Different thermal stimuli initiate hatching of <Emphasis Type="Italic">Daphnia</Emphasis> diapausing eggs originating from lakes and temporary waters

The simultaneous hatching of Daphnia resting eggs from a number of different lakes and ponds using one set of hatching cues can be difficult to achieve as environmental conditions differ considerably between water bodies. Therefore, optimal hatching conditions for ephippial eggs originating from shallow temporary waters may differ from those found in large lakes. The aim of our study was to compare the optimal thermal conditions for hatching ephippial eggs of Daphnia found in permanent lakes and those from shallow temporary ponds. We used ephippial eggs of Daphnia from the longispina species complex originating from two temporary city ponds and two deep lakes in Poland. The ephippia were protected against overheating at all stages of the field and laboratory work to prevent activation or killing of the eggs. After a refractory period (imposed storage in cool and dark conditions), ephippia were incubated at six different temperatures (6, 9, 12, 15, 18 and 21 °C) under a 16:8 L:D light regime. Our results indicate that hatching of resting eggs of Daphnia that inhabit lakes or ponds may require different thermal conditions. The hatching success of ephippial eggs originating from temporary waters was relatively high (30–56%) at all tested incubation temperatures, while for the ephippial eggs from the deep lakes it was lower (7–37%) and inversely related to water temperature. The divergent hatching responses of the ephippial eggs originating from temporary pools and lakes may reflect the typical thermal conditions during hatching in their native habitats. While in the deep lakes of the temperate zone Daphnia hatching typically occurs during the low water temperatures of early spring, in shallow ponds Daphnia hatching may occur throughout the year at varying water temperatures, from a few to over a dozen degrees Celsius.     

Genetic versus environmental influence on pigment variation in the ephippia of Daphnia pulicaria

1. The dormant eggs of Daphnia (Crustacea: cladocera) are encased in a protective, chitonous casing known as an ephippium. Ephippia are pigmented with melanin, and the degree of pigmentation ranges from transparent to opaque. Variation in pigmentation exists within and across populations and species, raising questions about the factors that influence the natural distribution of pigmentation. 2. We used image analysis to quantify pigmentation in ephippial casings of Daphnia pulicaria that were produced both in the field and in the laboratory. The percentage of the surface area of ephippia that is darkly pigmented ranged from 0.5 to 99.5%. The range, mean and variance of ephippial pigmentation varied across our 11 study lakes. 3. Laboratory experiments compared clonal variation (five clones/lake) and population variation (five lakes) at three temperatures (15 °C, 20 °C and 25 °C). The degree of pigment variation between clones was much greater than pigment variation between temperatures, indicating that there is a genetic component to pigmentation in the ephippia which is stronger than the effect of temperature. 4. Comparisons of pigment levels and lake characteristics were used to identify physical and biological factors associated with ephippial pigmentation. Investment in ephippial production was the strongest predictor with darker ephippia occurring more often in lakes with the highest ephippial production.     

Disentangling environmental,spatial, and historical effects on ostracod communities in shallow lakes

Planktonic cladocerans of the genus Daphnia are an example of organisms whose ability to disperse among aquatic habitats is limited to dispersal of dormant eggs, encapsulated in protective structures called ephippia. In the present study, we aimed to quantify the production of floating and sinking ephippia in lakes to test the hypothesis that, even if they eventually sink, most of them are first floating at the surface. In addition, we checked the egg content status of the ephippia. The results of this study revealed numbers of ephippia floating at lakes surface reaching thousands per square metre at the time of ephippia production, and constituting substantial share of that production. In studied shallow lowland lakes and in a deep mountain lake, most ephippia were first floating at the water surface, while in deep lowland lakes the proportion was reversed. Approximately half of ephippia that appeared initially at the water surface sank during the six-week sampling period. The egg content status did not differ between floating and sinking ephippia. High numbers of ephippial females staying in the surface layers at night in a mountain lake, and a laboratory experiment with ephippial females indicated that ephippia are actively oviposited to the water surface by gravid females.     

