Local adaptation of sex induction in a facultative sexual crustacean: insights from QTL mapping and natural populations of Daphnia magna

Dormancy is a common adaptation in invertebrates to survive harsh conditions. Triggered by environmental cues, populations produce resting eggs that allow them to survive temporally unsuitable conditions. Daphnia magna is a crustacean that reproduces by cyclical parthenogenesis, alternating between the production of asexual offspring and the sexual reproduction of diapausing eggs (ephippia). Prior to ephippia production, males (necessary to ensure ephippia fertilization) are produced parthenogenetically. Both the production of ephippia and the parthenogenetic production of males are induced by environmental factors. Here, we test the hypothesis that the induction of D. magna resting egg production shows a signature of local adaptation. We postulated that Daphnia from permanent ponds would produce fewer ephippia and males than Daphnia from intermittent ponds and that the frequency and season of habitat deterioration would correlate with the timing and amount of male and ephippia production. To test this, we quantified the production of males and ephippia in clonal D. magna populations in several different controlled environments. We found that the production of both ephippia and males varies strongly among populations in a way that suggests local adaptation. By performing quantitative trait locus mapping with parent clones from contrasting pond environments, we identified nonoverlapping genomic regions associated with male and ephippia production. As the traits are influenced by two different genomic regions, and both are necessary for successful resting egg production, we suggest that the genes for their induction co‐evolve.     

Two-egged ephippia in the chydorid Cladocera

SUMMARY. With a single reported exception, the ephippia of all specialized chydorid cladocerans hitherto described contain a single egg. Ephippia the European Chydorus ovalis and the North American species currently listed as C. cf. sphaericus have been found persistently, and so far invariably, to contain two eggs. The alternative strategies of producing 1-and 2-egged ephippia are briefly considered. Anomalous 1-egged ephippia of Daphnia (family Daphniidae) are also mentioned.     

Females from resting eggs and parthenogenetic eggs in the rotifer Brachionus calyciflorus: lipid droplets, starvation resistance and reproduction

1. In the heterogonic life cycle of monogonont rotifers, amictic (female‐producing) females develop from two types of eggs: fertilised resting (diapausing) eggs and parthenogenetic subitaneous eggs. Females hatched from resting eggs initiate clonal populations by female parthenogenesis and are called stem females. This study compares females from resting and parthenogenetic eggs that were produced under identical culture conditions and were of similar birth order. 2. Newborn stem females had many more lipid droplets in their tissues than similar‐sized, newborn females from parthenogenetic eggs. When neonates were stained with Nile Red and viewed under epifluorescent illumination, these droplets were shown to be sites of neutral‐lipid storage products. 3. Stem females had no posterolateral spines and short anterior spines, while their mothers and offspring in subsequent, parthenogenetic generations typically had long posterolateral spines and elongated anterior spines. 4. Newborn stem females survived starvation significantly longer than newborn females from parthenogenetic eggs. 5. When females from resting and parthenogenetic eggs were cultured from birth to death at a high food concentration, the reproductive potential (r day^?1) of the stem females was significantly higher (0.82–0.88 versus 0.70), primarily because of egg production at an earlier age. The mean lifetime fecundity (R_o) of stem females was significantly greater than that of females from parthenogenetic eggs. 6. Extensive lipid reserves should increase the ability of stem females to colonise new habitats. Firstly, compared with females from parthenogenetic eggs, stem females are more likely to experience starvation or food limitation. Resting eggs hatch in response to physical and chemical factors that are not directly related to food availability, and from sediments that may be far from food‐rich surface waters. Secondly, when food is abundant, stem females have a greater reproductive potential.     

Calculating egg volume when shape differs: when are equations appropriate?

