Experimental assessment of the fecundity of Eucypris virens (Ostracoda, Crustacea) under natural sex ratios

1. The adaptive significance of the observed variations in sex ratios in non‐marine ostracods is unclear. This study quantified the fecundity of females taken from a presumed fully sexual Eucypris virens population that were experimentally combined with different proportions of males (male : female sex ratios: 1 : 1, 1 : 2, 1 : 4, 1 : 8 and 0 : 1). 2. The results yielded no indications that female fecundity is altered by short‐term variations in the proportion of males, at least not within the range of sex ratios that are common in natural ostracod populations. Complete removal of males, however, did strongly reduce hatching success of dried eggs. This suggests the need for multiple mating events during the reproductive lifetime of the female. It also emphasizes the need for a minimum number of males, although this minimum number evidently may be rather low, as we found a high number of spermatozoa in the seminal receptacles after a single mating event. 3. The sex ratio in the source population was strongly female biased (1 : 3.4; n = 514), whereas in the hatchling assemblages reared in the laboratory, males and females were found in equal proportions (1 : 1.0; n = 1516), irrespective of the prevailing sex ratio. This clear discrepancy is intriguing, and points to the importance of epigenetic factors for the determination of field sex ratios.     

Production of Potato Spindle Tuber Viroid (PSTV) by Large Scale Fermentation of PSTY-Infected Potato Cell Suspension Cultures

Berlin, J., Wray, V., Forche, E., Reng, H.–G , Schler,H, Luckinger, R. and Mhlbach, H.–P. 1985. Production ofpotato spindle tuber viroid (PSTV) by large scale fermentationof PSTV–infected potato cell suspension cultures.—J.exp. Bot 36: 1985–1995. Cell suspension cultures of Solatiumdemissum, infected with the potato spindle tuber viroid (PSTV),were scaled up to volumes of up to 800 dm³ to provide sufficientand uniform plant material for subsequent studies on viroidbiosynthesis. Here we describe the technological aspects ofproducing the required amounts of biomass and viroid. The cells,which had been maintained on a medium containing expensive coconutmilk, were first adapted to rapid growth on the less expensiveB5–medium. The physiological state of the cells was monitoredby in vivo ³¹P–NMR spectroscopy Under the chosen conditionsthe scale–up from 10 dm³ inoculum from shake flasks tothe harvest of the 800 dm³ stirred fermenter lasted 38 d andprovided 112 kg biomass. Growth characteristics and viroid productionin shake flasks and large bioreactors were rather similar. Gelelectrophoretic analysis of isolated nucleic acids using silverstaining and Northern blot hybridization revealed a PSTV–contentof approximately 700 µg PSTV per kg fresh mass of culturedcells. Key words: Solanum demissum, plant cell cultures, potato spindle tuber viroid, biomass production, fermentation, in vivo ³¹P-NMR     

Mate guarding in relation to seasonal changes in the energy reserves of two freshwater amphipods (Gammarus fossarum and G. pulex)

1. We assessed sex‐specific seasonal changes in major energy storage compounds (triglycerides, glycogen) in Gammarus fossarum and Gammarus pulex collected from the field, with respect to their reproductive activity. 2. The dynamics of stored energy followed a seasonal pattern in both species and sexes. Moreover, over a 4‐year period, these changes were independent of the year in which they were investigated. Stored energy reached a peak in late winter, but was depleted in late summer and early autumn, coinciding with the reproductive periods. 3. Triglyceride (annual mean ± SD) accounted for 79.7 ± 11.9% of the total stored energy and was responsible for the seasonal pattern. In contrast, glycogen contributed a lesser percentage (20.3 ± 11.9%). Over the study period, the amount of stored energy ranged between 0.39 and 4.08 kJ g^?1 dry mass (triglyceride: 0.19–3.69 kJ g^?1 dry mass; glycogen: 0.14–0.80 kJ g^?1 dry mass). 4. In both species, the energy reserves of males were drastically depleted shortly before the cessation of precopulatory mate guarding in the field, thus offering a bioenergetic explanation for the reproductive period in these two widespread species.     

Extreme tolerance to environmental stress of sexual and parthenogenetic resting eggs of Eucypris virens (Crustacea,Ostracoda)

1. The freshwater ostracod (Ostracoda), Eucypris virens, is commonly found in European temporary pools, where its long‐term persistence completely relies on the build‐up of resting egg banks. Extreme tolerance of dormant eggs and seeds is widely assumed, but freshwater ostracod eggs are relatively poorly studied. The study of ostracod resting eggs is of particular relevance as it may yield the key to understanding the distribution of the sexes in many species capable of both sexual and asexual reproduction. 2. We assessed the tolerance of dried resting eggs produced by females originating from three populations with males and three all‐female E. virens populations. Hatching time and success was compared between control eggs and eggs exposed to one of seven ecologically relevant stressors: digestive enzymes, high salinity, deep freezing, hydration, UV‐B radiation, hypoxia and insecticide treatment. 3. None of the stressors reduced significantly the viability of either sexual or asexual eggs. When compared with the reproductive mode–specific controls, exposure to UV‐B radiation had a mild impact on the survival of sexual and asexual eggs (?16.8 and ?22.4%, respectively), but this was only significant for asexual eggs. These results point to an extreme tolerance of E. virens resting eggs and have important implications for the ecology and evolution of the species. 4. The timing of hatching was not affected by the stress treatment, except for UV‐B radiation. A marginally significant delay in hatching response was observed for UV‐B‐radiated eggs when compared to the overall mean, but this treatment effect was absent when compared with the reproductive mode–specific controls. 5. The populations with males produced eggs that hatched on average earlier (?1.5 days at 17 °C) and were more successful (+26%) than asexual eggs. Due to the limited number of populations and the population‐specific origin and age of the eggs, the possibility due to the differences in age and origin of the resting eggs, or to variations in local conditions, cannot be ruled out.     