The effect of food availability, female culture-density and photoperiod on ephippia production in Daphnia magna Straus (Crustacea: Cladocera)

SUMMARY. A method is described whereby ephippia (sexual eggs plus protective membranes) are experimentally induced at low food levels (≤ 0.05 mg spinach D. magna ^-1), high female culture-densities (≤ 0.4 D. magna ml^-1) and in short-day photoperiods (L:D < 12:12, 12°C). The density-dependent ephippial response was related to an increased encounter rate between females at the higher densities. External metabolites had no significant effect on ephippia production. Ephippia were formed in the second generation after exposure to short day-lengths. Five clones from the same population exhibited genetic variation in their critical photoperiods for induction of sexual reproduction (≥= 10% ephippia). There was evidence of a genetic difference in photoresponse between two populations of D. magna. The evolution of timing mechanisms for ephippia production are discussed in relation to the predictability of environmental change.     

Maternal effects after sexual reproduction in Daphnia magna

In this study we investigated the relationship between maternalsize and size of ephippial hatchlings in Daphnia magna. We observedthat larger females produce larger ephippia; these larger ephippiahave a higher hatching probability, yield larger neonates, whichin turn yield larger adults, producing more eggs. We interpretthese observations as an explanation why older and larger femalesof D.magna continue to produce ephippia despite a higher predationrisk.     

Resting eggs of lake- Daphnia I. Distribution, abundance and hatching of eggs collected from various depths in lake sediments

SUMMARY. 1. Resting eggs (ephippia) of Daphnia galeata, D. hyalina, D. cucullata and D. galeata X D. hyalina hybrids were collected from the upper 8 cm of sediments at various water depths from two glacial lakes in the Northern Federal Republic of Germany.
2. The horizontal distribution of 'intact' ephippia (i.e. those containing two apparently healthy eggs) was extremely patchy, with mean densities in the upper 8 cm ranging from 1325 to 113,907 m⁻² (coefficient of variation, C=65.5%; Schöhsee, Holstein) and 25,343–159,143 m⁻² (C=58.5%; Kel-lersee. Holstein). Ephippial densities were greatest in sediments from deep waters. The highest densities of intact ephippia were in the upper 4 cm of sediment, with progressively fewer with increasing depth.
3. A proportion of eggs obtained from sediments during autumn (September-November 1985) and spring (March-May 1986) were exposed to continuous light (approximately 18 W m⁻²) at three temperatures (6, 12, 20°C), and the incidence of hatches recorded.
4. Hatching success was consistently low (maximum 14.4%), with most hatching occurring at 12°C. There was no significant difference between the hatching success of ephippia collected during autumn and spring. Decapsulation of ephippial eggs inhibited hatching. Hatching was highly synchronous within each treatment, and the pattern did not differ significantly with sediment depth, though hatching success was lower in deep sediments. A huge reservoir of ephippia is present in lake sediments, of which a proportion may remain viable for many years. Periodic recruitment of hatchlings to the pelagic population may provide an important mechanism for the maintenance of genetic diversity in Daphnia populations, as well as affecting rates of evolutionary change.     

The effect of diapause on the genetic structure of Daphnia magna populations

For intermittent Daphnia magna populations embryonic diapause following bisexual reproduction is the means of surviving in harsh environmental conditions. Another advantage of bisexuality is genetic recombination which produces a variety of genotypes. Crowding and short-day photo-periods promote bisexual reproduction, but photoperiod, at least, can not be an obligatory factor in D. magna since some bisexual females have been found also during the periods of long daylength. Because the catastrophe dates vary considerably, the most advantageous reproductive strategy must be one with some flexibility in reacting to irregular catastrophes by producing ephippial resting eggs whenever the probability of a catastrophe exceeds a certain threshold value. Extensive genetic changes occurring during diapause suggest that there are considerable genetic differences in ephippial egg production and egg survival. Genetic drift alone can not account for the great genetic changes observed, although in populations reacting slowly to changing conditions the number of diapausing ephippia can be relatively low. If the ephippia hatch over a long period of time, they can act as a stabilizing mechanism for genotype frequencies.     

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.     