ABSTRACT Egg volume is often calculated from length and breadth, assuming little variation in egg shape within species. However, egg shape may vary among females, over time, or within clutches. If this variation is not predictable, volume should be estimated using another method. For example, in some species, egg shape changes consistently with laying order and volume can be estimated using laying‐order specific equations. We measured the length, breadth, volume, and mass of 249 Magellanic Penguin (Spheniscus magellanicus) eggs and regressed volume on length and breadth for first and second eggs separately. Differences between measured and calculated volume averaged <1% for both eggs. Using linear measurements, we calculated egg volume for 7085 clutches from 1983 to 2006. First and second eggs had similar calculated volumes (P > 0.05) in 19 of 24 yr; when they differed, the second egg was larger. In contrast, using single equations typically employed to compute egg volume, we erroneously concluded that first eggs were usually (20 of 24 yr) significantly larger than second eggs. Our regression equations should be applicable to other Spheniscus penguins because ratios of elongation coefficients (length divided by breadth) of first and second eggs are similar among these species. In other species of birds where egg shape changes with laying order, the volume of a sample of eggs should be measured to develop regression equations specific to the species and laying order. Slight variation in egg volume can have important evolutionary and ecological implications that would be erroneously interpreted without field tests to measure volume.     

Effect of population bottlenecks on the egg morphology of introduced birds in New Zealand

Hatching failure is inversely correlated with population bottleneck size among exotic birds introduced to New Zealand, but the mechanism for this is unclear. We assess whether the bottlenecks these species experienced during their introduction have affected hatching failure through deleterious changes in egg morphology. We measured egg size and shape of 13 species that passed through bottlenecks of 11–800 individuals during their establishment in the 19th century. Eggs were also measured in the source populations ('pre-bottleneck') of each species to compare pre- and post-bottleneck egg morphology directly. Significant changes in egg volume were found in six of 13 species, with most laying smaller eggs in New Zealand. Egg shape changed in four of 13 species but there was no directional bias; two species developed more elongated eggs and two species broader eggs. There was no relationship between bottleneck size and change in egg volume, but species passing through severe bottlenecks had greater variability in egg volume and were more likely to have eggs that deviated in shape from their source populations. There was no relationship between changes in either egg volume or shape and rates of hatching failure. Further work is needed to assess whether changes in egg morphology have negative consequences on offspring fitness and whether the observed changes are the result of differing environmental conditions in the introduced range.     

Inter-genotype variation in reproductive response to crowding among Daphnia pulex

Crowding is known to have a major influence on reproduction in the freshwater microcrustacean Daphnia pulex. We analyzed reproductive output of six different D. pulex genotypes under two different density regimes in the laboratory. Four of these genotypes reproduce via obligate parthenogenesis, allowing thorough analysis of the life history strategies of some asexual lines. Among 30,109 neonate offspring and 1041 resting egg ephippia collected, several trends were evident. Crowding induced increased resting egg production and reduced neonate offspring production among all genotypes. Offspring sex ratios grew more male-biased with maternal age. The extent, but not direction, of each of these trends varied among genotypes. Offspring sex ratios, and the very direction in which they changed in response to crowding, differed significantly among genotypes with some genotypes producing more and others fewer males in response to crowding. Obligately parthenogenetic genotypes seemed to respond to the crowding stimulus in similar ways as the facultatively parthenogenetic genotypes, as expected from the sexual origins of their genomes. The inter-genotype variation in life-history traits observed in this and other investigations calls into question the common practice of extrapolating results from a single Daphnia genotype to an entire species. Our findings are considered in the context of other research in the field of environmental influences on Daphnia reproduction with a review of representative literature.     

Sex, parthenogenesis and genetic structure of rotifers: microsatellite analysis of contemporary and resting egg bank populations

Cyclically parthenogenetic rotifers are a valuable model for investigating the relationship between reproductive mode and population structure, although advances in this field have been hindered by low allozyme variability in these organisms. A high genotypic diversity is predicted after population establishment, which would be eroded by clonal selection during the parthenogenetic phase. The resting egg bank, produced sexually, is presumed to store high levels of genetic diversity, with subsequent effects on planktonic population structure. Here, we provide the first application of microsatellite markers to a rotifer planktonic population and its associated resting egg bank. Seven polymorphic microsatellite loci were screened in populations of the rotifer Brachionus plicatilis in a temporary pond to analyse: (i) the genetic structure of the resting egg bank; (ii) the changes in the genetic structure of rotifer populations during the parthenogenetic phase; and (iii) the population structure after its initiation from resting eggs. Microsatellites proved to be a useful tool for clone identification, revealing a surprisingly high clonal diversity in rotifer populations. The last sample in the parthenogenetic phase showed evidence of clonal selection, as indicated by a low observed clonal diversity and the appearance of linkage disequilibria. The resting egg bank, analysed comprehensively for the first time in any zooplankter, is in Hardy-Weinberg and linkage equilibrium, and contains a high genotypic diversity. Unexpectedly, the resting egg bank differed from the planktonic population in its allelic composition, suggesting that resting egg hatching is biased.     