On the 13C/12C isotopic signal of day and night respiration at the mesocosm level

While there is currently intense effort to examine the ¹³C signal of CO₂ evolved in the dark, less is known on the isotope composition of day‐respired CO₂. This lack of knowledge stems from technical difficulties to measure the pure respiratory isotopic signal: day respiration is mixed up with photorespiration, and there is no obvious way to separate photosynthetic fractionation (pure c_i/c_a effect) from respiratory effect (production of CO₂ with a different δ¹³C value from that of net‐fixed CO₂) at the ecosystem level. Here, we took advantage of new simple equations, and applied them to sunflower canopies grown under low and high [CO₂]. We show that whole mesocosm‐respired CO₂ is slightly ¹³C depleted in the light at the mesocosm level (by 0.2–0.8‰), while it is slightly ¹³C enriched in darkness (by 1.5–3.2‰). The turnover of the respiratory carbon pool after labelling appears similar in the light and in the dark, and accordingly, a hierarchical clustering analysis shows a close correlation between the ¹³C abundance in day‐ and night‐evolved CO₂. We conclude that the carbon source for respiration is similar in the dark and in the light, but the metabolic pathways associated with CO₂ production may change, thereby explaining the different ¹²C/¹³C respiratory fractionations in the light and in the dark.     

Characterization of microsatellite markers for the invasive ant species Anoplolepis gracilipes

We present primer sequences for eight polymorphic microsatellite loci for the formicine ant Anoplolepis gracilipes, a serious pest species in South‐East Asia and Pacific islands and still spreading on all continents. Microsatellite loci were isolated with a highly efficient method of enrichment. The number of alleles ranged from two to 19 with an observed heterozygosity ranging from 0.842 to 1.0. The markers were designed for a sociogenetic study as well as for population genetics.     

Concordant phylogeography and cryptic speciation in two Western Palaearctic oak gall parasitoid species complexes

Little is known about the evolutionary history of most complex multi‐trophic insect communities. Widespread species from different trophic levels might evolve in parallel, showing similar spatial patterns and either congruent temporal patterns (Contemporary Host‐tracking) or later divergence in higher trophic levels (Delayed Host‐tracking). Alternatively, host shifts by natural enemies among communities centred on different host resources could disrupt any common community phylogeographic pattern. We examined these alternative models using two Megastigmus parasitoid morphospecies associated with oak cynipid galls sampled throughout their Western Palaearctic distributions. Based on existing host cynipid data, a parallel evolution model predicts that eastern regions of the Western Palaearctic should contain ancestral populations with range expansions across Europe about 1.6 million years ago and deeper species‐level divergence at both 8–9 and 4–5 million years ago. Sequence data from mitochondrial cytochrome b and multiple nuclear genes showed similar phylogenetic patterns and revealed cryptic genetic species within both morphospecies, indicating greater diversity in these communities than previously thought. Phylogeographic divergence was apparent in most cryptic species between relatively stable, diverse, putatively ancestral populations in Asia Minor and the Middle East, and genetically depauperate, rapidly expanding populations in Europe, paralleling patterns in host gallwasp species. Mitochondrial and nuclear data also suggested that Europe may have been colonized multiple times from eastern source populations since the late Miocene. Temporal patterns of lineage divergence were congruent within and across trophic levels, supporting the Contemporary Host‐tracking Hypothesis for community evolution.     

A microsatellite linkage map for Drosophila montana shows large variation in recombination rates,and a courtship song trait maps to an area of low recombination

Current advances in genetic analysis are opening up our knowledge of the genetics of species differences, but challenges remain, particularly for out‐bred natural populations. We constructed a microsatellite‐based linkage map for two out‐bred lines of Drosophila montana derived from divergent populations by taking advantage of the Drosophila virilis genome and available cytological maps of both species. Although the placement of markers was quite consistent with cytological predictions, the map indicated large heterogeneity in recombination rates along chromosomes. We also performed a quantitative trait locus (QTL) analysis on a courtship song character (carrier frequency), which differs between populations and is subject to strong sexual selection. Linkage mapping yielded two significant QTLs, which explained 3% and 14% of the variation in carrier frequency, respectively. Interestingly, as in other recent studies of traits which can influence speciation, the strongest QTL mapped to a genomic region partly covered by an inversion polymorphism.