Reproduction strategies of <Emphasis Type="Italic">Daphnia pulicaria</Emphasis> population in a high mountain lake of Southern Spain

For three consecutive years, a population study of Daphnia pulicaria was undertaken in Río Seco Lake, a small high-mountain lake, in order to elucidate the reproductive strategies adopted by Daphnia in this system. Daphnia appears to colonize this lake every spring by hatching from ephippia and reproduce by means of subitaneous (non-diapausing) and ephippial (diapausing) eggs. D. pulicaria in this lake is an obligate parthenogenetic population. There is a short time period for subitaneous egg production and a much longer period for ephippial egg production. The contribution of subitaneous eggs to Daphnia population density and structure appears to be low. Diapause onset showed a high temporal synchronization in the three studied years in Río Seco Lake, and day-length emerged as the main cue triggering diapause onset and the main explanatory factor for the proportion of ephippial females observed. The development and reproduction of D. pulicaria in Río Seco Lake involves taking a gamble on resting forms to guarantee inter-annual Daphnia persistence in the lake, giving priority to investment in future generations. Guest editor: Piet Spaak Cladocera: Proceedings of the 7th International Symposium on Cladocera     

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.     

To sink or float: the fate of dormant offspring is determined by maternal behaviour in Daphnia

• 1 As the ephippia (chitinous shells enclosing diapausing eggs) of pelagic crustaceans of the genus Daphnia have been occasionally reported to float at the water surface, we considered that this might be an adaptation promoting their passive dispersal. We investigated the mechanisms by which ephippia appear at the water surface.
• 2 While field surveys revealed that floating Daphnia ephippia are often numerous in various freshwater habitats, laboratory tests showed that newly formed ephippia are not buoyant initially. Once transferred to the surface by whatever means, however, they may remain there due either to surface tension or gas absorption.
• 3 Video recordings showed that all ephippia at the water surface in laboratory vessels were shed there by ephippial females when moulting (despite the attendant risk of exposure to UV radiation). This implies that the moulting behaviour of female Daphnia may determine the fate of their dormant offspring, predetermining whether they remain in the natal environment (when the ephippium is released into the water column) or disperse (when it is deposited at the water surface).
• 4 Our findings reveal a potential mechanism underlying the high dispersal capacity of freshwater cladocerans inhabiting island‐like aquatic habitats.

     

DNA extraction and amplification methods for ephippial cases of Daphnia resting eggs in lake sediments: a novel approach for reconstructing zooplankton population structure from the past

This study describes a method of DNA extraction and amplification for ephippial cases of Daphnia resting eggs from lake sediment. Recent studies have reconstructed succession records of Daphnia species by genetically analyzing Daphnia resting eggs stored in lake sediments and revealed changes in dominant Daphnia species that correspond well with environmental changes. However, this approach is not applicable to lakes where most of the resting eggs in the sediment have already hatched out. We modified conventional methods for DNA extraction and amplification to enable genetic analyses of the ephippial case that envelops and protects the resting eggs, and we compared the performance of the modified method to the conventional one. We confirmed that we could efficiently analyze the sequences of the ephippial cases collected in a sediment core using our modified method. It enables us to reconstruct changes in genetic structure of Daphnia populations regardless of hatching rates of the resting eggs.     

Studies on the photoactivation of the water-oxidizing enzyme : I. Processes limiting photoactivation in hydroxylamine-extracted leaf segments

In weak yet optimal light intensity, complete photoactivation of the water-oxidizing enzyme in NH₂OH-extracted wheat (Triticum aestivum, var Oasis) leaf segments could be obtained only after long dark preincubation. Photoactivation was not affected by ethylenediaminetetraacetate or inhibitors of photophosphorylation and protein synthesis, but was partially inhibited by a divalent cation ionophore. Complete photoactivation required ligation of ~4 Mn by the water oxidizing enzyme.

Without dark preincubation, photosystem II (PSII) was susceptible to weak light photoinhibition resulting in: (a) 50% maximum decrease in photooxidation of artificial electron donors by PSII: (b) increased times for the variable fluorescence rise (with 3-(3,4-dichlorophenyl)-1,1-dimethyl urea): (c) abolishment of photoactivation: and (d) the imposition of sensitivity to inhibitors of photophosphorylation and 70S but not 80S protein synthesis on subsequent light-dependent recovery from photoinhibition and recovery of O₂ evolution. Decrease in susceptibility to photoinhibition and increase in rates of photoactivation resulting from dark preincubations proved closely correlated. Neither protein synthesis nor increases in abundances of thylakoid Mn²⁺ and Ca²⁺ were required for escape from photoinhibition. However, photoactivation of the wateroxidizing enzyme in NH₂OH-extracted Chlamydomonas occurred in absence of dark preincubation and protein synthesis. Results are discussed in the context of disassembly/reassembly/resynthesis of specific PSII polypeptides.