Are they still viable? Physical conditions and abundance of <Emphasis Type="Italic">Daphnia pulicaria</Emphasis> resting eggs in sediment cores from lakes in the Tatra Mountains

All species of Daphnia (Cladocera) produce, at some stage in their life cycle, diapausing eggs, which can remain viable for decades or centuries forming a “seed bank” in lake sediments. Because of their often good preservation in lake sediment, they are useful in paleolimnology and microevolutionary studies. The focus of this study was the analysis of cladoceran resting eggs stored in the sediment in order to examine the ephippial eggs bank of Daphnia pulicaria Forbes in six mountain lakes in the High Tatra Mountains, the Western Carpathians (northern Slovakia and southern Poland). Firstly, we analyzed distribution, abundance and physical condition of resting eggs in the sediment for their later used in historical reconstruction of Daphnia populations by genetic methods. To assess changes in the genetic composition of the population through time, we used two microsatellite markers. Although DNA from resting eggs preserved in the High Tatra Mountain lake sediments was extracted by various protocols modified for small amounts of ancient DNA, DNA from eggs was not of sufficient quality for microsatellite analyses. Distribution curves of resting eggs from sediment cores correspond to the environmental changes that have occurred in the High Tatra Mountains area during last two centuries (atmospheric acid deposition, fish introduction) and demonstrate their influence on natural populations. Evaluation of ephippia physical condition (the most common category was empty ephippial covers) suggests that the majority of resting eggs hatched to produce a new generation of Daphnia or may be due to failed deposition of resting eggs by Daphnia to the chitinous case. In conclusion, age, low quantity and poor physical condition of resting eggs from these Tatra lake sediments proved to be unsuitable not just for use in genetic analyses, but also the possibilities of autogenous restoration of Daphnia populations from the resting egg banks in the Tatra sediments are negligible.     

Summer diapause in Daphnia as a reaction to the presence of fish

A chemical signal, released by a fish predator under summer-likehigh water temperature and long-day photoperiod, caused theformation of resting eggs in a clone of Daphnia magna. No ephippialfemales were recorded and no ephippia were released in the controltreatment during 45 days of the experiment. When exposed tofish water, the fraction of ephippial females reached a maximumof 3.7%, a value comparable to that registered in summer inthe GroBer Binnensee (Northern Germany), a hypertrophic lakeinhabited by fish, which was the source lake for our experimentalclone. The number of ephippia released within 45 days was onaverage 34 ± 22. Ephippia formation could not resultfrom the between-treatment differences in population density,and related patterns of food depletion, since no substantialdifference between control and fish treatment was observed.Instead, specific information on the presence of a predatorprovided a cue which induced the formation of resting eggs inDaphnia. Under heavy predation and very low survival probabilityof parthenogenetic females, ephippia formation in summer canbe adaptive, i.e. higher fitness can be achieved through survivalin the diapausing state than through the immediate reproductivegain via parthenogenesis.     

从白符虫兆(弹尾纲)活性卵的形状变化探讨沃尔巴克氏体的共生作用(英文)

Wolbachia属共生菌的侵染是引起跳虫——白符虫兆孤雌生殖的原因。对带有正常沃尔巴克氏体菌群的白符虫兆卵和通过利福平处理剔除沃尔巴克氏体菌群的白符虫兆卵的胚胎发育进行实验观察。白符虫兆的活性卵产出3到4天后,卵体大小显著性地增大,并伴随卵体形状从球形到圆饼形的变化。这些变化在利福平处理的或者是7%自然失活的非活性卵中都没有出现。推测沃尔巴克氏体在白符虫兆卵产出后的3天之内或者3天之前的受精卵发育或胚胎发育中发挥着重要作用;同时根据目前已有的研究结果推断沃尔巴克氏体对白符虫兆卵发育可能的影响机制。     

