidy on cell function are not well resolved. Here, we examine if endoreduplication may play a role in body size and/or caste differentiation in ants. Nuclear ploidy was measured by flow cytometry of whole individuals (providing the basis for overall body size patterns) and individual body segments for multiple polymorphic ant species. We used cell cycle values, interpreted as the mean number of endocycles performed by each cell in the sample, as our measure of overall endoreduplication. Among females of four polymorphic ant species, endoreduplication was positively related with size within the worker caste, but was not related to caste generally in two species where we also examined queens. Additionally, abdomens had the greatest endoreduplication of all body parts regardless of caste or size. We also found that males, having derived from haploid unfertilized eggs, had the highest rates of endoreduplication and may compensate for their haploid origin by performing an additional endocycle relative to females. These results suggest that endoreduplication may play a role in body size variation in eusocial insects and the development of some segment‐specific tissues.     

Development of non-reproductive castes in the neotropical termite generaCornitermes,Embiratermes andRhynchotermes (Isoptera,Nasutitermitinae)

Summary The developmental pattern of the neuter castes was studied in the mandibulate nasute generaCornitermes, Embiratermes andRhynchotermes. InCornitermes walkeri, all the workers and soldiers are male. There are two larval and a single worker instar. Workers can molt into presoldiers. InEmbiratermes chagresi andRhynchotermes perarmatus, both sexes are present among the neuters. A slight sexual dimorphism (males > females) is discernible among both larval instars and among workers ofE. chagresi; female workers can molt into presoldiers. InR. perarmatus, the sexual dimorphism is conspicuous from the first larval instar on. Male larvae go through two instars, then give rise to workers, which do not molt. InR. perarmatus, there is no worker stage in females, but a third larval instar, preceding the presoldier. Hypotheses are proposed as to the evolution of these caste patterns, attempting to conciliate present knowledge of Nasutitermitinae phylogeny and known evolutionary trends affecting termite caste patterns, according to the species' ecology.Research Associate: National Fund for Scientific Research (Belgium).     

柑橘愈伤组织倍性变化及其与体细胞胚胎发生能力的关系

柑橘愈伤组织是细胞融合、遗传转化等生物技术研究的良好材料。本实验室通过近20年的工作,先后诱导和保存了近80份不同种类和品种的柑橘愈伤组织。这些愈伤组织在MT基本培养基上继代保存。在长期保存过程中,部分材料仍然保持较好的胚胎发生能力。为研究愈伤组织在继代过程中的染色体变异趋势以及这种变异对胚胎发生能力的影响,本文选择有代表性的35份材料作为试材,在4年中连续3次使用流式细胞仪,对细胞DNA含量进行测定。结果发现,待测的35种愈伤组织均存在部分细胞DNA含量加倍的现象,而且,随着时间的推移,除佩奇橘柚、沙漠蒂甜橙、鲁斯脐橙、印度酸橘等4种愈伤组织的DNA含量加倍的细胞有减少的趋势外,其他愈伤组织的DNA含量加倍的细胞均有增加的趋势。数据分析表明,在待测的35种愈伤组织中,有71．4％的基因型的愈伤组织细胞出现了多倍体细胞增加的趋势。对这35种柑橘愈伤组织进行体细胞胚诱导,发现只有9种愈伤组织具有不同程度的胚胎发生能力,其他26种愈伤组织不能再生。相关性分析表明,柑橘愈伤组织细胞DNA含量增加的细胞比例与体细胞胚胎发生能力之间的相关性较小,相关系数为忙-0．10（P〈0．01）,未达到显著水平。另外,柑橘胚性愈伤组织继代时间与体细胞胚胎发生能力之间的相关性也不显著。     

白蚁表皮碳氢化合物研究进展

近年来, 固相微萃取等现代技术的使用显著促进了白蚁表皮碳氢化合物研究的开展。至今, 已有约29种白蚁的表皮碳氢化合物组分得到鉴定, 分属于木白蚁科、 鼻白蚁科、 原白蚁科和白蚁科, 其组分主要为正烷烃、 含有不同数量甲基的支链烷烃及少量烯烃。白蚁表皮碳氢化合物不仅具有一定的科、 属特异性, 大多数种类还具备特有组分, 表明其可作为种间识别的指标。表皮碳氢化合物组分在种内个体识别方面的作用, 在低等白蚁中多获得了支持性结果, 但也有研究认为在这些种类中表皮碳氢化合物不是种内个体识别（同巢个体识别）的唯一指标。发现其与品级分化的相关是近年来白蚁表皮碳氢化合物研究的重要进展。有些种类表皮碳氢化合物的年消长与生殖蚁的分化有关; 而另一些种类生殖蚁含有表皮碳氢化合物特有组分, 其含量与生殖蚁的生殖状态有关, 提示其可能在品级分化中发挥重要作用。作为研究白蚁品级分化和维持机理的新方向, 表皮碳氢化合物值得进一步研究探索。     

Relationship Between Ploidy Variation of Citrus Calli and Competence for Somatic Embryogenesis

This study focuses on the relationship between the genetic variation of calli and the competence for somatic embryogenesis in citrus. The DNA content of 35 citrus calli of different genotypes was measured three times by flow cytometry during a period of four years. The results showed that 71.4% of the genotypes had a progressive increase of varied cells, while those of Page tangelo, Shamouti sweet orange, Russ navel orange and Cleopatra decreased; significant difference in the variation degree (percentages) existed among genotypes. Studies carried out on the induction of somatic embryogenesis revealed that 9 out of the 35 genotypes had still kept the competence of somatic embryogenesis, and the rest 26 had lost the competence. Correlation analysis indicated that there was no significant relationship between the variation degree and the embryogenesis competence r=−0.10 (P<0.01), neither for the relationship between the subculture duration and the regeneration capacity.