     

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.     

Light Affects the Chloroplast Ultrastructure and Post-Storage Photosynthetic Performance of Watermelon (Citrullus lanatus) Plug Seedlings

Watermelon [Citrullus lanatus (Thunb.) Matsum. and Nakai] plug seedlings were stored at 15°C in the light at a photosynthetic photon flux density of 15 µmol·m⁻²·s⁻¹ or in darkness for 6 days, to evaluate their chloroplast ultrastructure, and associated photosynthetic characteristics. Storage in the dark caused swelling, disordered granal arrangement, and starch grain disappearance in the chloroplasts. In contrast, the chloroplasts stored in the light were relatively normal. As a result, the light-stored seedlings had a significantly higher chlorophyll content, Fv/Fm, and Pn than did dark-stored seedlings. Regardless of whether the seedlings were stored in light or darkness, the Gs and Ls of the seedlings significantly decreased, while the Ci obviously increased when the Pn decreased after 6 days of storage. This result suggests that the decreased Pn is not solely a stomatal effect, as the effects on the chloroplasts contributed to this photosynthetic inhibition. Six days after transplanting, seedlings that were stored in the light or darkness for 2 or 4 days showed complete recovery of chloroplast ultrastructure, chlorophyll content, Fv/Fm, Gs and Pn. When the storage period increased to 6 days, the dark-stored seedlings had a significantly lower Fv/Fm and Pn than the light-stored and control seedlings 6 days after transplanting, which was mainly ascribed to incomplete recovery of chloroplast ultrastructure. Furthermore, the light-stored seedlings exhibited a significantly higher shoot dry weight during storage and a higher percentage dry weight increase after transplanting than the dark-stored seedlings. These effects were enhanced by prolonged storage (4 to 6 days). This study demonstrated that dim light during storage is beneficial for maintaining chloroplast ultrastructure as well as photosynthetic efficiency in watermelon seedlings, thus contributing to the rapid recovery of post-storage photosynthetic performance, which ensures the transplant quality of the seedlings after removal from storage.     

The influence of temperature on light enhanced glycoalkaloid synthesis in potato

The influence of temperature on total glycoalkaloid (TGA) synthesis in tubers exposed to light (250 jumol m“² s”² PAR, Photosynthetically Active Radiation) or dark environments for 96 h was examined in three potato cultivars. Following 96 h light or dark the tubers were stored without light at 5°C or 24°C and TGA concentrations monitored over the subsequent 30 and 90 days. Exposure to light and cultivar were found to be major factors influencing TGA concentrations; temperature had no significant effect. TGA content in illuminated tubers of cvs ‘Pentland Hawk’ and ‘Kerrs Pink’ were significantly higher (P < 0.01) compared with tubers placed in the dark. TGA concentrations in cv. ‘Desiree’ increased significantly only following exposure to light at low temperatures (P < 0.05). Removal of tubers from storage at 5°C and immediate illumination at 24°C altered the ratio of glycoalkaloids in cvs ‘Pentland Hawk’ and ‘Kerrs Pink’. Regardless of cultivar and storage temperature TGA concentrations were higher at the end of the storage period compared with initial TGA concentrations. During storage TGA concentrations fluctuated widely and gradual accumulation of glycoalkaloids with time was rarely demonstrated except in cv. ‘Desiree’. Tubers stored at 24°C accumulated higher TGA concentrations than those stored at 5°C in cv. ‘Kerrs Pink’ but not in cvs ‘Pentland Hawk’ and ‘Desiree’. Tubers of cv. ‘Kerrs Pink’ exposed to light prior to storage accumulated glycoalkaloids more rapidly than unexposed tubers during storage at 24°C and occasionally at 5°C. Light enhanced glycoalkaloids are not degraded over time.     

Mechanism of photoactivation of electron transport in intact bryopsis chloroplasts

The mechanism of photoactivation of photosystem I electron transport was studied in intact Bryopsis corticulans chloroplasts. The evidence from chemical and photochemical studies suggests that photoactivation is a consequence of a reduction of an electron transport component, presumably ferredoxin-NADP⁺ reductase. O₂ does not act as a mediator of the process but rather acts as an electron acceptor after photoactivation has occurred. We suggest that the initial function of the chloroplasts in a transition from dark to light is to initiate pseudocyclic electron flow.     