Polymorphism in stem females and successive parthenogenetic generations in Brachionus calyciflorus Pallas

We have described the polymorphism in the hatchlings of resting eggs and the morphological variations between the stem females hatched from resting eggs and their successive parthenogenetic generations in the rotifer Brachionus calyciflorus. We hatched resting eggs of B. calyciflorus in two different culture mediums: unconditioned medium (IOM) and Asplanchna-conditioned (ACM). The hatching rate of resting eggs in IOM and ACM were 32.5% and 28.5%, respectively, and showed no significant difference. Stem females hatching from these resting eggs had three morphotypes (unspined, single short-spined, and two short-spined) and over 80% of these females were spineless ones. Moreover, the frequency of each morphotype stem females showed similar tendency in IOM and ACM. The production of a variety of morphotypes among stem mothers—rather than all unspined as has been previously reported for this species—may be regarded as a form of bet-hedging in this population of B. calyciflorus. Phenotypic changes in morphology between the stem females hatched from resting eggs and their successive parthenogenetic generations in B. calyciflorus were found. The possible mechanisms, responsible for high proportion of spineless phenotype at early generations from resting eggs and increased spined phenotype in successive parthenogenetic generations, were discussed in the article.     

'Ancient' DNA in the resting egg bank of a microcrustacean can serve as a palaeolimnological database

Recent work on the diapausing egg banks of zooplankton, such as Daphnia (Crustacea: Anomopoda), indicates that these eggs can remain viable for decades while, theoretically, DNA can remain intact for even longer periods (i.e. centuries or millennia). We isolated diapausing eggs of Daphnia from a 30 m long sediment core taken from a hypereutrophic, northern German lake (Belauer See), with some eggs found in dated core material as old as 4500 years. Using microsatellite markers, we analysed the genetic structure of the resting eggs dated as old as ca. 200 years, and found that, although levels of heterozygosity remained remarkably stable, significant genetic differentiation (Nei's D = 0.36; F(ST) = 0.15) between recent and 'ancient' resting eggs (including allele frequency shifts and private alleles) was detected. These shifts represent either species-level changes in this complex (i.e. species-specific characters of ephippia are not always robust), or intraspecific shifts in genetic variation, or a combination of both. This study demonstrates that the egg banks of aquatic zooplankton can serve as repositories of both genetic (intrapopulational) and ecological (interspecific) information. The use of molecular markers, such as microsatellites, on diapausing egg/seed banks may open new avenues of enquiry related to tracking the long-term genetic (and/or species) shifts that are associated with long-term environmental changes.     

Ecological and evolutionary significance of resting eggs in marine copepods: past, present, and future studies

The occurrence of a resting egg phase in the life cycle of marine and freshwater planktonic copepods is well documented and receiving increasing attention by investigators. The species generally occur in coastal marine waters, freshwater ponds and lakes in areas that undergo strong seasonal fluctuations, though examples have been reported for tropical and sub-tropical areas not subject to such extreme fluctuations. Typically such species disappear from the water column for portions of the year, but remain in the region as benthic resting eggs. Studies to date have focused on the conditions that promote the occurrence of resting eggs, the factors that affect their survival and hatching from sediments, the existence of egg banks in sediments, and the impact of resting eggs on plankton community structure. Benthic resting eggs of copepods include diapause eggs as well as subitaneous (non-diapause) eggs that are quiescent due to conditions in the sediments. As with other groups of organisms the resting egg phase is viewed as being critical for the perpetuation of species year after year, especially those that disappear from the water column for portions of the year. Some data indicate that eggs can survive for many years in sediments which would expand their influence to evolutionary time scales. This paper summarizes our understanding of embryonic dormancy in marine copepods.     