Identifying century-old long-spined <Emphasis Type="Italic">Daphnia</Emphasis>: species replacement in a mountain lake characterised by paleogenetic methods

Mountain lakes often harbour morphologically or genetically unique populations of zooplankton species, including cladocerans. Daphnia lacustris Sars, predominantly found in Fennoscandia but also known from two Central European lakes in the Tatra Mountains, is one of such taxa. This Daphnia species often forms morphotypes with extremely long tailspines. Historical literature from a century ago documented similar morphs from another lake in the Tatra mountain range, presently inhabited by the phenotypically very different D. galeata. Using a paleogenetic approach (partial sequencing of the mitochondrial gene for 12S rRNA from preserved ephippial eggs in the lake sediment), we tested the hypothesis that Daphnia species composition changed in the lake due to anthropogenic disturbances, and that long-spined morphs were actually another relict population of currently extinct D. lacustris. Ephippia with extremely long spines were successfully retrieved from sediment cores. Despite being morphologically very well preserved, intact eggs were found in less than 2% of analysed ephippia. Genetic analyses, benefiting in most cases from amplification of short 12S fragments using internal primers, proved that long-spined ephippia belonged to D. longispina, which apparently coexisted with D. galeata in the mid-twentieth century. Our results confirm that paleogenetic methods are useful for studying the recent population structures of zooplankton species forming dormant egg banks but lacking reliably identifiable remains in sediments, and show that the extreme development of tailspines in mountain-lake Daphnia is associated with as-yet unclear environmental factors rather than taxonomic status.     

Effect of Storage Environment on Hatching of the Cyst Nematode Globodera ellingtonae

Globodera spp. eggs go through a diapause, which remains dormant until favorable hatching conditions are reached. Because of the regulatory concerns with cyst nematodes, it is often only possible to rear eggs for research in the greenhouse. However, hatch is often lower for greenhouse-produced eggs than for eggs obtained from the field. The goal of this research was to determine storage conditions for Globodera ellingtonae eggs produced in the greenhouse that would increase percentage hatch. Over 3 yr, G. ellingtonae greenhouse-produced eggs were stored in different environments (−20°C, 4°C, room temperature, and the field) in either dry or moist soil. Percentage hatch after exposure to the different environments was determined in potato root diffusate. Across two experiments, field-produced eggs had higher hatch rates (65.2%) than greenhouse-produced eggs (10.4%). Temperature did not have an appreciable influence on hatch of eggs stored dry in two experiments (2.8% to 8.4% and 3.8% to 8.6%), but hatch of eggs stored in moist soil was significantly higher than in dry soil at all temperatures except −20°C (26.8% and 28.7%). However, the ability of G. ellingtonae greenhouse-, microplot-, and field-produced eggs to reproduce on potato in field microplots was not different. Although it may not be possible to produce G. ellingtonae eggs in the greenhouse that have the magnitude of hatch as those produced in the field, hatching can be greatly increased by storing eggs in moist soil at either 4°C or room temperature.     

The Duplex Nature of the Retina of the Nocturnal Gecko as Reflected in the Electroretinogram

The effect of light and dark adaptation on the electrical activity in two species of nocturnal gecko, Hemidactylus turcicus and Tarentola mauritanica was studied. The electroretinogram of both species changes from the scotopic type in the dark-adapted state to the photopic type after strong light adaptation. For the scotopic response fusion frequencies up to 18 flashes per sec. are obtained in both species. For the photopic response fusion frequencies up to 50 flashes per sec. are seen in Tarentola, and up to 25 flashes per sec. in Hemidactylus. Proceeding from dark to light adaptation the increment threshold (dI) is measured at different levels of adaptive illumination (I). At low levels of illumination the dI/I ratio is found to be small and at high levels of illumination to be large. No difference in the dI/I ratio is obtained for test lights of 462 and 605 mµ. During dark adaptation the change of threshold after exposure to moderate and weak lights (up to 10³ times dark threshold) is rather fast. After light adaptation to strong light (10⁶ times dark threshold) duplex dark adaptation curves are seen with a break separating a fast and a slow phase of dark adaptation. The significance of these results from a retina which possesses sense cells of only one type is discussed.