Covariation between clutch size, egg weight and egg shape: comparative evidence for chelonians

The variation in reproductive variables is documented both within and across species of chelonians. At both the generic and family levels, egg weight and clutch size show positive, significant correlations with adult carapace length. There is a negative correlation across both genera and families between clutch size and egg weight after removing the effects of body size, suggesting an evolutionary trade-off between these two life-history characteristics. However, the trade-off is not complete, since clutch size is positively correlated with clutch weight after removing the effects of body size. Terrestrial species lay fewer and larger eggs for their size than freshwater or marine species, but this association is statistically confounded by the fact that chelonian families form ecological groups. There is no significant association between habitat and clutch weight after removing the effects of body size, nor between latitude and either egg weight or clutch size, but temperate species have a heavier clutch weight after removing the effects of body size. Larger species lay eggs that are more spherical, but after controlling for body size, egg shape is not associated with clutch size. The patterns of covariation between adult weight, egg weight and clutch size contrast with those reported for birds and mammals, and some reasons for these differences are discussed.     

The role of parthenogenesis in the biology of two species of mayfly (Ephemeroptera)

1. From a single founder virgin female of each species, nine and seven successive parthenogenetic generations of Cloeon dipterum and Cloeon simile, respectively, were reared: all offspring were female. Eggs were removed from final instar nymphs, subimagos and virgin imagos of each generation and their development assessed. The life cycles of 487 individual C. dipterum and 315 C. simile were recorded, together with details of growth rate, egg production and adult size of these and other representatives of each generation. In order to distinguish between the effects of parthenogenesis and the effects of culture conditions, fertilized eggs of both species were reared under the same laboratory conditions. 2. Fewer unfertilized than fertilized eggs hatched. There was no consistent trend in the percentage hatch in successive parthenogenetic generations. In both species the time taken for parthenogenetic individuals to complete a generation varied considerably, and no relationship was found between generation length and the number of parthenogenetic generations that had passed, the developmental stage of the donor, or the time taken for the eggs to hatch. After 50 weeks there was, in both species, an overlap of up to six generations. The number of eggs produced by both species was significantly greater in the first parthenogenetic generation than in later generations but the numbers for later generations lie within the recorded range of egg number in imagos captured in the field. 3. To find if parthenogenetic individuals occur in the field, the sex ratios of last instar nymphs and subimagos of C. simile were measured in samples collected over 13 successive years from a set of small isolated pools. There was a female bias both in winter and at the start of the emergence season. It is possible that, when mature nymphs and emerging subimagos are damaged by predators, the eggs which are released from them survive and develop. 4. Mature, egg-containing nymphs of both species were present in the field throughout the year, although their numbers were small at the beginning of winter. 5. Oviposition by imagos captured from mating swarms and by virgin imagos, of both species, was recorded for the first time, showing that C. dipterum is not always ovoviviparous. It is suggested that early instar nymphs occurring in mid-winter come not only from unfertilized eggs released from injured nymphs and subimagos, but also from fertilized eggs laid in autumn.     

Egg characteristics are unreliable in determining maternity in communal clutches of guira cuckoos Guira guira

Egg characteristics have been used to determine egg maternity for birds in situations where two or more females lay eggs in a single nest (as in communal breeders and intraspecific parasites). We assessed the applicability of egg morphometry and eggshell appearance in ascribing egg ownership in communal clutches of guira cuckoos Guira guira , a species where up to seven females may lay eggs in a joint nest. We used both combined variables (including egg mass, length, width, shape and two eggshell variables), and a shape index to test whether eggs laid by each female were similar but different from eggs laid by other females. Also, we conducted discriminant function analyses to verify if eggs could be correctly classified to their mothers based on their characteristics. The correct maternity was determined by yolk protein electrophoresis of freshly-laid eggs. Individual female chutches were separated through egg characteristics or shape alone in 29% and 41% of the groups tested, respectively. Differences were mostly due to a single female that differed from her nest-mates in a unique egg variable. On average, 55% of the eggs analyzed were not assigned to the correct mother using egg dimensions and eggshell speckling pattern. In conclusion, the egg characteristics used do not reliably indicate maternity in guira cuckoo communal clutches. We therefore recommend a protein-based verification of egg appearance characteristics for assigning maternity in other species in which multiple female laying occurs.     

A simple tool for calculating egg shape,volume and surface area from digital images

The geometrical properties of eggs – such as volume and surface area – have uses ranging from ecological, physiological and morphological studies in birds, to predictions of chick condition in the poultry industry. Although measurements of an egg's length and breadth can be used to approximate its geometry, the coefficients used in these models are specific to the original test population, and intraspecific variation in egg shape means these methods cannot be used reliably beyond that population. Here I present a novel mathematical formula to describe the curvature of a bird's egg that can be used to calculate the shape, volume and surface area of an egg precisely from digital images. Using data from several species I demonstrate that the model has a greater level of accuracy than length‐ and breadth‐based methods, and release the user‐friendly tool for others to use for measuring eggs from digital images.     

水蚤滞育的数量遗传学研究:遗传与环境控制及遗传作用

在单性生殖循环水蚤群体中,滞育卵由有性生殖产生．在一系列实验中用到了不同种群和种类的水蚤,通过这些实验来观察：1)有性生殖和滞育卵复苏的遗传和环境控制;2)生活在相同区域中,但有不同生存微环境的近缘种类的有性生殖的光周期反应;3)在群体遗传结构上有性生殖的遗传效应(基因型均值和生活史性状遗传方差)．结果发现：1)遗传作用和环境作用,以及两者的相互作用都对有性生殖和滞育卵的孵化有显著的影响．GE显著的相互作用对环境中观察到的有性生殖来说,有助于维持其较高的遗传方差;2)在相同区域中,不同生存微环境的近缘种类的有性生殖的光周期反应有所不同．这有助于进一步区别近缘的水蚤种类,这也可能是一个水环境中同素异形的物种形成的例子;3)在有性生殖上,生活史性状平均值和遗传方差变化与前代选择造成的均值和遗传方差相反(遗传滑阻),这会造成暂时的适应不良(遗传滑阻和隐藏的遗传变异的表达),应对它补偿滞育的进化优势．     

Resting eggs in rotifers

The biology of resting eggs of monogonont rotifers is reviewed, covering literature published since the last major review by Gilbert (1974). The topics examined include resting egg production, morphology and species specificity, hatching, and evolutionary significance. Four major determinants of resting egg production are identified: mictic female production, male activity and fertility, female susceptibility to fertilization, and fertilized female fecundity. Recent work in these four areas is discussed as well as resting egg production in natural populations. Resting egg morphology, particularly shell structure and internal organization, is compared among species. Recent reports on the control of resting egg hatching in the laboratory are examined and the importance of temperature, light, diet, and salinity is reviewed. Two hatching patterns are contrasted, the first where eggs hatch at regular intervals over extended periods and the second where hatching is synchronized to some environmental cue. A latent period after resting egg formation, during which no hatching occurs, is defined for several species. The adaptive features of resting eggs are outlined including their contribution to genetic variability through recombination, their provision for environmental escape by dormancy, and their colonizing function resulting from their ease of dispersal. The type of cue utilized to initiate mictic female production as well as the pattern of resting egg hatching is related to environmental predictability.     

Status of Daphnia Resting Egg Banks in Bohemian Forest Lakes Affected by Acidification

We studied the egg banks of Daphnia gr. longispina in four acidified lakes in the Bohemian Forest (Czech Republic, Europe). Daphnia had become extinct in three of the studied lakes due to anthropogenic acidification and we assessed the possibility of autochthonous recovery of the population after recent amelioration of the lake environment. We determined the ephippia distribution in eleven cores 10–30 cm long, and the state of Daphnia resting eggs in over 13 000 ephippia from eight cores. Apparently well-preserved eggs were used for hatching experiments and for DNA amplification. Vertical profiles of ephippia densities in several cores did not agree with historical data on the Daphnia presence in the lakes, as the sediment had been repeatedly disturbed by human activities in the past. Ephippia are present near the surface of the sediment in all lakes, and they might therefore receive hatching cues. We were not able, however, to prove that viable eggs are present in the ephippia banks of the lakes. The percentage of the well-preserved eggs in all but one core was below 1%. In addition, no egg hatched in the hatching experiments and we were not able to amplify DNA from the preserved eggs of lakes where Daphnia is extinct, although amplification from relatively young eggs from the fourth lake was successful in 60% of the cases. We conclude that the recovery of Daphnia populations in studied lakes from autochthonous sources is unlikely.